Objective To explore the relationship between pathogenic microorganism types,age and season in 2 188 children with respiratory tract infections.Methods A total of 2 188 children with respiratory tract in-fections admitted to the Department of Pediatrics,962 Hospital,Joint Logistic Support Force of PLA from June 2023 to May 2024 were selected as the study subjects.Targeted next generation sequencing(tNGS)tech-nology was used to detect 107 common pathogenic microorganism in children with respiratory tract infections,including Haemophilus influenzae,rhinovirus,Moraxella catarrhalis,Mycoplasma pneumoniae,Staphylococcus aureus,Streptococcus pneumoniae,human parainfluenza virus,human respiratory syncytial virus,etc.The re-spiratory tract infection situation and epidemiological characteristics of children in Harbin were analyzed.Re-sults Among 2 188 pediatric patients,98.5%(2 156/2 188)tested positive for pathogenic microorganism,with Haemophilus influenzae accounting for the highest proportion of 33.5%(732/2 188),followed by rhino-virus of 25.0%(547/2 188)and Moraxella catarrhalis of 24.8%(543/2 188).The positive rates of Hae-mophilus influenzae and human adenovirus in male children were higher than those in female children(P<0.05),while there were no statistically significant differences in the positive positive rates of other pathogenic microorganism between male and female children(P>0.05).Except for human adenovirus and influenza A virus,which showed no statistically significant differences in positive rates among different age groups(P>0.05),there were statistically significant differences in the positive rates of other pathogenic microorganism a-mong different age groups(P<0.05).The positive rates of pathogenic microorganism in preschool children were relatively high.There were no statistically significant differences in the positive rates of Streptococcus and Staphylococcus aureus in different seasons(P>0.05),while there were statistically significant differences in the positive rates of other pathogenic microorganism in different seasons(P<0.05).The positive rates of Haemophilus influenzae,Streptococcus pneumoniae,human metapneumovirus,human parainfluenza virus and SARS-Cov-2 were the highest in summer(P<0.05).Conclusion 2 188 children with respiratory tract infec-tions were mainly caused by pathogenic microorganism such as Haemophilus influenzae,rhinovirus,and Moraxella catarrhalis,etc.Preschool children is a susceptible group,and the prevalence of pathogenic microor-ganism varies seasonally.In clinical practice,relevant prevention and control measures should be developed based on this characteristic to reduce the incidence of diseases.

Objective To elucidate the effects and mechanisms of Biejiajian pill(BJJP)-containing serum on the malignant biological behaviors of hepatocellular carcinoma Huh7 cells.Methods This research knocked down CKLF-like MARVEL transmembrane domain containing 6(CMTM6)expression using a CMTM6-specific small interfering RNA(siRNA).Healthy Sprague-Dawley rats were used to prepare normal rat serum and low-(0.55 g/kg),medium-(1.1 g/kg),and high-(2.2 g/kg)BJJP-containing.Huh7 cells were cultured with normal fetal bovine serum(BC),normal rat serum(NC),and low-,medium-,and high-dose BJJP serum(LBJJP,MBJJP,and HBJJP,respectively).BJJP-containing serum and si-CMTM6 were applied to Huh7 cancer cells,and the proliferation,migration,and invasion abilities were evaluated by CCK-8 and Transwell assays,respectively.Protein expression levels of proliferating cell nuclear antigen(PCNA),epithelial-mesenchymal transition(EMT)markers,and CMTM6 were detected by Western blot.Results CMTM6 knockdown significantly reduced the mRNA and protein expression level of CMTM6 in Huh7 cells(P＜0.05).There were no significant differences between the BC and NC groups in terms of cell proliferation,migration,invasion,expression levels of PCNA,EMT markers,and CMTM6(all P＞0.05).BJJP-containing serum markedly inhibited Huh7 cell proliferation,migration,and invasion(P＜0.05),downregulated PCNA,CMTM6,N-cadherin,and Vimentin expression,and upregulated E-cadherin compared with the NC group(all P＜0.05).CMTM6 knockdown suppressed malignant behaviors,with reduced PCNA,Vimentin,and N-cadherin and elevated E-cadherin expression(all P＜0.05).Conclusions BJJP-containing serum can significantly inhibit Huh7 cell growth,invasion,migration,and EMT progression,potentially mediated via CMTM6 suppression.

