1.Correlation of PLA1A expression level with clinicopathological features and immune infiltration in colorectal cancer
Cheng XUTAO ; Xi YANFENG ; Guo JIANGHONG ; Cui WEI ; Zhang ZHEN ; Bu PENG ; Wu JIAYI ; Liu JING
Chinese Journal of Clinical Oncology 2025;52(3):115-121
Objective:This study investigated the expression level of phosphatidylserine-specific phospholipase A1(PLA1A)in colorectal can-cer(CRC)and analyzed its correlations with clinicopathological features,prognosis,and immune infiltration.Methods:The expression level of PLA1A in CRC was screened,and the influence of this expression level on patient prognosis was analyzed using bioinformatics methods.A cohort of 192 patients diagnosed with CRC at Shanxi Province Cancer Hospital from January to December 2020 were selected.The PLA1A ex-pression level in those with CRC was determined using immunohistochemistry(IHC)and real-time quantitative reverse transcription PCR(RT-qPCR).The relationship between PLA1A level and the clinicopathological features of the patients with CRC was analyzed using the chi-square test.The expression levels of immune cell markers CD4 and CD8 as well as immunosuppressive checkpoints PD-1,TIM-3,and CTLA-4 in CRC were detected via IHC,and their correlations with PLA1A level were analyzed using the chi-square test.Results:The results of bioinformatics analysis showed that the expression level of PLA1A in CRC tissue was higher than paracancerous tissue,which correlated with overall surviv-al(OS)(P<0.05).The IHC and RT-qPCR results showed that PLA1A expression level was significantly upregulated in CRC tissiue(P<0.05).High PLA1A level was closely associated with the TNM stage,degree of differentiation,and lymph node metastasis(P<0.05).The IHC results demonstrated that PLA1A positively correlated with the infiltrating CD8+T cell level(P<0.05).In addition,the elevated PLA1A levels upregu-lated the expressions of immunosuppressive checkpoints PD-1,TIM-3,and CTLA-4(P<0.05).Conclusions:PLA1A is highly expressed in CRC,which is closely related to immune infiltrating cells and immunosuppressive checkpoints,suggesting that PLA1A plays an important role in immune infiltration in CRC,a finding that provides guidance in the treatment of CRC.
2.Transcriptomics of the Lieber-DeCarli mouse model of alcoholic liver injury
Tianyin RUAN ; Siyuan WANG ; Xutao LI ; Hao ZHANG ; Yuan PENG ; Chenghai LIU ; Yanyan TAO
Acta Laboratorium Animalis Scientia Sinica 2025;33(2):204-215
Objective To investigate the characteristics of liver injury in the Lieber-DeCarli alcoholic liver disease(ALD)mouse model and to analyze its transcriptomic profile.Methods Eighteen male C57BL/6J mice were randomly divided into an alcohol-fed group(n = 10)and a control group(n = 8).The alcohol-fed group received a Lieber-DeCarli ethanol diet,starting with an adaptive one-week phase using incremental concentrations of ethanol(10~57.3 mL/L),followed by 2 weeks of a 57.3 mL/L concentration of 95%ethanol,for a total of 3 weeks.The control group was provided with an isocaloric control diet for 3 weeks.At the end of the study,mice were sacrificed,and serum and liver tissue samples were collected.Serum liver function markers(ALT,AST),hepatic lipids(TC,TG),reduced glutathione(GSH),total superoxide dismutase(T-SOD),and malondialdehyde(MDA)were measured using biochemical assays.The levels of inflammatory cytokines(IL-6,IL-10,TNF-α,TGF-β1)in liver tissue were assessed by ELISA.Histopathological changes in liver tissue were examined using hematoxylin-eosin(HE)and Oil Red O staining.Immunohistochemical staining using the F4/80 antibody was employed to assess changes in macrophage expression.RNA-seq analysis was conducted to identify differentially expressed genes between the two groups of liver tissues,followed by GO and KEGG pathway enrichment analysis.qRT-PCR was used to validate the expression of these differentially expressed genes.Results Compared with the control group,the alcohol-fed mice exhibited a significant decrease in body weight(P<0.01).Serum ALT and AST levels were significantly elevated(P<0.01),while liver tissue levels of TC,TG,and MDA were significantly increased(P<0.05).Conversely,GSH and T-SOD levels were significantly reduced(P<0.05).The levels of inflammatory factors IL-6,TNF-α,and TGF-β1 were increased,which was consistent with the qRT-PCR validation results(P<0.05).Histological examination revealed disrupted hepatic lobular structure,with macrovesicular steatosis,microvesicular steatosis,and ballooning degeneration.Additionally,fat droplets in liver tissue were significantly increased,and macrophage expression was upregulated.Differential gene expression analysis,using a threshold of|log2 FC|>1 and q<0.05,identified 2063 differentially expressed genes,of which 1236 were upregulated and 827 downregulated.Enriched pathways included xenobiotic metabolism via cytochrome P450,cytokine-cytokine receptor interaction,chemokine signaling,steroid hormone biosynthesis,glutathione metabolism,and retinol metabolism.