Quantitative hepatitis B core antibody and quantitative hepatitis B surface antigen:Novel viral biomarkers for chronic hepatitis B management

The management of hepatitis B virus(HBV)infection now involves regular and appropriate monitoring of viral activity,disease progression,and treatment response.Traditional HBV infection biomarkers are limited in their ability to predict clinical outcomes or therapeutic effectiveness.Quantitation of HBV core antibodies(qAnti-HBc)is a novel non-invasive biomarker that may help with a variety of diagnostic issues.It was shown to correlate strongly with infection stages,hepatic inflammation and fibrosis,chronic infection exacerbations,and the presence of occult infection.Furthermore,qAnti-HBc levels were shown to be predictive of spontaneous or treatment-induced HBeAg and HBsAg seroclearance,relapse after medication termination,re-infection following liver transplantation,and viral reactivation in the presence of immunosuppression.qAnti-HBc,on the other hand,cannot be relied on as a single diagnostic test to address all problems,and its diagnostic and prognostic potential may be greatly increased when paired with qHBsAg.Commercial qAnti-HBc diagnostic kits are currently not widely available.Because many methodologies are only semi-quantitative,comparing data from various studies and defining universal cut-off values remains difficult.This review focuses on the clinical utility of qAnti-HBc and qHBsAg in chronic hepatitis B management.

Precore/basal core promoter mutants and hepatitis B viral DNA levels as predictors for liver deaths and hepatocellular carcinoma

AIM： To conduct a retrospective study in 400 chronic hepatitis B patients in order to identify hepatitis B viral factors associated with complications of liver disease or development of hepatocellular carcinoma. METHODS： The mean follow-up time was 83.6 ± 39.6 mo. Alpha-fetoprotein test and abdominal ultrasound were used for cancer surveillance. Hepatitis B basal core promoter mutants, precore mutants, genotypes, hepatitis B viral DNA （HBV DNA） level and hepatitis B e antigen （HBeAg） were measured. Univariate analysis and logistic regression were used to assess odds ratios for viral factors related to liver deaths and hepatocellular carcinoma development. RESULTS： During follow-up, 38 patients had liver deaths not related to hepatocellular carcinoma. On multivariate analysis, older age [odds ratio： 95.74 （12.13-891.31）, P 〈 0.0001], male sex [odds ratio： 7.61 （2.20-47.95）; P = 0.006], and higher Iogzo HBV DNA [odds ratio： 4.69 （1.16-20.43）; P 〈 0.0001] were independently predictive for these liver related deaths. Also, 31 patients developed hepatocellular carcinoma. Multivariate analysis showed that older age [odds ratio： 26.51 （2.36-381.47）; P = 0.007], presence of precore mutants [odds ratio： 4.23 （1.53-19.58）, P = 0.02] and presence of basal core promoter mutants [odds ratio： 2.93 （1.24-7.57）; P = 0.02] were independent predictors for progression to hepatocellular carcinoma. CONCLUSION： Our results show that high levels of baseline serum HBV DNA are associated with non- hepatocellular carcinoma-related deaths of liver failure, while genetic mutations in the basal core promoter and precore regions are predictive for development of hepatocellular carcinoma.

Serum hepatitis B core-related antigen as a surrogate marker of hepatitis B e antigen seroconversion in chronic hepatitis B

BACKGROUND Quantitative hepatitis B core-related antigen(qHBcrAg)has a better correlation with intrahepatic hepatitis B virus(HBV)covalently closed circular DNA(cccDNA)than HBV DNA or hepatitis B e antigen(HBeAg),but data are still lacking for its clinical application.AIM The aim was to investigate serum qHBcrAg levels in patients with chronic hepatitis B and assess the correlation of serum qHBcrAg with pregenomic RNA(pgRNA),cccDNA,and HBeAg seroconversion.METHODS This study was a secondary analysis of patients who underwent percutaneous liver biopsy between July 2014 and June 2019 in two multicenter randomized controlled clinical trials of peginterferon vs nucleos(t)ide analog(NUC)-based therapy(NCT03509688 and NCT03546530).Serum qHBcrAg,pgRNA,HBV DNA,hepatitis B core antigen,HBeAg,liver cccDNA,and HBV DNA were measured.The correlations of serum qHBcrAg with other biomarkers were analyzed.RESULTS A total of 139 patients were included.The mean qHBcrAg levels were 5.32±1.18 log10 U/mL at baseline and decreased during treatment(all P<0.0001).Serum qHBcrAg levels were positively correlated with pgRNA(r=0.597,P<0.0001)and cccDNA(r=0.527,P<0.0001)levels.The correlation of serum qHBcrAg level and intrahepatic HBV DNA levels at baseline was weak but significant(r=0.399,P<0.0001).HBcrAg predicted HBeAg seroconversion,with areas under the receiver operating characteristics curve of 0.788 at 24 wk and 0.825 at 48 wk.Log HBcrAg at wk 24 and 48 was independently associated with HBeAg seroconversion[odds ratio(OR)=2.402,95%confidence interval(CI):1.314-4.391,P=0.004;OR=3.587,95%CI:1.315-9.784,P=0.013].CONCLUSION Serum HBcrAg levels were correlated with HBV virological markers and could be used to predict HBeAg seroconversion.