OBJECTIVE:Premature ovarian failure has manifested a trend of younger,and stem cell therapy has been progressively implemented in clinical practice in recent years.Nevertheless,given the extensive range of sources and variegated existence of stem cells in diverse tissues,certain disparities prevail in their biological characteristics and functions.In this paper,the therapeutic efficacies of dissimilar sources of mesenchymal stem cells on animal models of premature ovarian failure were contrasted,with the aim of providing a basis for the clinical application of stem cells.METHODS:The animal model experiments of mesenchymal stem cell therapy for premature ovarian failure were retrieved from PubMed,The Cochrane Library,and EMbase,as well as Chinese databases such as CNKI,WanFang,VIP,and China Biomedical Literature Service.The search period extended from the inception to December 31,2023.Two researchers independently screened the literature,extracted and analyzed the data.The quality of the included studies was evaluated by means of the SYRCLE animal experiment bias risk assessment table.Main outcome measures:Follicle stimulating hormone,estradiol,luteinizing hormone,the quantity of follicles at all levels.Secondary outcome measure:Pregnancy rate.Network meta-analysis,mapping,and tabulation were executed using Stata 17.0 software after assessing the risk of bias in the included studies.RESULTS:Totally 24 animal experiment studies were incorporated,and the overall quality of the literature was mediocre,encompassing 7 distinct sources of mesenchymal stem cells.They were umbilical cord-derived mesenchymal stem cells,menstrual blood-derived mesenchymal stem cells,placenta-derived mesenchymal stem cells,human cord blood-derived mesenchymal stem cells,bone marrow-derived mesenchymal stem cells,adipose-derived mesenchymal stem cells,and amnio-derived mesenchymal stem cells.The network meta-analysis demonstrated that(1)in contrast to the blank group,mesenchymal stem cells from various sources were effective in enhancing the pregnancy rate and estradiol,reducing follicle-stimulating hormone and luteinizing hormone,augmenting the number of follicles at all levels,and diminishing the number of atretic follicles.(2)According to the area map under the cumulative sequencing curve,the three stem cells with the most prominent efficacy in improving estradiol levels were umbilical cord-derived mesenchymal stem cells(72.7％)＞adipose-derived mesenchymal stem cells(72.6％)＞menstrual blood-derived mesenchymal stem cells(71.7％).(3)The three kinds of stem cells with the highest efficacy in reducing follicle-stimulating hormone levels were the adipose-derived mesenchymal stem cells(96.3％)＞human cord blood-derived mesenchymal stem cells(65.4％)＞umbilical cord-derived mesenchymal stem cells(63.9％).(4)The three kinds of stem cells with the highest efficacy in reducing luteinizing hormone levels were adipose-derived mesenchymal stem cells(100.0％)＞umbilical cord-derived mesenchymal stem cells(51.6％)＞human cord blood-derived mesenchymal stem cells(46.8％).(5)The top three kinds of stem cells for increasing the number of primordial follicles were human cord blood-derived mesenchymal stem cells(76.3％)＞umbilical cord-derived mesenchymal stem cells(75.5％)＞menstrual blood-derived mesenchymal stem cells(57.5％).(6)The top three kinds of stem cells for increasing the number of primary follicles were umbilical cord-derived mesenchymal stem cells(75.3％)＞adipose-derived mesenchymal stem cells(53.0％)＞the placenta-derived mesenchymal stem cells(51.7％).(7)The top three kinds of stem cells for increasing the number of secondary follicles were adipose-derived mesenchymal stem cells(76.1％)＞menstrual blood-derived mesenchymal stem cells(66.8％)＞umbilical cord-derived mesenchymal stem cells(66.5％).(8)The top three kinds of stem cells in reducing the number of atretic follicles were adipose-derived mesenchymal stem cells(99.9％)＞bone marrow-derived mesenchymal stem cells(68.1％)＞umbilical cord-derived mesenchymal stem cells(53.4％).CONCLUSION:(1)For animal models of premature ovarian failure,the results of the network meta-analysis disclosed that various stem cell transplantation treatments were preponderant over the blank or placebo group to varying extents,and the efficacies were comparable.(2)The results indicated that umbilical cord-derived mesenchymal stem cells were the most frequently utilized and adipose-derived mesenchymal stem cells were the most potent.More high-quality experimental study data are requisite in the future for further validation.

