AIM:To investigate the peripheral T-lymphocyte subpopulation profile,and its correlations with hepatitis B virus(HBV) replication level in chronic HBV-infected(CHI) individuals with normal liver function tests(LFTs) ....AIM:To investigate the peripheral T-lymphocyte subpopulation profile,and its correlations with hepatitis B virus(HBV) replication level in chronic HBV-infected(CHI) individuals with normal liver function tests(LFTs) . METHODS:Frequencies of T-lymphocyte subpopu-lations in peripheral blood were measured by flow cytometry in 216 CHI individuals. HBV markers were detected with ELISA. Serum HBV DNA load was assessed with quantitative real-time PCR. Information of age at HBV infection,and maternal HBV infection status was collected. ANOVA linear trend test and linear regression were used in statistical analysis. RESULTS:CHI individuals had significantly decreased relative frequencies of CD3+,CD4+ subpopulationsand CD4+/CD8+ ratio,and increased CD8+ subset percentage compared with uninfected individuals(all P < 0.001) . There was a significant linear relationship between the load of HBV DNA and the parameters of T-lymphocyte subpopulations(ANOVA linear trend test P < 0.01) . The parameters were also significantly worse among individuals whose mothers were known to be HBV carriers,and those having gained infection before the age of 8 years. In multiple regressions,after adjustment for age at HBV infection and status of maternal HBV infection,log copies of HBV DNA maintained its highly significant predictive coefficient on T-lymphocyte subpopulations,whereas the effect of HBeAg was not significant. CONCLUSION:HBV DNA correlates with modification in the relative T-lymphocyte subpopulation frequencies. High viral load is more powerful than HBeAg in predicting the impaired balance of T-cell subsets.展开更多
Objective The clinical significance of differential distribution of hepatitis B virus(HBV)nucleocapsid antigen in hepatocytes remains unknown.The goal of this study is to determine the relationship between distinct HB...Objective The clinical significance of differential distribution of hepatitis B virus(HBV)nucleocapsid antigen in hepatocytes remains unknown.The goal of this study is to determine the relationship between distinct HBV core antigen distribution pattern and alanine transaminase(ALT),liver histological inflammatory activity grades,serum HBeAg status and HBV DNA level.Methods Total of 958 cases with chronic hepatitis B were recruited into this study.Liver function tests,serum HBV DNA level,serological HBV markers and liver immunohistochemistry were examined according to the conventional instructions.Chi Square tests were performed to analyze the differences among these groups.Results It was found that 552(58%)cases were tested positive for HBV core antigen by immunohistochemical staining.Cytoplasmic hepatitis B core antigen(HBcAg)expression correlated with ALT level and serum HBV DNA and liver inflammatory activity scores,however,nuclear HBcAg expression in hepatocytes was associated with normal ALT level,lower liver inflammatory activity score and higher serum HBV DNA level and rate of HBeAg positivity.Both nuclear and cytoplasmic HBcAg expression in hepatocytes associated with a middle ALT level and liver inflammatory activity score,higher rate of serum detectable HBeAg and a higher HBV DNA level.However,undetectable core antigen was related to a lower ALT level and histological inflammatory activity grade,lower positive HBeAg rate and HBV DNA level.Conclusions Undetectable liver HBcAg is associated with HBV clearance,ALT normalization and hepatitis B e antigen(HBeAg)seroconversion,and cytoplasmic HBcAg expression associated with higher hepatic inflammatory activity.However,nuclear HBcAg expression correlates with immune tolerance characterized with normal ALT and lower liver inflammatory activity,higher HBV replication level and higher rate of HBeAg positivity.展开更多