Progression and status of antiviral monitoring in patients with chronic hepatitis B:From HBsAg to HBV RNA

As alternative indexes of hepatitis B virus(HBV),co-valently closed circular DNA(cccDNA) transcriptional activity,hepatitis B surface antigen(HBsAg),hepatitis B core-related antigen(HBcrAg),and peripheral blood RNA known as pgRNA,have been advocated as novel serum markers for prediction of prognosis and treatment response in chronic hepatitis B(CHB). Since the availability of commercial quantitative assays of HBsAg in 2011,HBsAg has been widely used for predicting treatment response of patients with CHB. Patients who received interferon therapy have shown a sharper reduction of HBsAg level than those who received nucleoside drug(NAs) therapy. Upon peginterferon treatment,sustained responders have presented a larger reduction of HBsAg level than the non-responders. An absence of HBsAg decline,together with < 2 log reduction in HBV DNA at week 12,can serve as a stopping rule in HBsAg-negative patients infected with genotype D HBV. A sharp reduction of HBs Ag titer in the NAs therapy is a predictor of HBsAg clearance in long-term treatment. HBcrAg,which consists of three species of related proteins sharing an identical 149 amino acid sequence,including HbcAg,hepatitis B e antigen(HBeAg),and a truncated 22-kDa precore protein,is still detectable in situations where serum HBV DNA levels become undetectable or HBsAg loss is achieved. Therefore,HBcrAg remains a measurable serum marker to correlate with cccDNA in this situation. The decline in HBcrAg has been observed with NAs therapy and the pattern of decline might provide prognostic information on the risk of HBV posttreatment reactivation. Peripheral blood RNA,which is known as pgRNA,directly derives from cccDNA and reflects intrahepatic cccDNA level. Quantitative pgRNA has been suggested to be helpful in CHB management. However,commercial quantitative assays are lacking. Additionally,the use of simultaneous and continuous clearance of HBV RNA and HBV DNA in serum has been suggested to be a safe stopping rule of NAs therapy for patients with CHB. However,clinical studies of large sample sizes are needed to prove the feasibility andsignificance of using serum HBV RNA as the assessment standard of antiviral therapy in CHB and the safety of the stopping rule in clinics.

Association of core promoter mutations of hepatitis B virus and viral load is different in HBeAg(+) and HBeAg(-) patients

AIM:To identify the prevalence of hepatitis B e antigen (HBeAg) and to assess the association of hepatitis B virus (HBV) core promoter mutations and viral load in Indonesian patients.METHODS:Sixty-four patients with chronic hepatitis,65 with liver cirrhosis and 50 with hepatocellular carcinoma were included in this study.HBeAg and hepatitis B e antibody (HBeAb) tests were performed using enzyme-linked immunosorbent assay and the mutations were analyzed by sequencing.Viral load was measured by real-time polymerase chain reaction.RESULTS:Of 179 patients,108 (60.3%) were HBeAg(-) and 86 (79.6%) of these HBeAg(-) patients had been seroconverted.The A1896 mutation was not found in HBeAg(+) patients,however,this mutation was detected in 70.7% of HBeAg(-) patients.This mutation was frequently found when HBeAg was not expressed (87.7%),compared to that found in HBeAg seroconverted patients (65.1%).The A1899 mutation was also more prevalent in HBeAg(-) than in HBeAg(+) patients (P=0.004).The T1762/A1764 mutation was frequently found in both HBeAg(+) and HBeAg(-) patients,however,the prevalence of this mutation did not significantly differ among the two groups (P=0.054).In HBeAg(+) patients,the T1762/A1764 mutation was correlated with lower HBV DNA (P < 0.001).The A1899 mutation did not correlate with HBV DNA (P=0.609).In HBeAg(-) patients,the T1762/A1764 mutation alone was not correlated with HBV DNA (P=0.095),however,the presence of either the T1762/A1764 or A1896 mutations was associated with increased HBV DNA (P < 0.001).CONCLUSION:The percentage of HBeAg(-) patients is high in Indonesia,and most of the HBeAg(-) patients had been seroconverted.The A1896 mutation was most likely the major cause of HBeAg loss.The T1762/A1764 mutation alone was associated with lower viral loads in HBeAg(+) patients,but not in HBeAg(-) patients.

