Objective:To study the clinical value of hepatitis B virus pregenomic RNA(HBV-pgRNA)detection in the treatment of hepatitis B.Methods:60 patients with hepatitis B were included in the study.Serum HBV-pgRNA and HBV DNA...Objective:To study the clinical value of hepatitis B virus pregenomic RNA(HBV-pgRNA)detection in the treatment of hepatitis B.Methods:60 patients with hepatitis B were included in the study.Serum HBV-pgRNA and HBV DNA levels in different phases of infection and during treatment were detected,and serum hepatitis B surface antigen(HbsAg)titer was detected by chemiluminescent immunoassay.DNA was extracted from liver biopsy tissue,and covalently closed circular DNA was detected to predict the therapeutic value in patients.Results:At the initial stage of treatment,the level of HBV-pgRNA in phase I,II,III,and IV showed a gradual decrease.Comparing the levels of HBV-pgRNA before and after treatment,we found that the level of HBV-pgRNA was significantly lower after treatment(P<0.05).Among the indicators for predicting HBsAg seroconversion,the accuracy of HBV-pgRNA level was 85.0%(51/60).Conclusion:The clinical value of HBV-pgRNA detection in the treatment of hepatitis B is high.展开更多
Objective:To investigate the disparities and associations between HBV DNA and HBV RNA in various liver disease groups with respect to HBeAg status.Methods:Between September 2020 and September 2023,90 patients diagnose...Objective:To investigate the disparities and associations between HBV DNA and HBV RNA in various liver disease groups with respect to HBeAg status.Methods:Between September 2020 and September 2023,90 patients diagnosed with chronic hepatitis B(CHB),74 patients diagnosed with liver cirrhosis(LC),and 102 patients diagnosed with hepatocellular carcinoma(HCC)from the Department of Gastroenterology or Infection at the First Affiliated Hospital of Xi’an Jiaotong University were selected.HBV DNA,HBV RNA,and HBeAg quantitative tests were conducted using serum samples from the same patients.Results:In the three groups of cases,the HBV RNA load was higher when HBeAg was positive than when HBeAg was negative,and this difference was statistically significant.Only in the HCC group was the HBV DNA load significantly higher when HBeAg was positive than when HBeAg was negative.Additionally,there was a positive correlation between HBV DNA and HBV RNA regardless of HBeAg status.Conclusion:During HBeAg conversion,HBV RNA demonstrates a more sensitive response than HBV DNA.As CHB progresses to LC or HCC,HBV RNA exhibits better diagnostic value than HBV DNA.展开更多
AIMS To probe the effect of interferon in combination with rib- avirin on the plus and minus strands of hepatitis C virus RNA (HCV RNA). METHODS Twenty-three cases diagnosed as chronic hepatitis C (CHC) according to p...AIMS To probe the effect of interferon in combination with rib- avirin on the plus and minus strands of hepatitis C virus RNA (HCV RNA). METHODS Twenty-three cases diagnosed as chronic hepatitis C (CHC) according to positive HCV RNA/anti-HCV,fluctuating levels of aminotransferase activities (>1 year) and absence of other hepatitis virus marker,were studied. Among them,13 pa- tients received combined antiviral therapy (subcutaneous injection of 3MU of interferon-α three times per week for 3 months and intra- venous drip of 1 g of ribavirin per day during the first month of treatment with interferon) and 10 patients received single interfer- on therapy (the same as above-mentioned) as control. The plus and minus strands of HCV RNA in sera and peripheral blood mononuclear cells (PBMCs) of these patients were tested by means of nested reverse transcription-polymerase chain reaction (nested RT-PCR). RESULTS At the end of therapy,the abnormal ALT levels de- creased to normal range in 9 (69.23%) cases in the combined antiviral group. Of them,5 (55.56%)experienced post-therapy relapse and 4 (44.44%) were complete responders. In the inter- feron group,the ALT decreased to normal in 6 (60%) cases,of which,4 (66.67%) had post-therapy relapse and 2 (33.33%) were complete responders. The differences between the two groups were nonsignificant (P>0.05). At the end of therapy,the positive rate of the plus strand in sera decreased from 92.3% to 38.46% (P<0.05) and that of the minus strand in PBMCs,from 76.92% to 38.46% (P<0.05) in the combined antiviral group; and in the interferon group,the former decreased from 100% to 50% (P<0.05) and the latter,from 90% to 40% (P<0.05). Again,no significant differences were found between groups (P >0.05). The relapse occurred in patients whose plus strand HCV RNA in PBMCs remained positive before and after treatment. CONCLUSIONS Ribavirin could not enhance the antiviral effect of interferon. The absence of HCV RNA in serum does not mean complete clearance of HCV,and its value for evaluating the an- tiviral effect and prognosis is limited. Therefore,it is essential to measure the plus and minus strands of HCV RNA in sera and PBM- Cs simultaneously.展开更多
BACKGROUND Characteristics of alterations of serum hepatitis B virus(HBV) RNA in different chronic hepatitis B(CHB) patients still cannot be fully explained. Whether HBV RNA can predict HBeAg seroconversion is still c...BACKGROUND Characteristics of alterations of serum hepatitis B virus(HBV) RNA in different chronic hepatitis B(CHB) patients still cannot be fully explained. Whether HBV RNA can predict HBeAg seroconversion is still controversial.AIM To investigate whether HBV RNA can predict virological response or HBeAg seroconversion during entecavir(ETV) treatment when HBV DNA is undetectable.METHODS The present study evaluated 61 individuals who were diagnosed and treated with long-term ETV monotherapy at the Department of Infectious Diseases of Peking University First Hospital(China) from September 2006 to December 2007.Finally, 30 treatment-naive individuals were included. Serum HBV RNA were extracted from 140 μL serum samples at two time points. Then they were reverse transcribed to cDNA with the HBV-specific primer. The product was quantified by real-time quantitative PCR(RT-PCR) using TAMARA probes. Statistical analyses were performed with IBM SPSS 20.0.RESULTS Level of serum HBV RNA at baseline was 4.15 ± 0.90 log10 copies/mL. HBV RNA levels showed no significant difference between the virological response(VR)and partial VR(PVR) groups at baseline(P = 0.940). Serum HBV RNA significantly decreased among patients who achieved a VR during ETV therapy(P < 0.001). The levels of HBV RNA in both HBeAg-positive patients with seroconversion group and those with no seroconversion increased after 24 wk of treatment. Overall, HBV RNA significantly but mildly correlated to HBsAg(r =0.265, P = 0.041), and HBV RNA was not correlated to HBV DNA(r = 0.242, P =0.062). Furthermore, serum HBV RNA was an independent indicator for predicting HBeAg seroconversion and virological response. HBeAg seroconversion was more likely in CHB patients with HBV RNA levels below4.12 log10 copies/mL before treatment.CONLUSION The level of serum HBV RNA could predict HBeAg seroconversion and PVR during treatment. In the PVR group, the level of serum HBV RNA tends to be increasing.展开更多
