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Sarcoidosis is a complex disease with unknown etiology. Histologic hallmarks are nonnecrotizing and noncaseating granulomas, which usually affect the lungs but can grow widely. Computed tomography (CT) helps reveal typical features such as alveolar and ground-glass opacities and perivascular, subpleural nodules or active alveolitis, as well as atypical features such as bronchiectasis, pulmonary fibrosis and lung emphysema, and to differentiate with concomitant infection or neoplasms.
Fluorine-18-fluorodeoxyglucose (18F-FDG) is a glucose derivative that is highly concentrated in areas with intense glucose metabolic and high inflammatory activity, since inflammatory cells such as macrophages, lymphocytes, neutrophils have an elevated metabolic activity of this compound and positron emission tomography/computed tomography (PET-CT) with FDG has proven to be a non-invasive technique which is useful to assess several infectious, inflammatory and neoplastic disorders, such as toxoplasmosis, prosthetic joint infections, spinal osteomyelitis, fever of unknown origin (FUO), vasculitis and sarcoid lesions in sarcoidosis. The molecular mechanisms of FDG PET-CT, its role in sarcoidosis and its recent advances are discussed in this review. Information about its role in assessing the disease activity, treatment response, prognosis and pitfalls are also reported.
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Many conventional and adjuvant anticancer therapies can have serious adverse effects or yield subpar therapeutic results, and thus nanomedicine has begun to gain appeal for treating cancer. A family of nanocarriers called cubosomes exhibits promising theragnostic efficacy. They are self-assembling, honeycomb-like, three-dimensional structures in the bicontinuous cubic liquid crystalline phase. They have a large interfacial area, straightforward preparation, capacity to encapsulate hydrophobic, hydrophilic, and amphiphilic moieties, biodegradability, and targeted and controlled release of bioactive freight. Targeted drug delivery and a decrease in drug resistance have been accomplished by cubosomes. Cubosomes have also been employed in immunotherapy as a component of cancer medication therapy. Studies have revealed that the pharmacokinetics and safety characteristics of loaded medicines are enhanced by cubosomes. Cubosomes have been used in numerous preclinical trials to treat cancer and for theragnostic purposes. This review reports the most recent studies on the application of cubosomes in vitro and in vivo experimental models for the treatment of various neoplastic illnesses. The review highlights that cubosomes provide a flexible and promising platform for the administration of medicinal drugs.
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Chronic diseases such as cancer, diabetes, heart disease, and obesity are increasingly prevalent and well-researched. The use of traditional medicinal plants may lead to the discovery of new and potent medicinal compounds (phytochemicals) for treating various ailments. Modern medicine currently uses over 7000 phytochemicals, many of which have been utilized for generations by traditional healers. Medicinal plant products have a global market value of over $100 billion annually. Phytochemicals are compounds that are derived from plants and are rich in nutrients. They are often used for medicinal purposes. They are mostly secondary metabolites that can provide nutritional elements and promote long-term health outcomes. Phytochemicals found in herbs, nuts, legumes, grains, fruits, and vegetables include alkaloids, glycosides, tannins, resins, phenolic compounds, terpenoids, pigments, and natural antioxidants that may play a role in managing chronic diseases. These nutritional phytochemicals are encapsulated in various dosage forms and are designated as nutraceuticals. Natural plant diets, nutraceuticals, and nutritional treatments have gained a lot of attention for their potential to improve health and combat ailments. Further research is required to determine the safety, efficacy, and toxicity of plant-derived phytochemicals and nutraceuticals used to treat chronic diseases and meet nutritional needs. This review presents valuable information on the use of phytochemicals and nutraceuticals for treating cancer, autoimmune inflammatory disorders, diabetes, cardiovascular and metabolic disorders, and obesity. It explains the potential ways in which these natural compounds can provide therapeutic benefits in managing chronic illnesses and provides data on their safety and toxicity.
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Opioid growth factor (OGF) and its receptor, opioid growth factor receptor (OGFr), create a physiological axis that affects cellular growth by decreasing the G1/S phase of cell division in both normal and malignant cells. The OGF-OGFr axis is potent, rapid, reversible, stereospecific, receptor-mediated, serum- and anchorage-independent, dosage- and duration-dependent, and tonically active and is present at physiological doses. Cancer progression can be suppressed or accelerated when the OGF-OGFr pathway is altered by adding OGF externally or obstructing its receptor. Exogenous or endogenous antagonists of OGF and OGFr have been shown to modulate cancer cell propagation. OGF-OGFr pathway activation has the potential to reduce tumor development and metastasis and enhance the effectiveness of chemotherapy and radiation therapy while preserving healthy tissues, as shown by preclinical research conducted in animal models. Clinical trials in humans have also demonstrated promising results in treating various types of cancer, encompassing pancreatic, lung, and ovarian cancers. This review aims to summarize the recent developments that show the importance of the OGF-OGFr axis in cancer treatment.
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Sterol O-acyltransferase 1 (SOAT1), also known as Acyl-coenzyme A: cholesterol acyltransferase 1 (ACAT1), belongs to the membrane-bound O-acyltransferase enzyme family and plays a crucial role in maintaining intracellular cholesterol homeostasis. Numerous studies have documented the association between aberrant SOAT1 expression and various aspects of cancer biology, including cell proliferation, apoptosis, invasion, and metastasis. In this comprehensive review, we outline the multifaceted role of SOAT1 in tumor initiation, progression, and therapeutic responses. Furthermore, we elucidate its potential as a prognostic marker and highlight its clinical relevance as a promising therapeutic target for malignancies.
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The tumor microenvironment (TME) is closely related to the effect of radiotherapy on tumor cells. The changes in the TME after radiotherapy affect tumor growth, invasion, and metastasis. Tumor-associated macrophages (TAMs) are the most abundant immune cells in the TME. Their aggregation, phenotype change, and various cytokines released by macrophages after radiation therapy significantly affect the response of tumor cells to radiation and are important targets for improving the therapeutic effect of radiation therapy in the future. However, the effect of radiation on TAMs and the specific mechanism by which TAMs affect sensitivity to radiotherapy are still unclear. In this review, we highlight recent advances in the effects of radiotherapy on the phenotype of TAMs, the mechanism by which radiotherapy recruits TAMs, and the effect of TAMs on the radiosensitivity of tumor cells. The possible use of TAMs as a potential therapeutic target in combination with radiotherapy with a focus on improving radioresistance and enhancing anti-tumor immune activity has also been discussed. This provides a theoretical basis for the combined use of radiotherapy and targeted TAMs therapy in the future to improve the efficacy of radiotherapy.
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Tetraspanins are four transmembrane proteins that serve as a platform for interacting with a wide range of different molecules. It plays a crucial role in oncogenesis as well as in the metastasis in various cancer types. Numerous studies have investigated the use of various members of the tetraspanin family as prognostic markers and predictors of metastatic potential according to specific cancer types. This manuscript provides an overview of these advances, and their limitations, and suggests insights into future research areas for their clinical applications. Furthermore, the potential value of tetraspanin as a tumor-associated antigenic target will also be discussed in this review.
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Background: Interferon (IFN)-γ is a proinflammatory cytokine with a crucial role in intercellular communication during innate and acquired immune responses. IFN-γ interacts with various cell types, including endothelial cells. Here, we investigated the effects of pharmacological or low doses of IFN-γ in cultured endothelial cells.
Methods: Human endothelial cells were cultured in the presence of pharmacological or low-dose concentrations of IFN-γ. Signal transducer and activator of transcription (STAT) and Extracellular signal-regulated kinase (ERK) phosphorylation were investigated by enzyme linked immunosorbent assay (ELISA). Western blot for ERK was also performed. Transient ERK silencing was obtained by short interfering RNA (siRNA). Cell proliferation and migration were analysed by cell counting and wound assay, respectively.
Results: At pharmacological concentrations, IFN-γ activates the Janus kinases (JAK)/STAT pathway, leading to the overexpression of the cyclin-dependent kinase inhibitor 1A/p21 (CDKN1A/p21), which inhibits cell growth. In contrast, low-dose activated IFN-γ does not trigger the canonical JAK/STAT pathway and induces the phosphorylation of ERK. ERK activation is responsible for endothelial cell migration induced by low-dose activated IFN-γ.
Conclusions: We demonstrate that pharmacological and low-dose activated IFN-γ exert distinct effects on endothelial cells by triggering different signal transduction pathways. These findings shed light on the intricate signalling pathways employed by IFN-γ, and suggest that low-doses of IFN-γ might play a homeostatic role in endothelial cell during innate and acquired immune responses.
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Background: Infections caused by multi-resistant microorganisms are a public health issue. Synthetic products have been studied as a therapeutic alternative for these cases, with the aim of antibacterial activity or enhancing existing antibiotics.
Methods: This study aimed to evaluate the antibacterial activity upon modifying antibiotic actions and to perform a pharmacokinetic analysis of compounds derived from thiadiazine. The products were synthesised, and a chemical analysis was performed using thin-layer chromatography (TLC).
Results: After obtaining the compounds, minimum inhibitory concentration (MIC) and modified antibiotic activity tests were performed, in which norfloxacin and gentamicin antibiotics were used against multidrug-resistant (MDR) bacterial strains of Staphylococcus aureus 10 (S. aureus 10) and Pseudomonas aeruginosa 24 (P. aeruginosa 24) using the microdilution method. We also performed topological analyses, a similarity evaluation with drugs, and in-silico absorption, distribution, metabolism, excretion and toxicity (ADMET) analyses. Only two compounds showed antibacterial activity against the examined strains. However, all compounds showed activity against the S. aureus strain when associated with norfloxacin. The in-silico analysis showed that the compounds have high lipophilicity and non-toxic characteristics.
Conclusions: Therefore, the thiadiazine-derived compounds potentiate the antibiotic activity against S. aureus 10 bacteria and have ADMET properties, which are favourable for further studies on these compounds as potential low-toxicity sources for the MDR bacteria problem.
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Background: This research focused on evaluating the antioxidant properties and phenolic compound content of three different Chrysanthemum species from various regions in South Korea. Phenolic compounds play crucial roles in plant defense; they also attract pollinators and have applications in diverse industries, such as cosmetics, food supplements, and food packaging. Moreover, their radical-scavenging abilities make them promising in combating diseases like diabetes, Alzheimer's, and cancer.
Methods: The study encompassed a total of eight samples from three Chrysanthemum species, including Dendranthema indicum, Dendranthema boreale (D. boreale), and Aster spathulifolius (A. spathulifolius). These samples were collected from distinct regions in South Korea, namely Jeju Island, Pohang, Busan, and Gubong Island, and were extracted using methanol (MeOH). High-performance liquid chromatography (HPLC) analysis was conducted using a Waters Alliance system and a YMC Pack Pro C18 column. To prepare the samples, extracts were dissolved in MeOH, and stock solutions were created for standard compounds. Antioxidant activity was assessed using the 2,2-Diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-casino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assays, and HPLC was used to identify phenolic compounds in the plants.
Results: The study found strong antioxidant activity in all samples, with D. boreale samples from Jejudo Sanguk Flower (JSF) showing the highest potential for medicinal use. In the DPPH assay, JSF exhibited the most potent scavenging activity with a half-maximal inhibitory concentration (IC50) of 5.8 mg/mL. In the ABTS assay, JSF also scored highest at 2.8 mg/mL. Further research is needed to explore their medicinal applications due to their exceptional antioxidant properties. HPLC analysis successfully detected ten out of twelve standard phenolic compounds.
Conclusions: These findings underline the significant antioxidant potential of these Chrysanthemum samples, with JSF showing the most promise. The study underscores the need for further investigation into their potential medicinal and therapeutic applications, given their remarkable antioxidant properties. Overall, this study can help improve the market value of the Chrysanthemum family in various industries.
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Background: Immune infiltration-related genes have reported to play important roles in the prognosis of cholangiocarcinoma (CHOL). This study aimed to screen for prognostic markers of CHOL and construct a prognostic prediction model based on prognostic markers.
Methods: Immune cell infiltration was evaluated in CHOL tumor samples using the single sample Gene Set Enrichment Analysis (ssGSEA) algorithm, followed by immune clustering grouping. Based on immune grouping, differentially expressed genes (DEGs) were selected, and the prognostic markers of CHOL were screened from these DEGs through Cox regression analysis and the least absolute shrinkage and selection operator (LASSO) algorithm. A survival prognostic prediction model was constructed and validated based on the prognostic markers of CHOL.
Results: The samples were divided into two clusters, and 349 intersection DEGs were identified between the tumor and normal and cluster 2 groups and cluster 1 comparison groups, which were enriched in immune response, inflammatory response, and cytokine-cytokine receptor interaction-related functions and pathways. Based on these genes, six DEGs were screened to construct a prognostic risk prediction model. In the training and validation datasets, there was a significant correlation between the actual prognosis and the different risk groups of the samples divided based on the prediction model.
Conclusion: Our study established a prognostic signature associated with immune cell infiltration in patients with CHOL. This prognostic model may be used for diagnosis and prognosis of this disease.
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Background: Ferroptosis has been conceptualized as a unique form of iron-dependent cell death, which holds the potential to treat drug-resistant cancers. Erastin-based therapy is expected to circumvent the shortcomings of apoptosis-mediated therapies, but a variety of tumors are insensitive to erastin. Here, we examined the effect of Roxadustat, a hypoxia inducible factor (HIF)-prolyl hydroxylase (PHD) inhibitor, on erastin-induced ferroptosis in lung cancer cells and the underlying mechanisms.
Methods: Cytotoxicity of Roxadustat was investigated by lactate dehydrogenase (LDH) release assay in H1229 and A549 lung cancer cells. Ferroptosis-related activity of drug treatment was examined by the measurement of iron concentration, malondialdehyde (MDA), reactive oxygen species (ROS) and glutathione (GSH). Cell proliferation and migration abilities were analyzed by cell count kit (CCK)-8 and scratch assays. mRNA and protein expression levels were detected by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot. The ultrastructure of mitochondria was examined by transmission electron microscopy (TEM). The in vivo effect of Roxadustat on ferroptosis induction was evaluated in mouse xenograft model of lung cancer cells.
Results: Roxadustat could disrupt cellular redox homeostasis, increase lipid peroxidation and induce cellular features of ferroptosis at sub-toxic concentration in H1229 and A549 cells. The application of Roxadustat augmented erastin-induced ferroptosis in vitro and in vivo. Roxadustat treatment stabilized HIF-2α protein, which in turn promoted hepcidin expression. The interaction between hepcidin and ferroportin 1 (FPN1) reduced FPN1 protein level by promoting FPN1 degradation. Silencing hepcidin or over-expressing FPN1 could attenuate the intracellular iron level increase and dampen the effect of Roxadustat on erastin-induced ferroptosis.
Conclusions: These data demonstrated that Roxadustat enhances ferroptosis-inducing effect of erastin on lung cancer cells by regulating hepcidin/FPN1 axis through stabilizing HIF-2α, indicating the potential of Roxadustat as a ferroptosis-enhancing agent.
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Background: Gastric cancer (GC) is considered a significant health problem worldwide. The initial diagnosis of GC often occurs at an advanced disease stage and the prognosis is generally poor. The objective of this study is to investigate the role of neuropilin 1 (NRP1) in driving growth and metastasis of gastric cancer through the regulation of copper homeostasis. Our aim is to develop potential novel therapeutic strategies for GC.
Methods: Gene expression profiling and clinical data from gastric cancer patients were obtained from The Cancer Genome Atlas (TCGA). Differential analysis was performed using the limma package in R, and a prognostic model was constructed using least absolute shrinkage and selection operator (LASSO) regression. The diagnostic model was established using logistic regression. Cell culture experiments were conducted to validate the findings, including reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR), western blotting, cell counting kit (CCK)-8 assay, colony-forming assay, cell scratch test, transwell migration assay, immunofluorescence staining, and apoptosis analysis.
Results: NRP1 was found to be overexpressed in gastric cancer cells (p < 0.01). It was identified as a potential regulator of copper homeostasis, as its overexpression led to increased expression levels of ATPase copper transporting alpha (ATP7A; p < 0.01), a copper transporter. The phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT) signaling pathway was involved in mediating the effects of NRP1 on ATP7A expression. Functional assays demonstrated that NRP1 promoted cell proliferation (p < 0.05) and migration (p < 0.01), while inhibiting apoptosis (p < 0.05). Furthermore, treatment with a copper chelator sensitized NRP1-overexpressing cells and reduced their proliferation (p < 0.01).