(P<0.05).qRT-PCR validation confirmed the significant upregulation(e.g.,Mmp12,Gstm3,Cyp2a22)and downregulation(e.g.,Serpina1e,Acmsd,Mup3d)of 10 genes from each category,consistent with the transcriptome sequencing results.Conclusions The primary pathological mechanisms underlying alcoholic liver injury involve pathways related to xenobiotic metabolism and act via cytochrome P450,cytokine-cytokine receptor interaction,chemokine signaling,glutathione metabolism,and retinol metabolism.
3.Transcriptomics of the Lieber-DeCarli mouse model of alcoholic liver injury
Tianyin RUAN ; Siyuan WANG ; Xutao LI ; Hao ZHANG ; Yuan PENG ; Chenghai LIU ; Yanyan TAO
Acta Laboratorium Animalis Scientia Sinica 2025;33(2):204-215
Objective To investigate the characteristics of liver injury in the Lieber-DeCarli alcoholic liver disease(ALD)mouse model and to analyze its transcriptomic profile.Methods Eighteen male C57BL/6J mice were randomly divided into an alcohol-fed group(n = 10)and a control group(n = 8).The alcohol-fed group received a Lieber-DeCarli ethanol diet,starting with an adaptive one-week phase using incremental concentrations of ethanol(10~57.3 mL/L),followed by 2 weeks of a 57.3 mL/L concentration of 95%ethanol,for a total of 3 weeks.The control group was provided with an isocaloric control diet for 3 weeks.At the end of the study,mice were sacrificed,and serum and liver tissue samples were collected.Serum liver function markers(ALT,AST),hepatic lipids(TC,TG),reduced glutathione(GSH),total superoxide dismutase(T-SOD),and malondialdehyde(MDA)were measured using biochemical assays.The levels of inflammatory cytokines(IL-6,IL-10,TNF-α,TGF-β1)in liver tissue were assessed by ELISA.Histopathological changes in liver tissue were examined using hematoxylin-eosin(HE)and Oil Red O staining.Immunohistochemical staining using the F4/80 antibody was employed to assess changes in macrophage expression.RNA-seq analysis was conducted to identify differentially expressed genes between the two groups of liver tissues,followed by GO and KEGG pathway enrichment analysis.qRT-PCR was used to validate the expression of these differentially expressed genes.Results Compared with the control group,the alcohol-fed mice exhibited a significant decrease in body weight(P<0.01).Serum ALT and AST levels were significantly elevated(P<0.01),while liver tissue levels of TC,TG,and MDA were significantly increased(P<0.05).Conversely,GSH and T-SOD levels were significantly reduced(P<0.05).The levels of inflammatory factors IL-6,TNF-α,and TGF-β1 were increased,which was consistent with the qRT-PCR validation results(P<0.05).Histological examination revealed disrupted hepatic lobular structure,with macrovesicular steatosis,microvesicular steatosis,and ballooning degeneration.Additionally,fat droplets in liver tissue were significantly increased,and macrophage expression was upregulated.Differential gene expression analysis,using a threshold of|log2 FC|>1 and q<0.05,identified 2063 differentially expressed genes,of which 1236 were upregulated and 827 downregulated.Enriched pathways included xenobiotic metabolism via cytochrome P450,cytokine-cytokine receptor interaction,chemokine signaling,steroid hormone biosynthesis,glutathione metabolism,and retinol metabolism.(P<0.05).qRT-PCR validation confirmed the significant upregulation(e.g.,Mmp12,Gstm3,Cyp2a22)and downregulation(e.g.,Serpina1e,Acmsd,Mup3d)of 10 genes from each category,consistent with the transcriptome sequencing results.Conclusions The primary pathological mechanisms underlying alcoholic liver injury involve pathways related to xenobiotic metabolism and act via cytochrome P450,cytokine-cytokine receptor interaction,chemokine signaling,glutathione metabolism,and retinol metabolism.