乙型肝炎病毒(hepatitis B virus,HBV)e抗原(hepatitis B e antigen,HBeAg)阳性的慢性HBV感染依次经历非活动性肝炎(non-aggressive hepatitis,NAH)和活动性肝炎(aggressive hepatitis,AH)2个分期,但仍缺乏界定HBeAg阳性NAH与AH的可靠...乙型肝炎病毒(hepatitis B virus,HBV)e抗原(hepatitis B e antigen,HBeAg)阳性的慢性HBV感染依次经历非活动性肝炎(non-aggressive hepatitis,NAH)和活动性肝炎(aggressive hepatitis,AH)2个分期,但仍缺乏界定HBeAg阳性NAH与AH的可靠标准。本文根据179例患者的长期随访队列,以自发性HBeAg血清转换作为终点事件,采用Kaplan-Meier生存分析,指定了丙氨酸转氨酶(alanine transaminase,ALT)、HBV表面抗原(hepatitis B surface antigen,HBsAg)和HBV DNA识别HBeAg阳性NAH的功能截断值;在此基础上,评价了ALT串联HBsAg和串联HBV DNA识别HBeAg阳性NAH的性能。结果显示,ALT≤60 IU/L、HBsAg>4.602 log10IU/mL和HBV DNA>7.477 log10IU/mL为识别HBeAg阳性NAH的功能截断值。基于功能截断值,ALT串联HBsAg的患者中,病理学分级≤G1和“分级≤G1且分期≤S2”的构成比均为100%,病理学分期≤S1和“分级≤G2且分期≤S1”的构成比均为68.2%;ALT串联HBV DNA的患者中,病理学分级≤G1和“分级≤G1且分期≤S2”的构成比均为86.2%,病理学分期≤S1和“分级≤G2且分期≤S1”的构成比均为69.0%;ALT串联HBsAg识别病理学分级≤G1和“分级≤G1且分期≤S2”的阳性似然比均为+∞,识别病理学分期≤S1和“分级≤G2且分期≤S1”的阳性似然比均为2.034;ALT串联HBV DNA识别病理学分级≤G1和“分级≤G1且分期≤S2”的阳性似然比分别为3.000和3.068,识别病理学分期≤S1和“分级≤G2且分期≤S1”的阳性似然比均为2.106。以上结果提示,ALT串联HBsAg和串联HBV DNA均可有效识别HBeAg阳性NAH;且ALT串联HBsAg识别HBeAg阳性NAH的性能优于ALT串联HBV DNA。展开更多
Objective To assess on-treatment serum HBsAg and HBV DNA kinetics in HBeAg-positive CHB patients to predict the efficacy of pegylated interferon(PEG-IFN) in early phase of treatment. Methods Forty-one treatment-naive ...Objective To assess on-treatment serum HBsAg and HBV DNA kinetics in HBeAg-positive CHB patients to predict the efficacy of pegylated interferon(PEG-IFN) in early phase of treatment. Methods Forty-one treatment-naive HBeAg-positive patients treated with PEG-IFNα 2a at a dose of 180 μg/week for at least 24 weeks were evaluated. Their treatment response was assessed, including normalization of serum ALT, decline of serum HBV DNA and loss of HBeAg. Results We found that a decrease of HBV DNA level at the 4th week was positively correlated with the decrease of HBV DNA level at the 12th week and 24th week(r = 0.8202, P < 0.0001 and r = 0.6838, P < 0.0001, respectively). We observed that a decrease of HBsAg level at the 4th week was positively correlated with decrease of HBsAg level at the 12th week and 24th week(r = 0.4868, P = 0.0023 and r = 0.4251, P = 0.0109, respectively). A decrease of HBsAg level at the 24th week was positively correlated with the decrease of HBV DNA level at the 24th week(r = 0.5262, P = 0.0024). Serum level of IFN and IFN neutralizing antibody had no relationship with HBV DNA or HBsAg titers kinetics. Conclusions The decline of serum HBV DNA and hepatitis B surface antigen at the 4th week can be used to predict the response to PEG-IFNα 2a in patients with HBeAg positive chronic hepatitis B.展开更多
目的探讨乙肝病毒(Hepatitis B Virus,HBV)感染患者血清中乙型肝炎病毒脱氧核苷酸(HBV-DNA)载量与乙肝五项指标的相关性。方法选取沛县人民医院于2022年1月—2023年1月收治的100例HBV感染患者作为研究对象,所有患者均接受HBV-DNA载量与...目的探讨乙肝病毒(Hepatitis B Virus,HBV)感染患者血清中乙型肝炎病毒脱氧核苷酸(HBV-DNA)载量与乙肝五项指标的相关性。方法选取沛县人民医院于2022年1月—2023年1月收治的100例HBV感染患者作为研究对象,所有患者均接受HBV-DNA载量与乙肝五项指标检测,并按照其HBV-DNA载量进行分组,分别为<50 IU/mL组69例、50~<10^(4) IU/mL组10例、10^(4)~10^(10) IU/mL组21例,对比3组检测结果。结果10^(4)~10^(10) IU/mL组乙型肝炎e抗体(Hepatitis B Virus e Antibody,HBeAb)、乙型肝炎e抗原(Hepatitis B e Antigen,HBeAg)、乙型肝炎表面抗原(Hapatitis B Surface Antigen,HBsAg)指标高于50~<10^(4) IU/mL组、<50 IU/mL组,50~<10^(4) IU/mL组各指标水平高于<50 IU/mL组,差异有统计学意义(P均<0.05)。相关性分析后显示,HBeAb、HBeAg、HBsAg与HBV-DNA载量呈正相关(r=0.512、0.549、0.673,P均<0.05)。结论HBV-DNA载量与HBeAb、HBeAg、HBsAg指标之间有正性关联,可为评估HBV感染程度提供一定参考依据。展开更多
