Through molecular docking and experimental validation,24 SPF male KM mice were ran-domly divided into four groups:the control group(CON),model group(LPS),high-dose Yang Shuhua group(YSH-H),which was gavaged with drug solution containing 1 g/mL crude Yang Shuhua,and the low-dose Yang Shuhua group(YSH-L),which was gavaged with drug solution containing 0.5 g/mL crude Yang Shuhua for a 17-day experimental period.Mouse body weight was recorded during the experiment,and fecal scoring was done 1 h after intraperitoneal injection of LPS.At the end of the experiment,histopathological changes in the jejunum tissue were examined,and the expression of tight junction protein-related genes was detected.Compared to the CON group,mice in the LPS group showed significant decreases in villus height,villus height/crypt depth(V/C)ratio(P＜0.01),and significant increase in crypt depth(P＜0.01).The mRNA ex-pression levels of ZO-1 and claudin-1 were significantly decreased(P＜0.01).Compared to the LPS group,mice in the YSH-H group showed significant decrease in crypt depth(P＜0.05),significant increase in villus height(P＜0.05),and significant increase in V/C ratio(P＜0.01).The YSH-L group showed a significant increase in V/C ratio(P＜0.05),a trend of decreased crypt depth,and an increasing trend of villus height.The mRNA expression of ZO-1 in the YSH-H group showed a significant increase(P＜0.05)and claudin-1 showed an increasing trend.Luteolin,quercetin,kaempferol,eriodictyol,and the top 5 potential key targets TP53,IL-1β,TNF,AKT1,and IL-10 exhibited molecular docking scores less than-20.95 kJ/mol,indicating strong activity.GO and KEGG enrichment analysis suggested that Yang Shuhua compounds might act on enteritis through the TNF signaling pathway,IL-17 signaling pathway,and PI3K-Akt signaling pathway.The qPCR results showed an upward trend in the mRNA expression levels of TLR4 and AKT1,significant in-crease in Caspase-1 and ASC(P＜0.05),and significantly increased expression of NF-κB and NL-RP3(P＜0.01).The mRNA expression level of AKT1,TLR4,NLRP3,ASC,Caspase-1,and NF-κB decreased compared to the LPS group,with significant differences in Caspase-1 and NF-κB(P＜0.05).This study suggests that Yang Shuhua exerts anti-enteritis effects through multiple compo-nents and pathways,with blockade of the TLR4-AKT-NF-κB pathway possibly being a key thera-peutic mechanism for treating enteritis.

This study aims to investigate the medication rules of patented traditional Chinese medi-cine(TCM)compound formulas and molecular mechanisms of core drugs for treating sepsis using data mining and network pharmacology approaches.In the present study,we first searched the PubMed database,Web of Science database,and the China National Knowledge Infrastructure(CNKI)since the establishment of the library to April 30,2024 for the relevant literature on the treatment of sepsis by traditional Chinese medicine.The prescriptions were then statistically ana-lyzed for drug frequency and association analysis to obtain the core drugs.Then we screened the ef-fective active ingredients of the core drugs by TCMSP and other database platforms,obtained sep-sis-related genes in GeneCards and other databases,and statistically intersected targets,and predic-ted the mechanism of action of the core TCMs by subjecting the intersected targets to PPI analy-sis,GO function and KEGG pathway enrichment analysis.Finally,the relationship between key tar-gets and herbal components was examined in reverse by molecular docking method.The results showed that 64 compound formulas were obtained,with a total of 150 Chinese medicines,which were mostly sweet in taste,cold in nature,and belonged to the spleen,stomach and intestinal me-ridians.According to the association rules,the core drugs were identified as"mirabilite-peach ker-nel-rheum officinale".There were 79 intersecting targets between the core drugs and sepsis,with core targets such as IL-1β,EGFR and SRC.MAPK,TNF,IL-17 and other signaling pathways are involved to mediate inflammatory responses,apoptosis and other biological processes to exert ther-apeutic effects on sepsis.The molecular docking results indicated that the docking activity of the key targets with the main components of the drug,and sennoside E_qt has the lowest binding ener-gy and the best docking activity with SRC.In conclusion,this study showed that the prescription of Chinese medicine for sepsis is mostly based on tonifying the spleen and clearing heat.The mecha-nism of action of the core drug"mirabilite-peach kernel-rheum officinale"in the treatment of sep-sis is multilevel and multifaceted,which provides a certain theoretical basis for the treatment of sepsis by traditional Chinese medicine.