4.Correlation of PLA1A expression level with clinicopathological features and immune infiltration in colorectal cancer
Cheng XUTAO ; Xi YANFENG ; Guo JIANGHONG ; Cui WEI ; Zhang ZHEN ; Bu PENG ; Wu JIAYI ; Liu JING
Chinese Journal of Clinical Oncology 2025;52(3):115-121
Objective:This study investigated the expression level of phosphatidylserine-specific phospholipase A1(PLA1A)in colorectal can-cer(CRC)and analyzed its correlations with clinicopathological features,prognosis,and immune infiltration.Methods:The expression level of PLA1A in CRC was screened,and the influence of this expression level on patient prognosis was analyzed using bioinformatics methods.A cohort of 192 patients diagnosed with CRC at Shanxi Province Cancer Hospital from January to December 2020 were selected.The PLA1A ex-pression level in those with CRC was determined using immunohistochemistry(IHC)and real-time quantitative reverse transcription PCR(RT-qPCR).The relationship between PLA1A level and the clinicopathological features of the patients with CRC was analyzed using the chi-square test.The expression levels of immune cell markers CD4 and CD8 as well as immunosuppressive checkpoints PD-1,TIM-3,and CTLA-4 in CRC were detected via IHC,and their correlations with PLA1A level were analyzed using the chi-square test.Results:The results of bioinformatics analysis showed that the expression level of PLA1A in CRC tissue was higher than paracancerous tissue,which correlated with overall surviv-al(OS)(P<0.05).The IHC and RT-qPCR results showed that PLA1A expression level was significantly upregulated in CRC tissiue(P<0.05).High PLA1A level was closely associated with the TNM stage,degree of differentiation,and lymph node metastasis(P<0.05).The IHC results demonstrated that PLA1A positively correlated with the infiltrating CD8+T cell level(P<0.05).In addition,the elevated PLA1A levels upregu-lated the expressions of immunosuppressive checkpoints PD-1,TIM-3,and CTLA-4(P<0.05).Conclusions:PLA1A is highly expressed in CRC,which is closely related to immune infiltrating cells and immunosuppressive checkpoints,suggesting that PLA1A plays an important role in immune infiltration in CRC,a finding that provides guidance in the treatment of CRC.
5.Epidemiological burden of glaucoma in China based on Global Burden of Dis-ease database
Tianyu GAO ; Xutao GUAN ; Ruibao LIU ; Xirui YANG
Recent Advances in Ophthalmology 2024;44(5):382-386,390
Objective To analyze the epidemiological burden of glaucoma in China based on data from the Global Burden of Disease 2019(GBD 2019)database to provide a theoretical basis for the prevention and control of glaucoma in China.Methods Analysis of the glaucoma epidemic trend and age,period,and gender trends in China from 1990 to 2019 and the correlation between glaucoma and the Socio-Demographic Index(SDI)were conducted according to relevant data on the number of patients,prevalence,disability-adjusted life years(DALYs)and DALY rates downloaded from the GBD 2019 database.Results In 2019,China was ranked globally first for the number of people with glaucoma and second for DALYs due to glaucoma.In 2019,China's glaucoma prevalence was among the highest globally,with the number of pa-tients being 1.92 times that of 1990.Both the prevalence and the DALY rate demonstrated a persistent upward trajectory.The demographic distribution of patients with glaucoma in China revealed a curve which peaked among middle-aged individ-uals,with higher prevalence observed in males than females.An inverse correlation was identified between SDI and the age-standardized prevalence and DALY rates,which exhibited a negative average annual percentage change,indicative of a pro-gressive decline with rising SDI.Conclusion From 1990 to 2019,China experienced a significant increase in the burden of glaucoma,ranking among the highest globally,with a positive correlation observed between the disease burden and the level of social development.