目的探讨丙型肝炎病毒NS5A反式调节蛋白9(hepatitis C virus NS5Atransactivated protein 9,NS5ATP9)在乙型肝炎病毒(hepatitis B virus,HBV)共价闭合环状DNA(covalently closed circular DNA,cccDNA)形成与转录中的作用机制。方法利用...目的探讨丙型肝炎病毒NS5A反式调节蛋白9(hepatitis C virus NS5Atransactivated protein 9,NS5ATP9)在乙型肝炎病毒(hepatitis B virus,HBV)共价闭合环状DNA(covalently closed circular DNA,cccDNA)形成与转录中的作用机制。方法利用1.3拷贝HBV表达质粒转染Huh7和HepG2细胞、整合有4拷贝HBV基因组的HepG2.2.15细胞、在诱导型四环素启动子控制下表达HBV的HepAD38细胞构建NS5ATP9过表达或干扰的HBV细胞模型,收集样品和细胞上清液,提取RNA、HBV核心DNA(coreDNA)、cccDNA和蛋白,利用酶联免疫吸附试验、实时荧光定量聚合酶链反应(polymerase chain reaction,PCR)、Southern blot和Western blot技术检测HBV总RNA、前基因组RNA(pregenomic RNA,pgRNA)、乙型肝炎病毒s抗原(hepatitis B virus s antigene,HBsAg)、乙型肝炎病毒e抗原(hepatitis B virus e antigene,HBeAg)、松弛环状DNA(relax circular DNA,rcDNA)以及cccDNA水平。在HepG2细胞中转染乙型肝炎病毒x蛋白(hepatitis B virus x protein,HBx),通过免疫荧光成像及免疫共沉淀方法检测NS5ATP9与HBx的结合情况。双荧光素酶报告基因实验检测NS5ATP9对HBx启动子活性的影响。利用Huh7细胞转染HBV1.3及HBV稳定表达细胞株HepG2.2.15和HepAD38转染NS5ATP9过表达/干扰质粒,通过Western blot技术检测DDB1和SMC6的蛋白水平。结果在HBV病毒活跃的细胞中,NS5ATP9 mRNA水平[HepG2.2.15细胞:1.891±0.567比1.00±0.034,t=2.87,P=0.0351;HepAD38 tet+细胞:1.978±0.399比1.00±0.034,t=4.131,P=0.0091;HepAD38 tet-细胞:2.642±0.672比1.00±0.034,t=4.127,P=0.0091]和蛋白水平均显著增加。过表达NS5ATP9后可显著增加HBeAg[(5.402±0.327)S/COV比(2.68±0.552)S/COV,t=7.35,P=0.0018]、HBsAg[(2.846±0.185)S/COV比(1.512±0.221)S/COV,t=8.02,P=0.0013]、HBV pgRNA及rcDNA的表达水平,而干扰NS5ATP9后此增加作用消失[HBeAg:(2.029±0.09)S/COV比(3.733±0.445)S/COV,t=6.501,P=0.0029;HBsAg:(1.501±0.105)S/COV比(1.878±0.174)S/COV,t=3.216,P=0.0324)]。机制研究显示,NS5ATP9和HBx蛋白主要位于细胞核核仁内,并具有共定位信号,且NS5ATP9可显著提高HBx启动子(1071.06±79.44比488.47±40.12,t=13.09,P=0.00012)的转录活性。另外,过表达NS5ATP9可显著降低DDB1和SMC6的蛋白水平,而沉默NS5ATP9则可显著提高DDB1和SMC6的蛋白水平。结论HBV上调NS5ATP9的表达,形成HBV-NS5ATP9-HBV cccDNA-HBV的正反馈环路,NS5ATP9通过与HBx相互作用上调肝细胞中HBV cccDNA的形成与转录,进而促进慢性乙型肝炎的发生发展。展开更多
文摘AIM:To investigate the peripheral T-lymphocyte subpopulation profile,and its correlations with hepatitis B virus(HBV) replication level in chronic HBV-infected(CHI) individuals with normal liver function tests(LFTs) . METHODS:Frequencies of T-lymphocyte subpopu-lations in peripheral blood were measured by flow cytometry in 216 CHI individuals. HBV markers were detected with ELISA. Serum HBV DNA load was assessed with quantitative real-time PCR. Information of age at HBV infection,and maternal HBV infection status was collected. ANOVA linear trend test and linear regression were used in statistical analysis. RESULTS:CHI individuals had significantly decreased relative frequencies of CD3+,CD4+ subpopulationsand CD4+/CD8+ ratio,and increased CD8+ subset percentage compared with uninfected individuals(all P < 0.001) . There was a significant linear relationship between the load of HBV DNA and the parameters of T-lymphocyte subpopulations(ANOVA linear trend test P < 0.01) . The parameters were also significantly worse among individuals whose mothers were known to be HBV carriers,and those having gained infection before the age of 8 years. In multiple regressions,after adjustment for age at HBV infection and status of maternal HBV infection,log copies of HBV DNA maintained its highly significant predictive coefficient on T-lymphocyte subpopulations,whereas the effect of HBeAg was not significant. CONCLUSION:HBV DNA correlates with modification in the relative T-lymphocyte subpopulation frequencies. High viral load is more powerful than HBeAg in predicting the impaired balance of T-cell subsets.
