Objective To explore the effects of combined exposure to bisphenol A (BPA) and high-fat diet on liver lipid metabolism and hepatocyte senescence in mice, and to elucidate the potential mechanisms of the onset and development of non-alcoholic fatty liver disease (NAFLD). Methods Specific pathogen free C57BL/6J mice were randomly divided into six groups, with 10 mice with equal numbers of each sex in each group. The mice in the control group and the simple BPA group were fed with regular diet, while others four groups of mice were fed with high-fat diet. At the same time, the mice in the simple BPA group were intragastric administered with BPA at a dose of 50 μg/kg body weight, while the mice in the low-, medium- and high-dose BPA+high-fat groups were intragastric administered with BPA at doses of 5, 50 and 500 μg/kg body weight respectively. The mice in the control group and the high-fat group were intragastric administered with the same volume of corn oil once per day for 90 consecutive days. Liver tissues were subjected to hematoxylin-eosin (HE) and Oil Red O staining. Liver coefficients and lipid-stained area ratios were calculated. Serum level of total cholesterol, triglycerides, high-density lipoprotein, low-density lipoprotein, and the activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were determined using an automatic biochemical analyzer. The hepatic tumor necrosis factor-α (TNF-α), interleukin (IL)-6, and IL-10 levels were quantified by enzyme-linked immunosorbent assay. The relative expression of cholesterol regulatory element binding protein 1 (SREBP1), CCAAT enhancer binding protein α, P16, and phosphorylated histone H2AX (γ-H2AX) in liver tissues was detected using Western blotting. The interaction effect of the combined exposure to BPA and high-fat diet was observed based on the result of mice in the control group, the simple high-fat group, the simple BPA group, and the medium-dose BPA group+high-fat group (the combined exposure group) using a 2×2 factorial design. The results of mice in the simple high-fat group and the low-, medium-, and high-dose BPA+high-fat groups were used to observe the effect of BPA exposure dose under high-fat diet conditions. Results i) The interactive effect of combined exposure to BPA and high fat. The HE and Oil Red O staining results indicated that the combined exposure to BPA and high-fat diet successfully established NAFLD in mice. The interactive effect of combined exposure to BPA and high-fat diet on serum ALT activity and the relative expression of P16 in the liver tissue of female mice, as well as the serum ALT and AST activities and the relative expression of SREBP1 in the liver tissue of male mice was significant (all P<0.05). Specifically, the serum ALT activity of male mice in the combined exposure group was higher than that in the simple high-fat group (P<0.05), while the ALT activity in the serum of female mice in the combined exposure group was lower than that in the simple BPA group (P<0.05). The relative expression of SREBP1 protein in the liver tissue of male mice in the combined exposure group was higher than that in the control group, the simple high-fat group, and the simple BPA group (all P<0.05). For the other indicators, there were no significant differences in the interactive effect of combined exposure to BPA and high-fat diet (all P>0.05). ii) Dose effects of BPA exposure. The HE and Oil Red O staining result showed that the degree of vacuolar steatosis in the liver of female and male mice of medium- and high-dose BPA + high-fat groups was aggravated, and the range of inflammatory cell infiltration was expanded when compared with same-sex mice in the simple high-fat group. The serum ALT activity and the fat stained area ratio, as well as the relative expression of P16 in liver tissue of female mice in high-dose BPA + high-fat group increased (all P<0.05), while the level of IL-10 in liver tissue decreased (P<0.05), compared with the female mice in simple high-fat group. The serum ALT activity, the TNF-α level in liver tissue, and the relative expression of SREBP1, P16 and γ-H2AX proteins in liver tissue of male mice in high-dose BPA + high-fat group increased (all P<0.05), while the IL-6 level in liver tissue decreased (P<0.05), compared with the male mice in simple high-fat group. For the female or male mice in the low- and medium-dose BPA + high-fat groups, only some of the above indicators showed significant changes (all P<0.05). Conclusion The combined exposure to BPA and high-fat diet has a synergistic effect on the onset and development of NAFLD. The mechanism may be related to inducing cellular senescence and modulation of lipid synthesis pathways, thereby affecting liver steatosis. The exposure dose of BPA may affect the synergistic effect.