Objective The aim of this study was to detect the expression of miR-383-5p in bladder cancer tissues and bladder cancer 5637 cells,BIU-87 cells,TCCSUP cells and HT-1376 cells,and to explore the effects of miR-383-5p on the prolif-eration,invasion,migration and apoptosis of bladder cancer cells by targeting CIP2A.Methods The expression of miR-383-5p was detected by qRT-PCR in human bladder cancer tissues and their corresponding adjacent tissues,5637 cells,BIU-87 cells,TCCSUP cells,HT-1376 cells,human bladder transitional epithelial cells.BIU-87 cells with low miR-383-5p expression were selected for subsequent experiments.BIU-87 cells were divided into the blank group(normal culture),miR-383-5p NC group(negative control,transfected with miR-383-5p negative control),miR-383-5p mimic group(transfected with miR-383-5p mimic),and miR-383-5p mimic+pc-CIP2A group(co-transfected with miR-383-5p mimic and CIP2A overexpression plasmid pc-CIP2A).CCK-8 kit was used to detect the viability of BIU-87 cells in each group;Flow cytometry was used to detect apoptosis of BIU-87 cells;Transwell assay was used to measure cell invasion ability of BIU-87 cells;Scratch assay was used to measure cell migration ability of BIU-87 cells;Western blot was used to determine the expression of proteins related to apoptosis,invasion(MMP-2,MMP-9),and CIP2A/PP2A in BIU-87 cells;The dual luciferase assay was used to verify the targeting relationship between miR-383-5p and CIP2A in BIU-87 cells.Results The expression of miR-383-5p was low in bladder cancer tissues and bladder cancer cells.Compared with the blank group,BIU-87 cells in the miR-383-5p mimic group showed a significant increase the level of miR-383-5p(0.91±0.10 vs.1.67±0.24,P<0.01)and a significant decrease in the expression of CIP2A protein(1.32±0.17 vs.0.45±0.03,P<0.001),the cell viability,invasion,migration abilities,the expression of proteins related to invasion(MMP-2,MMP-9),and the expression of Bcl-2 protein[(100.00±4.36)% vs.(32.15±2.65)% ,(150.20±12.95)vs.(82.35±7.01),(77.91±3.63)% vs.(46.12±2.54)% ,1.02±0.11 vs.0.22±0.04,1.03±0.18 vs.0.21±0.04,1.01±0.14 vs.0.27±0.05,P<0.001];The apoptosis rate,the expression of caspase-3 and Bax proteins related to apoptosis,and PP2A expression were significantly increased[(14.02±2.29)% vs.(38.21±3.20)% ],0.81±0.11 vs.1.78±0.24,0.83±0.12 vs.1.72±0.24,0.27±0.02 vs.0.95±0.16,P<0.001].Compared with the miR-383-5p mimic group,BIU-87 cells in the miR-383-5p mimic+pc-CIP2A group significantly increased the cell viability,invasion,migration abilities,the expression of proteins related to invasion,and the expression of Bcl-2 protein[(32.15±2.65)% vs.(50.18±3.77)% ,(82.35±7.01)% vs.(116.30±13.70),(46.12±2.54)% vs.(58.43±3.15)% ,0.22±0.04 vs.0.60±0.08,0.21±0.04 vs.0.5 8±0.06,0.27±0.05 vs.0.64±0.08,P<0.05];The apoptosis rate,the expression of caspase-3,Bax,and PP2A was signifi-cantly reduced in the miR-383-5p mimic+pc-CIP2A group[(38.21±3.20)% (23.15±2.74)% ,1.78±0.24 vs.1.25±0.21,1.72±0.24 vs.1.23±0.18,0.95±0.16 vs.0.60±0.13,P<0.05].The results of dual luciferase experiments showed a corresponding tar-geting relationship between miR-383-5p and CIP2A.Conclusion Increasing the expression of miR-383-5p can inhibit the prolif-eration,invasion and migration of bladder cancer BIU-87 cells,and enhance the ability of apoptosis,which may be achieved by targe-ted regulation of CIP2A.