OBJECTIVE:Premature ovarian failure has manifested a trend of younger,and stem cell therapy has been progressively implemented in clinical practice in recent years.Nevertheless,given the extensive range of sources and variegated existence of stem cells in diverse tissues,certain disparities prevail in their biological characteristics and functions.In this paper,the therapeutic efficacies of dissimilar sources of mesenchymal stem cells on animal models of premature ovarian failure were contrasted,with the aim of providing a basis for the clinical application of stem cells.METHODS:The animal model experiments of mesenchymal stem cell therapy for premature ovarian failure were retrieved from PubMed,The Cochrane Library,and EMbase,as well as Chinese databases such as CNKI,WanFang,VIP,and China Biomedical Literature Service.The search period extended from the inception to December 31,2023.Two researchers independently screened the literature,extracted and analyzed the data.The quality of the included studies was evaluated by means of the SYRCLE animal experiment bias risk assessment table.Main outcome measures:Follicle stimulating hormone,estradiol,luteinizing hormone,the quantity of follicles at all levels.Secondary outcome measure:Pregnancy rate.Network meta-analysis,mapping,and tabulation were executed using Stata 17.0 software after assessing the risk of bias in the included studies.RESULTS:Totally 24 animal experiment studies were incorporated,and the overall quality of the literature was mediocre,encompassing 7 distinct sources of mesenchymal stem cells.They were umbilical cord-derived mesenchymal stem cells,menstrual blood-derived mesenchymal stem cells,placenta-derived mesenchymal stem cells,human cord blood-derived mesenchymal stem cells,bone marrow-derived mesenchymal stem cells,adipose-derived mesenchymal stem cells,and amnio-derived mesenchymal stem cells.The network meta-analysis demonstrated that(1)in contrast to the blank group,mesenchymal stem cells from various sources were effective in enhancing the pregnancy rate and estradiol,reducing follicle-stimulating hormone and luteinizing hormone,augmenting the number of follicles at all levels,and diminishing the number of atretic follicles.(2)According to the area map under the cumulative sequencing curve,the three stem cells with the most prominent efficacy in improving estradiol levels were umbilical cord-derived mesenchymal stem cells(72.7％)＞adipose-derived mesenchymal stem cells(72.6％)＞menstrual blood-derived mesenchymal stem cells(71.7％).(3)The three kinds of stem cells with the highest efficacy in reducing follicle-stimulating hormone levels were the adipose-derived mesenchymal stem cells(96.3％)＞human cord blood-derived mesenchymal stem cells(65.4％)＞umbilical cord-derived mesenchymal stem cells(63.9％).(4)The three kinds of stem cells with the highest efficacy in reducing luteinizing hormone levels were adipose-derived mesenchymal stem cells(100.0％)＞umbilical cord-derived mesenchymal stem cells(51.6％)＞human cord blood-derived mesenchymal stem cells(46.8％).(5)The top three kinds of stem cells for increasing the number of primordial follicles were human cord blood-derived mesenchymal stem cells(76.3％)＞umbilical cord-derived mesenchymal stem cells(75.5％)＞menstrual blood-derived mesenchymal stem cells(57.5％).(6)The top three kinds of stem cells for increasing the number of primary follicles were umbilical cord-derived mesenchymal stem cells(75.3％)＞adipose-derived mesenchymal stem cells(53.0％)＞the placenta-derived mesenchymal stem cells(51.7％).(7)The top three kinds of stem cells for increasing the number of secondary follicles were adipose-derived mesenchymal stem cells(76.1％)＞menstrual blood-derived mesenchymal stem cells(66.8％)＞umbilical cord-derived mesenchymal stem cells(66.5％).(8)The top three kinds of stem cells in reducing the number of atretic follicles were adipose-derived mesenchymal stem cells(99.9％)＞bone marrow-derived mesenchymal stem cells(68.1％)＞umbilical cord-derived mesenchymal stem cells(53.4％).CONCLUSION:(1)For animal models of premature ovarian failure,the results of the network meta-analysis disclosed that various stem cell transplantation treatments were preponderant over the blank or placebo group to varying extents,and the efficacies were comparable.(2)The results indicated that umbilical cord-derived mesenchymal stem cells were the most frequently utilized and adipose-derived mesenchymal stem cells were the most potent.More high-quality experimental study data are requisite in the future for further validation.