AIM To investigate the existence of hepatitis C virus (HCV) RNA in serum and in peripheral blood mononuclear cells (PBMC) of patients with hepatitis C and its clinical significance. METHODS HCV RNA was detected by ...AIM To investigate the existence of hepatitis C virus (HCV) RNA in serum and in peripheral blood mononuclear cells (PBMC) of patients with hepatitis C and its clinical significance. METHODS HCV RNA was detected by nested polymerase chain reaction (Nested PCR) in serum and in PBMC of 46 patients with acute hepatitis C (AHC) and in 42 with chronic hepatitis C (CHC). RESULTS The positive rate of HCV RNA in PBMC of patients with CHC was markedly higher than that of patients with AHC ( P <0 01). The positive rates of HCV RNA in serum of patients with AHC and CHC and in PBMC of patients with CHC were significantly higher than those of anti HCV positive patients with normal ALT level ( P <0 01). HCV RNA was negative in serum of 2 patients, but could be detected in PBMC. In 12 patients anti HCV was negative while HCV RNA was positive in serum. CONCLUSION ① detection of serum HCV RNA by nested PCR might be helpful in the early diagnosis of anti HCV negative hepatitis C; ② liver damage in patients with hepatitis C might be correlated with HCV viremia; ③ infection of PBMC by HCV might play an important role in the chronic liver damage in patients with HC and the chronicity of its clinical course; ④ PBMC might be considered as a “reservoir” for HCV.展开更多
AIM: To investigate the inhibitory effects of hepatitis B virus (HBV) replication and expression by transfecting artificial microRNA (amiRNA) into HepG2.2.15 cells. METHODS: Three amiRNA-HBV plasmids were constructed ...AIM: To investigate the inhibitory effects of hepatitis B virus (HBV) replication and expression by transfecting artificial microRNA (amiRNA) into HepG2.2.15 cells. METHODS: Three amiRNA-HBV plasmids were constructed and transfected into HepG2.2.15 cells. HBV antigen secretion was detected in the cells with transient and stable transfection by time-resolved fluoroimmunoassays (TRFIA). HBV DNA replication was examined by ? uorescence quantitative PCR, and the level of HBV S mRNA was measured by semi- quantitative RT-PCR. RESULTS: The efficiency of transient transfection of the vectors into 2.2.15 cells was 55%-60%. All the vectors had significant inhibition effects on HBsAg and HBeAg at 72 h and 96 h after transfection (P < 0.01 for all). The secretion of HBsAg and HBeAg into the supernatant was inhibited by 49.8% ± 4.7% and 39.9% ± 6.7%, respectively, at 72 h in amiRNA- HBV-S608 plasmid transfection group. The copy of HBV DNA within culture supernatant was also significantly decreased at 72 h and 96 h after transfection (P <0.01 for all). In the cells with stable transfection, the secretion of HBsAg and HBeAg into the supernatant was significantly inhibited in all three transfection groups (P < 0.01 for all, vs negative control). The copies of HBV DNA were inhibited by 33.4% ± 3.0%, 60.8% ± 2.3% and 70.1% ± 3.3%, respectively. CONCLUSION: In HepG2.2.15 cells, HBV replication and expression could be inhibited by artif icial microRNA targeting the HBV S coding region. Vector-based artificial microRNA could be a promising therapeutic approach for chronic HBV infection.展开更多
AIM： To observe the inhibition of hepatitis B virus replication and expression by transfecting vector-based small interference RNA （siRNA） pGenesiI-HBV X targeting HBV X gene region into HepG2.2.15 cells. METHODS：...AIM： To observe the inhibition of hepatitis B virus replication and expression by transfecting vector-based small interference RNA （siRNA） pGenesiI-HBV X targeting HBV X gene region into HepG2.2.15 cells. METHODS：pGenesil-HBV X was constructed and transfected into HepG2.2.15 cells via lipofection. HBV antigen secretion was determined 24, 48, and 72 h after transfection by time-resolved immunofluorometric assays （TRFIA）. HBV replication was examined by fluorescence quantitative PCR, and the expression of cytoplasmic viral proteins was determined by immunohistochemistry. RESULTS： The secretion of HBsAg and HBeAg into the supernatant was found to be inhibited by 28.5% and 32.2% （P 〈 0.01）, and by 38.67% （P 〈 0.05） and 42.86% （P 〈 0.01） at 48 h and 72 h after pGenesil-HBV X transfection, respectively. Immunohistochemical staining for cytoplasmic HBsAg showed a similar decline in HepG2.2.15 cells 48 h after transfection. The number of HBV genomes within culture supernatants was also significantly decreased 48 h and 72 h post-transfection as quantified by fluorescence PCR （P 〈 0.05）. CONCLUSION： In HepG2.2.15 cells, HBV replication and expression is inhibited by vector-based siRNA pGenesil- HBV X targeting the HBV X coding region.展开更多