Thirty SPF Kunming male mice aged 6-8 weeks were randomly divided into five groups:the control group,the model group,and the low,medium,and high dose groups of Dilong.Except for the control group,the other mice were continuously injected intraperitoneally with 80 mg/kg cyclophosphamide for 3 days to prepare an immunosuppressive model.After the model-ing was completed,the control group and the model group were given an equal amount of physio-logical saline by gavage,while the low,medium,and high dose Dilong groups were given 100,200,and 400 mg/kg of Dilong fermented material by gavage,respectively.The treatment lasted for 14 days,and thymus,spleen,blood,and intestinal contents were collected 2 hours after the last admin-istration.The results showed that cyclophosphamide caused a weight loss in mice during the exper-iment,reduced thymus index,white blood cells,lymphocytes,granulocytes,immunoglobulin levels in the blood,and disrupted gut microbiota.Dilong fermented material can effectively improve the decrease in body weight and thymus index of mice,increase the levels of immune cells and immu-noglobulin in the blood,improve the diversity and richness of intestinal microbiota,and increase the abundance of beneficial bacteria.Dilong fermented can effectively alleviate the immune suppres-sion and intestinal microbiota disorder caused by cyclophosphamide in mice.Considering all fac-tors,the optimal dosage is 200 mg/kg.

Based on network pharmacology,through molecular docking and experimental validation,the study explored the mechanism of the Hypericum japonicum-Rehmannia glutinosa-Salvia ple-beian compound(HRS)in the treatment of liver injury.Mice were randomly divided into a control group(CON group),a model group(CCL4 group),a high-dose drug group(HRS-H group),and a low-dose group(HRS-L group).A mouse liver injury model was established using CCL4 induction,liver tissue pathological morphology was observed,and the relative expression levels of liver in-flammatory cytokine genes was measured.Active ingredients of traditional Chinese medicine and targets related to Chinese medicine and diseases were obtained from databases such as Herb,TCM-SP,PubChem,Swiss Target Prediction,Gene Cards and DisGeNET.The intersection of targets was used to obtain potential drug targets.The potential targets were analyzed for protein-protein inter-action(PPI)using the string database and a network diagram of"drug-active component-intersec-tion target"was constructed using Cytoscape.DAVID database was used for GO and KEGG path-way analysis,and Auto Dock Tools software was used for molecular docking.Finally,the results of molecular docking by examining the expression of key target genes and downstream genes such as those related to the PI3K-AKT pathway and the autophagy pathway were experimentally valida-ted.Results:Animal experiment results showed that compared to the CON group,the CCL4 group of mice exhibited disrupted liver architecture,hepatocyte steatosis,vacuolization,and extensive in-flammatory cell infiltration.These characteristics were ameliorated by drug treatment groups with the HRS-H group demonstrating superior effects compared to the HRS-L group.RT-qPCR results from mouse livers showed significantly increased relative expression of TNF-α and INOS mRNA compared to the CON group in the CCL4 group(P＜0.01),and significantly increased IL-1β mR-NA relative expression(P＜0.05).Compared to the CCL4 group,the HRS-H group showed signifi-cantly decreased TNF-α,INOS,and IL-1β mRNA relative expressions(P＜0.01).155 potential tar-gets for HRS in alleviating liver damage were identified through network pharmacology,with top-ranked key target points including STAT3,SRC,PIK3R1,PIK3CA,AKT1,HSP90A11,EGFR,and ESR.Key active ingredients included Tetramethoxyluteolin,Hispidulin,Eupafolin,Kaempferol,and Eupaformonin.GO enrichment analysis yielded 940 entries,and KEGG enrichment analysis yielded 177 biological pathways.Molecular docking results showed a strong binding ability between the main components of HRS and key target points.RT-qPCR results showed increasing trends for EGFR,PI3KCA,HSP90A11,and NF-κB mRNA compared to the CON group in the CCL4 group,significantly increased AKT1 mRNA relative expression(P＜0.05),significant decreases in ULK1,ATG5,LC3B,and ATG7 mRNA relative expressions(P＜0.05),and extremely significant decreases in PTEN,ATG13,BECLIN-1,ATG16L1,ATG12,ATG4B,and ATG3 mRNA relative expressions(P＜0.01).Compared to the CCL4 group,the HRS-H group showed significantly de-creased PI3KCA,HSP90A11,and NF-κB mRNA relative expressions(P＜0.05),extremely signif-icantly decreased EGFR,AKT1,and mTOR mRNA relative expressions(P＜0.01),increased ULK1 relative expression trends,significantly increased PTEN,ATG16L1,ATG5,LC3B,and ATG7 mRNA relative expressions(P＜0.05),extremely significantly increased ATG13,BECLIN-1,ATG12,ATG4B,and ATG3 mRNA relative expressions(P＜0.01).Conclusion:The HRS ex-erts hepatoprotective effects through multi-component,multi-pathway approaches,with alleviating inflammation and promoting hepatocyte autophagy through PI3K-AKT pathway likely being im-portant mechanisms for its protective effects.