Objective: To explore the molecular mechanism of resveratrol (RES) in the treatment of oral squamous cell carcinoma (OSCC) through the use of biological information methods such as network pharmacology and molecular docking and to provide a theoretical reference for the clinical application of RES in the treatment of OSCC. Methods: The Swiss Target Prediction(http://www.swisstargetprediction.ch), SEA (http://sea.bkslab.org)database, and Pharm mapper database(http://lilab-ecust.cn) were used to retrieve RES-related targets, and the DISGENET (www.disgenet.org), OMIM (https://omim.org) and GeneCards (https://www.genecards.org) databases were used to screen OSCC disease targets. The intersection of drugs and disease targets was determined, and Cytoscape 3.7.2 software was used to construct a "drug-diseasetarget pathway" network. The Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) database was used to construct a target protein interaction network, and the DAVID database was used for enrichment analysis of key proteins. Finally, molecular docking validation of key proteins was performed using AutoDock and PyMOL. The enrichment analysis and molecular docking results were integrated to predict the possible molecular mechanisms of RES treatment in OSCC; western blot was used to determine the effect of resveratrol at different concentrations (50, 100) μmol/L on the expression of Src tyrosine kinase (SRC), epidermal growth factor receptor (EGFR), estrogen receptor gene 1 (ESR1), and phosphatidylinositol 3 kinase/protein kinase B (PI3K/AKT) signaling pathway proteins in OSCC HSC-3 cells. Results: A total of 243 targets of RES drugs and 6 094 targets of OSCC were identified. A total of 116 potential common targets were obtained by intersecting drugs with disease targets. These potential targets mainly participate in biological processes such as in vivo protein self-phosphorylation, peptide tyrosine phosphorylation, transmembrane receptor protein tyrosine kinase signaling pathway, and positive regulation of RNA polymerase Ⅱ promoter transcription, and they interfere with the PI3K/AKT signaling pathway to exert anti-OSCC effects. The docking results of resveratrol with OSCC molecules indicated that key targets, such as EGFR, ESR1, and SRC, have good binding activity. The results of cell-based experiments showed that resveratrol inhibited the protein expression of SRC, EGFR, ESR1, p-PI3K, and p-AKT in HSC-3 cells in a dose-dependent manner. Conclusion RES can inhibit the expression of its targets EGFR, ESR1, SRC, p-PI3K, and p-AKT in OSCC cells.

Objective The prediction of RR intervals in hypertensive patients can help clinicians to analyze and warn patients'heart condition.Methods Using 8 patients'data as samples,the RR intervals of patients were predicted by long short-term memory network(LSTM)and gradient lift tree(XGBoost),and the prediction results of the two models were combined by the inverse variance method to overcome the disadvantage of single model prediction.Results Compared with the single model,the proposed combined model had a different degree of improvement in the prediction of RR intervals in 8 patients.Conclusion LSTM-XGBoost model provides a method for predicting RR intervals in hypertensive patients,which has potential clinical feasibility.

Objective To investigate the molecular mechanism through which calenduloside E inhibits hepatocellular carcinoma(HCC)cell proliferation and migration.Methods HCC cell lines HepG2 and Huh7 treated with calenduloside E were examined for changes in cell viability using CCK-8 assay and expressions of GPX4,SLC7A11,LC3,P62 and phosphorylation of Akt/mTOR using Western blotting.The effects LY294002 and Rapamycin(the inhibitor and activator of autophagy,respectively)on proliferation and migration of calenduloside E-treated HCC cells were evaluated using EdU and Transwell assays.The TCGA database was used to explore the expression levels of GPX4 and SLC7A11 in HCC and normal liver tissues and their correlation with the patients'survival outcomes.GPX4 and SLC7A11 expressions were also detected in HCC cells and normal hepatocytes using RT-qPCR and Western blotting.Results Calenduloside E obviously inhibited the viability of HCC cells.GPX4 and SLC7A11 were highly expressed in HCC tissues and cell lines,and their expression levels were negatively correlated with the patients'survival.In HCC cell lines,calenduloside E significantly inhibited the expressions of GPX4 and SLC7A11 proteins,activated the Akt-mTOR pathway,and enhanced the expression of LC3 II.The inhibitory effect of calenduloside E on GPX4 and SLC7A11 expressions was significantly enhanced by rapamycin but attenuated by LY294002.Inhibiting the autophagy pathway obviously diminished the inhibitory effect of calenduloside E on proliferation and migration of HCC cells,while activating this pathway produced the opposite effect.Conclusion Calenduside E inhibits the proliferation and migration of HCC cells by down-regulating GPX4 and SLC7A11 expression via the autophagy pathway.