AIM： To observe the inhibition of hepatitis B virus （HBV） replication and expression in HepG2.2.15 cells by combination of small interfering RNAs （siRNAs）. METHODS： Recombinant plasmid psiI-HBV was constructed a...AIM： To observe the inhibition of hepatitis B virus （HBV） replication and expression in HepG2.2.15 cells by combination of small interfering RNAs （siRNAs）. METHODS： Recombinant plasmid psiI-HBV was constructed and transfected into HepG2.2.15 cells. At 48 h, 72 h and 96 h after transfection, culture media were collected and cells were harvested for HBV replication assay. HBsAg and HBeAg in the cell culture medium were detected by enzyme-linked immunoadsorbent assay （ELISA）. Intracellular viral DNA and covalently closed circular DNA （cccDNA） were quantified by real-time polymerase chain reaction （PCR）. HBV viral mRNA was reverse transcribed and quantified by reverse-transcript PCR （RT-PCR）. RESULTS： siRNAs showed marked anti-HBV effects. siRNAs could specifically inhibit the expression of HBsAg and the replication of HBV DNA in a dosedependent manner. Furthermore, combination of siRNAs, compared with individual use of each siRNA, exerted a stronger inhibition on antigen expression and viral replication. More importantlycombination of siRNAs significantly suppressed HBV cccDNA amplification. CONCLUSION： Combination of siRNAs mediates a stronger inhibition on viral replication and antigenexpression in HepG2.2.15 cells, especially on cccDNA amplification.展开更多
BACKGROUND Hepatitis B virus,together with hepatitis C virus,has been recognized as the leading causes of hepatocellular carcinoma(HCC).Long non-coding RNAs(lncRNAs)have been suggested in increasing studies to be the ...BACKGROUND Hepatitis B virus,together with hepatitis C virus,has been recognized as the leading causes of hepatocellular carcinoma(HCC).Long non-coding RNAs(lncRNAs)have been suggested in increasing studies to be the potential prognostic factors for HCC.However,the role of combined application of lncRNAs in estimating overall survival(OS)for hepatitis virus positive HCC(VHCC)is uncertain.AIM To construct an lncRNA signature related to the OS of VHCC patients to enhance the accuracy of prognosis prediction.METHODS The expression patterns of lncRNAs,as well as related clinical data were collected from 149 VHCC patients from The Cancer Genome Atlas database.The R package was adopted to obtain the differentially expressed lncRNAs(DElncRNAs).LncRNAs significantly associated with OS were screened by means of univariate Cox regression analysis,so as to construct a least absolute shrinkage and selection operator(LASSO)model.Subsequently,the constructed lncRNA signature was developed and validated.Afterwards,the prognostic nomogram was established,which combined the as-established lncRNA signature as well as the clinical features.Meanwhile,subgroup analysis stratified by the virus type was also performed.Finally,the above-mentioned lncRNAs were enriched to corresponding pathways according to the markedly coexpressed genes.RESULTS A total of 1420 DElncRNAs were identified,among which 406 were significant in univariate Cox regression analysis.LASSO regression confirmed 8 out of the 406 lncRNAs,including AC005722.2,AC107959.3,AL353803.1,AL589182.1,AP000844.2,AP002478.1,FLJ36000,and NPSR1-AS1.Then,the prognostic risk score was calculated.Our results displayed a significant association between the risk model and the OS of VHCC[hazard ratio=1.94,95%confidence interval(CI):1.61-2.34,log-rank P=2e-10].The inference tree suggested that the established lncRNA signature was useful in the risk stratification of VHCC.Furthermore,a nomogram was plotted,and the concordance index of internal validation was 0.763(95%CI:0.700-0.826).Moreover,the subgroup analysis regarding etiology confirmed this risk model.In addition,the Wnt signaling pathway,angiogenesis,the p53 pathway,and the PI3 kinase pathway were the remarkably enriched pathways.CONCLUSION An eight-lncRNA signature has been established to predict the prognosis for VHCC,which contributes to providing a novel foundation for the targeted therapy of VHCC.展开更多
Viruses can alter the expression of host microRNAs(miRNA s) and modulate the immune response during a persistent infection. The dysregulation of host miRNA s by hepatitis B virus(HBV) contributes to the proinflammator...Viruses can alter the expression of host microRNAs(miRNA s) and modulate the immune response during a persistent infection. The dysregulation of host miRNA s by hepatitis B virus(HBV) contributes to the proinflammatory and profibrotic changes within the liver. Multiple studies have documented the differential regulation of intracellular and circulating miRNA s during different stages of HBV infection. Circulating miRNA s found in plasma and/or extracellular vesicles can integrate data on viral-host interactions and on the associated liver injury. Hence, the detection of circulating miRNA s in chronic HBV hepatitis could offer a promising alternative to liver biopsy, as their expression is associated with HBV replication, the progression of liver fibrosis,and the outcome of antiviral treatment. The current review explores the available data on miRNA involvement in HBV pathogenesis with an emphasis on their potential use as biomarkers for liver fibrosis.展开更多
BACKGROUND: The resolution of hepatitis C, evidenced by normalization of liver function and disappearance of hepatitis C virus RNA from serum as determined by conventional laboratory assays, reflects virus eradication...BACKGROUND: The resolution of hepatitis C, evidenced by normalization of liver function and disappearance of hepatitis C virus RNA from serum as determined by conventional laboratory assays, reflects virus eradication. But in interferon treated patients the HCV RNA in serum sometimes could not show the virus in cells. Such factors as virus genotype, HCV RNA contents in serum, HCV specific cellular immunities after treatment were reported to predict the response to interferon therapy. In most patients, HCV RNA could detect the virus in peripheral blood mononucle-ar cell. The aim of this study was to investigate the predictive value of HCV RNA in PBMC of patients with chronic hepatitis C after interferon treatment. METHODS: Sixteen patients with chronic hepatitis C were treated with interferon for 24 weeks, and they all get complete responses at 12 weeks of treatment. At the end of treatment, the HCV RNA in PBMC and serum were detected by RT-PCR, and after stopping treatment, HCV RNA in serum was monitored continually. RESULTS: In 9 patients who were HCV RNA positive in their PBMC at the end of treatment, 8 showed serum HCV RNA positive after 24 weeks and another 1 after 1 year. In 7 patients with negative HCV RNA in their PBMC, only 2 patients relapsed in serum HCV RNA after 1-year follow-up, and others remained viral response after 3.5 years. CONCLUSION: HCV RNA in PBMC at the end of IFN treatment is a predictor of durable response to antiviral therapy in patients with chronic hepatitis C.展开更多