Conclusions: This study reveals that NRP1 plays a crucial role in driving the growth and metastasis of gastric cancer through the regulation of copper homeostasis. Targeting NRP1 and copper homeostasis may provide potential therapeutic strategies for the treatment of gastric cancer.
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Background: Exogenous Nel-like molecule type 1 (NELL1) is a viable alternative in orthodontics and other fields due to its osteogenic and angiogenic properties. This study aimed to elucidate the role of NELL1 in the inflammatory response elicited by orthodontic tooth movement (OTM), providing insights into the mechanisms behind this response and establishing a theoretical foundation for the utilizing NELL1 as a therapeutic option.
Methods: Human periodontal ligament (hPDL) cells obtained from clinical samples were divided into three groups: control, compressive forces (CF), and CF+ NELL1. Inflammatory factors (cyclooxygenase 2 (COX2), interleukin-6 (IL-6), interleukin-8 (IL-8), tumor necrosis factor-α (TNF-α), inducible nitric oxide synthase (iNOS) and interleukin-1β (IL-1β)) were measured using real-time quantitative PCR (RT-qPCR) and enzyme-linked immunosorbent assay (ELISA). Culture supernatants were co-cultured with THP-1 cells to assess macrophage differentiation. Immunoblotting and Co-Immunoprecipitation (Co-IP) assays were employed to investigate the molecular mechanism of NELL1 in hPDL cells.
Results: In comparison to the CF group, the CF+ NELL1 group exhibited reduced mRNA levels and production of inflammatory factors, including COX2, IL-6, IL-8, TNF-α, iNOS, and IL-1β (p < 0.05, p < 0.01, p < 0.001, and p < 0.0001) and reduced macrophage activation (p < 0.05). Immunoblotting assays showed that NELL1 could inhibit the activation of the nuclear factor-kappa B (NF-κB) signaling pathway, consequently regulating the expression of inflammatory factors (p < 0.0001, p < 0.05, p < 0.05, and p < 0.01). Co-Immunoprecipitation (Co-IP) results confirmed the intermolecular interaction between NELL1 and inhibitor of kappa B kinase (IKK), and overexpression of IKK in hPDL cells could reduce the anti-inflammatory effect of NELL1 following CF treatment (p < 0.01, p < 0.001, and p < 0.0001).
Conclusions: NELL1 presents a promising therapeutic option to inhibit the activation of the NF-κB signaling pathway by preventing phosphorylation and reducing the inflammatory response during the orthodontic tooth movement.
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Background: Death leads to significant biochemical and morphological changes in the tissues. The lung is crucial in forensic investigations for its unique decay characteristic. DNA denaturation and caspases are vital components of the postmortem apoptotic pathway. The study aims to evaluate the role of DNA fragmentation, caspase-3 expression, and autolytic changes in the determination of postmortem interval (PMI) in the lungs of male albino rats at 0, 6, 24, 48, and 72 hours (h) after death. The study also addresses the potential implications and challenges of applying these markers in real-life forensic cases.
Methods: In the present research, 35 male albino rats were equally allocated into five groups. The sacrificed bodies were stored in a chamber at 25 ± 2 °C for 0 h, 6 h, 24 h, 48 h, and 72 h after death. At the planned schedule, a biochemical examination of lung tissue DNA content and caspase-3 expression was validated by immunohistochemical analysis and histological assessment.
Result: A statistically significant time-dependent increase in lung DNA fragmentation and caspase-3 expression (p < 0.05) with prolonged PMI time starting from 6 h postmortem to 72 h.
Conclusion: DNA fragmentation, caspase-3, and histological autolysis scoring are reliable markers for estimating time passing since death.
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Background: The diffuse large B-cell lymphoma (DLBCL) is the most prevalent lymphoma worldwide. Cyclophosphamide (CP) is an important anticancer drug used in various cancer types, including DLBCL, but its use in clinical settings is primarily limited by its dose-related cardiotoxicity. This study aimed to explore prognostic biomarkers of DLBCL in germinal center B cell (GCB) subtype, intending to develop personalized treatment regimens, monitor and assess post-treatment outcome, and improve survival rate. Furthermore, this study aimed to explore the underlying mechanism of CP and devise a more appropriate dosage plan for achieving maximum clinical benefits while minimizing toxicity.
Method: GSE87371 dataset was obtained from Gene Expression Omnibus (GEO) to screen prognostic key genes for DLBCL using univariate Cox analysis, Least Absolute Shrinkage and Selection Operation (LASSO), Kaplan-Meier survival analysis, multivariate Cox regression analysis and operating characteristic curve (ROC) with area under the curve (AUC). Differentially expressed genes (DEGs) between the high and low risk scores groups were screened and analyses of Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Set Enrichment Analysis (GSEA) were performed to obtain key pathways involved in progression of DLBCL. Furthermore, the GCB DLBCL cell lines OCI-Ly1 and VAL were cultured and treated with different concentrations of CP. Moreover, cell proliferation was determined using Cell Counting Kit (CCK)-8 and colony-forming assay, cell migration activity was assessed using cell scratch and transwell, and cell apoptosis was evaluated using flow cytometry. Additionally, RT-qPCR and Western blot analyses were used to determine the expression levels of key genes and pathways involved in DLBCL.
Results: The expressions of leucine zipper motif (APPL) isoform 2 (APPL2), histone cell cycle regulator (HIRA), zinc finger protein 814 (ZNF814), ZYG11A, leucine-tyrosine-arginine motif-containing protein 1 (LYRM1) and valosin-containing protein-interacting protein 1 (VCPIP1) were found to be correlated with the prognosis of DLBCL and were used in constructing prognostic model. It was found that patients in the low-risk group survived longer as compared to those in the high-risk group (p < 0.05). However, the nuclear factor-kappaB (NF-κB) and interleukin (IL)-17 pathways were enriched through functional analysis. Furthermore, CP induced the apoptosis in GCB DLCBL cells (p < 0.05) and inhibited their proliferation (p < 0.05) and migration (p < 0.05). Moreover, CP regulated the expression of 6 key genes. It was observed that CP treatment elevated the expression levels of APPL2, HIRA, ZNF814, and ZYG11A and decreased LYRM1 and VCPIP1, at both mRNA (p < 0.05) and protein (p < 0.05) levels. Additionally, CP decreased the activity of NF-κB (phosphorylation of p65 (p-p65) protein; p < 0.05) and IL-17 (IL-17 mRNA, p < 0.05; IL-17 protein, p < 0.05) pathways.
Conclusion: APPL2, HIRA, ZNF814, ZYG11A, LYRM1 and VCPIP1 were identified as key genes involved in the prognosis of DLBCL. Moreover, CP could up-regulate the expressions of APPL2, HIRA, ZNF814, ZYG11A, down-regulate the expressions of LYRM1 and VCPIP1, restrain both NF-κB and IL-17 pathways, suppress cell proliferation, migration while promoting cell apoptosis, thus inhibiting the progression of GCB DLBCL.
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Background: Dysregulation of glycometabolism is considered an important marker for cancer, and the glycolytic components may be targets for cancer therapy. Hence, we aimed to explore the mechanism by which miR-199a-3p influences breast cancer (BC) glycolysis.
Methods: Cell glucose and phosphofructokinase (PFK) levels in miR-199a-3p over-expressing BC cells were determined using the PFK assay kit. Glucose consumption and adenosine-triphosphate (ATP) levels were measured in miR-199a-3p over-expressing BC cells with a Glucose assay kit and ATP detection kit. The expressions of miR-199a-3p, 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3 (PFKFB3) and GATA binding protein 3 (GATA3) were measured using qPCR in GATA3 over-expressing or knockdown BC cell lines. The expressions of glucose transporter type 5 (GLUT5), lactate dehydrogenase A (LDHA), pyruvate kinase M2 (PKM2) and PFKFB3 were examined using western blotting in two miR-199a-3p over-expressing BC cell lines. Chromatin immunoprecipitation quantitative PCR (ChIP-qPCR) was used to confirm that GATA3 bound to the specific region of the miR-199a-3p promoter in BC cells.
Results: Our results showed that miR-199a-3p suppressed PFKFB3 mRNA and protein expression in BC cells (p < 0.05). Mechanistically, luciferase assays demonstrated that miR-199a-3p reduced PFKFB3 by targeting the PFKFB3 3′ untranslated region (3′-UTR) directly. Furthermore, a recovery assay indicated that PFKFB3 overexpression reversed the suppressing effects of miR-199a-3p on aerobic glycolysis (AG) (p < 0.05). More importantly, miR-199a-3p was modulated by the GATA3 transcription element by binding to the promoter.
Conclusions: The GATA3/miR-199a-3p axis might be involved in BC cell glycometabolism through regulating PFKFB3 expression, which may provide a good prognosis for breast cancer.
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Background: Pistacia atlantica Desf. oleo-gum-resin as a herbal medicine has been claimed to be effective for treating bone disorders in traditional Persian medicine and has been used by traditional practitioners and regional healers, especially in the west of Iran, to manage osteoporosis and fractures for a long time. Since no study has been performed on the effects of P. atlantica (Pistacia atlantica) oleo-gum-resin on bone and bone-related disorders, the present study aimed to investigate the therapeutic effects of standardized resin in the ovariectomy-induced osteoporosis rat model.
Materials and Methods: The essential oil of P. atlantica oleo-gum-resin was analyzed by gas chromatography. Osteoporosis was induced in rats by ovariectomy. For eight weeks, three doses of gum and its encapsulated form (50, 100 and 200 mg/kg) were given through gavage to ovariectomized rats. The body weight, serum biochemical parameters including Calcium (Ca) and serum alkaline phosphatase (ALP), Hounsfield Unit (HU) of bone, and bone biomechanical properties were measured and compared with positive control (0.1 mg/kg/day alendronate).
Results: Alpha-pinene was detected as the main constituent of oleo-gum-resin with a concentration of 127.479 mg/mL. All three doses of gum and its encapsulated form significantly improved serum calcium and biomechanical properties of bone and decreased alkaline phosphatase compared to the control group (p < 0.05). Only encapsulated form at 200 mg/kg doses resulted in Hounsfield's improvement. The nanocapsulated form showed better results on the investigated parameters than the fresh gum.
Conclusions: P. atlantica demonstrated osteoprotective effects, probably due to the high content of Alpha-pinene. Clinical trials are required to confirm its effectiveness in managing postmenopausal osteoporosis.
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Background: The enhanced proliferation and invasiveness of cancer cells have been associated with the pathophysiology of gastric cancer (GC). However, the precise role of the regulatory system has yet to be identified. The primary objective of this investigation was to ascertain the potential involvement of LOC100507144, a long non-coding RNA, in the pathogenesis of GC.
Method: A total of 370 gastric cancer cases were collected for this study. GC cell lines were cultured and authenticated. Gene knockdown and overexpression were achieved using siRNAs and lentivirus. RT-qPCR was performed to assess gene expression. Clonogenic assay, immunohistochemistry, invasion assay, and Western blot were conducted. The dual-luciferase reporter gene assay was utilized to investigate mRNA target binding. Moreover, to simulate the aggressive in vivo behavior of GC, the cell lines BGC-823 and MGC-803 were used. Gain and loss of function studies examined cell proliferation and invasion.
Results: LOC100507144 expression was substantially higher in GC tissues than in neighboring normal tissues, and this elevation was linked with tumor pathogenesis. In vitro studies demonstrated that LOC100507144 overexpression significantly increased cell proliferation and invasion, whereas LOC100507144 knockdown suppressed these effects. The expression of the enzyme phospholysine phosphohistidine inorganic pyrophosphate phosphatase (LHPP) was considerably lower in adjacent tissues, which was inversely related to the stage of GC. The LOC100507144, as per the in vitro assay, regulated LHPP expression and encouraged the GC onset and progression.
Conclusions: In summary, the present investigation has determined that the upregulation of LOC100507144 leads to the downregulation of LHPP expression, promoting the advancement of GC.
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Background: Osimertinib, a third-generation tyrosine kinase inhibitor (TKI), is associated with a complex array of resistance pathways. Epithelial-mesenchymal transition (EMT) represents a mechanism contributing to resistance development in epidermal growth factor receptor (EGFR)-TKI treatment. Previous research suggests that the downregulation of Snail, a transcription factor linked to EMT, can delay the onset of osimertinib resistance. Slug, another member of the Snail family, remains underexplored in its impact on EMT-related osimertinib resistance. Dihydroartemisinin (DHA) has demonstrated efficacy in inhibiting the proliferation, invasion, and metastasis of various types of tumor cells, including lung cancer. This study confirmed that DHA could downregulate Slug expression and inhibit the metastasis and invasion of tumor cells.
Methods: The cultured osimertinib-resistant lung adenocarcinoma cell line was named H1975/OE. Real-time polymerase chain reaction (RT-PCR), western blotting, and Transwell assays were employed to confirm EMT occurrence in H1975/OE cells. To investigate the mechanism of EMT in osimertinib resistance, we evaluated EMT-related markers in H1975 parental cells and H1975/OE-resistant cells. Complete silencing of the regulator Slug in H1975/OE cells using small interfering RNA (siRNA) was followed by an examination of alterations in the expression levels of EMT-associated markers. Finally, DHA, a natural inhibitor, was used to assess its interference with osimertinib resistance linked to EMT.
Results: Evaluation of EMT-specific markers in the H1975 parental cell line and H1975/OE resistant cell line revealed upregulated N-cadherin expression and downregulated E-cadherin expression in H1975/OE cells. The enhanced invasiveness of H1975/OE indicated that EMT occurred in the resistant cell line. Subsequent experiments confirmed that Slug expression was upregulated in the resistant cell line and that knockout of Slug facilitated a delay in the occurrence of osimertinib resistance in H1975/OE cells. Further investigation demonstrated that DHA could inhibit Slug expression, alleviating osimertinib resistance associated with EMT.
Conclusions: Slug plays a pivotal role in osimertinib resistance associated with EMT. DHA effectively overcomes osimertinib resistance linked to EMT by inhibiting Slug expression.
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Background: The primary reason for the development of chemotherapeutic intestinal mucositis induced by 5-fluorouracil (5-FU) is the activation of macrophages in the mucosa. This study aimed to explore how berberine can alleviate inflammation in macrophages and the resulting intestinal mucosal inflammation in mice induced by 5-FU.
Methods: Tohoku Hospital Pediatrics-1 (THP-1) cell inflammatory response and mouse intestinal mucositis were induced by 5-FU and treated with berberine. The levels of inflammation-related factors and autophagy related proteins in THP-1 cells were detected by reverse transcription quantitative polymerase chain reaction (RT-qPCR) and western blotting. The concentrations of double-stranded DNA (dsDNA) and interleukin-1β (IL-1β) in mice serum and the small intestine hematoxylin-eosin (HE) staining were used to evaluate intestinal mucosal damage. Immunoglobulin A (IgA), an indicator of mucosal immunity, was detected in mice serum by enzyme-linked immunosorbent assay (ELISA). We used quantitative polymerase chain reaction (qPCR) to detect the relative contents of four important strains (Bifidobacterium, Lactobacillus, Escherichia coli, and Enterococcus) in the colon contents of mice. Additionally, we employed liquid chromatograph-mass spectrometer/mass spectrometer (LC-MS/MS) technique to measure the concentrations of three main short-chain fatty acids (acetic acid, propionic acid, and butyric acid) in the mice's plasma. Furthermore, we employed ultra-performance liquid chromatography-quadrupole-time of flight-mass spectrometer (UPLC-Q Tof-MS) technique to analyze the non-targeted metabolomics of mouse serum.
Results: Berberine has been found to inhibit the expressions of NOD-Like Receptor Thermal Protein Domain Associated Protein 3 (NLRP3), Caspase-1, and IL-1β in THP-1 cells (p < 0.05). Additionally, it effectively suppresses the expression of autophagy related proteins LC3 and Beclin-1 in THP-1 cells (p < 0.05). Furthermore, in a mouse model study, berberine significantly enhances the levels of beneficial bacteria Bifidobacterium and Lactobacillus, as well as the concentrations of three main short-chain fatty acids (SCFAs) in the plasma of mice (p < 0.05). Moreover, it reduced the concentration of pro-inflammatory factor dsDNA and increased the mucosal immunity index IgA concentration in blood (p < 0.05). The untargeted metabolomics results demonstrated that berberine could regulate the inflammatory response of mice by impacting the metabolism of taurine, glycerol phospholipid, arachidonic acid, and primary bile acid biosynthesis.