HBV pgRNA联合HBcrAg对慢性乙型肝炎患者停药后复发的预测价值

目的分析乙型肝炎病毒(hepatitis B virus,HBV)前基因组RNA(pregenomic RNA,pgRNA)水平联合HBV核心相关抗原(hepatitis B virus core-related antigen,HBcrAg)定量对慢性乙型肝炎(chronic viral hepatitis B,CHB)患者停药后复发风险的预测价值。方法选取中国人民解放军联勤保障部队第926医院2020年6月至2021年6月收治的113例CHB患者为研究对象,所有患者均已给予足疗程的正规抗病毒治疗,停药前均检测批pgRNA与HBcrAg。根据患者停药1年内复发情况分为复发组(38例)和未复发组(70例),比较两组患者的一般资料、肝功能、肾功能、甲胎蛋白(alphafetoprotein,AFP)、pgRNA及HBcAg水平等指标。应用多因素Logistic回归分析CHB患者停药后复发的影响因素。应用受试者工作特征(receiver operator characteristic,ROC)曲线分析pgRNA联合HBcrAg对CHB患者停药后复发风险的预测价值。结果复发组患者饮酒史比例[47.37%(18/38)比22.86%(16/70)]、AFP[(29.64±7.18)μg/L比(20.38±6.46)μg/L]、pgRNA[(7.97±1.99)lg拷贝/ml比(4.97±1.24)lg拷贝/ml]和HBcrAg[(7.04±1.76)lg IU/ml比(5.11±1.28)lg IU/ml]水平均显著高于未复发组(P均<0.05)。多因素Logistic回归分析表明,饮酒史(OR=5.354,95%CI:1.055~68.858,P=0.046)、AFP(OR=1.189,95%CI:1.036~1.468,P=0.015)、pgRNA(OR=1.047,95%CI:1.117~8.109,P=0.007)和HBcrAg(OR=2.152,95%CI:1.154~4.308,P=0.021)是CHB患者停药后复发的独立危险因素。pgRNA与HBcrAg联合预测CHB患者停药后复发的ROC曲线下面积为0.954,最佳截点为>0.128,此时敏感度为98.9%,特异度为97.1%。结论pgRNA和HBcrAg与CHB患者停药后复发风险密切相关,早期监测两者水平有助于发现停药后复发高风险的患者,早期调整治疗方案。

Humoral and cellular immunogenecity of DNA vaccine based on hepatitis B core gene in rhesus monkeys

INTRODUCTIONHepatitis B virus (HBV) is the most commonetiologic agent for infectious liver diseases. It isestimated that there are more than 250 millionchronic HBV carriersin the world today and thereis a significant association among persistentinfection, liver cirrhosis and hepatocellularcarcinoma[1-3].

Serum concentration of sFas and sFasL in healthy HBsAg carriers,chronic viral hepatitis B and C patients