To explore the effect of Macleaya cordata extract(MCE)on growth performance and serum biochemistry of Sichuan White Geese,and to analyze its mechanism of action by network pharmacology and molecular docking technology combined with animal experiments verification.A total of 90 1-day-old healthy goslings were randomly divided into 5 groups with 18 goslings per group after 5 d of adaptive feeding.The control group(CON)was fed with basal diet,the antibiotic group(CTC)was supplemented with 450 mg/kg chlortetracycline premix,and the low MCE group(MCEL),medium MCE group(MCEM)and high MCE group(MCEH)were supple-mented with 200,300 and 400 mg/kg MCE,respectively.The experimental period was 21 d.On the basis of the above experiments,Network pharmacology and molecular docking technology were used to predict the core targets and signaling pathways of MCE to promote geese growth from the levels of antioxidant,immune and apoptosis,and the expression levels of the core target genes were detected by Real-time fluorescence quantitative PCR.The results showed that compared with the CON group,MCE supplementation could increase the average daily weight gain,decrease the ratio of feed to gain,significantly increase the contents of serum GH,T3,T4,TP and ALB(P＜0.05),and significantly decrease the contents of serum AST and TG(P＜0.05).Network pharmacology analysis predicted 2 active ingredient and 237 active ingredient targets,and concluded that the mechanism of MCE promoting the growth of Sichuan White Geese may be related to the role of 5 key targets such as SRC,HSP90AA1,CASP3,ESR1 and MAPK14,and the action of MAPK,apop-tosis and other signaling pathways.Molecular docking results showed that the active ingredients of sanguinarine and chelerythrine in MCE could act through MAPK14.The validation results of core targets showed that MCE could significantly reduce the mRNA expression levels of CytC,CASP2,CASP3 and CASP9 in spleen(P＜0.05)and significantly increase the mRNA expression levels of Mcl-1(P＜0.05).These results indicated that MCE could promote the growth performance of Si-chuan White Geese by regulating apoptosis,promoting the secretion of serum growth-related hor-mones and improving biochemical indicators.