Conclusions: Berberine has demonstrated its ability to effectively suppress the inflammatory reaction of THP-1 cells induced by 5-FU. Furthermore, it also influences the “intestinal flora-metabolite-inflammation” pathway by regulating the composition of the intestinal flora, increasing the production of SCFAs, reducing the expression of inflammatory factors, and preserving the structural integrity of the intestinal mucosa.
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Background: Placenta-associated 8 protein (PLAC8) has been documented to play a role in tumorigenesis and tumor progression in various malignancies. However, there is still limited understanding of its function in Osteosarcoma (OS). This study aimed to analyze the potential effects and understand the underlying molecular mechanism of PLAC8 in OS.
Methods: The expression patterns of PLAC8 were investigated in OS cell lines and clinical tissue samples. The effects of PLAC8 on cell proliferation and movement were assessed by Cell Counting Kit-8 (CCK-8), colony formation, Transwell®, and wound-healing assays. Additionally, qRT-PCR, Western blotting, and luciferase reporter assays were performed to examine the expression and explore the molecular mechanisms behind PLAC8. Finally, to further determine the effect of PLAC8 in vivo, tumorigenesis and metastasis assays were employed in a mouse xenograft tumor model.
Results: The current study revealed that PLAC8 was significantly overexpressed in clinical OS samples (p < 0.01) and in indicated cell lines (p < 0.05). In vitro experiments uncovered that PLAC8 could enhance the proliferative and metastatic probability of OS (p < 0.05). In addition, PLAC8 knockdown attenuated OS tumorigenesis in vivo. Moreover, PLAC8 was positively associated with the levels of C-X-C motif chemokine ligand 5 (CXCL5) in osteosarcoma, and PLAC8 overexpression upregulated CXCL5 mRNA (p < 0.01) and protein (p < 0.05). Mechanistically, microRNA-363-3p (miR-363-3p), suppressed by PLAC8, could directly target CXCL5 mRNA, leading to a decrease in CXCL5. Functionally, rescue and blocking assays validated that PLAC8 could promote the growth and mobility of osteosarcoma cells by releasing the targeting of CXCL5 by miR-363-3p.
Conclusions: In summary, the oncogenic role of PLAC8 contributed to the development of osteosarcoma by mediating the miR-363-3p/CXCL5 axis. The finding may serve as a valuable reference for precise treatment and therapeutic development in osteosarcoma.
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Background: Tumor-associated macrophage (TAM) is pivotal in cancer progression. Esculentoside A (EsA) is a saponin extracted from plants, which has anti-cancer and anti-inflammatory effects. However, its anti-cancer effects through inhibiting TAM activation are still unclear and need further research.
Methods: Human myeloid leukemia mononuclear cells (THP-1) differentiate into M2 macrophages under induction. The cultivation of A549 and H1299 cells was performed in a conditioned medium from M2 macrophages (M2-CM) to fathom the effects of EsA on viability of cancer cells and their abilities to migrate and invade. The macrophage markers were measured by quantitative reverse transcription polymerase chain reaction (qRT-PCR) and flow cytometry. Investigations on how EsA impacts interleukin (IL)-6/signal transducer and activator of transcription 3 (STAT3) signaling pathway were conducted by means of western blot and immunofluorescence.
Results: Adherent and rounded THP-1 cells were observed through propidium monoazide (PMA) treatment. THP-1 cells expressed M1 macrophage markers with induction of lipopolysaccharide/interferon-γ and expressed M2 macrophage markers after induction of IL-13/IL-4 (p < 0.01). EsA may dampen IL-13/IL-4-induced M2 macrophage polarization as indicated by downregulation of the mRNA levels of CD206 (M2 macrophage marker) and peroxisome proliferator-activated receptor γ (PPARγ) (p < 0.001), and inhibit IL-6 expression in cells and STAT3 phosphorylation (p < 0.05). Moreover, EsA reversed M2-CM-induced promotion of lung cancer cell migration and invasion (p < 0.05).
Conclusions: EsA inhibits lung cancer cell migration and invasion by mediating macrophage polarization through IL-6/STAT3 signaling pathway.
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Background: Breast cancer and cervical cancer are the leading causes of mortality in women worldwide, necessitating ongoing efforts to develop effective treatments. Anticancer drug development is an important aspect of cancer research, with the goal of discovering and designing compounds that target specific aspects of cancer biology while causing minimal harm to normal cells. Compounds that interact with DNA are an intriguing class of chemotherapeutic drugs used to treat cancer. This study investigated the anticancer effect of 5,6,7-Trihydroxyflavone (THF) on breast cancer and cervical cancer cells.
Methods: The anticancer potential of 5,6,7-Trihydroxyflavone (THF) against two human breast cancer cell lines (the MDA-MB231 cell line (HTB-26™) and the MCF-7 cell line (HTB-22™)) and one human cervical cancer cell line (the HeLa cell line and the CRM-CCL-2™) was explored using in-vitro cell based assays including 3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di-phenytetrazoliumromide (MTT) assay, Lactate dehydrogenase release and Caspase-3 activity followed by the Herring sperm DNA (HS-DNA) binding UV-visible spectrophotometric assay. For in-silico investigations, Molecular docking studies (using P53, Caspase, and nuclear factor-kappa B (NF-κB)) and molecular dynamic simulation studies were performed followed by the density functional theorys (DFTs) calculations.
Results: Significant growth inhibition (GI50) values of 11.1 ± 1.06, 3.23 ± 0.81, and 6.14 ± 0.96 μM were found against HeLa, MDA-MB231, and MCF-7 cells, respectively. From the LDH-release assay and Caspase-3 assay, THF was found more potential towards MDA-MB231 cells. The results showed that, compared with control, the percentage LDH release and Caspase-3 activities of THF were found statistically significant (increased) when compared between the two different concentrations (p < 0.05) and were found highly significant as p < 0.001, compared to control. THF was also discovered to bind to DNA via groove binding, with both electrostatic and non-electrostatic interactions contributing to the free energy of the binding. At the end, THF's remarkable cytotoxic potential was demonstrated by molecular docking and molecular dynamics simulations.
Conclusion: These findings suggest the potential application of THF that this could be the hypothetical compound, which would need extensive preclinical and clinical testing before being considered a viable treatment option.
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Background: Chronic low-level cadmium exposure causes nephrotoxicity in rats. The microRNA-21 (miR-21) is highly expressed in cadmium-exposed rats. The aim of this study is to investigate the mechanism of miR-21 in regulation of Transforming Growth Factor-beta 1 (Tgf-β1)/Mothers Against Decapentaplegic Homolog (SMAD) Family Member 3 (Smad3) signaling through Phosphatase and Tensin Homolog (Pten) to cause nephrotoxicity of long-term low-level cadmium exposure in rats.
Method: A rat model of long-term low-level cadmium exposure was established, and si-miR-21 lentivirus and Pten agonist (MT-141) were added for grouping. The serum and kidney tissues were extracted for pathological changes by Hematoxylin and eosin (HE) staining. The miR-21, Pten, Tgf-β1, and Smad3 were detected with PCR. Enzyme-linked immunosorbent assay (ELISA) was performed for Superoxide Dismutase (SOD), Malondialdehyde (MDA), Hydroxyproline (Hyp), Tumor Necrosis Factor-α (TNF-α), and Interleukin-10 (IL-10). The tissue apoptosis was observed by Terminal deoxynucleotidyl transferase dUTP Nick End Labeling (TUNEL).
Results: The results proved that miR-21, Tgf-β1 and Smad3 were highly expressed in cadmium-exposed rats. After miR-21 inhibition, the Pten expression increased, while Tgf-β1 and Smad3 expression and oxidative stress decreased. After Pten activation, the expression of miR-21, Tgf-β1 and Smad3 decreased, which also inhibited nephrotoxicity in cadmium-exposed rats.
Conclusions: Inhibition of miR-21 could ameliorate nephrotoxicity in rats caused by cadmium exposure, and the mechanism may be involved in the regulation of Pten.
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Background: Sophoridine has been reported to suppress multiple cancer types. However, its exact underlying mechanism on the epithelial-mesenchymal transition (EMT) in colorectal cancer (CRC) is yet to be elucidated. Therefore, this study was aimed to reveal its role in colorectal cancer (CRC).
Methods: Potential targets of sophoridine and their relevant biological functions were predicted through bioinformatics tools. To investigate the effects of sophoridine on the EMT, migration, and invasion of cells, and the Wnt signaling pathway in CRC, the CRC cells, HCT116 and SW480, were treated with sophoridine and transfected with short hairpin RNA against Zonula occludens-1 (ZO-1) (sh-ZO-1) and Cyclin D1 (CCND1) overexpression plasmid. Furthermore, the quantification analyses, including quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and Western blot, were performed to determine the expression levels of EMT-associated factors such as ZO-1, vimentin, snail, alpha-smooth muscle actin (α-SMA), and E-cadherin and proteins associated with the Wnt signaling pathway (CCND1 and β-catenin). Moreover, wound healing and Transwell assays were used to evaluate the migration and invasion within CRC cells. Additionally, xenograft assay was applied to examine tumorigenesis in-vivo.
Results: CCND1 and ZO-1 were predicted as the potential targets of sophoridine. Furthermore, sophoridine was observed to reduce the rate of migration and invasion, and decreased the levels of vimentin, snail, α-SMA, CCND1, and β-catenin while elevating the levels of E-cadherin and ZO-1 in CRC cells (p < 0.5). Moreover, ZO-1 knockdown and CCND1 overexpression reversed the inhibitory effects of sophoridine on the cell migration and invasion, as well as on the expressions of vimentin, snail, α-SMA, ZO-1, β-catenin, E-cadherin, and CCND1 (p < 0.5) in CRC cells. Additionally, ZO-1 knockdown and CCND1 overexpression were found to suppress the tumor growth in mice treated with sophoridine (p < 0.01).
Conclusion: Sophoridine exhibits inhibitory effects on the EMT, migration, and invasion and reduces tumor growth in CRC by regulating the expression levels of ZO-1 and CCND1.
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Background: Treating glioma with triptolide (TP) produces unsatisfactory outcomes. Previous studies showed that TP combined with succinic acid could sustain antitumor activity during cancer treatment. However, this activity of TP linked with succinic acid has been less investigated during the treatment of glioma. In this study, triptolide-succinic acid ester was synthesized, and its antitumor activity in glioma cells in vitro was determined.
Methods: TP was coupled with succinic anhydride (SA) to obtain triptolide-succinic acid ester (TP-SAE). Cell counting kit-8 (CCK-8), transwell, wound healing assays, and flow cytometry analysis of apoptosis were used to evaluate the antitumor activity of TP-SAE in vitro.
Results: Results from the cell counting kit-8 assay revealed that TP-SAE rapidly reduced proliferation of glioma cells compared with TP and TP + SA. Transwell and wound healing assays revealed that TP-SAE significantly decreased the invasion and migration of glioma cells compared with TP and TP + SA. The flow cytometry apoptosis assay indicated that apoptosis in glioma cells treated with TP-SAE was significantly higher than in those treated with TP and TP + SA.
Conclusions: Triptolide-succinic acid ester could inhibit the proliferation, invasion, and migration activity of glioma cells and promote apoptosis of glioma cells in vitro.
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Background and Objective: Rheumatoid arthritis (RA) is a systemic autoimmune disease that causes joint deformities and even complete loss of joint function. Artesunate (ART) is an active ingredient in Chinese herbal medicines, usually used to treat malaria. This study aimed to investigate the effect of Artesunate on collagen-induced arthritis (CIA) in rats, along with its efficacy and underlying mechanism.
Methods: The CIA rat model was created using bovine type II collagen and incomplete F-style adjuvants. The rats with CIA were divided into three groups: the Model group, the tripterygium hypoglaucum hutch (THH) group, and the ART group. The model group received 0.9% normal saline, the THH group was given a daily dose of 150 mg·kg-1 THH, and the ART group was given a daily dose of 10 mg·kg-1 artesunate for 30 days. The negative control (NC) group received only an equal volume of 0.9% normal saline. The body weight, arthritis index, and paw thickness of the rats were recorded throughout the treatment. The levels of tumor necrosis factor α (TNF-α) and interleukin 17A (IL-17A) in the serum of rats with CIA were assessed using enzyme-linked immunosorbent assay (ELISA). Furthermore, the indices of vital immune organs, including the thymus and spleen, were calculated. The hindpaw of each group was collected and subjected to histopathological examinations using hematoxylin-eosin (H&E) staining, safranin O-fast green staining and X-ray methods.
Results: The arthritis index, the thickness of the foot, and the contents of IL-17A in the CIA model group were significantly increased compared to the NC group (p < 0.05). Moreover, synovial hyperplasia and cartilage damage were significantly decreased in the ART group (p < 0.05). However, the immune organ index and serum TNF-α level did not reach statistical significance (p > 0.05). Additionally, the serum IL-17A level was significantly lower than the model group (p < 0.05).
Conclusion: Artesunate exerts a protective effect on joint damage in rats with CIA, which may reduce inflammation and delay the development of the disease by inhibiting the secretion of IL-17A.
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Background: Obesity is a polyetiological condition characterized by excessive adiposity, resulting from a disequilibrium between energy intake and energy expenditure. High-fat diets (HFDs) have been identified as key drivers of the obesity epidemic and its comorbidities. Butin, a natural flavonoid with antioxidant and oxidative stress-modulating properties, has been proposed as a potential therapeutic agent for obesity. This study investigated the efficacy of butin against HFD-induced obesity in rats.
Methods: Thirty Wistar rats were orally administered butin diluted with 0.5% sodium carboxymethyl cellulose for 30 days. The rats were randomly divided into five groups: typical diet, HFD control, HFD + 10 mg/kg butin, HFD + 20 mg/kg butin, and 20 mg/kg butin alone. At the end of the study, biochemical parameters including lipid profile [total cholesterol (TC), triglycerides (TG), high-density lipoprotein (HDL)], liver function test [alanine aminotransferase (ALT), aspartate aminotransferase (AST)], kidney test [creatinine, blood urea nitrogen (BUN)], free fatty acid (FFA), as well as antioxidant activity [superoxide dismutase (SOD), glutathione (GSH), catalase (CAT), and malonaldehyde (MDA)], were performed. Furthermore, pro-inflammatory cytokines [tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6)], concentrations of serum insulin, leptin, ghrelin, adiponectin (ADP), and fibroblast growth factor 15 (FGF15) were also evaluated.
Results: Butin exhibited substantial efficacy in mitigating HFD-induced obesity in rats. Butin administration resulted in a statistically significant reduction in the levels of TG, TC, TNF-α, IL-1β, IL-6, MDA, ALT, AST, BUN, and creatinine, while simultaneously elevating the levels of HDL, SOD, GSH, and CAT. Moreover, butin significantly modulated serum insulin, leptin, ghrelin, ADP, FFA, and FGF15 suggesting its potential for managing obesity-related metabolic and inflammatory markers. Analysis of the data revealed that butin treatment could regulate potential biomarkers and other metabolic pathways associated with HFD-induced obesity in rats.
Conclusion: Butin may attenuate hyperlipidemia and protect against HFD-induced obesity in rats through a pleiotropic mechanism of action. Additionally, butin may serve as an alternative clinical supplement to ameliorate obesity complications.
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Background: Since non-SMC condensin I complex subunit G (NCAPG) was implicated in glioblastoma (GBM), the aim of our study is to further elucidate the underlying mechanism.
Methods: The data on NCAPG expression in GBM were retrieved using bioinformatics. The assays of cell counting kit-8, scratch, transwell, tube formation, and flow cytometry were conducted to test the malignant phenotypes of GBM cells. An intradermal angiogenesis assay was additionally performed using male BALB/c nude mice to quantify new blood capillaries and to determine the formation of vessel wall. Western blot was used to detect proteins related to Phosphatidylinositol 3-Kinases/Protein Kinase B (PI3K/AKT) signaling pathway, fas-associated protein with death domain (FADD), and caspase 8/9 expression. Both quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot were used to detect NCAPG, Cyclin D1, vascular endothelial growth factor (VEGF), B-cell lymphoma-2 (Bcl-2), and Bcl-2 associated X (Bax) expression.