6.Research on the value of double closed-loop management mode in maintenance and fault control of hemodialysis machine
Xutao CAI ; Junfeng LIU ; Chen ZHANG ; Jun XU
China Medical Equipment 2024;21(1):172-177
Objective:To construct a double-closed-loop management model for medical equipment and explore its application value in hemodialysis machine maintenance and fault management.Methods:Based on the closed-loop management of clinical operation of medical equipment and the closed-loop management of technical support,a dual closed-loop management model of equipment was constructed.65 hemodialysis machines in clinical use in Huashan Hospital Fudan University from January 2021 to January 2023 were selected and divided into a conventional mode and double-closed-loop mode according to different management modes.The conventional mode adopted conventional equipment management methods,and the double-closed-loop mode adopted a double-closed-loop management model.The cost-effectiveness,social benefit,failure occurrence,effective management quality matters,disinfection status,average patient waiting time and satisfaction were compared between the two groups.Results:The hemodialysis machine operating profit growth rate,diagnosis and treatment cost growth rate,service life index,scientific research service growth rate,diagnosis and treatment service growth rate,startup rate and operation rate of the double-closed-loop mode were(3.95±1.04)%,(3.80±0.58)%,(1.58±0.31)%,(4.30±0.95)%,(7.91±1.58)%,(96.58±2.76)%and(89.90±5.58)%,which were higher than those of the Conventional mode,the difference was statistically significant(t=5.418,10.070,7.490,17.570,11.820,8.849,6.840,P<0.05).The technical support expenses growth rate,bacterial colony count,bacterial endotoxin content and average patient waiting time of patients in the Double-closed-loop mode were(2.60±0.33)%,(0.370±0.008)cfu/ml,(0.0063±0.0011)EU/ml and(0.76±0.13)h,which were less than those of the conventional mode,the difference was statistically significant(t=23.040,82.985,14.482,19.530,P<0.05).The incidence rate of hemodialysis machine failure in the double-closed-loop mode was 9.23%(6/65),which was lower than that in the conventional mode,the difference was statistically significant(x2=6.392,P<0.05);among the 120 items of management data collected,quality control testing,maintenance and repair,clinical operation,information data,and scrap processing effectiveness rates were 95.83%(115/120),89.17%(107/120),96.67%(116/120),95.00%(114/120),and 97.50%(117/120),respectively,which were higher than those of the conventional mode,the difference was statistically significant(x2=15.238,16.596,9.808,15.585,16.119,P<0.05).Conclusion:The application of closed-loop management model to hemodialysis machine maintenance and fault management can effectively improve equipment cost-effectiveness,social benefits,management quality and patient satisfaction,and reduce the incidence of faults.
7.Research Progress in TCM for the Improvement of Nasal Mucosa Remodeling after Endoscopy in Chronic Sinusitis
Jiling LIU ; Ling ZHOU ; Xutao YAO ; Yan WU
Chinese Journal of Information on Traditional Chinese Medicine 2024;31(1):185-190
TCM for the improvement of nasal mucosa remodeling after chronic sinusitis endoscopy is the key to the effective prevention of new lesions and recurrence in the surgical cavity mucosa.This article reviewed the mechanism of the nasal mucosa remodeling phenomenon after chronic sinusitis endoscopy and the unique advantages of TCM treatment.Deficiency,phlegm and blood stasis leading to"lung loss"is the perennial root of nasal mucosa remodeling.The treatment is"tonifying deficiency and removing phlegm,invigorating qi and promoting blood circulation",using acupuncture and medicine,and both internal and external treatment to regulate Th17/Treg balance and reduce the inflammatory infiltration;blocking the phosphorylation of MAPK and NF-κB pathway to inhibit TGF-β1-induced myofibroblasts,reduce collagen deposition and inhibit their remodeling,and promote the process of mucosal epithelialization,which can provide ideas for the theoretical and clinical treatment of TCM prevention and treatment of nasal mucosa remodeling.