Objective To elucidate the effects and mechanisms of Biejiajian pill(BJJP)-containing serum on the malignant biological behaviors of hepatocellular carcinoma Huh7 cells.Methods This research knocked down CKLF-like MARVEL transmembrane domain containing 6(CMTM6)expression using a CMTM6-specific small interfering RNA(siRNA).Healthy Sprague-Dawley rats were used to prepare normal rat serum and low-(0.55 g/kg),medium-(1.1 g/kg),and high-(2.2 g/kg)BJJP-containing.Huh7 cells were cultured with normal fetal bovine serum(BC),normal rat serum(NC),and low-,medium-,and high-dose BJJP serum(LBJJP,MBJJP,and HBJJP,respectively).BJJP-containing serum and si-CMTM6 were applied to Huh7 cancer cells,and the proliferation,migration,and invasion abilities were evaluated by CCK-8 and Transwell assays,respectively.Protein expression levels of proliferating cell nuclear antigen(PCNA),epithelial-mesenchymal transition(EMT)markers,and CMTM6 were detected by Western blot.Results CMTM6 knockdown significantly reduced the mRNA and protein expression level of CMTM6 in Huh7 cells(P＜0.05).There were no significant differences between the BC and NC groups in terms of cell proliferation,migration,invasion,expression levels of PCNA,EMT markers,and CMTM6(all P＞0.05).BJJP-containing serum markedly inhibited Huh7 cell proliferation,migration,and invasion(P＜0.05),downregulated PCNA,CMTM6,N-cadherin,and Vimentin expression,and upregulated E-cadherin compared with the NC group(all P＜0.05).CMTM6 knockdown suppressed malignant behaviors,with reduced PCNA,Vimentin,and N-cadherin and elevated E-cadherin expression(all P＜0.05).Conclusions BJJP-containing serum can significantly inhibit Huh7 cell growth,invasion,migration,and EMT progression,potentially mediated via CMTM6 suppression.