Results: NCAPG exhibited a high expression pattern in GBM. Overexpressed NCAPG drove the malignant phenotypes in vitro and angiogenesis in vivo. This was accompanied by increased Cyclin D1, Bcl-2, VEGF, procaspase-8/9, phosphorylation (p) of PI3K, and phosphorylated-Protein Kinase B (p-AKT) levels, while concurrently diminishing the Bax, cleaved caspase-3 and p-forkhead box O4 (FOXO4). In contrast, silencing the NCAPG produced opposite results. Further, the effects of NCAPG overexpression and knockdown were reversed by PI3K/AKT suppressor LY294002 and activator insulin-like growth factor 1 (IGF-1) (p < 0.01), respectively.
Conclusions: NCAPG plays an oncogenic role in GBM, and its silencing presents a potentially therapeutic option for patients with GBM.
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Background: Type 2 diabetes mellitus (T2DM) is a widespread metabolic disorder characterized with various multifaceted complications, among which cardiac autonomic neuropathy (CAN) stands out as a significant concern. The CAN affects the autonomic nerves that regulate heart function, thereby exposing individuals with T2DM to considerable risks. The study aims to investigate the influencing factors of T2DM combined with CAN and their predictive value based on Ewing test.
Methods: This study was conducted between March and November 2021. The clinical data of 119 T2DM patients were analyzed retrospectively. These patients were categorized into the diabetes cardiac autonomic neuropathy positive (DCAN+) and negative (DCAN-) groups based on Ewing test results. The results from venous blood test of the two groups were compared respectively. Various biochemical parameters, including fasting plasma glucose, insulin levels, and lipid profiles were determined. Logistic regression was employed to identify factors influencing T2DM combined with CAN, and the predictive performance of the model was subsequently evaluated.
Results: Patients in the DCAN+ group exhibited higher age, body mass index, diabetic duration, prevalence of coronary heart disease, and peripheral neuritis compared to the DCAN- group (p < 0.05). Additionally, DCAN+ individuals had elevated levels of serum uric acid (SUA), fasting insulin (FINS), and homeostasis model assessment-β (HOMA-β) (p < 0.05). Multivariate logistic regression highlighted age, diabetic duration, peripheral neuritis, SUA, FINS, and HOMA-β as significant risk factors for DCAN. The predictive model demonstrated good diagnostic accuracy, with HOMA-β exhibiting particularly high clinical value (p < 0.01).
Conclusions: Age, diabetic duration, peripheral neuritis, SUA, FINS, and HOMA-β emerged as influential risk factors for DCAN and demonstrated high diagnostic value with particular emphasis on the contribution of the HOMA-β. Establishing a model based on risk factors effectively predicts the risk of DCAN. Targeted measures should be taken to reduce the incidence of DCAN and improve the prognosis and quality of life of T2DM patients.
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Background: Ephedra sinica extract (ESE) exhibits anti-inflammatory, antioxidant, and neuroprotective effects. Previous studies have reported the efficacy of ESE against Alzheimer's disease (AD). Nevertheless, its mechanism is still a mystery. This research aimed to explore the effect of ESE on AD from the perspective of neuroinflammation and oxidative damage.
Methods: An AD rat model was built by injecting 10 μg of amyloid-β peptide (Aβ1-42) into the bilateral hippocampus CA1 region. Subsequently, the AD rats received intragastric administration of 100, 200, or 300 mg/kg of ESE for 4 weeks. The effects of ESE on the cognitive performance of AD rats were studied through passive avoidance and Morris water maze tests. The concentrations of inflammatory factors (Interleukin-1 β (IL-1β), IL-6, and tumor necrosis factor-α (TNF-α)) and oxidative factors [glutathione (GSH), malondialdehyde (MDA) and nitric oxide (NO)] in rat hippocampal tissue were measured with corresponding biochemical kits, and NOD-like receptor family pyrin domain containing 3 (NLRP3) inflammasome-associated protein expression levels in rat hippocampal tissue were measured by Western blot.
Results: The behavioral tests confirmed that ESE ameliorated the cognitive dysfunction of AD rats (p < 0.05 and p < 0.01). ESE treatment significantly lowered the concentration of all inflammatory factors (IL-1β, IL-6, and TNF-α) and oxidative factors (MDA and NO) (p < 0.05 and p < 0.01), whereas it increased the levels of GSH in the hippocampus of the AD rats (p < 0.05). Additionally, inhibition of NLRP3 inflammasome-associated protein expression was observed after ESE administration (p < 0.05 and p < 0.01).
Conclusion: ESE ameliorates AD rats' cognitive dysfunction by reducing neuroinflammation and oxidative stress.
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Background: Hypermethylation and downregulation of transcription factor 21 (TCF21) are frequently observed in non-small cell lung cancer (NSCLC). DNA methyltransferase 1 (DNMT1) is highly expressed in lung squamous cell carcinoma (LUSC). Accordingly, this study examines the biological roles of TCF21 and DNMT1 in LUSC cells.
Methods: Analyses concerning TCF21 expression in LUSC and the correlation between DNMT1 and TCF21 were performed using bioinformatics. After transfection of DNMT1/TCF21 overexpression plasmids, or small interfering RNAs targeting TCF21 and DNMT1 (siTCF21 and siDNMT1) into LUSC cells, the regulatory effects of TCF21 and DNMT1 on the malignant behavior of LUSC cells were verified by cell counting kit-8 assay, flow cytometry, and transwell assay. Quantitative real-time reverse transcription polymerase chain reaction or western blot was used for quantification.
Results: TCF21 expression was downregulated in LUSC tissues and cells (p < 0.05), and expressions of DNMT1 and TCF21 were negatively correlated (r = –0.09, p = 0.047). TCF21 silencing enhanced viability, migration, and invasion, reduced apoptosis, upregulated expressions of paired box 8 (PAX8), baculoviral IAP repeat containing 3 (BIRC3), B-cell lymphoma-2 (Bcl-2), X-linked inhibitor of apoptosis (XIAP), matrix metallopeptidase 9 (MMP9), N-cadherin, Vimentin, phosphorylated-protein kinase B (p-Akt) and phosphorylated-p65 (p-p65), downregulated expressions of BCL2 associated X protein (Bax) and E-cadherin, and elevated p-Akt/Akt and p-p65/p65 ratios in LUSC cells (p < 0.05). DNMT1 overexpression exerted the same effects in LUSC cells as TCF21 silencing, while DNMT1 silencing did the opposite effect. DNMT1 overexpression decreased, while DNMT1 silencing increased TCF21 expression. Additionally, DNMT1 overexpression reversed the effects of TCF21 overexpression in LUSC cells (p < 0.05).
Conclusion: DNMT1 overexpression promotes LUSC cell migration and invasion by downregulating TCF21 through activation of the Akt/p65 pathway.
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Background: Costunolide can inhibit liver fibrosis and the progression of hepatocellular carcinoma. Therefore, this study intends to explore whether Costunolide can interfere with hepatocytes simple steatosis by inducing autophagy.
Methods: Initially, Transformed Human Liver Epithelial-2 (THLE-2) cells were treated with different concentrations (0, 5, 10, 20, 30, 40, and 50 μM) of Costunolide. Moreover, a simple steatosis model of THLE-2 cells was induced by treating them with free fatty acids. Furthermore, Rapamycin was used to activate, and 3-methyladenine was used to inhibit the autophagy pathway in THLE-2 cells. Autophagy flux was measured using tandem confocal microscopy. Cell viability, lipid deposition, and triglyceride content were evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, oil red O staining, and the triglyceride content assay kit, respectively. Additionally, the protein expressions of autophagy-related genes, such as Beclin-1 and microtubule-associated protein 1 light chain 3-II and -I, as well as mitogen-activated protein kinase pathway were assessed using Western blot analysis.
Results: Costunolide treatment increased the number of autophagosomes and autolysosomes, and significantly reduced deposition of lipid and triglyceride content in free fatty acids-induced fatty liver cells (p < 0.001). However, free fatty acids inhibited Beclin-1 expression and decreased microtubule-associated protein 1 light chain 3-II/I ratio, while Costunolide treatment reversed this effect and induced autophagy in THLE-2 cells (p < 0.05). Furthermore, free fatty acids activated mitogen-activated protein kinase 14 and extracellular-signal-regulated kinase phosphorylation, thereby activating the mitogen-activated protein kinase pathway, while Costunolide treatment inhibited the activation of this pathway (p < 0.001). Additionally, Costunolide and rapamycin exhibited similar effects on autophagy in free fatty acids-treated THLE-2 cells, and 3-methyladenine reversed the effects of Costunolide (p < 0.05).
Conclusions: Costunolide interferes with simple steatosis of hepatocytes by inducing autophagy.
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Background: Acute lung injury (ALI) is an important cause of death in patients with sepsis. The imbalance of M1-M2 polarization of macrophages (MACs) is a momentous factor leading to the progression of sepsis-induced ALI. Moreover, Yes-associated protein 1 (YAP1)/Hypoxia-inducible factor-1α (HIF-1α) plays a pivotal role in regulating inflammatory response, oxidative stress, and autophagy. Therefore, this study aimed to explore whether YAP1/HIF-1α regulates sepsis-induced ALI through autophagy and polarization of macrophages.
Methods: Initially, THP-1 cells were cultured in vitro and the phenotype transformation of M1 and M2 was induced by treating them with Lipopolysaccharide (LPS) + Interferon-γ (IFN-γ) or Interleukin-4 (IL-4), respectively. The expression levels of YAP1, HIF-1α, M1 markers, and M2 markers were assessed using Western blot analysis. The impacts of YAP1 and HIF-1α overexpression on M1 markers (inducible Nitric Oxide Synthase (iNOS) and Cluster of Differentiation (CD)80), M2 markers (Arginase-1 (ARG1) and CD206), autophagy markers (Microtubule-associated protein 1A/1B-light chain 3B-II/I (LC3BII/I), Beclin1, and p62), and inflammatory factors (Tumor Necrosis Factor-alpha (TNF-α), IL-6, and IL-10) were evaluated in LPS-induced sepsis ALI cell model in vitro using cell transfection, quantitative real-time polymerase chain reaction (qRT-PCR), and Western blot analysis. Finally, the expression of LC3B in macrophages was determined using the immunofluorescence method.
Results: The expression level of YAP1 was found to be up-regulated in M2 macrophages, while the expression of HIF-1α was up-regulated in M1 macrophages. Furthermore, YAP1 overexpression promoted the transformation of LPS-induced ALI macrophages into M2 phenotype, increased the expression of autophagy factors (LC3BII/I and Beclin1), and reduced the activity of inflammatory factors (TNF-α and IL-6). However, HIF-1α overexpression could balance the M2 polarization of macrophages induced by YAP1 overexpression. Additionally, HIF-1α overexpression elevated the expression of macrophage autophagy factors (p62) and inflammatory factors (TNF-α and IL-6) and reduced the expression of macrophage autophagy factors (LC3BII/I and Beclin1) and the viability of IL-10.
Conclusions: In summary, YAP1/HIF-1α can improve sepsis-induced ALI, thereby regulating the level of macrophage autophagy and balancing M1-M2 polarization.
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Background: Despite the coexpression of Lysophosphatidic Acid-1 (LPA1) and purinergic receptor P2X, ligand-gated ion channel 3 (P2X3) receptors in vesicle-associated primary sensory neurons, their potential interaction contributing to bladder pain pathophysiology remains obscure. In this study, we aimed to investigate whether the interaction between LPA1 and P2X3 receptors in dorsal root ganglion (DRG) neurons drives the development of cyclophosphamide (CYP) -induced bladder pain syndrome.
Methods: We induced bladder pain syndrome in male C57BL/6J mice via intraperitoneal administration of CYP (300 mg/kg). The experimental mice are grouped into saline (i.t.), CYP (i.p.), LPA (i.t.), Ki16198 (i.t.), A317491 (i.t.). A combination of behavioral, neuropharmacological, molecular biological, neuroretrograde tracing, and electrophysiological techniques was employed to investigate the mechanisms of LPA1 and the P2X3 receptor in bladder pain syndrome.
Results: Administration of CYP resulted in bladder pain-like nociceptive behavior and a significant surge in urinary bladder LPA content and triggers the release of inflammatory factors Nuclear Factor Kappa-Light-Chain-Enhancer of Activated B Cells (NF-κB) and Interleukin-1 beta (IL-1β), resulting in pain-like nociceptive behavior. Nociceptive behavior was emulated through intrathecal injection of LPA (2.5 μg, 5 μL), which was significantly mitigated by the LPA1 receptor antagonist, Ki16198. α,β-meATP (100 μM) induced a significant increase in inward currents and action potential frequency in mouse DRG neurons, which was effectively counteracted by Ki16198. Direct application of LPA under physiological conditions also amplified the function of P2X3 receptors. While Ki16198 did not influence the upregulation of phosphorylated Extracellular Signal-Regulated Kinase (p-ERK) protein, it notably inhibited p38 protein. The selective p-38 antagonist SB202190 inhibited the upregulation of P2X3 recepor (P2X3R) as well as the levels of NF-κB and IL-1β, both of which were suppressed by the application of Ki16198 (i.t.).
Conclusions: In summary, our study suggests that targeting the p38 mitogen-activated protein kinase (MAPK) signaling pathway may offer therapeutic potential for alleviating bladder pain in mice by mitigating the sensitization of LPA-LPA1 receptor signaling to the nociceptive receptor P2X3R in DRG neurons.
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Background: Diabetic foot ulcer (DFU) is one of the major chronic complications of diabetes mellitus and is difficult to cure using existing conventional wound dressings. With the aim of finding more therapeutic targets for DFU, this study examined whether curcumin could improve DFU by altering the expression of C-reactive protein (CRP).
Methods: DFU rat models were successfully constructed and treated with curcumin and different CRP mediators. The effect on DFU progression was then studied by evaluating the levels of fasting blood glucose (FBG) and serum CRP, the expression of CRP in wound tissue was detected by immunohistochemical (IHC) staining. The extent of granulation tissue formation and re-epithelialization was observed by Hematoxylin-Eosin (H&E) staining, the relative density of microvessels was detected by CD31 immunofluorescence staining, and collagen deposition and tissue fibrosis were observed by Masson staining.
Results: The DFU animal model was successfully constructed and showed increased levels of FBG (p < 0.01) and serum CRP content (p < 0.01), and less wound healing (p < 0.001) on day 40. Treatment of DFU rats with 40 mg/kg curcumin (medium concentration) showed the best effect for the promotion of wound healing (p < 0.01), reducing the FBG level (p < 0.01), and reducing CRP (p < 0.01), with results even better than administration of sesame oil. Curcumin promoted the re-epithelialization of wound skin and new epidermis formation (p < 0.01), increased vascular density (p < 0.001), and increased collagen deposition (p < 0.01). These were all further improved by CRP knockdown (p < 0.05), and reversed by CRP overexpression (p < 0.01).
Conclusion: Curcumin can affect the reconstruction of granulation tissue, angiogenesis, collagen formation and deposition, re-epithelialization and formation of tissue fibrosis in the damaged skin tissue of a rat model of DFU. It does this by reducing the expression of CRP, suggesting that curcumin and CRP may have clinical value in the treatment of DFU.
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Background: Pakistan faced five distinct waves of COVID-19 caused by unique SARS-CoV-2 strains. Of these, the third wave, which started in March 2021 and ended in June 2021, proved worst causing 335,728 reported infections and 7849 deaths. During the third wave SARS-CoV-2 alpha variant remained dominant causing more than seventy percent of infections. Hence, the current study aimed to investigate the entropy rate in SARS-CoV-2 through pan-virome analysis during the third wave of COVID-19 in Pakistan.
Methods: Whole-genome sequencing of five viral isolates (three from female and two from male patients) was performed and in-silico analysis was carried out by taking into account other reported worldwide sequences. Bioinformatics analysis was done, and a phylogenetic tree was constructed to find out the clustering. Prokka was utilized to predict the genes and annotate all the viromes. The pan-genome of SARS-CoV-2 was inferred with Snippy.
Results: The phylogenetic analysis reveals that most of the genomes were identical. It was discovered that the most common change was C > T, whereas T > A substitutions were the rarest. There was a total of nine different types of mutations and four types of mutations had a significant impact on phenotypes. The four specific types of mutations identified in our study have been observed to lead to notable changes in the genetic makeup of the SARS-CoV-2 virus. Such mutations suggested a high entropy in the pan-virome of third wave SARS-CoV-2 strains.
