Based on network pharmacology,molecular docking,and experimental validation,this study explored the mechanisms of Geranium wilfordii Maxim(GWM)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(GWM-H group),and a low-dose group(GWM-L group).The liv-er injury model in mice was induced by CCl4,and liver tissue pathological morphology was ob-served,along with the measurement of the relative gene expression levels of liver inflammatory factors.Active ingredients of traditional Chinese medicine and target information of Chinese medi-cine and diseases were obtained through databases such as TCMSP,PubChem,Swiss Target Pre-diction,Super-PRED,Gene Cards,and DisGeNET.Intersecting the targets of liver injury,necropto-sis,and drugs yielded potential drug targets.String database was used for protein-protein interac-tion(PPI)analysis of the potential targets.Furthermore,Cytoscape was utilized to construct a net-work diagram of"drug-disease-active ingredient-intersection target,"Wei Sheng Xin was used for GO and KEGG pathway analysis.Molecular docking was performed using MOE software,and the results of molecular docking were experimentally validated to detect the expression of key targets in the RIPK1/RIPK3/MLKL signaling pathway.Animal experiments showed that compared to the CON group,the CCl4 group of mice exhibited a significant increase in liver organ index(P＜0.05),markedly elevated serum AST activity(P＜0.05),and a highly significant increase in ALT activity(P＜0.01).Pathological examination revealed chaotic liver lobules,severe hepatocyte steatosis,ex-tensive hepatocyte necrosis,and inflammatory cell infiltration in the livers of mice in the CCl4 group.In comparison to the CCl4 group,the GWM-H group showed a significant decrease in liver organ index(P＜0.05),while the GWM-L group displayed a downward trend.The GWM-H group exhibited a significant reduction in serum AST activity(P＜0.05),the GWM-L group showed a decreasing trend in serum AST activity,the GWM-H group demonstrated a highly significant de-crease in serum ALT activity(P＜0.01),and the GWM-L group displayed a significant decrease in serum ALT activity(P＜0.05).Histopathological examination revealed that the drug treatment groups could improve CCl4-induced liver injury,with the GWM-H group showing better efficacy than the GWM-L group.RT-qPCR results of liver tissues showed that compared to the CON group,the CCl4 group exhibited a highly significant increase in the relative expression of IL-1βand PGE2 mRNA(P＜0.01),while the mRNA relative expression of COX2 showed an increasing trend.In contrast,compared to the CCl4 group,the GWM-H group showed a remarkably significant decrease in the relative expression of IL-1βmRNA(P＜0.01),a significant decrease in PGE2 mR-NA expression(P＜0.05),and a decreasing trend in COX2 mRNA expression.Through network pharmacology,56 potential targets related to GWM in ameliorating necroptosis-induced liver injury were identified.Key targets,based on degree value,include TNF,Bcl2,HSP90AA1,and Caspase8,while the key components are quercetin,luteolin,kaempferol,and ellagic acid.Functional enrich-ment analysis yielded 2 173 entries for GO and 146 biological pathways for KEGG.Molecular doc-king results indicated a strong binding capacity between the main components of GWM and key targets.RT-qPCR experimental results showed that compared to the CON group,the CCl4 group exhibited a extremely significantly increase in the mRNA relative expression of TNF-α,TNFR1,MLKL(P＜0.01),significantly increase in the mRNA relative expression of FAS,RIPK1,RIPK3 mRNA(P＜0.05),and a significant decrease in Caspase 8 mRNA expression(P＜0.05).The addi-tion of GWM successfully reversed this trend;compared to the CCl4 group,the GWM-H group showed a highly significant decrease in the mRNA relative expression of TNF-α,TNFR1,FAS and MLKL mRNA(P＜0.01),significant decrease in RIPK1,RIPK3 mRNA expression(P＜0.05),and an increasing trend in CASPASE8 mRNA expression.GWM exerts hepatoprotective effects through multiple components and pathways,among which inhibition of the RIPK1/RIPK3/MLKL pathway reduces hepatocyte necroptosis,potentially serving as one of the essential mechanisms for its protective effects.