Chronic infection with the hepatitis B virus(HBV) is the leading risk factor for the development of hepatocellular carcinoma(HCC). With nearly 750000 deaths yearly, hepatocellular carcinoma is the second highest cause...Chronic infection with the hepatitis B virus(HBV) is the leading risk factor for the development of hepatocellular carcinoma(HCC). With nearly 750000 deaths yearly, hepatocellular carcinoma is the second highest cause of cancer-related death in the world. Unfortunately, the molecular mechanisms that contribute to the development of HBV-associated HCC remain incompletely understood. Recently, micro RNAs(mi RNAs), a family of small non-coding RNAs that play a role primarily in post-transcriptional gene regulation, have been recognized as important regulators of cellular homeostasis, and altered regulation of mi RNA expression has been suggested to play a significant role in virus-associated diseases and the development of many cancers. With this in mind, many groups have begun to investigate the relationship between mi RNAs and HBV replication and HBV-associated disease. Multiple findings suggest that some mi RNAs, such as mi R-122, and mi R-125 and mi R-199 family members, are playing a role in HBV replication and HBV-associated disease, including the development of HBV-associated HCC. In this review, we discuss the current state of our understanding of the relationship between HBV and mi RNAs, including how HBV affects cellular mi RNAs, how these mi RNAs impact HBV replication, and the relationship between HBV-mediated mi RNA regulation and HCC development. We also address the impact of challenges in studying HBV, such as the lack of an effective model system for infectivity and a reliance on transformed cell lines, on our understanding of the relationship between HBV and mi RNAs, and proposepotential applications of mi RNA-related techniques that could enhance our understanding of the role mi RNAs play in HBV replication and HBV-associated disease, ultimately leading to new therapeutic options and improved patient outcomes.展开更多
BACKGROUND Diffusion-weighted magnetic resonance imaging has shown promise in the detection and quantification of hepatic fibrosis. In addition, the liver has numerous endogenous micro-RNAs(miRs) that play important r...BACKGROUND Diffusion-weighted magnetic resonance imaging has shown promise in the detection and quantification of hepatic fibrosis. In addition, the liver has numerous endogenous micro-RNAs(miRs) that play important roles in the regulation of biological processes such as cell proliferation and hepatic fibrosis.AIM To assess diffusion-weighted magnetic resonance imaging and miRs in diagnosing and staging hepatic fibrosis in patients with chronic hepatitis C.METHODS This prospective study included 208 patients and 82 age-and sex-matched controls who underwent diffusion-weighted magnetic resonance imaging of the abdomen, miR profiling, and liver biopsy. Pathological scoring was classified according to the METAVIR scoring system. The apparent diffusion coefficient (ADC) and miR were calculated and correlated with pathological scoring.RESULTS The ADC value decreased significantly with the progression of fibrosis, from controls(F0) to patients with early fibrosis(F1 and F2) to those with late fibrosis(F3 and F4)(median 1.92, 1.53, and 1.25 × 10^(-3) mm^2/s, respectively)(P = 0.001).The cut-off ADC value used to differentiate patients from controls was 1.83 × 10^(-3) mm^2/s with an area under the curve(AUC) of 0.992. Combining ADC and miR-200 b revealed the highest AUC(0.995) for differentiating patients from controls with an accuracy of 96.9%. The cut-off ADC used to differentiate early fibrosis from late fibrosis was 1.54 × 10^(-3) mm^2/s with an AUC of 0.866. The combination of ADC and miR-200 b revealed the best AUC(0.925) for differentiating early fibrosis from late fibrosis with an accuracy of 80.2%. The ADC correlated with miR-200 b(r =-0.61, P = 0.001), miR-21(r =-0.62, P = 0.001), and miR-29(r = 0.52,P = 0.001).CONCLUSION Combining ADC and miRs offers an alternative surrogate non-invasive diagnostic tool for diagnosing and staging hepatic fibrosis in patients with chronic hepatitis C.展开更多
AIM: To examine the effect of hepatitis C virus (HCV) structural mimics of regulatory regions of the genome on HCV replication.METHODS: HCV RNA structural mimics were constructed and tested in a HCV genotype 1b aBB7 r...AIM: To examine the effect of hepatitis C virus (HCV) structural mimics of regulatory regions of the genome on HCV replication.METHODS: HCV RNA structural mimics were constructed and tested in a HCV genotype 1b aBB7 replicon,and a Japanese fulminant hepatitis-1 (JFH-1) HCV genotype 2a infection model.All sequences were computer-predicted to adopt stem-loop structures identical to the corresponding elements in full-length viral RNA.Huh7.5 cells bearing the BB7 replicon or infected with JFH-1 virus were transfected with expression vectors generating HCV mimics and controls.Cellular HCV RNA and protein levels were quantified by real-time polymerase chain reaction and Western blotting,respectively.To evaluate possible antisense effects,complementary RNAs spanning a mimic were prepared.RESULTS: In the BB7 genotype 1b replicon system,mimics of the polymerase (NS-5B),X and BA regions inhibited replication by more than 90%,50%,and 60%,respectively.In the JFH-1 genotype 2 infection system,mimics that were only 74% and 46% identical in sequence relative to the corresponding region in JFH-1 inhibited HCV replication by 91.5% and 91.2%,respectively,as effectively as a mimic with complete identity to HCV genotype 2a.The inhibitory effects were confirmed by NS3 protein levels.Antisense RNA molecules spanning the 74% identical mimic had no significant effects.CONCLUSION: HCV RNA structural mimics can inhibit HCV RNA replication in replicon and infectious HCV systems and do so independent of close sequence identity with the target.展开更多
基金supported by the grant from SPPH Foundation for Development of Science and Technology(2021BJ-26)International Science and Technology Cooperation Projects of Shaanxi Province(2022KW-14).