Objective To investigate the effect of Biejiajian Pill(BJJP)on malignant biological behavior of hepatocellular carcinoma Hep3B cells and its regulatory mechanism.Methods A total of 72 healthy SD rats were randomly divided into blank control(BC)group,low(0.55 g/kg),medium(1.10 g/kg)and high(2.20 g/kg)BJJP experimental group,and drug-containing serum was prepared.Hep3B cells were divided into BC group,normal rat serum treatment(NC)group,low dose BJJP(LBJJP)group,medium dose BJJP(MBJJP)group and high dose BJJP(HBJJP)group,empty plasmid(pcDNA3.1)group,and CKLF-like MARVEL transmembrane domain containing 6(CMTM6)overexpression(pcDNA3.1-CMTM6)group,and the NC+pcDNA3.1 group,MBJJP+pcDNA3.1 group,NC+pcDNA3.1-CMTM6 group and MBJJP+pcDNA3.1-CMTM6 group.The proliferation of hepatoma Hep3B cells was detected by CCK-8.The migration and invasion of hepatoma Hep3B cells were detected by Transwell assay.The expression levels of proliferation-related proteins proliferating cell nuclear antigen(PCNA),epithelial-mesenchymal transition(EMT)related proteins(E-cadherin,N-cadherin and Vimentin),and CMTM6 proteins in hepatoma Hep3B cells were detected by Western blotting experiments.Results Compared with those in BC group,there were no significant differences in the proliferation,migration and invasion abilities of Hep3B cells,or the expression levels of PCNA,EMT related proteins(E-cadherin,N-cadherin and Vimentin)and CMTM6 protein in NC group(P＞0.05).Compared with NC group,LBJJP,MBJJP and HBJJP drug-containing serum inhibited the proliferation,migration and invasion of Hep3B cells,downregulated the protein expression of PCNA;MBJJP and HBJJP upregulated the protein expression of E-cadherin.The protein expressions of N-cadherin,Vimentin and CMTM6 were downregulated,with significant differences(P＜0.05).Compared with pcDNA3.1 group,the protein expression of CMTM6,cell proliferation,migration,invasion,PCNA protein expression,N-cadherin protein expression,and Vimentin protein expression in Hep3B cells in pcDNA3.1-CMTM6 group were significantly upregulated,while the protein expression of E-cadherin was significantly downregulated(P＜0.05).Compared with NC+pcDNA3.1 group,the proliferation,migration and invasion abilities of Hep3B cells in MBJJP+pcDNA3.1 group were decreased,the expression levels of PCNA,Vimentin and N-cadherin protein were decreased,while the expression level of E-cadherin protein was increased.Compared with NC+pcDNA3.1-CMTM6 group,MBJJP+pcDNA3.1-CMTM6 group had the same results in the proliferation,migration and invasion abilities of Hep3B cells and the protein expression levels of PCNA,Vimentin,N-cadherin and E-cadherin.The differences were statistically significant(all P＜0.05).Conclusion BJJP may inhibit the proliferation,migration,invasion and EMT of hepatoma Hep3B cells by regulating the expression of CMTM6.

Objective To investigate the effect of Biejiajian Pill(BJJP)on malignant biological behavior of hepatocellular carcinoma Hep3B cells and its regulatory mechanism.Methods A total of 72 healthy SD rats were randomly divided into blank control(BC)group,low(0.55 g/kg),medium(1.10 g/kg)and high(2.20 g/kg)BJJP experimental group,and drug-containing serum was prepared.Hep3B cells were divided into BC group,normal rat serum treatment(NC)group,low dose BJJP(LBJJP)group,medium dose BJJP(MBJJP)group and high dose BJJP(HBJJP)group,empty plasmid(pcDNA3.1)group,and CKLF-like MARVEL transmembrane domain containing 6(CMTM6)overexpression(pcDNA3.1-CMTM6)group,and the NC+pcDNA3.1 group,MBJJP+pcDNA3.1 group,NC+pcDNA3.1-CMTM6 group and MBJJP+pcDNA3.1-CMTM6 group.The proliferation of hepatoma Hep3B cells was detected by CCK-8.The migration and invasion of hepatoma Hep3B cells were detected by Transwell assay.The expression levels of