文摘Objective The clinical significance of differential distribution of hepatitis B virus(HBV)nucleocapsid antigen in hepatocytes remains unknown.The goal of this study is to determine the relationship between distinct HBV core antigen distribution pattern and alanine transaminase(ALT),liver histological inflammatory activity grades,serum HBeAg status and HBV DNA level.Methods Total of 958 cases with chronic hepatitis B were recruited into this study.Liver function tests,serum HBV DNA level,serological HBV markers and liver immunohistochemistry were examined according to the conventional instructions.Chi Square tests were performed to analyze the differences among these groups.Results It was found that 552(58%)cases were tested positive for HBV core antigen by immunohistochemical staining.Cytoplasmic hepatitis B core antigen(HBcAg)expression correlated with ALT level and serum HBV DNA and liver inflammatory activity scores,however,nuclear HBcAg expression in hepatocytes was associated with normal ALT level,lower liver inflammatory activity score and higher serum HBV DNA level and rate of HBeAg positivity.Both nuclear and cytoplasmic HBcAg expression in hepatocytes associated with a middle ALT level and liver inflammatory activity score,higher rate of serum detectable HBeAg and a higher HBV DNA level.However,undetectable core antigen was related to a lower ALT level and histological inflammatory activity grade,lower positive HBeAg rate and HBV DNA level.Conclusions Undetectable liver HBcAg is associated with HBV clearance,ALT normalization and hepatitis B e antigen(HBeAg)seroconversion,and cytoplasmic HBcAg expression associated with higher hepatic inflammatory activity.However,nuclear HBcAg expression correlates with immune tolerance characterized with normal ALT and lower liver inflammatory activity,higher HBV replication level and higher rate of HBeAg positivity.
基金supported by Major Science and Technology Special Project of China Eleventh Five-year Plan(2008ZX10002-004)Major Science and Technology Special Project of China Twelfth Five-year Plan (2012ZX10002003)Natural Science Foundation of China (81101240) to NL
文摘Objective To assess on-treatment serum HBsAg and HBV DNA kinetics in HBeAg-positive CHB patients to predict the efficacy of pegylated interferon(PEG-IFN) in early phase of treatment. Methods Forty-one treatment-naive HBeAg-positive patients treated with PEG-IFNα 2a at a dose of 180 μg/week for at least 24 weeks were evaluated. Their treatment response was assessed, including normalization of serum ALT, decline of serum HBV DNA and loss of HBeAg. Results We found that a decrease of HBV DNA level at the 4th week was positively correlated with the decrease of HBV DNA level at the 12th week and 24th week(r = 0.8202, P < 0.0001 and r = 0.6838, P < 0.0001, respectively). We observed that a decrease of HBsAg level at the 4th week was positively correlated with decrease of HBsAg level at the 12th week and 24th week(r = 0.4868, P = 0.0023 and r = 0.4251, P = 0.0109, respectively). A decrease of HBsAg level at the 24th week was positively correlated with the decrease of HBV DNA level at the 24th week(r = 0.5262, P = 0.0024). Serum level of IFN and IFN neutralizing antibody had no relationship with HBV DNA or HBsAg titers kinetics. Conclusions The decline of serum HBV DNA and hepatitis B surface antigen at the 4th week can be used to predict the response to PEG-IFNα 2a in patients with HBeAg positive chronic hepatitis B.