文摘Objective:To study the clinical value of hepatitis B virus pregenomic RNA(HBV-pgRNA)detection in the treatment of hepatitis B.Methods:60 patients with hepatitis B were included in the study.Serum HBV-pgRNA and HBV DNA levels in different phases of infection and during treatment were detected,and serum hepatitis B surface antigen(HbsAg)titer was detected by chemiluminescent immunoassay.DNA was extracted from liver biopsy tissue,and covalently closed circular DNA was detected to predict the therapeutic value in patients.Results:At the initial stage of treatment,the level of HBV-pgRNA in phase I,II,III,and IV showed a gradual decrease.Comparing the levels of HBV-pgRNA before and after treatment,we found that the level of HBV-pgRNA was significantly lower after treatment(P<0.05).Among the indicators for predicting HBsAg seroconversion,the accuracy of HBV-pgRNA level was 85.0%(51/60).Conclusion:The clinical value of HBV-pgRNA detection in the treatment of hepatitis B is high.
文摘Objective:To investigate the disparities and associations between HBV DNA and HBV RNA in various liver disease groups with respect to HBeAg status.Methods:Between September 2020 and September 2023,90 patients diagnosed with chronic hepatitis B(CHB),74 patients diagnosed with liver cirrhosis(LC),and 102 patients diagnosed with hepatocellular carcinoma(HCC)from the Department of Gastroenterology or Infection at the First Affiliated Hospital of Xi’an Jiaotong University were selected.HBV DNA,HBV RNA,and HBeAg quantitative tests were conducted using serum samples from the same patients.Results:In the three groups of cases,the HBV RNA load was higher when HBeAg was positive than when HBeAg was negative,and this difference was statistically significant.Only in the HCC group was the HBV DNA load significantly higher when HBeAg was positive than when HBeAg was negative.Additionally,there was a positive correlation between HBV DNA and HBV RNA regardless of HBeAg status.Conclusion:During HBeAg conversion,HBV RNA demonstrates a more sensitive response than HBV DNA.As CHB progresses to LC or HCC,HBV RNA exhibits better diagnostic value than HBV DNA.
基金Supported by the Science Foundation of the Education Committee of China (1990) 360.
文摘AIMS To probe the effect of interferon in combination with rib- avirin on the plus and minus strands of hepatitis C virus RNA (HCV RNA). METHODS Twenty-three cases diagnosed as chronic hepatitis C (CHC) according to positive HCV RNA/anti-HCV,fluctuating levels of aminotransferase activities (>1 year) and absence of other hepatitis virus marker,were studied. Among them,13 pa- tients received combined antiviral therapy (subcutaneous injection of 3MU of interferon-α three times per week for 3 months and intra- venous drip of 1 g of ribavirin per day during the first month of treatment with interferon) and 10 patients received single interfer- on therapy (the same as above-mentioned) as control. The plus and minus strands of HCV RNA in sera and peripheral blood mononuclear cells (PBMCs) of these patients were tested by means of nested reverse transcription-polymerase chain reaction (nested RT-PCR). RESULTS At the end of therapy,the abnormal ALT levels de- creased to normal range in 9 (69.23%) cases in the combined antiviral group. Of them,5 (55.56%)experienced post-therapy relapse and 4 (44.44%) were complete responders. In the inter- feron group,the ALT decreased to normal in 6 (60%) cases,of which,4 (66.67%) had post-therapy relapse and 2 (33.33%) were complete responders. The differences between the two groups were nonsignificant (P>0.05). At the end of therapy,the positive rate of the plus strand in sera decreased from 92.3% to 38.46% (P<0.05) and that of the minus strand in PBMCs,from 76.92% to 38.46% (P<0.05) in the combined antiviral group; and in the interferon group,the former decreased from 100% to 50% (P<0.05) and the latter,from 90% to 40% (P<0.05). Again,no significant differences were found between groups (P >0.05). The relapse occurred in patients whose plus strand HCV RNA in PBMCs remained positive before and after treatment. CONCLUSIONS Ribavirin could not enhance the antiviral effect of interferon. The absence of HCV RNA in serum does not mean complete clearance of HCV,and its value for evaluating the an- tiviral effect and prognosis is limited. Therefore,it is essential to measure the plus and minus strands of HCV RNA in sera and PBM- Cs simultaneously.