Objective To investigate the mechanism mediating the regulatory effect of lncRNA MAGI2-AS3 on cisplatin(DDP)resistance in non-small cell lung cancer(NSCLC).Methods MAGI2-AS3 and miR-1269a expression levels were detected by qRT-PCR in DDP-sensitive lung cancer cell lines(A549 and H1299)and their resistant counterparts(A549/DDP and H1299/DDP).In A549 and H1299 cells with MAGI2-AS3 silencing and A549/DDP and H1299/DDP cells overexpressing MAGI2-AS3,the effects of 20 μmol/L DDP on cell viability and apoptosis were examined with CCK-8 assay,colony formation assay,flow cytometry and Western blotting,and the changes in epithelial-mesenchymal transition(EMT)were assessed with wound healing and Transwell assays.The interaction between MAGI2-AS3,miR-1269a and PTEN was predicted using GEPIA,StarBase and miRDB and verified with luciferase reporter gene assay and radioimmunoprecipitation(RIP)assay.A miR-1269a mimic and pcDNA3.1-PTEN plasmid were used to perform the rescue assay.Results MAGI2-AS3 expression was significantly downregulated in lung cancer tissues(P<0.05)in association with a poor prognosis(P<0.05).In the two DDP-resistant lung cancer cell lines,MAGI2-AS3 expression was significantly lowered as compared with the sensitive cells.Silencing MAGI2-AS3 significantly enhanced cell viability and promoted EMT of A549 and H1299 cells irrespective of DDP treatment,and also decreased DDP-induced apoptosis of the cells.In A549/DDP and H1299/DDP cells,MAGI2-AS3 overexpression strongly repressed cell viability and EMT irrespective of DDP treatment and promoted DDP-induced cell apoptosis.Luciferase reporter gene and RIP assays confirmed the binding of MAGI2-AS3 with miR-1269a and the binding of miR-1269a with 3'-UTR domain of PTEN.The rescue assay demonstrated that MAGI2-AS3 acted as a sponge for miR-1269a to promote PTEN expression and downregulate AKT phosphorylation,thus inhibiting EMT and promoting DDP-induced apoptosis of A549/DDP cells.Conclusion MAGI2-AS3 enhances DDP sensitivity of NSCLC by targeted regulation of the miR-1269a/PTEN/AKT signaling axis.

Objective To investigate the molecular mechanism through which calenduloside E inhibits hepatocellular carcinoma(HCC)cell proliferation and migration.Methods HCC cell lines HepG2 and Huh7 treated with calenduloside E were examined for changes in cell viability using CCK-8 assay and expressions of GPX4,SLC7A11,LC3,P62 and phosphorylation of Akt/mTOR using Western blotting.The effects LY294002 and Rapamycin(the inhibitor and activator of autophagy,respectively)on proliferation and migration of calenduloside E-treated HCC cells were evaluated using EdU and Transwell assays.The TCGA database was used to explore the expression levels of GPX4 and SLC7A11 in HCC and normal liver tissues and their correlation with the patients'survival outcomes.GPX4 and SLC7A11 expressions were also detected in HCC cells and normal hepatocytes using RT-qPCR and Western blotting.Results Calenduloside E obviously inhibited the viability of HCC cells.GPX4 and SLC7A11 were highly expressed in HCC tissues and cell lines,and their expression levels were negatively correlated with the patients'survival.In HCC cell lines,calenduloside E significantly inhibited the expressions of GPX4 and SLC7A11 proteins,activated the Akt-mTOR pathway,and enhanced the expression of LC3 II.The inhibitory effect of calenduloside E on GPX4 and SLC7A11 expressions was significantly enhanced by rapamycin but attenuated by LY294002.Inhibiting the autophagy pathway obviously diminished the inhibitory effect of calenduloside E on proliferation and migration of HCC cells,while activating this pathway produced the opposite effect.Conclusion Calenduside E inhibits the proliferation and migration of HCC cells by down-regulating GPX4 and SLC7A11 expression via the autophagy pathway.