文摘目的探讨乙肝病毒(Hepatitis B Virus,HBV)感染患者血清中乙型肝炎病毒脱氧核苷酸(HBV-DNA)载量与乙肝五项指标的相关性。方法选取沛县人民医院于2022年1月—2023年1月收治的100例HBV感染患者作为研究对象,所有患者均接受HBV-DNA载量与乙肝五项指标检测,并按照其HBV-DNA载量进行分组,分别为<50 IU/mL组69例、50~<10^(4) IU/mL组10例、10^(4)~10^(10) IU/mL组21例,对比3组检测结果。结果10^(4)~10^(10) IU/mL组乙型肝炎e抗体(Hepatitis B Virus e Antibody,HBeAb)、乙型肝炎e抗原(Hepatitis B e Antigen,HBeAg)、乙型肝炎表面抗原(Hapatitis B Surface Antigen,HBsAg)指标高于50~<10^(4) IU/mL组、<50 IU/mL组,50~<10^(4) IU/mL组各指标水平高于<50 IU/mL组,差异有统计学意义(P均<0.05)。相关性分析后显示,HBeAb、HBeAg、HBsAg与HBV-DNA载量呈正相关(r=0.512、0.549、0.673,P均<0.05)。结论HBV-DNA载量与HBeAb、HBeAg、HBsAg指标之间有正性关联,可为评估HBV感染程度提供一定参考依据。
文摘目的探讨丙型肝炎病毒NS5A反式调节蛋白9(hepatitis C virus NS5Atransactivated protein 9,NS5ATP9)在乙型肝炎病毒(hepatitis B virus,HBV)共价闭合环状DNA(covalently closed circular DNA,cccDNA)形成与转录中的作用机制。方法利用1.3拷贝HBV表达质粒转染Huh7和HepG2细胞、整合有4拷贝HBV基因组的HepG2.2.15细胞、在诱导型四环素启动子控制下表达HBV的HepAD38细胞构建NS5ATP9过表达或干扰的HBV细胞模型,收集样品和细胞上清液,提取RNA、HBV核心DNA(coreDNA)、cccDNA和蛋白,利用酶联免疫吸附试验、实时荧光定量聚合酶链反应(polymerase chain reaction,PCR)、Southern blot和Western blot技术检测HBV总RNA、前基因组RNA(pregenomic RNA,pgRNA)、乙型肝炎病毒s抗原(hepatitis B virus s antigene,HBsAg)、乙型肝炎病毒e抗原(hepatitis B virus e antigene,HBeAg)、松弛环状DNA(relax circular DNA,rcDNA)以及cccDNA水平。在HepG2细胞中转染乙型肝炎病毒x蛋白(hepatitis B virus x protein,HBx),通过免疫荧光成像及免疫共沉淀方法检测NS5ATP9与HBx的结合情况。双荧光素酶报告基因实验检测NS5ATP9对HBx启动子活性的影响。利用Huh7细胞转染HBV1.3及HBV稳定表达细胞株HepG2.2.15和HepAD38转染NS5ATP9过表达/干扰质粒,通过Western blot技术检测DDB1和SMC6的蛋白水平。结果在HBV病毒活跃的细胞中,NS5ATP9 mRNA水平[HepG2.2.15细胞:1.891±0.567比1.00±0.034,t=2.87,P=0.0351;HepAD38 tet+细胞:1.978±0.399比1.00±0.034,t=4.131,P=0.0091;HepAD38 tet-细胞:2.642±0.672比1.00±0.034,t=4.127,P=0.0091]和蛋白水平均显著增加。过表达NS5ATP9后可显著增加HBeAg[(5.402±0.327)S/COV比(2.68±0.552)S/COV,t=7.35,P=0.0018]、HBsAg[(2.846±0.185)S/COV比(1.512±0.221)S/COV,t=8.02,P=0.0013]、HBV pgRNA及rcDNA的表达水平,而干扰NS5ATP9后此增加作用消失[HBeAg:(2.029±0.09)S/COV比(3.733±0.445)S/COV,t=6.501,P=0.0029;HBsAg:(1.501±0.105)S/COV比(1.878±0.174)S/COV,t=3.216,P=0.0324)]。机制研究显示,NS5ATP9和HBx蛋白主要位于细胞核核仁内,并具有共定位信号,且NS5ATP9可显著提高HBx启动子(1071.06±79.44比488.47±40.12,t=13.09,P=0.00012)的转录活性。另外,过表达NS5ATP9可显著降低DDB1和SMC6的蛋白水平,而沉默NS5ATP9则可显著提高DDB1和SMC6的蛋白水平。结论HBV上调NS5ATP9的表达,形成HBV-NS5ATP9-HBV cccDNA-HBV的正反馈环路,NS5ATP9通过与HBx相互作用上调肝细胞中HBV cccDNA的形成与转录,进而促进慢性乙型肝炎的发生发展。