8.Application of egg yolk oil combined with Qingrejiedu decoction for external washing to promote wound healing after perianal abscess surgery
Shixiong WU ; Lihui WEN ; Manjun LIU ; Lei ZHANG ; Bo LIU ; Xutao QI
Journal of China Medical University 2024;53(5):446-450,457
Objective To explore the promoting effect of egg yolk oil combined with heat-clearing and detoxi-fying traditional Chinese medicine external washing on postoperative wound healing in patients with perianal abscesses based on growth factors in the wound granu-lation tissue.Methods Eighty patients with perianal abscesses admitted to our hospital from January 2023 to May 2023 were randomly divided into a control group and an observation group,with 40 patients in each group.Three days after surgery,the control group received routine treatment,such as external washing with heat-clearing and detoxifying Chinese herbal medicine and dressing changes with vase-line gauze.The observation group received additional treatment with egg yolk oil,and both groups were treated continuously for 7 days.The clinical efficacy,improvement in clinical symptoms,and wound recovery at different time points;serum inflammatory cytokines;and expression levels of wound granulation tissue-related factors were compared between the two groups.Results The total effective rate was higher in the observation group than the control group(95%vs.75%,P<0.05).The time of pruritus disappearance,decaying flesh,emer-gence of new epithelium,and healing were shorter in the observation group than in the control group(P<0.05).The pain degree,wound secretion,granulation morphology and surrounding tissue edema score of the observation group were significantly decreased at 7 and 10 days after surgery compared with the control group(P<0.05).Serum levels of interleukin(IL)-1β,IL-8,and tumor necrosis factor-α were lower in the observation group than the control group at 7 and 10 days after surgery(P<0.05).The levels of basic fibroblast growth factor,vascular endothelial cell growth factor,and transforming growth factor-βin wound granulation tissue were higher in the observation group than the control group at 7 and 10 days after surgery(P<0.05).Conclusion The external application of egg yolk oil combined with heat-clearing and detoxifying Chinese herbs can significantly improve the clinical symptoms of patients with perianal abscesses and promote wound healing,with significant therapeutic effects.This may be related to the inhibition of inflammatory cytokine expression and the promotion of wound granulation tissue growth factor expression.
9.Transcriptomics-based analysis of the effect of compound Dancao granules on hepatocyte apoptosis in mice with non-alcoholic steatohepatitis
Xutao LI ; Siyuan WANG ; Tianyin RUAN ; Hao ZHANG ; Yuan PENG ; Chenghai LIU ; Yanyan TAO
Acta Laboratorium Animalis Scientia Sinica 2024;32(12):1543-1555
Objective Transcriptome sequencing technology(RNA-seq)was used to analyze the mechanism of compound Dancao granules as an intervention for high-fat feed combined with carbon tetrachloride(CCl4)-induced non-alcoholic steatohepatitis.Methods 45 male C57BL/6J mice were split into two groups at random:normal control group,model control group,obeccholic acid group 10 mg/(kg·d),and compound Dancao granules low-and high-dose groups 3.74 g/(kg·d)and 7.48 g/(kg·d),with 9 mice in each group.Normal diet was made available to the control group,and the mice in the model group were given a high-fat diet combined with the subcutaneous injection of CC14,with 100%CC14 solution(4 mL/kg)in the first application,and 40%CC14-olive oil solution(2 mL/kg)in the second application,twice