Based on network pharmacology,molecular docking,and experimental validation,this study explored the mechanisms of Geranium wilfordii Maxim(GWM)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(GWM-H group),and a low-dose group(GWM-L group).The liv-er injury model in mice was induced by CCl4,and liver tissue pathological morphology was ob-served,along with the measurement of the relative gene expression levels of liver inflammatory factors.Active ingredients of traditional Chinese medicine and target information of Chinese medi-cine and diseases were obtained through databases such as TCMSP,PubChem,Swiss Target Pre-diction,Super-PRED,Gene Cards,and DisGeNET.Intersecting the targets of liver injury,necropto-sis,and drugs yielded potential drug targets.String database was used for protein-protein interac-tion(PPI)analysis of the potential targets.Furthermore,Cytoscape was utilized to construct a net-work diagram of"drug-disease-active ingredient-intersection target,"Wei Sheng Xin was used for GO and KEGG pathway analysis.Molecular docking was performed using MOE software,and the results of molecular docking were experimentally validated to detect the expression of key targets in the RIPK1/RIPK3/MLKL signaling pathway.Animal experiments showed that compared to the CON group,the CCl4 group of mice exhibited a significant increase in liver organ index(P＜0.05),markedly elevated serum AST activity(P＜0.05),and a highly significant increase in ALT activity(P＜0.01).Pathological examination revealed chaotic liver lobules,severe hepatocyte steatosis,ex-tensive hepatocyte necrosis,and inflammatory cell infiltration in the livers of mice in the CCl4 group.In comparison to the CCl4 group,the GWM-H group showed a significant decrease in liver organ index(P＜0.05),while the GWM-L group displayed a downward trend.The GWM-H group exhibited a significant reduction in serum AST activity(P＜0.05),the GWM-L group showed a decreasing trend in serum AST activity,the GWM-H group demonstrated a highly significant de-crease in serum ALT activity(P＜0.01),and the GWM-L group displayed a significant decrease in serum ALT activity(P＜0.05).Histopathological examination revealed that the drug treatment groups could improve CCl4-induced liver injury,with the GWM-H group showing better efficacy than the GWM-L group.RT-qPCR results of liver tissues showed that compared to the CON group,the CCl4 group exhibited a highly significant increase in the relative expression of IL-1βand PGE2 mRNA(P＜0.01),while the mRNA relative expression of COX2 showed an increasing trend.In contrast,compared to the CCl4 group,the GWM-H group showed a remarkably significant decrease in the relative expression of IL-1βmRNA(P＜0.01),a significant decrease in PGE2 mR-NA expression(P＜0.05),and a decreasing trend in COX2 mRNA expression.Through network pharmacology,56 potential targets related to GWM in ameliorating necroptosis-induced liver injury were identified.Key targets,based on degree value,include TNF,Bcl2,HSP90AA1,and Caspase8,while the key components are quercetin,luteolin,kaempferol,and ellagic acid.Functional enrich-ment analysis yielded 2 173 entries for GO and 146 biological pathways for KEGG.Molecular doc-king results indicated a strong binding capacity between the main components of GWM and key targets.RT-qPCR experimental results showed that compared to the CON group,the CCl4 group exhibited a extremely significantly increase in the mRNA relative expression of TNF-α,TNFR1,MLKL(P＜0.01),significantly increase in the mRNA relative expression of FAS,RIPK1,RIPK3 mRNA(P＜0.05),and a significant decrease in Caspase 8 mRNA expression(P＜0.05).The addi-tion of GWM successfully reversed this trend;compared to the CCl4 group,the GWM-H group showed a highly significant decrease in the mRNA relative expression of TNF-α,TNFR1,FAS and MLKL mRNA(P＜0.01),significant decrease in RIPK1,RIPK3 mRNA expression(P＜0.05),and an increasing trend in CASPASE8 mRNA expression.GWM exerts hepatoprotective effects through multiple components and pathways,among which inhibition of the RIPK1/RIPK3/MLKL pathway reduces hepatocyte necroptosis,potentially serving as one of the essential mechanisms for its protective effects.

Objectives The function of the semicircular canal receptors and the pathway of the vestibulo-ocular reflex (VOR) can be diagnosed with the clinical head impulse test.The aim of the study was to investigate the horizontal VOR by means of video head impulse test in peripheral vestibular disorders.Methods Using the vHIT,we examined horizontal semicircular canal VOR in a group of 55 patients and a control group of 20 healthy subjects.The group of patients included 10 cases of vestibular neuritis (VN),6 cases of vestibular schwannoma (VS),12 cases of Meniere's disease (MD),and 15 cases of bilateral vestibulopathy (BV),as well as 13 cases of idiopathic sudden hearing loss with vertigo(ISHL).Results Instantaneous gains of 40 ms,60 ms and 80 ms of horizontal VOR were 0.88 ± 0.17,0.94 ± 0.13 and 0.96 ±0.13,respectively.Regression gain at 60 ms was 0.99 ± 0.11,and asymmetry was 5.6 ± 3.5.Normal range of 60 ms instantaneous gain was ＞ 0.73,normal range of regression gain was ＞ 0.80.AbnormalvHIT was found in VS (100％),VN (90.9％),BV (86.7％),MD (40.0％) and ISHL (38.5％).Three conditions of refixation saccades occurred in cases with abnormal VOR:isolated covert saccades (12.5％),isolated overt saccades (45.0％) and the combination of overt and covert saccades (42.5％).Conclusions The vHIT detects abnormal VOR changes in the combination of gain assessment and refixation saccades.Since isolated covert saccades in VOR changes can only be seen with vHIT,peripheral vestibular disorders are likely to be misdiagnosed with the HIT.

Curcumin can induce cell apoptosis in human chondrosarcoma cells through extrinsic death receptor pathway,inhibit proliferation of lung cancer cells by inducing cell cycle arrest,and can suppress proliferation and induce apoptosis in cholangiocarcinoma cells through blocking multiple signaling pathways,suppress the formation of tumor blood vessels by reducing the expression of vascular endothelial growth factor in Ehrlich ascites cancer cells,inhibit breast cancer cell motility and invasiveness by regulating the expression of adhesion molecules and increase the sensitivity of chemotherapy drugs to cancer cells by regulating the expression of multidrug resistance related genes.Curcumin has explored new approaches for the treatment of tumor.

Purpose:To investigate the effect of in vitro antisense oligodeoxynucleotide of cyclin D 1 on the cyclin D 1 gene expression and cell proliferation of human stomach adenocarcinoma cell BGC 823 cell line.Methods:Phosphorothioate cyclin D 1 ASODN and random oligodeoxynucleotide (RODN) were synthesized and transfected into BGC 823 Cells. Their effects on cell proliferation were examined by MTT method,RT PCR method,immunohistochemical study.Results:Cyclin D 1 ASODN could significantly inhibit the growth of BGC 823 Cell lines.The RODN showed no such effect.The inhibition peaked at 48 hour after transfection by MTT method and was dose dependent.ASODN could downregulate the expression levels of cyclin D 1 mRNA and protein by RT PCR method and immunohistochemical study respectively.Conclusions:The data suggested that ASODN could specifically inhibit the expression of cyclin D 1 mRNA and protein and regulate cell cycle and cell proliferation of BGC 823 cells. [