Conclusion: This research provides valuable insights into unique SARS-CoV-2 strains and dominant variants during the third wave in the country. The phylogenetic tree and virome annotation revealed in tracking transmission patterns. These research efforts contribute to global endeavors in controlling the COVID-19 pandemic and preparing for future viral outbreaks.
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Background: Prostate cancer (PC) ranks as the fifth deadliest tumor in the world and has been regarded as a serious threat for the life and health of middle-aged and elderly men. Some studies have shown that LINC00861 regulated the anti-tumor immune response, and may have a certain impact on tumor immunotherapy. Therefore, to explore the related role of LINC00861 in PC, we detected the expression of LINC00861 in PC, and regulated the expression of LINC00861 after confirming its expression. This study aimed to explore the potential of LINC00861 in promoting the immunity of cytotoxic T cells and influencing the biological behavior of PC cells and tumor growth by regulating programmed cell death 1 ligand 1 (PD-L1).
Methods: Expression of LINC00861 and PD-L1 was detected in 10 cases of PC and adjacent tissues. Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) analysis was performed to assess the expression level of LINC00861 in cancer cells. Cell lines with differential expression were selected for interference and overexpression of LINC00861, followed by grouping. The vitality of PC cells was observed, and measurements were taken for cell proliferation, migration, invasion, PD-L1, B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X protein (Bax), and expression of inflammatory factors such as Interleukin (IL)-6, IL-8, and tumor necrosis factor (TNF)-α in the cells. To establish the subcutaneous PC tumor model, PC cells from each group were subcutaneously implanted in the right flank of BALB/c mice using 100 μL of phosphate-buffered saline (PBS). Mouse tumor tissues were collected to observe tumor size, assess tumor-infiltrating lymphocytes, and evaluate the infiltration of cytotoxic T cells (CD3+ and CD8+).
Results: In PC tissues and cells, PC-3 cells were selected for subsequent experiments through cell screening. Successful transfection of LINC00861 inhibition and overexpression vectors was achieved in PC cells. Inhibiting the expression of LINC00861 upregulated the expression of PD-L1, enhanced cell growth vitality, proliferation, migration, invasion, and cytokine production capabilities, and reduced apoptosis. Overexpression reversed these effects. In in vivo experiments, inhibiting LINC00861 expression resulted in rapid tumor growth, accompanied by increased tumor volume and dense tumor tissue cells. Overexpression of LINC00861 produced the opposite results for these effects. The findings also indicated that inhibiting LINC00861 led to a significant increase in PD-L1 expression in tumor tissues and a significant decrease in CD3+ T and CD8+ T levels (p < 0.05). When LINC00861 was overexpressed, PD-L1 levels were reduced, while CD3+ T and CD8+ T levels were improved.
Conclusions: LINC00861 is down-regulated in PC tissues. The inhibition of LINC00861 increases the expression of PD-L1, inhibits cytotoxic T cells immunity, and enhances the growth of PC cells and tumor growth and metastasis. Overexpression inhibits the development of PC.
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Background: Nitrogen permease regulator-like 2 (NPRL2) acts as an effective gene for diverse tumor suppression and its expression is up-regulated in castration-resistant prostate cancer (CRPC). However, potential mechanisms of NPRL2 in bladder cancer (BCa) progression remain to be discovered. To explore the function and molecular mechanism of NPRL2 in the development and progression of BCa, bioinformatics analysis and verification were performed.
Methods: Extensive bioinformatic analysis explored NPRL2 expression associated with BCa using data from the Cancer Genome Atlas (TCGA) database. BCa cell lines (T24 and 5637 cells) were used to verify the function and molecular mechanism of NPRL2 in BCa, and they were divided into four groups: NC-CMV-NPRL2 (transfected with PDS237_pL-CMV-GFP vector), CMV-NPRL2 (transfected with PDS237_pL-CMV-GFP-TUSC4), NC-sh-NPRL2 (transfected with pl6.3-SHRNA-BSD vector), and sh-NPRL2 (transfected with pl6.3-SHRNA-GFP-NPRL2). The proliferation, invasion, migration, cell cycle and apoptosis of T24 and 5637 cells were further evaluated by Cell Counting Kit-8 (CCK-8), Transwell, flow cytometry and Western blot assays. Nude mice were used to establish xenotransplantation models, and three groups (control, sh1-NPRL2, sh2-NPRL2) were developed. The control group was subcutaneously injected with T24 cell suspension. The sh-NPRL2 groups were subcutaneously injected with two groups of T24 cells that had been stably transduced with NPRL2 knockout genes.
Results: NPRL2 gene expression exhibited remarkable upregulation in BCa (p = 0.004). Overexpression of NPRL2 enhanced the proliferation and invasion of T24 and 5637 cells, while simultaneously inhibiting apoptosis. Furthermore, NPRL2 deficiency in the xenotransplantation model exhibited inhibition of tumorigenesis in vivo. Additionally, Immunoprecipitation and Shotgun Liquid Chromatography-Mass Spectrometry (LC-MS) analysis revealed a protein interaction between NPRL2 and alpha-enolase (ENO1) which was highly expressed in BCa. ENO1 had a noticeable effect on BCa stage and survival by promoting cell proliferation, epithelial-mesenchymal transition (EMT), small mothers against decapentaplegic (SMAD)2/3 phosphorylation, and ENO1's stability was enhanced by NPRL2.
Conclusions: The present study reveals that NPRL2 is upregulated in BCa and can enhance SMAD2/3 phosphorylation by upregulating ENO1. NPRL2 is a potential biomarker for BCa diagnosis and prognosis.
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Background: Licoricidin (LCD) is a bioactive compound isolated from licorice with anticancer properties. This study aimed to investigate the effect of LCD on the biological behaviors of hepatocellular carcinoma (HCC) and to explore its underlying molecular mechanisms.
Methods: The effect of LCD, at gradient concentrations (0, 5, 10, 15 and 20 μM), on the viability of human normal liver epithelial cells (THLE-3) and HCC cells was assessed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Moreover, the impacts of LCD, at different concentrations (0, 5 and 10 μM), on the HCC cell cycle, apoptosis, proliferation, and migration were evaluated using flow cytometry, colony formation experiment, and Transwell assay. Furthermore, the levels of cell cycle-related proteins, protein kinase B (AKT)/p21 pathway-related proteins, and pyruvate dehydrogenase kinase isoform 1 (PDK1)-related proteins were assessed using western blot assay and quantitative real-time polymerase chain reaction (qRT-PCR). Rescue experiments were performed to conform whether the LCD works through PDK1.
Results: It was found that cell viability remained unaffected in THLE-3 cells but reduced in HCC cells (p < 0.001) following LCD treatment. Furthermore, LCD hampered the proliferation and migration, boosted apoptosis, and induced cell cycle arrest in the G0/G1 phase of HCC cells (p < 0.05). Moreover, LCD reduced the expression levels of B-cell lymphoma-2 (Bcl-2), cyclin A, cyclin-dependent kinase 2 (CDK2), and phosphorylated-AKT (p-AKT) while elevating Bcl-2 associated X protein (Bax) and p21 levels (p < 0.05). Additionally, LCD decreased PDK1 expression level, but overexpression of PDK1 reversed its regulatory impacts on the AKT/p21 pathway (p < 0.001). Moreover, overexpressed PDK1 counteracted the impacts of LCD on repressing the proliferation and migration and boosting cell cycle arrest in the G0/G1 phase and apoptosis of HCC cells (p < 0.05).
Conclusion: LCD hinders the proliferation and migration of HCC cells and enhances the apoptosis and cycle arrest at the G0/G1 phase by modulating the PDK1/AKT/p21 pathway.
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Background: Acute kidney injury (AKI) is a major public health concern, leading to high morbidity and mortality. Despite decades of research, the pathogenesis of AKI remains incompletely clear, and effective therapies and predictive biomarkers are lacking. Ferroptosis is a novel regulated cell death associated with many diseases. This study aims to identify ferroptosis-related microRNA (miRNA) regulatory networks in AKI patients.
Method: Single-cell RNA (scRNA)-sequencing data obtained from the kidney and urine of AKI patients underwent bioinformatics analysis to determine the ferroptosis-related miRNA regulatory networks in patients with AKI. Pseudotime trajectory analysis was performed to assess the relationship between the developmental trajectory of the proximal tubule (PT) cells and AKI cells. The competing endogenous RNA (ceRNA) network was constructed using the ferroptosis-associated messenger RNA (mRNA), microRNA (miRNA), and long non-coding RNA (lncRNA).
Results: The pseudotime trajectory analysis showed that the developmental trajectory of PT cells exhibited the potential for AKI cell transformation. Ferroptosis pathway enrichment was co-expressed with the developmental trajectory of PT cells. Finally, the identified candidate miRNA screened from three different models (has-miR-3144-3p) was associated with the ferroptosis related ceRNA network with related mRNAs and lncRNAs.
Conclusions: In this study, the single-cell data of PT cells derived from urine in AKI patients effectively simulates the developmental trajectory of AKI. Meanwhile, the pseudotime trajectory analysis demonstrated ferroptosis pathway enrichment is co-expressed with the pseudotime trajectory analysis. Therefore, the ferroptosis-associated ceRNA network constructed is helpful for further research.
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Background: Tissues surrounding ruptured aneurysms exhibit heightened inflammatory activation. Oxidative stress is a pivotal factor in endothelial injury, where moderate and high concentrations of reactive oxygen species induce apoptosis via oxidative stress. Furthermore, oxidative stress and reactive oxygen species have been implicated in vascular smooth muscle cells (VSMC) phenotypic switch and apoptosis. This study aimed to investigate the impact of N-acetyl-L-cysteine (NAC) in a rat aneurysm model subjected to a high salt diet and surgical intervention.
Methods: Rats were randomly divided into four groups: normal control, surgery alone, surgery with low-dose NAC, and surgery with high-dose NAC. A cerebral aneurysm model was induced in the latter three groups by ligating the posterior branches of bilateral renal arteries and the left common carotid artery and administering a high salt (8% NaCl) diet for 3 months. The control group received a high salt (8% NaCl) diet only. Inflammatory factor levels were assessed using enzyme-linked immunosorbent assay (ELISA), and related protein expression was examined via western blot.
Results: Treatment with NAC significantly delayed the progression of cerebral aneurysms, exerting an inhibitory effect on inflammation. Expression levels of nucleotide-binding oligomerization domain, leucine-rich repeats and pyrin domain-containing protein 3 (NLRP3), apoptosis-associated speckled protein (ASC), Caspase-1, interleukin-6 (IL-6), and tumor necrosis factor (TNF) proteins in cerebral aneurysm tissues were markedly reduced. NAC treatment led to a significant decrease in reactive oxygen species (ROS) levels, particularly in the high-dose group. Transmission electron microscopy of the ultrastructure of the cells in each group revealed that cells in the aneurysms of the model group were charred and dead, while those in the arterial tissues of the NAC group exhibited intact nuclei and cytoplasm with rounded edges.
Conclusion: NAC can diminish ROS production in cerebral aneurysm model rats, attenuating the oxidative stress and inflammatory response in cerebral aneurysms. This effect contributes to the reduction and delayed onset of cerebral aneurysms, ultimately improving the prognosis of cerebral aneurysms. Additionally, NAC may partially inhibit the expression process induced by ROS-mediated NLRP3 inflammasome.
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Background: Cisplatin-based therapy remains a critical treatment for patients with advanced lung adenocarcinoma (LUAD). However, the development of resistance to cisplatin hampers the sustained benefit to patients. Recent studies have shown that molecules secreted by stromal tissue play a pivotal role in tumor chemoresistance. This study aimed to validate the role of CXC-chemokine ligand 14 (CXCL14), a stromal-secreted molecule, and its functional mechanism in cisplatin resistance.
Methods: We analyzed The Cancer Genome Atlas (TCGA) database using bioinformatics methods, leading to the selection of CXCL14 for validation on a LUAD tissue microarray (TMA) of 120 patients. The protein expression of CXCL14 was assessed through immunohistochemistry (IHC). A549 and H3255 cells were cultured in two-dimensional (2D) or three-dimensional (3D) patterns. In this study, we evaluated cell viability, cell apoptosis and death, molecular expression, signaling pathways, and xenograft growth using in vitro methods such as Cell Counting Kit-8 (CCK8) assays, colony formation assays, flow cytometry, cell live/dead double staining, quantitative PCR, Western blot, RNA sequencing, and in vivo mouse tumor-bearing models.
Results: CXCL14 was found to be highly expressed in the stroma of lung adenocarcinoma in both the TCGA training cohort and the TMA validation cohort, suggesting its potential role in the interaction between stromal and tumor cells. Cisplatin inhibited cell viability, induced cell apoptosis in 2D or 3D cultured A549 and H3255 cells in vitro, and suppressed tumor growth in vivo. These effects were significantly alleviated by adding CXCL14. Further investigation revealed that CXCL14 promoted cisplatin resistance by upregulating Angiopoietin Like Protein 4 (ANGPTL4), activating the downstream extracellular regulated protein kinases (ERK) signaling pathways.
Conclusion: CXCL14, a stromal marker, promotes cisplatin resistance in LUAD by activating ANGPTL4 and the downstream ERK signaling pathway. Identifying the downstream effectors, or the use of neutralizing/non-neutralizing antibodies targeting this pathway may provide valuable insights and serve as a reference for developing strategies to effectively control cisplatin resistance, a topic that warrants further investigation.
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Background: The comprehensive role of salmonella pathogenicity island 1 (SPI1) in neural stem cells (NSCs) remains unclear. Therefore, this study aimed to explore the effect of SPI1 on the differentiation of NSCs.
Methods: NSCs were administered with lipopolysaccharides (LPS, 100 ng/mL) to construct an NSC model in vitro. Relative mRNA expression levels of SPI1 and fat mass and obesity-associated protein (FTO) were assessed using quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Protein levels of phosphor (p)-janus kinase 1 (JAK1), p-signal transducer and activator of transcription 3 (STAT3), and interleukin-6 receptor (IL6R) were determined using Western blot analysis. Furthermore, the viability of NSCs was assessed using a cell counting kit-8 assay. NSC identification and neurite outgrowth were observed using immunofluorescence. The N6-methyladenosine (m6A) modification sites of IL6R were determined using the RNA Modification Base Database (RMBase). Additionally, the correlation between FTO and IL6R was confirmed through RNA-binding protein immunoprecipitation-qRT-PCR.
Results: NSCs exhibited an increase in mRNA level of SPI1 over time, reaching to peak at day 12 (p < 0.001). Moreover, LPS treatment significantly reduced the expression of SPI1 and neurite growth, and inhibited the JAK1/STAT3 pathway, while enhancing the expression of FTO (p < 0.01). The inhibitory impact of LPS was enhanced by silencing SPI1, while SPI1 overexpression reversed its inhibitory effects on NSCs (p < 0.05). Furthermore, IL6R was predicted to contain m6A modification sites. The increased FTO expression in LPS-treated NSCs inhibited the growth of neurite, the expression levels of the JAK1/STAT3 pathway, and IL6R (p < 0.01). Moreover, silencing SPI1 enhanced the effect of FTO, while overexpression of SPI1 reversed this effect (p < 0.05).
Conclusions: SPI1 negatively regulates FTO expression, thereby elevating m6A modification of IL6R, hence promoting NSC differentiation. This finding presents a novel strategy for propelling the clinical application of NSCs.
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Background: Pyroptosis has been proved to play a role in vision impairment of acute glaucoma, a type of optic neuropathy caused by an abrupt increase in intraocular pressure, leading to irreversible visual loss. The objective of this study was to probe into the protective effect of pyroptosis inhibitor necrosulfonamide (NSA) on vision loss in acute glaucoma and its potential mechanism.