a week for a total of 6 weeks.Each drug group was administered the respective drug from week 3 for a total of 4 weeks.12 h after the last administration,the serum and liver tissues of mice in each group were collected,and a biochemical kit was used to detect serum liver function.Hematoxylin-eosin(HE),sirius scarlet,and oil red O staining were used to examine histopathological changes to the liver.The levels of IL-6,IL-10,TNF-α and TGF-β in mice liver were detected via ELISA,and the expression of α-SMA was observed by immunohistochemistry.Differential gene expression was analyzed by RNA-seq and functional enrichment analysis.To verify the differential expression of mRNA,quantitative reverse transcription PCR(qRT-PCR)was used.TDT-mediated dUTP nick-end labeling(TUNEL)staining was employed to identify apoptosis.Results The model control groups had significantly higher levels of serum alanine aminotransferase(ALT),aspartate aminotransferase(AST),total cholesterol(TC),and triglycerides(TG)than normal control group(P<0.01).Additionally,there was obvious inflammatory cell infiltration in the liver tissue,collagen deposition in the sink and interlobule areas,and a significant increase in lipid droplet area(P<0.01).The levels of IL-6 and TNF-α in liver tissue were significantly increased(P<0.01),the levels of IL-10 and TGF-β were decreased(P<0.01),and the expression of α-SMA was significantly increased(P<0.01).The levels of TC,TG,ALT,and AST were significantly lower in groups that received compound Dancao granules and obeccholic acid than the model control group(P<0.01),and inflammatory cell infiltration,collagen deposition,and fat accumulation in the sink and interlobule areas were improved(P<0.01).The levels of IL-6 and TNF-α in liver tissue were significantly decreased(P<0.01),the levels of IL-10 and TGF-β were increased(P<0.05,P<0.01),and the expression of α-SMA was significantly decreased(P<0.01).RNA-seq sequencing result showed that 2819 genes in the normal control group were differentially expressed compared with the model control group,with 543 up-regulated and 2276 down-regulated genes.In a comparison of the model control group and compound Dancao granules group,240 genes were differentially expressed,including 206 up-regulated genes and 34 down-regulated genes.There were 221 genes with overlapping expression in the 2 groups and functional enrichment highlighted cell cycle(Cdt1,Plk1,Bub1b,Ttk,Knl1,Esco2,Cdc6,Ndc80,Cdc25b,Sgo1,Ccnb2,Espl1,Ccne1,Mcm4,Mcm5,Fbxo5,Bub1,Mcm2),apoptosis(Caspase3,Bax,P53,Apaf1,Bak,Caspase8),the P53 signaling pathway(P53,Ccnb2,Apaf1,Bak,Bax,Gtse1,Caspase3,Ccne1),arachidonic acid metabolism(Hpgds,Cyp2c54,Cyp2b10,Tbxas1,Cyp2c50),galactose metabolism(Hk3,Gla,Hk2,Akr1b7)and other signaling pathway genes.RNA-seq sequencing analysis showed that compound Danicao granules mainly regulated the apoptosis signaling pathway,and qRT-PCR confirmed that the mRNA expression of Caspase3,Bax,P53,Apaf1,Bak and Caspase8 in the liver tissue of the model control group was increased compared with that of the normal control group(P<0.01).Compared with the model control group,the compound Dancao granules group showed decreased mRNA expression of Caspase3,Bax,P53,Apaf1,Bak and Caspase8 in liver tissue(P<0.01).TUNEL staining showed that the number of cells showing nuclear shrinkage and apoptotic bodies decreased in the compound Dancao granule administration group.Conclusions Compound Dancao granules had a significant protective effect against non-alcoholic steatohepatitis induced by high-fat feed combined with CCl4,and its mechanism might be connected to the control of genes linked to apoptosis.