文摘BACKGROUND Characteristics of alterations of serum hepatitis B virus(HBV) RNA in different chronic hepatitis B(CHB) patients still cannot be fully explained. Whether HBV RNA can predict HBeAg seroconversion is still controversial.AIM To investigate whether HBV RNA can predict virological response or HBeAg seroconversion during entecavir(ETV) treatment when HBV DNA is undetectable.METHODS The present study evaluated 61 individuals who were diagnosed and treated with long-term ETV monotherapy at the Department of Infectious Diseases of Peking University First Hospital(China) from September 2006 to December 2007.Finally, 30 treatment-naive individuals were included. Serum HBV RNA were extracted from 140 μL serum samples at two time points. Then they were reverse transcribed to cDNA with the HBV-specific primer. The product was quantified by real-time quantitative PCR(RT-PCR) using TAMARA probes. Statistical analyses were performed with IBM SPSS 20.0.RESULTS Level of serum HBV RNA at baseline was 4.15 ± 0.90 log10 copies/mL. HBV RNA levels showed no significant difference between the virological response(VR)and partial VR(PVR) groups at baseline(P = 0.940). Serum HBV RNA significantly decreased among patients who achieved a VR during ETV therapy(P < 0.001). The levels of HBV RNA in both HBeAg-positive patients with seroconversion group and those with no seroconversion increased after 24 wk of treatment. Overall, HBV RNA significantly but mildly correlated to HBsAg(r =0.265, P = 0.041), and HBV RNA was not correlated to HBV DNA(r = 0.242, P =0.062). Furthermore, serum HBV RNA was an independent indicator for predicting HBeAg seroconversion and virological response. HBeAg seroconversion was more likely in CHB patients with HBV RNA levels below4.12 log10 copies/mL before treatment.CONLUSION The level of serum HBV RNA could predict HBeAg seroconversion and PVR during treatment. In the PVR group, the level of serum HBV RNA tends to be increasing.
文摘AIM To investigate the existence of hepatitis C virus (HCV) RNA in serum and in peripheral blood mononuclear cells (PBMC) of patients with hepatitis C and its clinical significance. METHODS HCV RNA was detected by nested polymerase chain reaction (Nested PCR) in serum and in PBMC of 46 patients with acute hepatitis C (AHC) and in 42 with chronic hepatitis C (CHC). RESULTS The positive rate of HCV RNA in PBMC of patients with CHC was markedly higher than that of patients with AHC ( P <0 01). The positive rates of HCV RNA in serum of patients with AHC and CHC and in PBMC of patients with CHC were significantly higher than those of anti HCV positive patients with normal ALT level ( P <0 01). HCV RNA was negative in serum of 2 patients, but could be detected in PBMC. In 12 patients anti HCV was negative while HCV RNA was positive in serum. CONCLUSION ① detection of serum HCV RNA by nested PCR might be helpful in the early diagnosis of anti HCV negative hepatitis C; ② liver damage in patients with hepatitis C might be correlated with HCV viremia; ③ infection of PBMC by HCV might play an important role in the chronic liver damage in patients with HC and the chronicity of its clinical course; ④ PBMC might be considered as a “reservoir” for HCV.
基金The National Natural Science Foundation ofChina, No. 30700698
文摘AIM: To investigate the inhibitory effects of hepatitis B virus (HBV) replication and expression by transfecting artificial microRNA (amiRNA) into HepG2.2.15 cells. METHODS: Three amiRNA-HBV plasmids were constructed and transfected into HepG2.2.15 cells. HBV antigen secretion was detected in the cells with transient and stable transfection by time-resolved fluoroimmunoassays (TRFIA). HBV DNA replication was examined by ? uorescence quantitative PCR, and the level of HBV S mRNA was measured by semi- quantitative RT-PCR. RESULTS: The efficiency of transient transfection of the vectors into 2.2.15 cells was 55%-60%. All the vectors had significant inhibition effects on HBsAg and HBeAg at 72 h and 96 h after transfection (P < 0.01 for all). The secretion of HBsAg and HBeAg into the supernatant was inhibited by 49.8% ± 4.7% and 39.9% ± 6.7%, respectively, at 72 h in amiRNA- HBV-S608 plasmid transfection group. The copy of HBV DNA within culture supernatant was also significantly decreased at 72 h and 96 h after transfection (P <0.01 for all). In the cells with stable transfection, the secretion of HBsAg and HBeAg into the supernatant was significantly inhibited in all three transfection groups (P < 0.01 for all, vs negative control). The copies of HBV DNA were inhibited by 33.4% ± 3.0%, 60.8% ± 2.3% and 70.1% ± 3.3%, respectively. CONCLUSION: In HepG2.2.15 cells, HBV replication and expression could be inhibited by artif icial microRNA targeting the HBV S coding region. Vector-based artificial microRNA could be a promising therapeutic approach for chronic HBV infection.
基金Supported by Natural Science Foundation of Shanxi Province, China, No.20051114
文摘AIM： To observe the inhibition of hepatitis B virus replication and expression by transfecting vector-based small interference RNA （siRNA） pGenesiI-HBV X targeting HBV X gene region into HepG2.2.15 cells. METHODS：pGenesil-HBV X was constructed and transfected into HepG2.2.15 cells via lipofection. HBV antigen secretion was determined 24, 48, and 72 h after transfection by time-resolved immunofluorometric assays （TRFIA）. HBV replication was examined by fluorescence quantitative PCR, and the expression of cytoplasmic viral proteins was determined by immunohistochemistry. RESULTS： The secretion of HBsAg and HBeAg into the supernatant was found to be inhibited by 28.5% and 32.2% （P 〈 0.01）, and by 38.67% （P 〈 0.05） and 42.86% （P 〈 0.01） at 48 h and 72 h after pGenesil-HBV X transfection, respectively. Immunohistochemical staining for cytoplasmic HBsAg showed a similar decline in HepG2.2.15 cells 48 h after transfection. The number of HBV genomes within culture supernatants was also significantly decreased 48 h and 72 h post-transfection as quantified by fluorescence PCR （P 〈 0.05）. CONCLUSION： In HepG2.2.15 cells, HBV replication and expression is inhibited by vector-based siRNA pGenesil- HBV X targeting the HBV X coding region.