Based on network pharmacology,molecular docking,and experimental validation,this study explored the mechanism by which Hypericum japonicum Thunb-Prunella vulgaris treat liver injury.Mice were randomly divided into four groups:a control group(CON),a model group(CCl4),a high-dose drug group(TXD-H),and a low-dose drug group(TXD-L).A mouse liver in-jury model was established using CCl4 induction.The pathological morphology of liver tissue was observed,and the serum levels of aspartate aminotransferase(AST)and alanine aminotransferase(ALT)were measured.Active ingredients of traditional Chinese medicine and targets related to these medicines and diseases were obtained from databases such as TCMSP,PubChem,Swiss Tar-get Prediction,GeneCards,and DisGeNET.The intersection of these targets was used to identify potential drug targets.A network diagram illustrating the relationships between"drug-active com-ponent-intersection target"was constructed using Cytoscape.Potential targets were analyzed using the STRING database for protein-protein interaction(PPI)analysis and the DAVID database for Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis.Molecular docking validation was performed using AutoDock Tools software.Subsequently,key target genes,including those related to the NLRP3 inflammasome and pyroptosis,were detected to validate the molecular docking results.Animal experimental results showed that compared to the CON group,serum AST and ALT activities in the CCl4 group mice were significantly increased(P＜0.01),while in the TXD-L group,serum AST and ALT activities were significantly decreased(P＜0.05)compared to the CCl4 group,and in the TXD-H group,AST and ALT activities were significantly decreased(P＜0.01).Through network pharmacology,135 potential targets were i-dentified,with key components found to be tetramethoxyluteolin,quercetin,kaempferol,luteolin and morin based on degree values,and key targets including TNF,SRC,AKT1,EGFR and ESR1.GO enrichment analysis yielded 304 entries,while KEGG enrichment analysis identified 91 biologi-cal pathways.Molecular docking results demonstrated strong binding between the main compo-nents of Hypericum japonicurn Thunb-Prunella vulgaris and key targets.qPCR results showed that compared to the CON group,the CCl4 group exhibited upregulated relative expression levels of SRC,EGFR,TNF-α,AKT1,and IL-18 mRNA,with significant increases in MyD88,NF-κB,IL-1β,NLRP3,Caspase-1,and ASC mRNA(P＜0.05),and significant upregulation of TLR4 and GS-DMD mRNA(P＜0.01).Compared to the CCl4 group,the TXD-H group displayed significant downregulation of EGFR,AKT1,TLR4,IL-1β,and GSDMD mRNA(P＜0.01),significant decrea-ses in TNF-α,MyD88,NF-κB,NLRP3,and ASC mRNA(P＜0.05),while SRC,IL-18,and Caspase-1 mRNA showed a downward trend.In conclusion,Hypericum japonicum Thunb-Prunel-la vulgaris exerts hepatoprotective effects through multiple components and pathways,among which inhibition of the NF-κB-NLRP3 pathway to reduce hepatocyte pyroptosis may be one of the important pathways for its protective effects.

Background: s/Aims: Non-invasive models stratifying clinically significant portal hypertension (CSPH) are limited. Herein, we developed a new non-invasive model for predicting CSPH in patients with compensated cirrhosis and investigated whether carvedilol can prevent hepatic decompensation in patients with high-risk CSPH stratified using the new model. Methods: Non-invasive risk factors of CSPH were identified via systematic review and meta-analysis of studies involving patients with hepatic venous pressure gradient (HVPG). A new non-invasive model was validated for various performance aspects in three cohorts, i.e., a multicenter HVPG cohort, a follow-up cohort, and a carvediloltreating cohort. Results: In the meta-analysis with six studies (n=819), liver stiffness measurement and platelet count were identified as independent risk factors for CSPH and were used to develop the new “CSPH risk” model. In the HVPG cohort (n=151), the new model accurately predicted CSPH with cutoff values of 0 and –0.68 for ruling in and out CSPH, respectively. In the follow-up cohort (n=1,102), the cumulative incidences of decompensation events significantly differed using the cutoff values of <–0.68 (low-risk), –0.68 to 0 (medium-risk), and >0 (high-risk). In the carvediloltreated cohort, patients with high-risk CSPH treated with carvedilol (n=81) had lower rates of decompensation events than non-selective beta-blockers untreated patients with high-risk CSPH (n=613 before propensity score matching [PSM], n=162 after PSM). Conclusions Treatment with carvedilol significantly reduces the risk of hepatic decompensation in patients with high-risk CSPH stratified by the new model.

Thirty SPF Kunming male mice aged 6-8 weeks were randomly divided into five groups:the control group,the model group,and the low,medium,and high dose groups of Dilong.Except for the control group,the other mice were continuously injected intraperitoneally with 80 mg/kg cyclophosphamide for 3 days to prepare an immunosuppressive model.After the model-ing was completed,the control group and the model group were given an equal amount of physio-logical saline by gavage,while the low,medium,and high dose Dilong groups were given 100,200,and 400 mg/kg of Dilong fermented material by gavage,respectively.The treatment lasted for 14 days,and thymus,spleen,blood,and intestinal contents were collected 2 hours after the last admin-istration.The results showed that cyclophosphamide caused a weight loss in mice during the exper-iment,reduced thymus index,white blood cells,lymphocytes,granulocytes,immunoglobulin levels in the blood,and disrupted gut microbiota.Dilong fermented material can effectively improve the decrease in body weight and thymus index of mice,increase the levels of immune cells and immu-noglobulin in the blood,improve the diversity and richness of intestinal microbiota,and increase the abundance of beneficial bacteria.Dilong fermented can effectively alleviate the immune suppres-sion and intestinal microbiota disorder caused by cyclophosphamide in mice.Considering all fac-tors,the optimal dosage is 200 mg/kg.