Objective To investigate the mechanism mediating the regulatory effect of lncRNA MAGI2-AS3 on cisplatin(DDP)resistance in non-small cell lung cancer(NSCLC).Methods MAGI2-AS3 and miR-1269a expression levels were detected by qRT-PCR in DDP-sensitive lung cancer cell lines(A549 and H1299)and their resistant counterparts(A549/DDP and H1299/DDP).In A549 and H1299 cells with MAGI2-AS3 silencing and A549/DDP and H1299/DDP cells overexpressing MAGI2-AS3,the effects of 20 μmol/L DDP on cell viability and apoptosis were examined with CCK-8 assay,colony formation assay,flow cytometry and Western blotting,and the changes in epithelial-mesenchymal transition(EMT)were assessed with wound healing and Transwell assays.The interaction between MAGI2-AS3,miR-1269a and PTEN was predicted using GEPIA,StarBase and miRDB and verified with luciferase reporter gene assay and radioimmunoprecipitation(RIP)assay.A miR-1269a mimic and pcDNA3.1-PTEN plasmid were used to perform the rescue assay.Results MAGI2-AS3 expression was significantly downregulated in lung cancer tissues(P<0.05)in association with a poor prognosis(P<0.05).In the two DDP-resistant lung cancer cell lines,MAGI2-AS3 expression was significantly lowered as compared with the sensitive cells.Silencing MAGI2-AS3 significantly enhanced cell viability and promoted EMT of A549 and H1299 cells irrespective of DDP treatment,and also decreased DDP-induced apoptosis of the cells.In A549/DDP and H1299/DDP cells,MAGI2-AS3 overexpression strongly repressed cell viability and EMT irrespective of DDP treatment and promoted DDP-induced cell apoptosis.Luciferase reporter gene and RIP assays confirmed the binding of MAGI2-AS3 with miR-1269a and the binding of miR-1269a with 3'-UTR domain of PTEN.The rescue assay demonstrated that MAGI2-AS3 acted as a sponge for miR-1269a to promote PTEN expression and downregulate AKT phosphorylation,thus inhibiting EMT and promoting DDP-induced apoptosis of A549/DDP cells.Conclusion MAGI2-AS3 enhances DDP sensitivity of NSCLC by targeted regulation of the miR-1269a/PTEN/AKT signaling axis.

Objective To explore the molecular mechanism of resveratrol(RES)in the treatment of oral squamous cell carcinoma(OSCC)through the use of biological information methods such as network pharmacology and molecular docking and to provide a theoretical reference for the clinical application of RES in the treatment of OSCC.Methods The Swiss Target Prediction(http://www.swisstargetprediction.ch),SEA(http://sea.bkslab.org)database,and Pharm map-per database(http://lilab-ecust.cn)were used to retrieve RES-related targets,and the DISGENET(www.disgenet.org),OMIM(https://omim.org)and GeneCards(https://www.genecards.org)databases were used to screen OSCC disease tar-gets.The intersection of drugs and disease targets was determined,and Cytoscape 3.7.2 software was used to construct a"drug-diseasetarget pathway"network.The Search Tool for the Retrieval of Interacting Genes/Proteins(STRING)data-base was used to construct a target protein interaction network,and the DAVID database was used for enrichment analy-sis of key proteins.Finally,molecular docking validation of key proteins was performed using AutoDock and PyMOL.The enrichment analysis and molecular docking results were integrated to predict the possible molecular mechanisms of RES treatment in OSCC;western blot was used to determine the effect of resveratrol at different concentrations(50,100)μmol/L on the expression of Src tyrosine kinase(SRC),epidermal growth factor receptor(EGFR),estrogen re-ceptor gene 1(ESR1),and phosphatidylinositol 3 kinase/protein kinase B(PI3K/AKT)signaling pathway proteins in OSCC HSC-3 cells.Results A total of 243 targets of RES drugs and 6 094 targets of OSCC were identified.A total of 116 potential common targets were obtained by intersecting drugs with disease targets.These potential targets mainly participate in biological processes such as in vivo protein self-phosphorylation,peptide tyrosine phosphorylation,trans-membrane receptor protein tyrosine kinase signaling pathway,and positive regulation of RNA polymerase Ⅱ promot-er transcription,and they interfere with the PI3K/AKT signaling pathway to exert anti-OSCC effects.The docking results of resveratrol with OSCC molecules indicated that key targets,such as EGFR,ESR1,and SRC,have good binding activi-ty.The results of cell-based experiments showed that resveratrol inhibited the protein expression of SRC,EGFR,ESR1,p-PI3K,and p-AKT in HSC-3 cells in a dose-dependent manner.Conclusion RES can inhibit the expres-sion of its targets EGFR,ESR1,SRC,p-PI3K,and p-AKT in OSCC cells.