基金The Youth Foundation of Heilongjiang Province,No.QC06C061the Foundation of Education Department,Heilongjiang Province,No.11521089
文摘AIM： To observe the inhibition of hepatitis B virus （HBV） replication and expression in HepG2.2.15 cells by combination of small interfering RNAs （siRNAs）. METHODS： Recombinant plasmid psiI-HBV was constructed and transfected into HepG2.2.15 cells. At 48 h, 72 h and 96 h after transfection, culture media were collected and cells were harvested for HBV replication assay. HBsAg and HBeAg in the cell culture medium were detected by enzyme-linked immunoadsorbent assay （ELISA）. Intracellular viral DNA and covalently closed circular DNA （cccDNA） were quantified by real-time polymerase chain reaction （PCR）. HBV viral mRNA was reverse transcribed and quantified by reverse-transcript PCR （RT-PCR）. RESULTS： siRNAs showed marked anti-HBV effects. siRNAs could specifically inhibit the expression of HBsAg and the replication of HBV DNA in a dosedependent manner. Furthermore, combination of siRNAs, compared with individual use of each siRNA, exerted a stronger inhibition on antigen expression and viral replication. More importantlycombination of siRNAs significantly suppressed HBV cccDNA amplification. CONCLUSION： Combination of siRNAs mediates a stronger inhibition on viral replication and antigenexpression in HepG2.2.15 cells, especially on cccDNA amplification.
文摘BACKGROUND Hepatitis B virus,together with hepatitis C virus,has been recognized as the leading causes of hepatocellular carcinoma(HCC).Long non-coding RNAs(lncRNAs)have been suggested in increasing studies to be the potential prognostic factors for HCC.However,the role of combined application of lncRNAs in estimating overall survival(OS)for hepatitis virus positive HCC(VHCC)is uncertain.AIM To construct an lncRNA signature related to the OS of VHCC patients to enhance the accuracy of prognosis prediction.METHODS The expression patterns of lncRNAs,as well as related clinical data were collected from 149 VHCC patients from The Cancer Genome Atlas database.The R package was adopted to obtain the differentially expressed lncRNAs(DElncRNAs).LncRNAs significantly associated with OS were screened by means of univariate Cox regression analysis,so as to construct a least absolute shrinkage and selection operator(LASSO)model.Subsequently,the constructed lncRNA signature was developed and validated.Afterwards,the prognostic nomogram was established,which combined the as-established lncRNA signature as well as the clinical features.Meanwhile,subgroup analysis stratified by the virus type was also performed.Finally,the above-mentioned lncRNAs were enriched to corresponding pathways according to the markedly coexpressed genes.RESULTS A total of 1420 DElncRNAs were identified,among which 406 were significant in univariate Cox regression analysis.LASSO regression confirmed 8 out of the 406 lncRNAs,including AC005722.2,AC107959.3,AL353803.1,AL589182.1,AP000844.2,AP002478.1,FLJ36000,and NPSR1-AS1.Then,the prognostic risk score was calculated.Our results displayed a significant association between the risk model and the OS of VHCC[hazard ratio=1.94,95%confidence interval(CI):1.61-2.34,log-rank P=2e-10].The inference tree suggested that the established lncRNA signature was useful in the risk stratification of VHCC.Furthermore,a nomogram was plotted,and the concordance index of internal validation was 0.763(95%CI:0.700-0.826).Moreover,the subgroup analysis regarding etiology confirmed this risk model.In addition,the Wnt signaling pathway,angiogenesis,the p53 pathway,and the PI3 kinase pathway were the remarkably enriched pathways.CONCLUSION An eight-lncRNA signature has been established to predict the prognosis for VHCC,which contributes to providing a novel foundation for the targeted therapy of VHCC.
基金Ministerul Cercetarii si Inovarii,Programul 1,subprogramul 1.2.Performanta institutionala,No.PFE_23/2018。
文摘Viruses can alter the expression of host microRNAs(miRNA s) and modulate the immune response during a persistent infection. The dysregulation of host miRNA s by hepatitis B virus(HBV) contributes to the proinflammatory and profibrotic changes within the liver. Multiple studies have documented the differential regulation of intracellular and circulating miRNA s during different stages of HBV infection. Circulating miRNA s found in plasma and/or extracellular vesicles can integrate data on viral-host interactions and on the associated liver injury. Hence, the detection of circulating miRNA s in chronic HBV hepatitis could offer a promising alternative to liver biopsy, as their expression is associated with HBV replication, the progression of liver fibrosis,and the outcome of antiviral treatment. The current review explores the available data on miRNA involvement in HBV pathogenesis with an emphasis on their potential use as biomarkers for liver fibrosis.
文摘BACKGROUND: The resolution of hepatitis C, evidenced by normalization of liver function and disappearance of hepatitis C virus RNA from serum as determined by conventional laboratory assays, reflects virus eradication. But in interferon treated patients the HCV RNA in serum sometimes could not show the virus in cells. Such factors as virus genotype, HCV RNA contents in serum, HCV specific cellular immunities after treatment were reported to predict the response to interferon therapy. In most patients, HCV RNA could detect the virus in peripheral blood mononucle-ar cell. The aim of this study was to investigate the predictive value of HCV RNA in PBMC of patients with chronic hepatitis C after interferon treatment. METHODS: Sixteen patients with chronic hepatitis C were treated with interferon for 24 weeks, and they all get complete responses at 12 weeks of treatment. At the end of treatment, the HCV RNA in PBMC and serum were detected by RT-PCR, and after stopping treatment, HCV RNA in serum was monitored continually. RESULTS: In 9 patients who were HCV RNA positive in their PBMC at the end of treatment, 8 showed serum HCV RNA positive after 24 weeks and another 1 after 1 year. In 7 patients with negative HCV RNA in their PBMC, only 2 patients relapsed in serum HCV RNA after 1-year follow-up, and others remained viral response after 3.5 years. CONCLUSION: HCV RNA in PBMC at the end of IFN treatment is a predictor of durable response to antiviral therapy in patients with chronic hepatitis C.