Methods: An acute glaucoma model in rats was established by anterior chamber perfusion. The expression of pyroptosis proteins was estimated through western blot and quantified real-time polymerase chain reaction (qRT-PCR), while the interleukin (IL)-1β and IL-18 levels in retinal vitreous tissues were determined using enzyme-linked immunosorbent assay (ELISA). The retinal tissues were examined by electron microscopy, and performed immunofluorescence staining. The rats were given different doses of NSA, and the visual function was assessed by flash visual evoked potentials and full field electroretinogram analysis. The retinal structure was evaluated by optical coherence tomography peripapillary analysis and hematoxylin and eosin (H&E) staining. In vitro, Müller cells were modeled by oxygen-glucose deprivation/reperfusion (OGD/R) and treated with NSA. The effects of NSA on cell proliferation and pyroptosis-related proteins were evaluated using cell counting kit-8 (CCK-8) assay, qRT-PCR, and western blot.
Results: After modeling, the acute glaucoma rats significantly increased the key proteins and inflammatory factors associated with pyroptosis (p < 0.05) and typical features of pyroptosis were observed. NSA treatment alleviated visual impairment and improved retinal damage in rats with acute glaucoma, and reduced the expression of IL-1β and IL-18 by inhibiting gasdermin-D (GSDMD). In the OGD/R model of Müller cells, the cell proliferation was inhibited, and the expressions of NOD-like receptor family pyrin domain containing 3 (NLRP3), absent in melanoma 2 (AIM2) inflammasome, Cleaved-GSDMD, IL-1β, and IL-18 were upregulated, while NSA increased cell proliferation and inhibited pyroptosis.
Conclusions: Pyroptosis was involved in the pathogenesis of acute glaucoma in rats, and NSA can protect visual function by inhibiting pyroptosis. Therefore, NSA has great potential in treating acute glaucoma.
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Background: CDP-diacylglycerol synthase 1 (CDS1), a functional protein regulating cell growth and lipid synthesis, is implicated in cancer development. Despite its potential role, the impact of CDS1 in hepatocellular carcinoma (HCC) remains elusive. This study aimed to explore the role of CDS1 on HCC progression. Finally, we elucidated whether CDS1 affects HCC pathogenesis and its associated signal transduction.
Methods: CDS1 expression was assessed in tissue samples, correlating with clinical characteristics and prognostic outcomes in HCC. Vector, CDS1 overexpression, si-CDS1-2, and si-CDS1-3 groups were established using primary liver cancer tissues, adjacent normal tissues, human HCC cells, liver epithelial cells, and RNA/proteins extracts. In vitro and in vivo experiments were performed to determine the biological function and molecular basis of CDS1, including Cell Counting Kit-8, cell cycle, and 5-ethynyl-2′-deoxyuridine assays for proliferation, Transwell and Boyden assays for invasion and metastasis. Moreover, apoptosis assay, quantitative real-time polymerase chain reaction, and western blotting were shown.
Results: Analysis of The Cancer Genome Atlas datasets revealed a significant increase in CDS1 expression in HCC, suggesting a potential influence on liver cancer cell growth, invasion, and apoptosis. Immunohistochemical analysis revealed elevated CDS1 protein levels in HCC samples compared to adjacent tissues. Furthermore, western blotting revealed a plausible carcinogenic role of CDS1 in HCC linked to the RAC-Alpha Serine/Threonine-Protein Kinase (AKT)-Mechanistic Target of Rapamycin (mTOR) signaling pathway. Finally, experimental findings confirmed that CDS1 promotes HCC proliferation, invasion, and metastasis while inhibiting apoptosis, potentially through the AKT-TSC2-mTOR pathway.
Conclusions: CDS1 modulates the proliferation, migration, and invasion of liver cancer cells, impeding normal apoptotic processes. The underlying mechanism includes the activation of the AKT-TSC2-mTOR signaling pathway.
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Background: Hepatitis C virus (HCV) is a single-stranded enveloped virus with a ribonucleic acid (RNA) genome of approximately 9.6 kb that predominantly infects the liver. The main objective of the current research was to determine the unknown HCV genotypes by analyzing the 5′-untranslated region (5′-UTR) to identify different genotypes and classify them by their phylogenetic relationships.
Methods: Serum samples (104) were taken from untyped chronic HCV isolates, and cDNA was synthesized using reverse transcriptase enzyme. Nested polymerase chain reaction (PCR) was used to amplify the 5′-UTR followed by agarose gel electrophoresis. The 5′-UTR amplified products were sequenced by the Sanger method. Evolutionary analysis was performed using Molecular Evolutionary Genetics Analysis Version 7 (MEGA7) software.
Results: 104 untyped chronic HCV patients were studied, representing the higher frequency of males (62%). Out of the total, nine samples of 5′-UTR products were sequenced for genotype confirmation. In the phylogenetic analysis, genotype 3 was reported to be 89% (3a 77%, 3b 11%) and 11% unknown genotype. Overall, in the phylogenetic tree, the 5′-UTR sequences were successfully clustered in a clad with the reference sequences of genotype 3a, and 3b, while one sequence showed an unknown genotype.
Conclusions: The study concludes that genotype 3 is predominant, and identification of genotypes by this procedure is an accurate and alternative method.
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Background: LINC01048 is a recognized oncogene. LINC01048 regulates cellular functions through various mechanisms, affecting gene expression, activating cellular signaling pathways, and modulating the cellular functions. The primary aim of this research is to explore how LINC01048 influences the biological function of bladder cancer (BC) and the extracellular matrix (ECM), along with its regulatory role in the yes-related protein 1 (YAP1)/Wnt/β-catenin pathway.
Method: Data from The Cancer Genome Atlas (TCGA) data was utilized to acquire lncRNA expression and clinical information for bladder cancer (BC). Differential analysis compared normal and tumor groups, followed by survival analysis for the target lncRNA. Gene Set Enrichment Analysis (GSEA) identified regulatory pathways associated with LINC01048. Clinical samples were collected to measure LINC01048 expression in BC patients, and its correlation with clinical features was analyzed. In BC cell lines and normal bladder cells, LINC01048 mRNA expression was measured by Real-Time Quantitative Polymerase Chain Reaction (RT-qPCR). Two groups of cells, with high and low LINC01048 expression, were transfected, and post-transfection levels were quantified. Cell proliferation, apoptosis, ECM receptor-related proteins (Collagen I, α-smooth muscle actin (α-SMA), Integrin alpha 6 (ITGA6), CD44), and pathway proteins (YAP1/Wnt/β-catenin) were assessed using Cell Counting Kit-8 (CCK-8), clonogenic formation, flow cytometry, and Western blot. Stable cell lines with high or low LINC01048 expression were established and injected into nude mice to create a BC model. Changes in tumor volume and mass were monitored among different transfection groups. LINC01048 expression in tumor tissues and pathway protein levels were measured.
Result: LINC01048 was significantly overexpressed in both BC tissues and cell lines, showing a strong association with the ECM receptor interaction pathway. Overexpression enhanced BC cell growth, increased ECM receptor interaction proteins, and elevated YAP1/Wnt/β-catenin pathway proteins while decreasing apoptosis. si-LINC01048 transfection yielded opposite effects (p < 0.05). In a nude mouse BC model, overexpression led to significantly larger tumor volumes, higher expression of ECM receptor interaction proteins, and upregulation of YAP1/Wnt/β-catenin pathway proteins compared to the control group. si-LINC01048 led to smaller BC volumes, reduced expression of ECM receptor interaction proteins, and downregulation of YAP1/Wnt/β-catenin pathway proteins (p < 0.05).
Conclusion: In bladder cancer, LINC01048 promotes the expression of ECM receptor interaction proteins and cellular behaviors by activating the YAP1/Wnt/β-catenin signaling pathway.
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Background: Head and neck squamous cell carcinoma (HNSCC) is associated with a substantial incidence and metastasis rate. G protein subunit alpha 13 (GNA13), a member of the G protein family, is closely associated with the development of multiple tumors. We intended to explore the function and mechanism of GNA13 on the malignant phenotype of HNSCC.
Methods: The biological effects of GNA13 on HNSCC cells were analyzed by immunoblotting and PCR. The effects of GNA13 on fibroblast activity under the action of exosomes were investigated by isolating exosomes from HNSCC cells and co-culturing them with fibroblasts. The molecular pathway by which GNA13 regulates the miR-26a-5p/STE20-related adaptor beta (STRADB) axis through exosomes on fibroblasts was confirmed by in vitro experiments. The influence that fibroblasts overexpressing STRADB have a malignant phenotype on HNSCC cells was confirmed using a co-culture system. The promotion of cancer cell growth in nude mice by GNA13 was verified by in vivo experiments.
Results: GNA13 was found to be an independent prognostic factor regulating the growth of HNSCC. GNA13 induced fibroblast transformation and promoted its growth and migratory activity through exosomal regulation of the miR-26a-5p/STRADB axis. Meanwhile, fibroblasts overexpressing STRADB reversed the effect of GNA13 deletion upon the malignant phenotype of HNSCC cells.
Conclusions: GNA13, as a key gene, significantly affects the malignant phenotype of HNSCC and influences fibroblast activity via the exosome/miR-26a-5p/STRADB axis, further altering the microenvironment in which the tumor resides. GNA13 may be a promising prognostic biomarker among patients with HNSCC.
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Background: Microsatellite instability (MSI) status may alter the tumor microenvironment (TME) and can affect the effectiveness of immune checkpoint blocking (ICB) immunotherapy in colorectal cancer (CRC) patients. This study aimed to determine the MSI-related genes as biomarkers for response to ICB therapy in CRC.
Methods: The role and signature of MSI-related genes in CRC were analyzed through the data acquirement (LinkedOmics database), copy number alteration (CNA) assay (cBioPortal database), Association immune cells and Troponin C2 (TNNC2) levels analysis (Immune Cell Abundance Identifier (immuCellAI)), Tumor Immune Estimation Resource (TIMER) and Gene Expression Profiling Interactive Analysis (GEPIA). Silencing of TNNC2 was constructed in vitro and used for cell count kit 8 (CCK-8). CCK-8 was used to detect cell growth after relevant treatments. The cell apoptosis was assessed using the Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) apoptosis detection kit. The wound healing assay was used to determine cell migration. The enrichment of the TNNC2 pathways was performed using Gene Set Enrichment Analysis (GSEA).
Results: The results showed that a total of 15 MSI-related genes were identified, including TNNC2 and ACSL6. Then, TNNC2 high-frequency amplification was found in the CRC. The expression of TNNC2 was lower in MSI-high group than MSI-low or microsatellite stability (MSS) group (p < 0.05). Compared to the high expression of TNNC2, the low expression of TNNC2 is more conducive to ICB response (p < 0.05). Additionally, TNNC2 regulated immune cell infiltration and ICB immunotherapy-related genes. Furthermore, TNNC2 was upregulated in CRC tissues and cell lines (p < 0.05). Cellular function assays showed that silence of TNNC2 decreased cell proliferation and migration, and induced apoptosis in CRC cells. Biological analysis also showed that TNNC2 plays a critical role in the activation of Wnt/β-catenin and PPAR in CRC.
Conclusion: Hence, it can be concluded that TNNC2 is an MSI-related gene in response to ICB therapy in colorectal cancer. This study provided a better understanding of potential biomarker of ICB response and underlying mechanism in CRC, which is beneficial for CRC diagnosis and treatment research.
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Background: Breast cancer has become the most common cancer worldwide. Early detection of metastatic events is vital for improving survival. Currently, the serum tumor markers for monitoring metastatic breast cancer in clinic practice are carbohydrate antigen (CA153) and carcinoembryonic antigen (CEA) based on the 2007 edition of American Society of Clinical Oncology (ASCO) guidelines. This study aims to identify more specific, sensitive, and cost-effective tumor markers to improve the accuracy of predicting distant metastasis of breast cancer.
Methods: A total of 468 breast cancer patients, including 170 patients with distant metastasis and 298 without metastasis, were retrospectively reviewed. The levels of the serum tumor markers ferritin, CA153, and CEA in both the metastasis and non-metastasis groups, in subgroups of different pathologic types, different metastatic sites, and different tumor burden, were analyzed via one-way analysis of variance and the receiver operating characteristics (ROC) curve. The differences between ROC curves were compared using the paired comparison method.
Results: High serum ferritin levels predict distant breast cancer metastasis with the highest specificity and sensitivity among all three markers, especially in hormone receptor (HR)-/ human epidermal growth factor receptor 2 (Her2)+ and HR-/Her2- molecular subtypes. The prediction accuracy for metastasis increased when a combination of two or all three markers, and the combination of ferritin and CA153 could be the optimal choice to maximize the efficiency and predictive ability for metastasis. The baseline levels of all three markers were associated with metastasis tumor burden and could be more sensitive in predicting patients with liver metastasis than patients with lung and bone metastases.
Conclusions: The findings obtained from this study suggest that serum ferritin is a putative candidate as a non-invasive biomarker for monitoring breast cancer metastasis that is even more sensitive and specific than the traditional breast tumor markers CA153 and CEA. However, further studies with a more extensive set of patient data are needed to validate the conclusions.
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Background: Berberine (BBR) is an isoquinoline alkaloid derived from Coptis chinensis. It exhibits various biochemical and pharmacological effects, including the improvement of hyperglycemia, attenuation of insulin resistance and inhibition of lipid synthesis. These properties have led to its widespread use in clinical practice.
Objective: This study was designed to investigate the effect of berberine (BBR) on cerebral ischemia/reperfusion injury (CIRI) and delineate the relevant mechanism involving Toll-like receptor 4 (TLR4)/nuclear factor kappa B (NF-κB) signaling pathway.
Materials and Methods: Fifty male Sprague Dawley (SD) rats were randomly divided into sham operation group (SG), model group (MG), berberine-low dose group (BBR-L), berberine-medium dose group (BBR-M) and berberine-high dose group (BBR-H). Ten rats were assigned to each group. The rats in BBR-L, BBR-M and BBR-H were given 25, 50, 100 mg/kg BBR, respectively, via intragastric administration, while the rats in SG and MG groups were given equal volume normal saline once a day for 5 days. CIRI rat model was constructed using the suture-occluded method. The rats in each group were scored for neurological function using the Zea Longa method. The changes in the serum levels of tumor necrosis factor alpha (TNF-α), interleukin (IL)-6 and IL-1β were detected with enzyme-linked immunosorbent assay (ELISA). Western blotting was employed to determine the expression levels of proteins.
Results: Compared with the MG, the BBR-treated groups exhibited a lower degree of neurological deficits, brain infarct volume and apoptosis rate of brain tissues. The BBR-treated groups exhibited a lower degree of neurological deficits in CIRI rats. The serum levels of TNF-α, IL-6, IL-1β and malonaldehyde, as well as the expressions of TLR4 and NF-κB/p65 in brain tissues, were reduced following BBR administration, while superoxide dismutase activity in serum was increased (p < 0.05).
Conclusion: In conclusion, BBR exerts a protective effect against CIRI in rats by reducing the production of inflammatory factors via the suppression of the TLR4 and NF-κB/p65 signaling pathway.
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Background: Asiaticoside has been affirmed to enhance the healing process of burn wounds, but the underlying molecular mechanism needs to be further elucidated. Therefore, this study aims to investigate the mechanism by which asiaticoside affects the burn serum-induced human dermal microvascular endothelial cell (HDMVEC) injury.
Methods: Sprague-Dawley (SD) rats (n = 20) were divided into two groups: the burn group and the Sham group. Rats in the burn group were exposed to an electrically heated brass rod. HDMVECs were transfected with small interfering RNA targeting alpha-actinin-4 (ACTN4) (siACTN4) and negative control of siACTN4 (siNC). Burn serum-induced HDMVEC injury was used to mimic microvascular injury in vivo. Furthermore, HDMVECs were treated with asiaticoside and SB203580 (p38 mitogen-activated protein kinase (MAPK) specific inhibitor). The monolayer permeability in HDMVECs was evaluated using transendothelial electrical resistance, and cell viability was assessed using cell counting kit-8 (CCK-8) assay. The localization of zonula occludens-1 (ZO-1) and filamentous actin (F-actin) was determined through immunofluorescence staining. Moreover, the mRNA/protein expression levels of ACTN4, phosphorylated (p)-p53, p53, p-p38, and p38 in treated HDMVECs were evaluated using quantitative real-time PCR (qRT-PCR) and Western blot analysis.