proliferation-related proteins proliferating cell nuclear antigen(PCNA),epithelial-mesenchymal transition(EMT)related proteins(E-cadherin,N-cadherin and Vimentin),and CMTM6 proteins in hepatoma Hep3B cells were detected by Western blotting experiments.Results Compared with those in BC group,there were no significant differences in the proliferation,migration and invasion abilities of Hep3B cells,or the expression levels of PCNA,EMT related proteins(E-cadherin,N-cadherin and Vimentin)and CMTM6 protein in NC group(P＞0.05).Compared with NC group,LBJJP,MBJJP and HBJJP drug-containing serum inhibited the proliferation,migration and invasion of Hep3B cells,downregulated the protein expression of PCNA;MBJJP and HBJJP upregulated the protein expression of E-cadherin.The protein expressions of N-cadherin,Vimentin and CMTM6 were downregulated,with significant differences(P＜0.05).Compared with pcDNA3.1 group,the protein expression of CMTM6,cell proliferation,migration,invasion,PCNA protein expression,N-cadherin protein expression,and Vimentin protein expression in Hep3B cells in pcDNA3.1-CMTM6 group were significantly upregulated,while the protein expression of E-cadherin was significantly downregulated(P＜0.05).Compared with NC+pcDNA3.1 group,the proliferation,migration and invasion abilities of Hep3B cells in MBJJP+pcDNA3.1 group were decreased,the expression levels of PCNA,Vimentin and N-cadherin protein were decreased,while the expression level of E-cadherin protein was increased.Compared with NC+pcDNA3.1-CMTM6 group,MBJJP+pcDNA3.1-CMTM6 group had the same results in the proliferation,migration and invasion abilities of Hep3B cells and the protein expression levels of PCNA,Vimentin,N-cadherin and E-cadherin.The differences were statistically significant(all P＜0.05).Conclusion BJJP may inhibit the proliferation,migration,invasion and EMT of hepatoma Hep3B cells by regulating the expression of CMTM6.

Objective To observe whether the IL-23/Th17 inflammatory pathway is involved in the development of calcific aortic valve disease,and whether secukinumab can delay the progression of calcific aortic valve disease by inhibiting this pathway.Methods Forty-seven mice were divided into a blank control group,model group,and secukinumab group according to the random number table method.The blank control group was fed normal chow,while the model group and secukinumab group were fed pro-calcification chow for 16 weeks to establish a calcific aortic valve disease model.After intervention with secukinumab for 4 weeks,peak flow velocity changes in the aortic valves were detected under Doppler ultrasonography in all mice.Relevant indexes were determined by hematoxylin and eosin staining,Von Kossa staining,immunohistochemical staining,ELISA,and qPCR.Results Compared with the model group,the secukinumab group showed significantly reduced peak flow velocity(P＜0.05)and serum IL-6,IL-17,and IL-23 levels(P＜0.05)in the aortic valve.Compared with the secukinumab group,the model group's leaflet thickness was significantly increased,and there were more calcium deposits.Immunohistochemical result showed that macrophage infiltration(P＜0.05),IL-17A(P＜0.05)and IL-23(P＞0.05)levels in the valve leaflets were reduced in the secukinumab group compared with the model group.PCR result suggested that the expression of STAT3,BMP-2,and α-SMA mRNA was significantly lower in the secukinumab group than the model group(P＜0.05).Conclusions The IL-23/Th17 inflammatory pathway is involved in the pathogenesis of calcific aortic valve disease.The inflammation,fibrosis,osteogenic differentiation,and calcification of mouse valves were alleviated after intervention with secukinumab,which may delay disease progression by inhibiting the IL-23/Th17 inflammatory pathway.