Objective To explore the molecular mechanism of resveratrol(RES)in the treatment of oral squamous cell carcinoma(OSCC)through the use of biological information methods such as network pharmacology and molecular docking and to provide a theoretical reference for the clinical application of RES in the treatment of OSCC.Methods The Swiss Target Prediction(http://www.swisstargetprediction.ch),SEA(http://sea.bkslab.org)database,and Pharm map-per database(http://lilab-ecust.cn)were used to retrieve RES-related targets,and the DISGENET(www.disgenet.org),OMIM(https://omim.org)and GeneCards(https://www.genecards.org)databases were used to screen OSCC disease tar-gets.The intersection of drugs and disease targets was determined,and Cytoscape 3.7.2 software was used to construct a"drug-diseasetarget pathway"network.The Search Tool for the Retrieval of Interacting Genes/Proteins(STRING)data-base was used to construct a target protein interaction network,and the DAVID database was used for enrichment analy-sis of key proteins.Finally,molecular docking validation of key proteins was performed using AutoDock and PyMOL.The enrichment analysis and molecular docking results were integrated to predict the possible molecular mechanisms of RES treatment in OSCC;western blot was used to determine the effect of resveratrol at different concentrations(50,100)μmol/L on the expression of Src tyrosine kinase(SRC),epidermal growth factor receptor(EGFR),estrogen re-ceptor gene 1(ESR1),and phosphatidylinositol 3 kinase/protein kinase B(PI3K/AKT)signaling pathway proteins in OSCC HSC-3 cells.Results A total of 243 targets of RES drugs and 6 094 targets of OSCC were identified.A total of 116 potential common targets were obtained by intersecting drugs with disease targets.These potential targets mainly participate in biological processes such as in vivo protein self-phosphorylation,peptide tyrosine phosphorylation,trans-membrane receptor protein tyrosine kinase signaling pathway,and positive regulation of RNA polymerase Ⅱ promot-er transcription,and they interfere with the PI3K/AKT signaling pathway to exert anti-OSCC effects.The docking results of resveratrol with OSCC molecules indicated that key targets,such as EGFR,ESR1,and SRC,have good binding activi-ty.The results of cell-based experiments showed that resveratrol inhibited the protein expression of SRC,EGFR,ESR1,p-PI3K,and p-AKT in HSC-3 cells in a dose-dependent manner.Conclusion RES can inhibit the expres-sion of its targets EGFR,ESR1,SRC,p-PI3K,and p-AKT in OSCC cells.

Objective To explore the molecular mechanism of resveratrol(RES)in the treatment of oral squamous cell carcinoma(OSCC)through the use of biological information methods such as network pharmacology and molecular docking and to provide a theoretical reference for the clinical application of RES in the treatment of OSCC.Methods The Swiss Target Prediction(http://www.swisstargetprediction.ch),SEA(http://sea.bkslab.org)database,and Pharm map-per database(http://lilab-ecust.cn)were used to retrieve RES-related targets,and the DISGENET(www.disgenet.org),OMIM(https://omim.org)and GeneCards(https://www.genecards.org)databases were used to screen OSCC disease tar-gets.The intersection of drugs and disease targets was determined,and Cytoscape 3.7.2 software was used to construct a"drug-diseasetarget pathway"network.The Search Tool for the Retrieval of Interacting Genes/Proteins(STRING)data-base was used to construct a target protein interaction network,and the DAVID database was used for enrichment analy-sis of key proteins.Finally,molecular docking validation of key proteins was performed using AutoDock and PyMOL.The enrichment analysis and molecular docking results were integrated to predict the possible molecular mechanisms of RES treatment in OSCC;western blot was used to determine the effect of resveratrol at different concentrations(50,100)μmol/L on the expression of Src tyrosine kinase(SRC),epidermal growth factor receptor(EGFR),estrogen re-ceptor gene 1(ESR1),and phosphatidylinositol 3 kinase/protein kinase B(PI3K/AKT)signaling pathway proteins in OSCC HSC-3 cells.Results A total of 243 targets of RES drugs and 6 094 targets of OSCC were identified.A total of 116 potential common targets were obtained by intersecting drugs with disease targets.These potential targets mainly participate in biological processes such as in vivo protein self-phosphorylation,peptide tyrosine phosphorylation,trans-membrane receptor protein tyrosine kinase signaling pathway,and positive regulation of RNA polymerase Ⅱ promot-er transcription,and they interfere with the PI3K/AKT signaling pathway to exert anti-OSCC effects.The docking results of resveratrol with OSCC molecules indicated that key targets,such as EGFR,ESR1,and SRC,have good binding activi-ty.The results of cell-based experiments showed that resveratrol inhibited the protein expression of SRC,EGFR,ESR1,p-PI3K,and p-AKT in HSC-3 cells in a dose-dependent manner.Conclusion RES can inhibit the expres-sion of its targets EGFR,ESR1,SRC,p-PI3K,and p-AKT in OSCC cells.