Results: Asiaticoside negated the inhibitory impacts of burn serum on viability and F-actin levels in HDMVECs (p < 0.05) as well as the promotive impacts of burn serum on monolayer permeability and ZO-1 levels. The expression of ACTN4 at both mRNA and protein was downregulated in burn serum-induced HDMVECs and upregulated by asiaticoside (p < 0.05). Asiaticoside reduced the protein expression levels of p-p53/p53 and p-p38/p38 and level of ZO-1, suppressed the monolayer permeability as well as promoted F-actin level and viability in burn serum-induced HDMVECs, all of which could be reversed by ACTN4 silencing (p < 0.05). Furthermore, SB203580 further offset the impacts of ACTN4 silencing on the protein expression levels of p-p53/p53 and p-p38/p38, levels of ZO-1 and F-actin as well as the monolayer permeability in burn serum-induced HDMVECs (p < 0.05).
Conclusions: These findings suggest that asiaticoside enhances the healing process in burn serum-induced HDMVEC injury by regulating the ACTN4-mediated p38-MAPK/p53 signaling pathway.
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Background: Heat shock proteins (HSPs) are essential for the growth of various cells. The development of automated and precise machine-learning tools for the quick prediction of HSPs is significant because conventional methods are expensive, and there is abundant protein sequence information accessible in the post-genomic era, which can be easily used to develop machine-learning based tools.
Methods: The proposed method utilized the already available dataset from the PredHSP tool. The Composition of k Spaced Amino Acid Pair feature was calculated using iFeature, and an enhanced sampling technique was proposed for the balanced dataset. Six machine learning models were developed to predict HSPs, and their robustness was assessed using a ten-fold cross-validation technique. The best model was finalized among six machine learning models and evaluated by the following metrics: accuracy, precision, recall, f1 score, and area under the curve (AUC). The command line utility for the use was available as a GitHub repository.
Results: Six machine learning models were assessed with ten-fold cross-validation, and support vector machine (SVM) outperformed with a higher overall accuracy (87%) compared to existing methods, which can predict all HSP types in one run. For usage, the model was deployed as a command line utility in GitHub.
Conclusions: Machine learning is a powerful method to predict the HSPs by identifying hidden patterns inside the sequence. HSPs are important chaperons in research, and their quick prediction will function in many important aspects of biological research. This model helps to predict the heat shock protein in eukaryotes.
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Background: Most significant wounds often exhibit poor healing. Hence, the exploration of novel approaches to enhance skin wound healing (SWH) and further investigate potential underlying mechanisms remains crucial. Our aim was to delineate the collective impact of concentrated growth factor (CGF) and hypoxia-pretreated adipose-derived stem cell (ADSC) sheets on SWH and investigate the underlying mechanism.
Methods: The optimal concentration of CoCl2, a hypoxia-inducing reagent, and CGF extract for cell proliferation and ADSC growth factor production in rats was determined using dose gradient experiments. Mice were categorized into six groups according to ADSC treatment: (A) control, (B) ADSC sheets, (C) ADSC sheets treated with CoCl2, (D) CGF particles, (E) CGF and ADSC sheets, and (F) CGF and ADSC sheet combination treated with CoCl2. The impact of CGF and hypoxia on ADSC proliferation was investigated through western blotting and Cell Counting Kit-8 (CCK-8) assay. Immunohistochemistry, Masson staining, and hematoxylin and eosin staining were conducted ten days after the surgical procedure to assess the effects. Key markers for skin wound healing were evaluated, and the underlying mechanism in human ADSCs was examined.
Results: In vitro studies demonstrated synergistic promotion of proliferation, transwell migration, and growth factor production through the combined application of 10% CGF extract and 50 μM CoCl2. This combination notably activated the Integrin-linked kinase (ILK)/protein kinase B (AKT)/mammalian target of rapamycin (mTOR)/hypoxia-inducible factor 1-alpha (HIF-1α) pathway in ADSCs derived from both rats and humans, leading to a significant acceleration of skin wound healing (SWH) in vivo.
Conclusion: The acceleration of skin wound healing (SWH) in nude mice was observed when using rat-derived ADSC sheets treated with the combination of CGF and CoCl2, which was achieved through the activation of the ILK/AKT/mTOR/HIF-1α signaling pathway.
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Background: Ginkgolic acid (GA) impedes the invasion of cancer cells, and reverses the drug resistance of carboplatin-resistant cell lines. This study was conducted to explore the influence of GA on the sensitivity of liver cancer cells to sorafenib (SOR) and the relevant mechanism.
Methods: Genes aberrantly expressed in GA-treated liver cancer cells and SOR-resistant liver cancer cells were analyzed by GEO2R, and cofilin 2 (CFL2) level in liver cancer cells was analyzed using the StarBase. Liver cancer cells were transfected with small interfering RNA targeting CFL2 (siCFL2). Subsequently, the cells were subjected to treatments with GA and SOR alone or together. Assessment of the viability, proliferation, and apoptosis of the treated cells was performed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), 5‐ethynyl‐2′‐deoxyuridine (EdU) fluorescent staining, and flow cytometry assays, respectively. Quantification of proliferating cell nuclear antigen (PCNA), B-cell lymphoma 2 (Bcl-2), BCL2 associated X protein (Bax), Cleaved-caspase 3, and CFL2 in cell samples was achieved through quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot.
Results: GA diminished cell viability and enhanced the effect of SOR on repressing the viability and proliferation, accelerating apoptosis, decreasing PCNA and Bcl-2 levels, and increasing Bax and Cleaved-caspase 3 levels in liver cancer cells. Low expression of CFL2 was observed in liver cancer cells, however, SOR elevated its level, and GA further enhanced this increment. Silencing CFL2 offset the aforementioned roles of GA in SOR-treated liver cancer cells.
Conclusions: GA has the potential to enhance the sensitivity of liver cancer cells to SOR by increasing CFL2 level.
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Background: Amentoflavone (AF) is a polyphenolic compound present in Selaginella tamariscina, exhibiting various activities. Nevertheless, the potential neuroprotective effects of AF in animal models of Parkinson's disease (PD) induced by lipopolysaccharide (LPS) remain unexplored.
Methods: In the PD animal model, induced by LPS, the experimental procedure involved establishing the model through LPS administration, followed by animal euthanasia. Brain samples were then collected for subsequent analysis. Protein and mRNA expressions of tumor necrosis factor-alpha (TNF-α), interleukin (IL)-1β, IL-6, inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), IL-4, transforming growth factor-beta (TGF-β), arginase-1 (Arg-1), and cluster of differentiation (CD)206 were assessed using Western blot and Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) assays, respectively. Immunohistochemical staining and confocal laser scanning microscopy were employed to detect the expression of tyrosine hydroxylase (TH), the key enzyme responsible for dopamine (DA) synthesis, and alpha-synuclein (α-syn), which serves as the pathological basis of PD.
Results: The LPS-induced PD animal model successfully replicated functional deficits resembling those observed in human PD. AF was shown to ameliorate the reduction in excitability, motor dysfunction, depression-like behavior, and anxiety-like behavior.
Conclusions: In LPS-induced PD animal models, AF demonstrates a neuroprotective effect on DA neurons by suppressing the microglia-mediated inflammatory response. In terms of effectiveness, early prophylactic administration of AF appears to yield better results compared to late therapeutic administration.
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Background: Patients with metastatic colorectal cancer (CRC) generally suffer from poor prognosis. Therefore, there is an urgent need to further delineate the mechanism of metastasis in CRC, which is of clinical significance for the development of more targeted therapy. This study aimed to explore the potential role of endothelial cells-derived interleukin-6 (IL-6) in CRC metastasis.
Methods: Human umbilical vein endothelial cell (HUVEC)-conditioned medium and CRC-conditioned medium were prepared, with the latter being the control. Enzyme-linked immunosorbent assay was employed to detect IL-6 levels in HUVEC-conditioned or CRC-conditioned culture media. CRC cells were separately cultured in both media to investigate the effect of HUVECs on CRC metastasis using cell counting kit-8 assay, scratch test and Transwell assay. The role of HUVEC-derived IL-6 in CRC cells was investigated using rescue assay. The expressions of the osteopontin protein and the proteins in glycogen synthase kinase-3β (GSK3β)-β-catenin signaling pathways were determined by Western blotting. To confirm the mediating role of osteopontin in the relationship between IL-6 and CRC, the expression of osteopontin in CRC cells was up-regulated.
Results: IL-6 level was higher in HUVEC-conditioned medium than in CRC-conditioned medium (p < 0.001). CRC cells in the HUVEC-conditioned medium showed enhanced viability, migration and invasion (p < 0.05). HUVEC-secreted IL-6 facilitated malignant phenotypes and activated osteopontin and GSK3β-β-catenin signaling pathway of CRC cells in a dose-dependent manner (p < 0.05). Moreover, osteopontin overexpression contributed to CRC metastasis in vitro and activated GSK3β-β-catenin signaling pathway (p < 0.001), while such effect was abrogated by IL-6 silencing (p < 0.001).
Conclusion: HUVEC-derived IL-6 facilitated the metastasis of CRC cells by regulating osteopontin in a paracrine manner.
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Objective: The mitochondrial coenzyme Q (MitoQ) has protective effects against oxidative stress and inflammatory responses. This study investigated the potential effect of MitoQ on alleviating sepsis-induced acute lung injury (ALI) by activating the adenosine monophosphate-activated protein kinase (AMPK)-sirtuin1 (SIRT1) pathway.
Methods: The in vivo sepsis-induced ALI rat model and the in vitro A549 ALI cell model were used in this study. However, the untreated rats or cells were used as the control group and the MitoQ-treated models were used as the experimental group. The hematoxylin and eosin (H&E) staining was used to stain lung tissues. The effects of MitoQ were observed by calculating lung weight and lung permeability index (LPI). Furthermore, the levels of occluding, claudin-1, zonula occludens-1 (ZO-1), tumour necrosis factor alpha (TNF-α), interleukin-1beta (IL-1β), monocyte chemoattractant protein-1 (MCP-1), caspase-3, and nuclear factor-kappaB (NF-κB) were assessed using quantitative real-time polymerase chain reaction (qRT-PCR), for evaluating the junction integrity, apoptosis, and inflammation. Moreover, to examine the effect of MitoQ on the AMPK-SIRT pathway, the levels of p-AMPK/AMPK, and SIRT1 were evaluated using Western blot analysis. Additionally, the levels of gasdermin D (GSDMD), cleaved caspase 1, cleaved caspase 11, and lactate dehydrogenase (LDH) were used to investigate the effect of MitoQ on cell pyroptosis and cell damage. Additionally, the role of AMPK and SIRT1 pathways on pyroptosis and cellular damage was determined.
Results: In the lung tissues of the ALI model group, the H&E-stained area (p < 0.01), the lung wet/dry weight ratio (p < 0.01), and the LPI were significantly increased (p < 0.01). Moreover, these values were counteracted with MitoQ treatment (p < 0.05). The ALI rats treated with MitoQ survived longer than those without treatment (p < 0.01). Furthermore, in the ALI model group (both in tissues and cells), a significant decrease was observed in the levels of occludin, claudin-1, ZO-1, caspase-3, and NF-κB (p < 0.01), while the levels of TNF-α, IL-1β, MCP-1, cleaved GSDMD, cleaved caspase-1, cleaved caspase-11, and LDH were significantly increased (p < 0.01). Interestingly, treatment with MitoQ reversed all of these factors (p < 0.05). In the ALI model group, the levels of p-AMPK/AMPK and SIRT1 proteins were significantly decreased (p < 0.01), and their levels were reversed following MitoQ treatment (p < 0.05). Furthermore, the compound C (CC) decreased the levels of p-AMPK/AMPK and SIRT1 (p < 0.01), while the SIRT1 antagonist Ex-527 decreased only the level of SIRT1 (p < 0.01). However, both the CC and Ex-527 reversed the levels of cleaved GSDMD (p < 0.01), cleaved caspase-1 (p < 0.05), cleaved caspase-11 (p < 0.01), and LDH (p < 0.01), which were inhibited by MitoQ in ALI cells.
Conclusion: Our findings suggest that MitoQ exerts protective effects on sepsis-induced ALI by activating the AMPK-SIRT1 pathway and inhibiting pyroptosis. These results offer a potential therapeutic application of MitoQ in treating sepsis-induced lung injury and provide insights into the underlying mechanisms involved.
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Background: Low-level mosaicism is a common trait of early human development. Although mosaic embryos may lead to healthy live births, the direct effects of mosaicism are unknown. While embryo self-correction was demonstrated in mouse models, humans can only study the effects of chromosomal aberrations and blastocyst mosaicism on the early peri-implantation period by extending in vitro embryo culture up to 12 days post-fertilization.
Methods: The established culture protocols were followed to generate embryonic stem cells. The 24 high-quality mosaicism/aneuploid mosaicism blastocysts were detected by preimplantation genetic testing for aneuploidy (PGT-A). The isolated inner cell masses (ICMs) were seeded onto feeder-free dishes, and after 10–12 days of culture, there were six blastocyst ICM-generated stem cell clones. The cells shed during the stem cell growth were harvested, and next-generation sequencing was performed on stem cells and the shed cells. To test pluripotency, a small number of stem cells were isolated and subcultured.
Results: The PGT-A status was confirmed from the results of next-generation sequencing of stem cell exfoliated cells and stem cells. The mosaics in five blastocysts were fully repaired while the mosaic in one blastocyst was partially repaired. Simultaneously, the cells were verified to have pluripotency and the ability to differentiate into three germ layers by immunofluorescence, flow cytometry, and in vitro differentiation analyses.
Conclusion: Human mosaicism/aneuploid mosaicism blastocysts have the potential for self-correction by eliminating mosaic cells.
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Background: The healing process in diabetic wounds is an emerging challenge, emphasizing the development of new pharmacologic strategies. Ferulic acid (FA) and astragaloside IV (AS-IV) have been recognized for their angiogenic potential, but how their combination impacts the angiogenesis under a high-glucose environment is yet to be explored. Therefore, this study was designed to assess the synergistic effects of FA and AS-IV and their underlying mechanisms on angiogenesis in a high-glucose environment.
Methods: Network pharmacology was utilized to screen the common targets associated with diabetic wounds, as well as FA and AS-IV. Subsequently, the signal pathways involved in these common targets were enriched using the Kyoto Encyclopedia of Genes and Genomes (KEGG) database. Furthermore, the angiogenesis-promoting effects of FA and AS-IV were validated using flow cytometry, scratch assay, and tube formation assay. Additionally, the molecular mechanism underlying the impacts of FA and AS-IV on angiogenesis was evaluated using si-RNA interference assay.
Results: Using bioinformatics analysis, 101 common target genes linked to diseases were identified. These target genes were enriched in various signaling pathways such as hydroxylated hypoxia-inducible factor 1 (HIF-1) signaling pathway and vascular endothelial growth factors (VEGFs) signaling pathway. Furthermore, cytological experiments revealed that FA and AS-IV promoted angiogenesis under a high-glucose condition by activating the HIF-1 signaling pathway. The combination of these two drugs significantly reduced the apoptosis level, increased cell migration, and promoted tube formation. Additionally, they increased the expression levels of VEGFs, vascular endothelial growth factor receptor 2 (VEGFR2), and HIF-1α proteins. Conversely, the protein expression level of the von Hippel-Lindau tumor-suppressor protein (VHL) significantly decreased. These effects were partially reversed when HIF-1α was down-regulated using specific si-HIF-1α.
Conclusions: FA in combination with AS-IV promotes angiogenesis within a high-glucose environment by activating the HIF-1 signaling pathway. Hence, FA+AS-IV might be a promising option for treating diabetic wounds. These findings provide a novel experimental and theoretical basis for managing diabetic wounds.
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Background: Diabetic retinopathy (DR), characterized by retinal microvascular dysfunction, is a common complication of diabetes. Plasminogen Activator Inhibitor 1 (PAI-1), a key regulator of fibrinolysis and inflammation, has been implicated in the pathogenesis of DR, its exact role and underlying mechanisms remain unclear. The objective of this study aimed to investigate the role of PAI-1 in regulating the function of retinal microvascular endothelial cells (RMECs) under high glucose (HG) conditions and to explore the involvement of Yes-associated protein 1 (YAP1) in this process.
Methods: RMECs were cultured and subjected to HG, PAI-1 overexpression, PAI-1 inhibitor (TM5441), YAP1 inhibitor (K-975) treatment, or their combinations. Cell viability was assessed using the cell counting kit-8 (CCK-8) assay. However, YAP1 expression was determined using western blot. The apoptosis rate in the RMECs was evaluated using flow cytometry and caspase-3 activity assay. Additionally, the levels of inflammatory cytokines and vascular endothelial growth factor (VEGF) were determined using enzyme-linked immunosorbent assay (ELISA).