Objective To explore the effect of miR-296-3p on hepatic fibrosis induced by bile duct ligation(BDL)in rats.Methods 25 SD rats were randomly divided into sham group,model(BDL)group,NC adv group,miR-296-3p adv group and miR-296-3p sponge adv group,with 5 rats in each group.The pathological changes were ob-served in rat liver tissue via HE,Masson and Sirius Red staining;the levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST)and total bilirubin(TBIL)in the serum of rat in each group were detected;the expression levels of miR-296-3p,interleukin(IL)-6,IL-1 β,tumor necrosis factor(TNF)-α and smooth muscle actin(α-SMA),type Ⅰ collagen(Col1A1),and connective tissue growth factor(CTGF)mRNA in rat liver tissue were detected by qRT-PCR;the expression levels of α-SMA,Col1A1 and CTGF proteins were detected by Western blot.Immunohistochemical staining(IHC)was performed to detect the expression of α-SMA in liver tissue.Target genes of miR-296-3p was predicted by bioinformatic analysis using the online database.Zinc finger and BTB do-main-containing protein20(ZBTB20)mRNA and protein expression levels were detected.Results The pathologi-cal staining results showed that compared with sham group,a large number of infiltrated inflammatory cells and col-lagen deposition were observed in the liver tissues of rats in the BDL group and NC adv group.Compared with NC adv group,the inflammatory cells and collagen deposition decreased in the liver tissues of miR-296-3p adv group.However,in miR-296-3p sponge adv group,collagen product and inflammatory reaction increased.Compared with sham group,the contents of ALT,AST and TBIL in serum of rats in BDL group and NC adv group increased,the expression level of miR-296-3p decreased,the mRNA expression levels of IL-6,IL-1β,TNF-α increased,and the mRNA and protein expression levels of α-SMA,Col1A1 and CTGF increased(all P＜0.05).Compared with the NC adv group,the contents of ALT,AST and TBIL in serum of rats in miR-296-3p adv group decreased,the ex-pression level of miR-296-3p increased,the mRNA expression levels of IL-6,IL-1 β and TNF-α,and the mRNA and protein expression levels of α-SMA,Col1A1 and CTGF in liver tissues decreased(all P＜0.05).The results of miR-296-3p sponge adv group were opposite to those of miR-296-3p adv group(all P＜0.05).The bioinformat-ics website predicted that ZBTB20 might be a candidate target gene of miR-296-3p.Compared with sham group,the expression of ZBTB20 mRNA and protein in the liver tissues of BDL group and NC adv group increased(P＜0.05),and the expression of ZBTB20 in the liver tissues of miR-296-3p adv group decreased compared with NC adv group(P＜0.05).However,the expression of ZBTB20 in liver tissues of miR-296-3p sponge adv group in-creased(P＜0.05).Conclusion miR-296-3p expression decreases in BDL-induced hepatic fibrosis in rats,and miR-296-3p may inhibit hepatic fibrosis in BDL rats by targeting ZBTB20.

Objective To investigate the effect of miR-199a-5p on common bile duct ligation(BDL)-induced liver fibrosis in rats by regulating intestinal flora.Methods The 25 SD rats were randomly divided into five groups:the Sham group,the BDL group,the negative control adenovirus(NC adv)group,the miR-199a-5p adv group and the miR-199a-5p sponge adv group.The pathological changes of liver tissue and the degree of liver fibrosis were ob-served by HE,Masson and Sirius Red staining.The levels of aspartate aminotransferase(AST),alanine amin-otransferase(ALT),total bilirubin(TBIL)and direct bilirubin(DBIL)in serum of rats were determined by a fully automatic biochemical analyzer.The mRNA expression level of miR-199a-5p in liver tissue of rats was detec-ted by qRT-PCR.The protein expression levels of α-smooth muscle actin(α-SMA)and collagen type 1 alpha 1(COL1A1)in liver tissue of rats were detected by double immunofluorescence staining and Western blot experi-ment.Rat feces were collected for 16S rRNA high-throughput sequencing.Results The expression of miR-199a-5p was up-regulated in the liver tissue of BDL rats(P＜0.01).Compared with the NC adv group,the degree of liver injury and collagen deposition were relatively serious,the levels of AST,ALT,TBIL and DBIL in serum and the expression levels of α-SMA and COL1A1 in liver tissue increased in the miR-199a-5p adv group(all P＜0.05).However,the results of miR-199a-5p sponge adv intervention were opposite(all P＜0.05).The 16S rRNA sequencing results showed that rats treated with miR-199a-5p adv were characterized by increased diversity and richness of intestinal microbiota,changed composition of intestinal microbiota,while the results of miR-199a-5p sponge adv interfering with the bacterial community were opposite(all P＜0.05).Conclusion miR-199a-5p promotes liver fibrosis of BDL rats,and its mechanism may be related to regulating the diversity and abundance of intestinal microbiota.