AIM:To estimate the amount of apoptosis among healthy HBsAg carriers,patients with chronic HBV infection treated wibh lamivudine and patients with chronic HCV infection treated with interferon alpha and ribavirin.Activity of apoptosis was evaluated by serum sFas/sFasL concentration measurement. Moreover dependence between apoptosis and HBV-DNA or HCV-RNA levels was studied. METHODS:Eighty-six persons were included into study:34 healthy HBsAg carders,33 patients with chronic HBV infecl^on and 19 patients with chronic HCV infection.Serum levels of sFas/sFasL were measured by ELISA assay.HBV-DNA and HCV-RNA were measured by RT-PCR assay.Levels of sFas/sFasL were determined before and 2 and 12 wk after therapy in patients with chronic hepatitis B and C infection. HBV-DNA or HCV-RNA was detected before treatment and 6 mo after treatment. RESULTS:Twenty-four (71%) healthy HBsAg carders showed HBV-DNA over 10~5/mL,which was comparable to the patients with chronic hepatitis B.independently from HBV-DNA levels, the concentration of sFas among healthy HBsAg carders was comparable to healthy persons.Among patients with chronic hepatitis B and C,the concentration of sFas was significantly higher in comparison to healthy HBsAg carriers and healthy persons.In chronic hepatitis B patients the concentration of sFas was decreased during lamivudine treatment.Among chronic hepatitis C patients the concentration of sFas was increased during IFN alpha and ribavirin treatment,sFasL was not detected in control group.Furbhermore sFasL occurred more frequently in chronic hepatitis C patients in comparison to chronic hepatitis B patients. CONCLUSION:There are no correlations between apoptosis and HBV-DNA levels.However ther is an association between apoptosis and activity of inflammation in patients with chronic HBV infection.Apoptosis can be increased in patients with chronic hepatitis C by effective treatment which may be a result of apoptosis stimulation by IFN-α.

质粒介导的RNA干扰对AD293细胞中HBcAg表达的抑制作用

目的研究质粒介导的RNA干扰效应对HBcAg基因表达的抑制作用。方法设计并构建针对HBcAg基因的小发夹RNA表达载体,将构建好的shRNA表达载体和HBcAg-增强型绿荧光蛋白融合蛋白表达载体共转染人胚肾细胞株AD293,以空载体组以及针对无关序列的shRNA表达载体组为阴性对照,流式细胞术和实时荧光定量PCR法检测RNAi的抑制效果。结果构建的特异性shRNA表达载体可以抑制HBcAg基因在AD293细胞中的表达,流式细胞仪检测抑制率可达76%,实时荧光定量PCR法检测HBcAg基因的mRNA,抑制率可达58.6%。结论质粒介导的RNAi可以有效抑制HBcAg基因的表达,为利用RNAi进行抗乙型肝炎病毒复制研究提供了一个可供选择的方法。

慢性乙型肝炎患者血清HBV pgRNA与HBV DNA和HBeAg水平变化关系研究

目的研究慢性乙型肝炎(CHB)患者血清乙型肝炎病毒前基因组RNA(HBV pgRNA)与乙型肝炎病毒基因(HBV DNA)和乙型肝炎e抗原(HBeAg)水平变化关系。方法选择2018年6月至2021年6月收治的72例CHB患者作为研究对象,以样本值/临界值(S/CO)作为HBeAg检测的结果,将1<S/CO≤5的患者纳入低浓度组(16例),5<S/CO≤10的患者纳入中浓度组(35例),>10的患者纳入高浓度组(21例)。定量检测三组HBV pgRNA、HBV DNA、HBeAg水平,分析HBV pgRNA、HBV DNA水平与HBeAg水平的相关性。结果三组HBV pgRNA水平分别为(5.85±0.64)拷贝/mL、(7.12±0.38)拷贝/mL、(7.75±0.59)拷贝/mL,三组HBV DNA水平分别为(5.34±0.49)拷贝/mL、(7.16±0.62)拷贝/mL、(9.35±0.54)拷贝/mL,三组HBV pgRNA、HBV DNA水平比较,均有低浓度组<中浓度组<高浓度组,P<0.05;经相关性分析,HBV pgRNA与HBeAg呈正相关(r=0.566,P=0.002),HBV DNA和HBeAg呈正相关(r=0.496,P=0.005)。结论随着HBeAg水平的增加,HBV pgRNA、HBV DNA水平逐渐提升,CHB患者血清HBV pgRNA、HBV DNA水平和HBeAg水平呈正相关,临床可结合三项指标对HBV的感染和复制、CHB患者的病情进行更加准确地诊断。

