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Objective To determine the predictive factors for antiviral therapy in hepatitis B e antigen (HBeAg)-negative chronic hepatitis B virus (HBV) infection [HBeAg(-) CHBI] patients with HBV DNA＜4.3 lg IU/mL.Methods A total of 179 HBeAg (-) CHBI patients were retrospective analyzed.Histology activity index (HAI) and fibrosis (S) were scored according to the Knodell HAI scoring system,and HAI＞3 and/or S≥3 was adopted as indications for treatment.Univariate and multiple regression analysis were used to assess factors associated with treatment indications.Receiver operating curves (ROC) and area under curve (AUC) were used to determine the predictive value of relevant factors.Results There were 81 cases with HAI＞3 (45.3％) and 72 with S≥3 (40.22％),and the proportion of patients with indications for treatment was 54.7 ％.Multiple regression analysis showed that age,γ-glutamyl transpeptadase (γ-GT),platelet (PLT) and albumin (Alb) were the predictive factors for the severity of liver damage and indication for treatment (all P＜0.05).The AUC for age,PLT,γ-GT and Alb were 0.655,0.657,0.726 and 0.65,respectively,and the corresponding Yoden index for age,PLT,γ-GT,and Alb were 0.297,0.426,0.03 and 0.012,respectively,the sensitivities of predicting HBeAg (-CHBI for treatment indications were 0.643,0.842,0.705 and 0.653,respectively.Conclusions This study shows that 54.7％ of HBeAg(-)CHBI patients with HBV DNA＜4.3 lg IU/ml have significant liver histological changes and require antiviral treatment.Older age,higher γ-GT,lower PLT and lower Alb levels are the predictive factors for treatment.
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Objective To investigate the differential expression of circular RNA ( circRNA ) in patients with chronic HBV infection of different stages.Methods Seven patients with chronic HBV infection admitted in Taizhou People's Hospital from October 2014 to October 2015 were enrolled, including 4 with chronic hepatitis B ( CHB ) and 3 chronic HBV carriers;3 healthy subjects served as controls. Peripheral blood mononuclear cells (PBMCs) were separated,and the expression of circRNA molecules in PBMCs were detected by new generation of circRNA microarray and validated by fluorescent quantitative PCR.The interaction sites between circRNA and miRNA were predicted with Arraystar miRNA target prediction software.Target genes regulated by the circRNA related to miRNA were analyzed by Gene oncology (Go) and Kyoto encyclopedia of genes and genomes (KEGG) analysis.SPSS 17.0 software was used for statistical analysis.Results Compared with the healthy controls , 137 circRNA molecules of differential expression were found in patients with chronic hepatitis B , of which 89 were up-regulated and 48 were down-regulated; while 444 circRNA molecules of differential expression , of which 130 were up-regulated (>5 fold in 34 ) and 314 down-regulated , were found in chronic HBV carriers.Compared with chronic HBV carriers , 1041 circRNA molecules of differential expression were found in CHB patients , including 663 up-regulated and 378 down-regulated (>5 fold in 54).There were many miRNA responsive elements which complementary with seed regions on miRNA in different circRNA molecules.Target gene analysis demonstrated that 533 target genes regulated by hsa_circ_0038646 were related to miRNAs , 249 target genes found in hsa_circ_0087354 were related to microRNAs.GO analysis showed that function of target genes regulated by hsa_circ_0038646 related to miRNA mainly enriched in activin binding.Function of target genes regulated by hsa_circ_0087354 related to miRNA mainly enriched in armadillo repeat domain binding.KEGG analysis showed that hsa_circ_0038646 molecules related to miRNA mainly involved in T cell receptor , estrogen receptor signaling pathway and so on.Hsa_circ_0087354 molecules related to miRNA mainly involved in adherens junction , MAPK signaling pathway and so on. Conclusion There are differential expressions of circRNA in patients at different clinical stages of chronic HBV infection , which might be involved in immune regulation of chronic HBV infection through the regulation of multiple target genes and signaling pathways.
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Objective To investigate the effect of microRNA (miRNA )‐548ah targeting histone deacetylase‐4 (HDAC4) on the replication and expression of hepatitis B virus (HBV) .Methods HepG2 , 2 ,15 cells were transfected by mimics and inhibitors . The expressions of miRNA‐548ah and HDAC4 before and after transfection were detected by fluorescent quantitative polymerase chain reaction (PCR) . The expression of HDAC4 protein in HepG2 ,2 ,15 cells was detected by Western blotting .The target gene of miRNA‐548ah was analyzed by bioinformatics methods .3′UTR dual‐luciferase expression vector containing candidate HDAC4 target genes was built to test the luciferase activity .The levels of hepatitis B e antigen (HBeAg) and hepatitis B surface antigen (HBsAg) in the supernatant of cultured HepG2 ,2 ,15 cells were detected by enzyme‐linked immunosorbent assay (ELISA) .The level of HBV DNA in the supernatant of HepG2 ,2 ,15 cells was detected by fluorescent quantitative PCR .The t‐test was used for comparison between two groups ,SNK‐q tests were used for multiple groups comparisons .Results The expressions of miRNA‐548ah in HepG2 ,2 ,15 and HepG2 cells were 5 .74 ± 0 .02 and 2 .96 ± 0 .40 , respectively (t= 11 .89 ,P< 0 .01) ,and the expressions of HDAC4 mRNA were 9 .38 ± 0 .39 and 18 .13 ±0 .34 ,respectively (t = 29 .39 , P < 0 .01) . The expression of miRNA‐548ah in HepG2 ,2 ,15 cells was inhibited by transfection of miRNA‐548ah inhibitors (1 .01 ± 0 .13 ,t= 15 .48 , P< 0 .01) .Compared with control group ,the levels of HBsAg ([6 .45 ± 0 .46 ] IU/mL vs [2 .60 ± 0 .20 ] IU/mL , t = 7 .48 , P <0 .01) ,HBeAg ([5 .49 ± 0 .27] NCU/mL vs [4 .15 ± 0 .34 ] NCU/mL , t = 3 .10 , P < 0 .05 ) and HBV DNA ([3 .93 ± 0 .06] lg copy/mL vs[2 .04 ± 0 .07] lg copy/mL ,t = 18 .89 , P< 0 .01) in the supernatant of cultured HepG2 ,2 ,15 cells significantly decreased in inhibitor group . The expression of HDAC4 in HepG2 ,2 ,15 cells significantly decreased after transfection of miRNA‐548ah mimics (2 .98 ± 0 .94) ,but significantly increased after transfection of miRNA‐548ah inhibitors (23 .77 ± 6 .74 ) , with statistical significance (F= 9 .34 , P< 0 .01) .The expression of HDAC4 protein was also significantly inhibited after transfection of miRNA‐548ah mimics (0 .53 ± 0 .14 vs 0 .23 ± 0 .02 , t = 3 .58 , P = 0 .02) .The activity of luciferase was significantly inhibited by transfection of miRNA‐548ah mimics (7 .62 ± 0 .45 vs 6 .65 ±0 .27 ,t = 3 .18 , P = 0 .03 ) .Conclusion miRNA‐548ah may promote the replication and expression of HBV through the regulation of target gene HDAC4 .