10.Transcriptomics-based analysis of the effect of compound Dancao granules on hepatocyte apoptosis in mice with non-alcoholic steatohepatitis
Xutao LI ; Siyuan WANG ; Tianyin RUAN ; Hao ZHANG ; Yuan PENG ; Chenghai LIU ; Yanyan TAO
Acta Laboratorium Animalis Scientia Sinica 2024;32(12):1543-1555
Objective Transcriptome sequencing technology(RNA-seq)was used to analyze the mechanism of compound Dancao granules as an intervention for high-fat feed combined with carbon tetrachloride(CCl4)-induced non-alcoholic steatohepatitis.Methods 45 male C57BL/6J mice were split into two groups at random:normal control group,model control group,obeccholic acid group 10 mg/(kg·d),and compound Dancao granules low-and high-dose groups 3.74 g/(kg·d)and 7.48 g/(kg·d),with 9 mice in each group.Normal diet was made available to the control group,and the mice in the model group were given a high-fat diet combined with the subcutaneous injection of CC14,with 100%CC14 solution(4 mL/kg)in the first application,and 40%CC14-olive oil solution(2 mL/kg)in the second application,twice a week for a total of 6 weeks.Each drug group was administered the respective drug from week 3 for a total of 4 weeks.12 h after the last administration,the serum and liver tissues of mice in each group were collected,and a biochemical kit was used to detect serum liver function.Hematoxylin-eosin(HE),sirius scarlet,and oil red O staining were used to examine histopathological changes to the liver.The levels of IL-6,IL-10,TNF-α and TGF-β in mice liver were detected via ELISA,and the expression of α-SMA was observed by immunohistochemistry.Differential gene expression was analyzed by RNA-seq and functional enrichment analysis.To verify the differential expression of mRNA,quantitative reverse transcription PCR(qRT-PCR)was used.TDT-mediated dUTP nick-end labeling(TUNEL)staining was employed to identify apoptosis.Results The model control groups had significantly higher levels of serum alanine aminotransferase(ALT),aspartate aminotransferase(AST),total cholesterol(TC),and triglycerides(TG)than normal control group(P<0.01).Additionally,there was obvious inflammatory cell infiltration in the liver tissue,collagen deposition in the sink and interlobule areas,and a significant increase in lipid droplet area(P<0.01).The levels of IL-6 and TNF-α in liver tissue were significantly increased(P<0.01),the levels of IL-10 and TGF-β were decreased(P<0.01),and the expression of α-SMA was significantly increased(P<0.01).The levels of TC,TG,ALT,and AST were significantly lower in groups that received compound Dancao granules and obeccholic acid than the model control group(P<0.01),and inflammatory cell infiltration,collagen deposition,and fat accumulation in the sink and interlobule areas were improved(P<0.01).The levels of IL-6 and TNF-α in liver tissue were significantly decreased(P<0.01),the levels of IL-10 and TGF-β were increased(P<0.05,P<0.01),and the expression of α-SMA was significantly decreased(P<0.01).RNA-seq sequencing result showed that 2819 genes in the normal control group were differentially expressed compared with the model control group,with 543 up-regulated and 2276 down-regulated genes.In a comparison of the model control group and compound Dancao granules group,240 genes were differentially expressed,including 206 up-regulated genes and 34 down-regulated genes.There were 221 genes with overlapping expression in the 2 groups and functional enrichment highlighted cell cycle(Cdt1,Plk1,Bub1b,Ttk,Knl1,Esco2,Cdc6,Ndc80,Cdc25b,Sgo1,Ccnb2,Espl1,Ccne1,Mcm4,Mcm5,Fbxo5,Bub1,Mcm2),apoptosis(Caspase3,Bax,P53,Apaf1,Bak,Caspase8),the P53 signaling pathway(P53,Ccnb2,Apaf1,Bak,Bax,Gtse1,Caspase3,Ccne1),arachidonic acid metabolism(Hpgds,Cyp2c54,Cyp2b10,Tbxas1,Cyp2c50),galactose metabolism(Hk3,Gla,Hk2,Akr1b7)and other signaling pathway genes.RNA-seq sequencing analysis showed that compound Danicao granules mainly regulated the apoptosis signaling pathway,and qRT-PCR confirmed that the mRNA expression of Caspase3,Bax,P53,Apaf1,Bak and Caspase8 in the liver tissue of the model control group was increased compared with that of the normal control group(P<0.01).Compared with the model control group,the compound Dancao granules group showed decreased mRNA expression of Caspase3,Bax,P53,Apaf1,Bak and Caspase8 in liver tissue(P<0.01).TUNEL staining showed that the number of cells showing nuclear shrinkage and apoptotic bodies decreased in the compound Dancao granule administration group.Conclusions Compound Dancao granules had a significant protective effect against non-alcoholic steatohepatitis induced by high-fat feed combined with CCl4,and its mechanism might be connected to the control of genes linked to apoptosis.

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