HCV-cAg与HCV-Ab、HCV-RNA及肝功能之间的关系探讨

目的:探讨丙型肝炎病毒核心抗原(hepatitis C virus core antigen,HCV-cAg)与丙型肝炎病毒RNA(hepatitis C virus RNA,HCV-RNA)关系以及丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)、谷氨酰转肽酶(GGT)水平与HCV-RNA病毒载量之间的相关性。方法:选取78例HCV抗体检测阳性的丙型肝炎或疑似丙型肝炎病人作为观察组,另选取同期健康体检者20名作为对照组。采用ELISA法和荧光定量PCR法等对2组血清样本进行HCV-cAg、HCV-RNA和ALT、AST、GGT水平测定,并比较各参数之间的关系。结果:观察组HCV-cAg阳性检出率为41.0%(32/78),HCV-RNA阳性检出率为53.8%(42/78),差异有统计学意义(P<0.01),且二者之间具有较好一致性(κ=0.747)。观察组ALT、AST和GGT水平均明显高于对照组(P<0.01),但与HCV-RNA病毒载量均无明显相关性(P>0.05)。结论:HCV-cAg和HCV-RNA在检测丙型肝炎方面有较好一致性和相关性,可为丙型肝炎的临床诊断提供有价值的依据。

慢性乙型肝炎患者停用核苷(酸)类似物后循环血清中HBV pgRNA、HBcrAg表达水平与复发的相关性分析

目的检测慢性乙型肝炎(CHB)患者停用核苷(酸)类似物(NUC)后循环血清中HBV前基因组RNA(pgRNA)以及乙型肝炎核心相关抗原(HBcrAg)的表达水平,分析停药后CHB患者循环血中不同时段HBV pgRNA、HBcrAg水平与复发之间的相关性。方法选取2019年12月—2022年7月川北医学院附属医院门诊就诊者中抗HBV治疗至少5年到达完全应答并满足2017年版欧洲肝病学会指南停药标准的CHB患者108例,根据停药时间分为停药后4、12、24周组;根据复发情况分为复发组与未复发组。运用实时荧光定量PCR法检测CHB患者循环血清中HBV pgRNA水平,运用ELISA检测研究对象静脉血中HBcrAg的表达水平,采用实时荧光定量PCR法高精度检测HBV DNA载量。计量资料两组间比较采用t检验;多组间比较采用单因素方差分析,进一步两两比较采用LSD-t检验。采用Pearson相关检验分析循环血中各指标间的相关性。结果CHB患者停药后在随访4~12周时复发率17.1%,24周后累积复发率达到29.3%,其中单独HBV DNA复阳者各占64.3%、60.0%,HBeAg单独复阳者各占28.6%、20.0%,HBV DNA、HBeAg同时复阳者各占7.1%、20.0%。停药24周时,CHB患者循环血清中HBV pgRNA、HBcrAg、HBV DNA表达水平较停药时及停药后4周时明显高表达,不同时段比较差异具有统计学意义(P值均<0.05)。复发组循环血清中HBV pgRNA、HBcrAg、HBV DNA水平较未复发组显著高表达(t值分别2.549、8.654、27.429,P值均<0.05);对复发组进一步分析发现,在12~24周时复发组患者循环血清中HBV pgRNA、HBcrAg与HBV DNA水平较4~12周时高表达(P值均<0.05)。复发组停药时循环血清中HBV pgRNA、HBcrAg的表达水平较未复发组停药时显著高表达(t值分别为18.561、6.152,P值均<0.001)。相关性分析显示,CHB患者停药后复发组循环血清中HBV pgRNA、HBcrAg与HBV DNA呈正相关(r值分别为0.82、0.66,P值均<0.001);未复发组循环血清中HBV pgRNA、HBcrAg与HBV DNA均无相关性(r值分别为0.14、0.04,P值均>0.05)。结论停药时复发组HBV pgRNA、HBcrAg水平较未复发组高表达,提示未复发组在停药时的HBV pgRNA、HBcrAg水平可能作为CHB患者可安全停药的参考临界值指标,HBV pgRNA、HBcrAg水平检测可能是未来抗HBV治疗终点选择的潜在参考指标之一。

HBV pgRNA在慢性乙型肝炎进程中的可能意义

HBV pgRNA是HBV cccDNA的直接转录产物,能够通过反映HBV cccDNA的转录活动状态进而反映慢性乙型肝炎病情进展,指导临床治疗和判断预后。与其他常用HBV感染血清标志物相比,HBV pgRNA在反映HBV复制活动和抗病毒治疗效果等方面更灵敏,对于抗病毒治疗阶段终点有预测价值。通过综合HBV pgRNA与HBcrAg、HBV cccDNA的相关性来阐述HBV pgRNA在反映慢性乙型肝炎治疗过程中病情变化的可能意义。