Results: It was observed that HG treatment and overexpression of PAI-1 significantly increased YAP1 expression in RMECs. Furthermore, HG and overexpressed PAI-1 significantly reduced cell viability, while promoting apoptosis, inflammatory response, and angiogenesis in RMECs. However, treatment with PAI-1 inhibitor or the YAP1 inhibitor reversed these effects, leading to significantly increased (p < 0.01) cell viability and decreased (p < 0.01) apoptosis, inflammatory response, and angiogenesis.
Conclusions: In conclusion, our findings suggest that PAI-1 upregulates YAP1 expression in RMECs under HG conditions, thereby inhibiting cell proliferation, and promoting apoptosis, inflammatory response, and angiogenesis. The use of PAI-1 inhibitor or YAP1 inhibitor can reverse these effects, highlighting the potential therapeutic value of targeting PAI-1 and YAP1 in the treatment of DR. These findings provide new insights into the underlying molecular mechanisms involved in the pathogenesis of DR and might contribute to the development of novel therapeutic strategies for this disease.
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Backgrounds and Objective: Retinal ischemia-reperfusion (I/R) injury is a main cause of retinal ganglion cell (RGC) damage, leading to visual impairment. In this study, we investigated the therapeutic potential of metformin, an antidiabetic drug, in alleviating RGC injury in a mouse model of retinal I/R injury. We aimed to explore the role of metformin in modulating the hypoxia inducible factor-1α (HIF-1α)-vascular endothelial growth factor (VEGF) pathway, which is known to be involved in the development of retinal ischemia.
Methods: Eight-week-old male C57BL/6 J mice (6 each group), anesthetized with pentobarbital, were treated with the intraperitoneal injection of metformin in the model + metformin group after one day by a retinal I/R procedure. Three days after the I/R injury, the therapeutic effect was evaluated by the staining of surviving RGCs, real-time quantitative PCR (qRT-PCR), and Western blot analysis.
Results: Our findings revealed that metformin treatment reduced RGC loss in retinal I/R injury model. Mechanistically, metformin administration effectively inhibited HIF-1α and vascular endothelial growth factor (VEGF), which are key components of the HIF-1α-VEGF pathway, and were upregulated (p < 0.01) in the I/R injury group. Furthermore, metformin treatment resulted in the restoration of visual proteins, including opsin and rhodopsin, as well as the upregulation of neurotrophic factors (nerve growth factor (NGF) and Brain Derived Neurotrophic Factor (BDNF)) (p < 0.01), which play crucial roles in RGC survival and function. These results highlight the neuroprotective effects of metformin in attenuating RGC injury by suppressing the HIF-1α-VEGF pathway in a mouse model of retinal I/R injury.
Conclusion: The findings suggest that metformin may be a potential therapeutic agent for the treatment of retinal ischemic disorders. Further investigations are needed to elucidate the underlying molecular mechanisms and validate the translational potential of metformin in clinical settings.
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Background: Esophageal adenocarcinoma (EAC), the major form of esophageal cancer, has the high incidence worldwide and patients with EAC have poor prognosis due to limited treatment. It is critical to develop a comprehensive understanding of progression of EAC cells for developing more effective and targeted therapies. Since the reduced form of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase 5 (NOX5)-S has been associated with the progression of EAC, we aimed to explore the effect of NOX5-S on the EAC cell progression and to elucidate the mechanisms underlying this effect.
Methods: The differential expression of NOX5-S in human normal esophageal epithelial cells Het1A and human EAC cells OE19 was evaluated using western blot and quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR). The cells were transfected with pCMV-tag5a-NOX5-S (overexpressing plasmids (OE)-NOX5-S), si-NOX5-S, and blank control (OE-NC) to assess the effects of NOX5-S on cell proliferation, migration, and apoptosis using cell counting kit-8, colony formation, transwell, and terminal deoxynucleotidyl transferase (TdT) dUTP nick-end labeling (TUNEL) assays. The western blot was used for evaluating phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT)/nuclear factor-kappaB (NF-κB) axis activation, the Recilisib was used for PI3K/AKT activation and the Prostratin was used for NF-κB pathway activation. The induced alterations of cell proliferation, migration and apoptosis were measured.
Results: The expression of NOX5-S was higher in EAC OE19 cells (p < 0.001). Transfection of small interfering (si)-RNA against si-NOX5-S successfully decreased the NOX5-S expression (p < 0.001), and transfection of NOX5-S overexpressing plasmids (OE)-NOX5-S successful increased the NOX5-S expression (p < 0.001). Increased/decreased NOX5-S expression promoted/inhibited EAC cell proliferation (p < 0.001) and migration (p < 0.001), induced less/more EAC cell apoptosis (p < 0.001), and increased/decreased all levels of p-PI3K/PI3K, p-AKT/AKT and p-NF-κB/NF-κB (p < 0.001). Recilisib elevated p-AKT/AKT, p-NF-κB/NF-κB and p-PI3K/PI3K in NOX5-S knockdown cells (p < 0.001) and Prostratin increased p-NF-κB/NF-κB (p < 0.001) in NOX5-S knockdown cells. Both Recilisib and Prostratin reversely increased the proliferation (p < 0.001) and migration (p < 0.001), and reversely decreased apoptosis (p < 0.001) in EAC cells.
Conclusions: The NOX5-S knockdown suppresses the progression of EAC cells by inactivating the PI3K/AKT/NF-κB pathway. Knockdown of NOX5-S and inactivation of the PI3K/AKT and NF-κB signaling pathways could be potential strategies for treating EAC.
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Background: Non-small cell lung cancer (NSCLC) represents 85% of all lung cancer (LC) cases that are malignant tumors, while lung adenocarcinoma (LUAD) is the commonest type of NSCLC. Family with sequence similarity 107 member A (FAM107A) is a suppressor gene in many cancers, but its mechanism in LUAD has been less studied. The objective of the experiment is to explore the role and potential mechanism of FAM107A in the progression of LUAD.
Methods: Expression levels of histone deacetylase 2 (HDAC2) and FAM107A in LUAD tissues/cells were determined using StarBase and quantitative reverse transcription polymerase chain reaction. In the first part, FAM107A specific short hairpin RNA (shFAM107A) was transfected into H1299 cells and FAM107A overexpression plasmid was transfected into PC9 cells. In the second part, shHDAC2 and shFAM107A were co-transfected into H1299 cells, and HDAC2 and FAM107A overexpression plasmids were co-transfected into PC9 cells. Then, chromatin immunoprecipitation assays were employed to determine HDAC2 enrichment in FAM107A promoter region. Loss- and gain-of-function experiments (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), colony formation assay, and flow cytometry) were conducted to verify the regulation of FAM107A on malignant phenotype of LUAD cells. The influence of HDAC2/FAM107A axis on LUAD cell biological behaviors was verified through rescue experiments.
Results: FAM107A mRNA exhibited a low expression in LUAD tissues/cells (p < 0.001). Its overexpression suppressed the viability, proliferation, and B-cell lymphoma-2 (Bcl-2) level in LUAD cells, and promoted the apoptosis as well as Bcl-2-Associated X (Bax) and Cleaved Caspase-3 protein levels (p < 0.01), while its silencing produced the opposite results. HDAC2 mRNA expression was highly expressed in LUAD tissues (p < 0.001). Its overexpression negated the aforementioned effects of FAM107A overexpression (p < 0.01), while its inhibition neutralized the effects of FAM107A silencing (p < 0.01). HDAC2 protein expression was enriched in FAM107A promoter region (p < 0.001), and FAM107A protein expression was inhibited by HDAC2 through its histone deacetylation.
Conclusions: Repression of FAM107A induced by HDAC2 promotes colony formation, while inhibiting the apoptosis of LUAD cells.
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Background: The immune response mediated by alveolar macrophages is critical in lung injury pathogenesis. Takeda G protein-coupled receptor 5 (TGR5) is implicated in the immune cell-mediated responses. This study explored the effects and underlying mechanisms of TGR5 in the lipopolysaccharide (LPS)-induced inflammatory responses in alveolar macrophages.
Methods: Mouse alveolar macrophage (MH-S) cells were exposed to LPS, and TGR5 knockdown cells were established using the Clustered Regularly Interspaced Short Palindromic Repeats/Cas9 (CRISPR/Cas9) method. The changes of inflammation responses and macrophage polarity after TGR5 knockdown in LPS-treated MH-S cells were analyzed by quantitative reverse transcriptase real-time polymerase chain reaction (qRT-PCR) and enzyme-linked immunosorbent assay (ELISA). The impact of TGR5 knockdown on protein expression related to the signal transducer and activator of the transcription 3 (STAT3) pathway was assessed through Western blot. Finally, MH-S cells were treated with chenodeoxycholic acid (CDCA, a TGR5 agonist) or cucurbitacin B (CuB, a STAT3 inhibitor), and changed in the expression of STAT3 pathway proteins and the macrophage activation markers inducible nitric oxide synthase (iNOS) and cluster of differentiation 206 (CD206) were detected using Western blot and qRT-PCR.
Results: LPS stimulation enhanced inflammation responses in MH-S cells, with TGR5 knockdown exacerbating LPS-induced inflammation. Additionally, TGR5 knockdown promoted LPS-induced M1 polarization of macrophages, increasing Interleukin (IL)-6 expression and the phospho (p)-STAT3/STAT3 ratio in LPS-induced MH-S cells. Treatment with CDCA or CuB reduced IL-6 expression and the p-STAT3/STAT3 ratio, inhibiting M1 polarization of macrophages in TGR5 knockdown MH-S cells.
Conclusions: TGR5 inhibits inflammation and promotes macrophage polarization toward the M2 phenotype in LPS-induced MH-S cells. Its effect is achieved, in part, by inhibiting the STAT3 signaling pathway.
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Background: Patients with diarrhea-type irritable bowel syndrome (IBS-D) often have symptoms of anxiety and depression. Several studies suggest that IBS-D patients may suffer from structural disturbances in the gut microbiota associated with psychiatric symptoms such as anxiety and depression. In this investigation, we employed 16S rRNA gene sequencing and non-targeted metabolomics analysis to delve into the distinct microorganisms and metabolites associated with the physical and psychological symptoms experienced by individuals with IBS-D. This endeavor offers valuable insights into the potential for targeted intestinal therapy in IBS-D patients.
Methods: 42 IBS-D patients and 20 healthy controls received a firm diagnosis of the condition between June 2020 and February 2022. 16S-rRNA gene amplicon sequencing on the Illumine platform and liquid chromatography tandem-mass spectrometry (LC-MS) were performed to detect and analyze the feces of the healthy control group and IBS-D group to obtain differential microorganisms and metabolites.
Results: In patients with IBS-D, there was a notable reduction in the abundance and diversity of their intestinal microbiota compared to the healthy control group. Additionally, the ratio of Firmicutes to Bacteroides in IBS-D patients was lower than that observed in the healthy control group. Furthermore, significant distinctions were observed in the fecal metabolites of IBS-D patients compared to those of the healthy control group. The metabolism of alanine, aspartic acid, glutamate, and bile secretions were disturbed. Based on clinical and psychological symptom analysis, as well as analysis of the microbiome and metabolome features, it was found that Blautia, Intestinibacter, and Romboutsia were positively correlated with the severity of irritable bowel syndrome (IBS), as well as levels of anxiety and depression. The intestinal microbiota metabolite 2-Dodecenal was closely related to the presence of IBS-D. The 3-hydroxy-aminobenzoic acid, one of the fecal metabolites involved in tryptophan metabolism, in IBS-D patients complicated with anxious-depressive states was significantly reduced.
Conclusion: This study has unveiled substantial disparities in fecal microbiota and metabolomic profiles between individuals afflicted with IBS-D and comorbid anxious-depressive conditions and their healthy counterparts. These findings provide valuable insights for prospective therapeutic interventions targeting the gut in this context.
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Background: Kawasaki disease (KD) is a common multi-system vascular inflammatory disease in childhood. The main complication is coronary artery lesions (CALs). Despite its widespread prevalence, the precise pathogenesis of KD remains unknown. The purpose of this study is to investigate the potential therapeutic role and mechanism of Oridonin (Ori) in improving vascular endothelial cell injury in KD.
Methods: The synthetic biotin coupled with Ori pulls down lysate protein products from Human umbilical vein endothelial cells (HUVECs). Subsequently, the pull-down proteins were detected using Liquid Chromatography-tandem Mass Spectrometry (LC-MS/MS), and bioinformatics analysis was utilized to screen for the key Ori binding protein Peptidylarginine Deiminase 2 (PADI2). HUVECs were then pretreated with various concentrations of Ori, including 0.5 μM, 1.0 μM, and 2.0 μM. The monocytic leukemia cell line (THP1) was treated with 15% serum obtained from KD patients and co-cultured with HUVECs to reconstruct the inflammatory environment of endothelial cells in KD. Moreover, by constructing the PADI2 interference plasmid for cell regulation experiments and in vivo experiments on mice injected with lactobacillus casei cell wall extract (LCWE) solution, the underlying mechanism of Ori treatment against KD was determined using Cell Counting Kit-8 (CCK-8), flow cytometry, Enzyme-Linked Immunosorbent Assay (ELISA), quantitative Real-Time Polymerase Chain Reaction (qRT-PCR), and Western blot techniques.
Results: The key Ori binding protein PADI2 was screened using LC-MS/MS detection of Ori pull-down proteins. Ori therapy improves KD-induced endothelial cell inflammatory damage, as evidenced by increased cell viability, reduced apoptosis, and decreased levels of pro-inflammatory cytokines like Interleukin-1β (IL-1β), Interleukin-6 (IL-6), and Tumor necrosis factor-α (TNF-α) (p < 0.05). Additionally, Ori greatly reduced the expression of PADI2, phosphorylated NF-kappaB p65 (p-p65), and phosphorylated inhibitor of nuclear factor-kappa B (NF-κB) (phosphorylated inhibitor of NF-κB (p-IκB)) in HUVECs (p < 0.05). Ori demonstrated the optimum efficiency at 2 μM concentration (p < 0.05). Interfering with PADI2 markedly increased KD-induced HUVEC cell viability and inhibited apoptosis (p < 0.01). This interference also reduced the levels of IL-1β, TNF-α, and IL-6 along with downregulating the expression of PADI2, p-p65, and p-IκB (p < 0.01). The results of in vivo experiments revealed that both Ori and interfering with PADI2 alleviated coronary endothelial cell injury and inflammation. They also significantly elevated Nitric Oxide (NO) levels (p < 0.01), and significantly reduced endothelin (ET)-1 and pro-inflammatory cytokine levels (p < 0.01), as well as the expression of p-p65 and p-IκB in coronary tissue of mice (p < 0.01). Interference with PADI2 based on Ori stimulation significantly reduced p-p65 and p-IκB expression (p < 0.01).
Conclusions: Ori exhibited the ability to improve the inflammatory damage of endothelial cells in KD by regulating the expression and viability of PADI2 to affect the activation of the NF-κB pathway and nuclear transport.
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Background: Leukaemia is a blood-related cancer that is the leading cause of death worldwide. Piperine, a spicy alkaloid found in the spice Piper nigrum, has multifunctional properties. However, its anticancer effects against human chronic myeloid leukemia (CML) cells are yet to be investigated. The present study aims to investigate the anticancer potential of piperine using the human CML cell line KCL22.
Methods: The anticancer effects of piperine were investigated through 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) assay, generation of reactive oxygen species (ROS), cellular apoptosis, and cell cycle arrest. Additionally, in silico molecular docking was utilized to predict cell death by detecting the binding interactions of piperine with surface proteins of leukemia cells, specifically death receptors 4 (DR4) and death receptors 5 (DR5).
Results: Data in the MTT assay showed that piperine treatment inhibited the cellular proliferation of KCL22 cells in a dose-dependent manner. Furthermore, nuclear condensation and ROS generation significantly increased when KCL22 cells were treated with piperine. Further study on cell cycle progression showed that piperine treatment inhibited cell cycle progression at G2/M and S phases. In silico molecular docking study showed that piperine has a good binding interaction with both DR4 and DR5. No violation of Lipinski's rule of five was observed. Piperine exhibited drug-likeness properties without toxicity.
Conclusions: These findings suggest that piperine may have potential as an anticancer drug against human chronic myeloid leukemia.