基金Supported by Pennsylvania state CURE grant,No.4100057658,[to Steel LF and Bouchard MJ(partially)]a Ruth L Kirschstein(F31)Predoctoral Fellowship,No.5F31CA171712-03,[to Lamontagne J(partially)]
文摘Chronic infection with the hepatitis B virus(HBV) is the leading risk factor for the development of hepatocellular carcinoma(HCC). With nearly 750000 deaths yearly, hepatocellular carcinoma is the second highest cause of cancer-related death in the world. Unfortunately, the molecular mechanisms that contribute to the development of HBV-associated HCC remain incompletely understood. Recently, micro RNAs(mi RNAs), a family of small non-coding RNAs that play a role primarily in post-transcriptional gene regulation, have been recognized as important regulators of cellular homeostasis, and altered regulation of mi RNA expression has been suggested to play a significant role in virus-associated diseases and the development of many cancers. With this in mind, many groups have begun to investigate the relationship between mi RNAs and HBV replication and HBV-associated disease. Multiple findings suggest that some mi RNAs, such as mi R-122, and mi R-125 and mi R-199 family members, are playing a role in HBV replication and HBV-associated disease, including the development of HBV-associated HCC. In this review, we discuss the current state of our understanding of the relationship between HBV and mi RNAs, including how HBV affects cellular mi RNAs, how these mi RNAs impact HBV replication, and the relationship between HBV-mediated mi RNA regulation and HCC development. We also address the impact of challenges in studying HBV, such as the lack of an effective model system for infectivity and a reliance on transformed cell lines, on our understanding of the relationship between HBV and mi RNAs, and proposepotential applications of mi RNA-related techniques that could enhance our understanding of the role mi RNAs play in HBV replication and HBV-associated disease, ultimately leading to new therapeutic options and improved patient outcomes.
基金Science and Technology Development Foundation(STDF),Project NO.3457(TC/4/Health/2010/hep-1.6)
文摘BACKGROUND Diffusion-weighted magnetic resonance imaging has shown promise in the detection and quantification of hepatic fibrosis. In addition, the liver has numerous endogenous micro-RNAs(miRs) that play important roles in the regulation of biological processes such as cell proliferation and hepatic fibrosis.AIM To assess diffusion-weighted magnetic resonance imaging and miRs in diagnosing and staging hepatic fibrosis in patients with chronic hepatitis C.METHODS This prospective study included 208 patients and 82 age-and sex-matched controls who underwent diffusion-weighted magnetic resonance imaging of the abdomen, miR profiling, and liver biopsy. Pathological scoring was classified according to the METAVIR scoring system. The apparent diffusion coefficient (ADC) and miR were calculated and correlated with pathological scoring.RESULTS The ADC value decreased significantly with the progression of fibrosis, from controls(F0) to patients with early fibrosis(F1 and F2) to those with late fibrosis(F3 and F4)(median 1.92, 1.53, and 1.25 × 10^(-3) mm^2/s, respectively)(P = 0.001).The cut-off ADC value used to differentiate patients from controls was 1.83 × 10^(-3) mm^2/s with an area under the curve(AUC) of 0.992. Combining ADC and miR-200 b revealed the highest AUC(0.995) for differentiating patients from controls with an accuracy of 96.9%. The cut-off ADC used to differentiate early fibrosis from late fibrosis was 1.54 × 10^(-3) mm^2/s with an AUC of 0.866. The combination of ADC and miR-200 b revealed the best AUC(0.925) for differentiating early fibrosis from late fibrosis with an accuracy of 80.2%. The ADC correlated with miR-200 b(r =-0.61, P = 0.001), miR-21(r =-0.62, P = 0.001), and miR-29(r = 0.52,P = 0.001).CONCLUSION Combining ADC and miRs offers an alternative surrogate non-invasive diagnostic tool for diagnosing and staging hepatic fibrosis in patients with chronic hepatitis C.
基金Supported by In part Grants from NIDDK DK042182 and the Herman Lopata Chair for Hepatitis Research (Wu GY)
文摘AIM: To examine the effect of hepatitis C virus (HCV) structural mimics of regulatory regions of the genome on HCV replication.METHODS: HCV RNA structural mimics were constructed and tested in a HCV genotype 1b aBB7 replicon,and a Japanese fulminant hepatitis-1 (JFH-1) HCV genotype 2a infection model.All sequences were computer-predicted to adopt stem-loop structures identical to the corresponding elements in full-length viral RNA.Huh7.5 cells bearing the BB7 replicon or infected with JFH-1 virus were transfected with expression vectors generating HCV mimics and controls.Cellular HCV RNA and protein levels were quantified by real-time polymerase chain reaction and Western blotting,respectively.To evaluate possible antisense effects,complementary RNAs spanning a mimic were prepared.RESULTS: In the BB7 genotype 1b replicon system,mimics of the polymerase (NS-5B),X and BA regions inhibited replication by more than 90%,50%,and 60%,respectively.In the JFH-1 genotype 2 infection system,mimics that were only 74% and 46% identical in sequence relative to the corresponding region in JFH-1 inhibited HCV replication by 91.5% and 91.2%,respectively,as effectively as a mimic with complete identity to HCV genotype 2a.The inhibitory effects were confirmed by NS3 protein levels.Antisense RNA molecules spanning the 74% identical mimic had no significant effects.CONCLUSION: HCV RNA structural mimics can inhibit HCV RNA replication in replicon and infectious HCV systems and do so independent of close sequence identity with the target.