Inhibition of hepatitis B virus by oxymatrine in vivo

AIM To investigate the anti-HBV effect ofoxymatrine (oxy) in vivo.METHODS HBV transgenic mice were producedby micro-injection of a 4.2kb fragmentcontaining the complete HBV genomes.Expression level of HBsAg and HBcAg in thetransgenic mice liver was determined byimmunohistochemical assay.RESULTS Four groups (6 mice in each group)were injected intraperitoneally with oxy at thedosage of 100,200, and 300 mg/kg or with salineonce a day for 30 days. Both HBsAg and HBcAgwere positive in livers of all the six mice in thecontrol group (injected with saline), and werepositive in livers of two mice in 100 mg/kg groupand 300mg/kg group. In 200mg/kg group,HBsAg and HBcAg were negative in livers of allthe six mice. Based on the results, 200 mg/kg isthe ideal dosage to explore the effect of oxy atdifferent time points. According to the oxytreatment time, mice were divided into fourgroups: 10 d, 20 d, 30 d and 60 d (4 mice in eachgroup). Each mouse underwent liver biopsy twoweeks before the treatment of oxy. Down-regulation of HBsAg and HBcAg appeared aftertreatment of oxymatrine for 10 d and 20 d, Dane-like particles disappeared after the treatment ofoxy for 20d under electron microscopy,however, the expression level of HBsAg andHBcAg returned to normal 60 d later after oxytreatment.CONCLUSION oxymatrine can reduce thecontents of HBsAg and HBcAg in transgenic miceliver, longer treatment time and larger dosagedo not yield better effects.

Establishment of transgenic mouse harboring hepatitis B virus (adr subtype) genomes

INTRODUCTIONHepatitis B virus (HBV) belongs to the group ofhepatovirus, a major pathogen of human acute andchronic hepatitis B[1 4], which has a very closeassociation with human hepatocellular carcinoma(HCC)[5-8], For example, a statistical data from ahospital in Shanghai showed that 80% of HCCpatients were positive for HBsAg ( personalcommunication).

血清HBV RNA在HBeAg阳性慢性乙型肝炎患者不同时期的表达水平及检测价值

目的探讨血清HBV RNA在HBeAg阳性慢性乙型肝炎(CHB)患者不同时期的表达水平及潜在临床价值。方法选取2019年8月—2020年12月于杭州市西溪医院肝病科门诊及住院部就诊的CHB患者61例,根据HBeAg阳性CHB患者的抗病毒治疗情况分为3组:HBeAg阳性CHB[HBeAg(+)、HBV DNA(+)]未治患者(A组),HBeAg血清学转换前[HBeAg(+)、HBV DNA(-)]经治患者(B组),HBeAg血清学转换后[HBeAg(-)、HBV DNA(-)]经治患者(C组),检测不同时期患者外周血HBV RNA载量,并分析其与HBsAg、HBV DNA的相关性。符合正态分布的计量资料2组间比较采用t检验;非正态分布的计量资料2组间比较采用Mann-Whitney U检验;计数资料2组间比较采用χ^(2)检验;采用Pearson或Spearman相关分析描述两变量间的相关性。结果3组患者HBV RNA阳性率分别为100%(22/22)、88.2%(15/17)、22.7%(6/22)。HBeAg阳性CHB未治患者HBV RNA与HBsAg、HBV DNA均呈正相关(r值分别为0.612、0.922,P值均<0.01),在HBeAg血清学转换前和HBeAg血清学转换后的经治患者中,HBV RNA与HBsAg无相关性。经治HBeAg阳性组的HBV RNA、HBsAg均高于HBeAg阴性组,差异均有统计学意义(Z值分别为-4.44、-2.41,P值均<0.05)。HBV DNA阳性组HBV RNA显著高于HBV DNA阴性组,差异有统计学意义(Z=-6.16,P<0.01)。结论CHB患者经核苷(酸)类药物抗病毒治疗HBV DNA阴转后仍有部分患者能检测出血清HBV RNA;且HBV RNA只能来自肝内cccDNA,因此HBV RNA比HBV DNA更能反映肝内病毒复制活性,对CHB人群管理有着一定的临床价值。