Based on network pharmacology,through molecular docking and experimental validation,the study explored the mechanism of the Hypericum japonicum-Rehmannia glutinosa-Salvia ple-beian compound(HRS)in the treatment of liver injury.Mice were randomly divided into a control group(CON group),a model group(CCL4 group),a high-dose drug group(HRS-H group),and a low-dose group(HRS-L group).A mouse liver injury model was established using CCL4 induction,liver tissue pathological morphology was observed,and the relative expression levels of liver in-flammatory cytokine genes was measured.Active ingredients of traditional Chinese medicine and targets related to Chinese medicine and diseases were obtained from databases such as Herb,TCM-SP,PubChem,Swiss Target Prediction,Gene Cards and DisGeNET.The intersection of targets was used to obtain potential drug targets.The potential targets were analyzed for protein-protein inter-action(PPI)using the string database and a network diagram of"drug-active component-intersec-tion target"was constructed using Cytoscape.DAVID database was used for GO and KEGG path-way analysis,and Auto Dock Tools software was used for molecular docking.Finally,the results of molecular docking by examining the expression of key target genes and downstream genes such as those related to the PI3K-AKT pathway and the autophagy pathway were experimentally valida-ted.Results:Animal experiment results showed that compared to the CON group,the CCL4 group of mice exhibited disrupted liver architecture,hepatocyte steatosis,vacuolization,and extensive in-flammatory cell infiltration.These characteristics were ameliorated by drug treatment groups with the HRS-H group demonstrating superior effects compared to the HRS-L group.RT-qPCR results from mouse livers showed significantly increased relative expression of TNF-α and INOS mRNA compared to the CON group in the CCL4 group(P＜0.01),and significantly increased IL-1β mR-NA relative expression(P＜0.05).Compared to the CCL4 group,the HRS-H group showed signifi-cantly decreased TNF-α,INOS,and IL-1β mRNA relative expressions(P＜0.01).155 potential tar-gets for HRS in alleviating liver damage were identified through network pharmacology,with top-ranked key target points including STAT3,SRC,PIK3R1,PIK3CA,AKT1,HSP90A11,EGFR,and ESR.Key active ingredients included Tetramethoxyluteolin,Hispidulin,Eupafolin,Kaempferol,and Eupaformonin.GO enrichment analysis yielded 940 entries,and KEGG enrichment analysis yielded 177 biological pathways.Molecular docking results showed a strong binding ability between the main components of HRS and key target points.RT-qPCR results showed increasing trends for EGFR,PI3KCA,HSP90A11,and NF-κB mRNA compared to the CON group in the CCL4 group,significantly increased AKT1 mRNA relative expression(P＜0.05),significant decreases in ULK1,ATG5,LC3B,and ATG7 mRNA relative expressions(P＜0.05),and extremely significant decreases in PTEN,ATG13,BECLIN-1,ATG16L1,ATG12,ATG4B,and ATG3 mRNA relative expressions(P＜0.01).Compared to the CCL4 group,the HRS-H group showed significantly de-creased PI3KCA,HSP90A11,and NF-κB mRNA relative expressions(P＜0.05),extremely signif-icantly decreased EGFR,AKT1,and mTOR mRNA relative expressions(P＜0.01),increased ULK1 relative expression trends,significantly increased PTEN,ATG16L1,ATG5,LC3B,and ATG7 mRNA relative expressions(P＜0.05),extremely significantly increased ATG13,BECLIN-1,ATG12,ATG4B,and ATG3 mRNA relative expressions(P＜0.01).Conclusion:The HRS ex-erts hepatoprotective effects through multi-component,multi-pathway approaches,with alleviating inflammation and promoting hepatocyte autophagy through PI3K-AKT pathway likely being im-portant mechanisms for its protective effects.