在大肠杆菌中反义RNA抑制乙型肝炎病毒核心抗原的表达

将乙型肝炎病毒核心抗原(HBcAg)基因的DNA序列分别按正、反方向置tac启动子的控制下,并克隆进入同一个质粒中,转化大肠杆菌JM103后,观察到HBcAg和HBeAg的合成均明显低于仅含有HBcAg基因的细菌,最大抑制率可达70％～80％,分子杂交可见反义RNA存在。

乙型肝炎病毒核心基因启动子变异对HBeAg表达及病情的影响

目的:研究乙型肝炎病毒核心基因启动子(HBVBCP)变异对HBeAg表达及病情的影响。方法:采用PCR微板核酸杂交结合ELISA检测显示技术,对176例HBV慢性感染者血清进行检测HBVBCP区核苷酸(nt)1762碱基A→T和1764G→A联合突变。采用ELISA检测技术,检测患者血清HBV标志物。结果:在176例HBV慢性感染者中检出HBVBCP区T1762A1764G变异者73例,HBVBCP变异的阳性率为41.5%,HBVBCP变异在HBeAg阴性病例的阳性率为49.4%,显著高于HBeAg阳性病例的阳性率33.3%。HBVBCP双变异在慢性乙型肝炎轻、中、重度组,肝炎肝硬化组,慢性重症肝炎组和原发性肝癌组的阳性率分别为28.0%、31.4%、34.2%、39.4%、60.0%和70.0%。各型肝病与HBVBCP变异的阳性或阴性进行相关分析,结果有显著性差异。结论:本组病例HBVBCP变异的阳性率为41.5%,变异可引起HBV感染者的HBeAg阴转。HBVBCP变异与HBV慢性感染的临床类型呈正相关,随着病情加重,BCP变异的阳性率有增高趋势。

miR-155对HBV蛋白表达的抑制作用及机制

目的观察mi R-155对SOCS1 m RNA及蛋白水平的影响,探究mi R-155对HBs Ag、HBe Ag表达的抑制作用。方法从Hep G2.2.15细胞中提取基因组,经PCR扩增得到mi R-155前体序列,纯化并连接入pm R-m Cherry载体,构建重组质粒pmi R-155,经去内毒素后转染至Hep G2.2.15细胞。将细胞分为重组组(转染pmi R-155质粒)、空载组(转染pm Rm Cherry质粒)、转染试剂组、空白组。采用荧光实时定量PCR检测各组细胞内mi R-155的表达,RT-PCR检测各组细胞SOCS1 m RNA的表达,Western blotting检测各组细胞SOCS1 蛋白的表达,ELISA检测各组细胞HBs Ag、HBe Ag的表达。结果荧光实时定量PCR结果显示,以空白组细胞内mi R-155的表达量为基准,重组组mi R-155表达量(519.43±52.10)明显高于空载组(24.24±16.70)及转染试剂组(35.04±26.09,P<0.05);RT-PCR结果显示,重组组SOCS1 m RNA表达量(0.63±0.91)显著低于空白组(P<0.05);Western blotting结果显示,重组组SOCS1蛋白表达量显著低于空白组。以空白组的表达量为基准,重组组HBs Ag和HBe Ag蛋白表达的抑制率分别为55.62%±3.77%和47.87%±2.46%(P<0.01)。结论过表达的mi R-155可抑制细胞内SOCS1和HBV蛋白的表达。

“Anti-HBc alone” in human immunodefi ciency virus-positive and immuno-suppressed lymphoma patients

Hepatitis B virus (HBV) infection is endemic in various parts of the world. A proportion of patients have resolved prior exposure to HBV, as evidenced by the clearance of circulating hepatitis B surface antigen and the appearance of antibody to hepatitis B core antigen (anti-HBc), which could produce protective antibody to hepatitis B surface antigen (anti-HBs). With time, anti-HBs in some patients may become negative. Such patients are described as having occult HBV infection or "anti-HBc alone". In the context of immunodef icient patients, such as HIV patients or lymphoma patients undergoing immunosuppressive immunotherapy, the lack of protective anti-HBs may increase the risk of hepatitis B reactivation. Serum HBV DNA testing may be necessary in "anti-HBc alone" patients, to detect patients at a high risk of developing HBV infection allowing appropriate prophylactic management.