慢性乙型肝炎患者血清白细胞介素10,12,18、干扰素-γ水平和前C/C区变异的关系

目的 :探讨慢性乙型肝炎患者血清白细胞介素 (IL) 10 ,12 ,18、干扰素 (IFN) γ含量与HBV前C/C区突变的关系。方法 :5 2例慢性乙型肝炎患者和 2 5例健康对照者均于清晨空腹采血 ,以双抗夹心ELISA法检测IL 10、IL 12、IL 18、IFN γ血清含量 ,并同时检测HBV DNA和乙肝病毒血清学标志 ,在HBV DNA阳性病例中应用微板核酸杂交技术检测前C/C区基因突变。结果 :33.3%的HBeAg阴性慢性乙肝患者是因HBV前C/C区变异所致 ,仍表现为HBV活动性复制 ;并且慢性乙肝重度组病人HBV前C/C区突变发生率显著高于轻中度组 (P <0 .0 5 )。前C/C区变异组IL 10、IL 12、IL 18、IFN γ含量显著高于正常对照组 (P <0 .0 1) ,非变异组IL 12、IL 18、IFN γ含量显著高于正常对照组 (分别为P <0 .0 1,P <0 .0 1和P <0 .0 5 ) ,前C/C区变异组IL 12、IL 18血清含量显著高于非变异组 (P <0 .0 1)。结论 :慢性乙型肝炎患者血清IL 10、IL 12、IL 18、IFN γ水平的变化和前C/C区突变关系密切 ,宿主免疫压力的增高可能是导致HBV前C/C区变异的重要原因之一。

消化道恶性肿瘤化疗与乙型肝炎病毒再激活的临床分析

目的乙型肝炎病毒(hepatitis B virus,HBV)感染者在接受细胞毒性化疗或免疫抑制治疗时存在HBV再激活的风险。文中探讨HBV感染的消化道恶性肿瘤术后化疗患者化疗后HBV再激活及肝功能损害发生情况。方法回顾性分析102例合并HBV感染的胃癌和结、直肠癌术后患者HBV再激活情况,所有患者进行HBV血清学和生物化学检测。结果 102例患者有46.1%(47/102)出现肝功能异常,有40.2%(41/102)出现HBV再激活;41例HBV再激活患者中35例出现肝功能异常(85.4%),其中16例化疗剂量调整或推迟化疗,8例终止化疗。HBV再激活肝功能异常发生率(85.4%)高于未出现HBV再激活患者(19.7%,12/61),两者比较差异有统计学意义(P<0.01);化疗前检测HBV-DNA>103拷贝/ml患者HBV再激活发生率高于HBV-DNA<103拷贝/ml患者(60.0%vs 27.4%,P<0.01)。结论 HBV感染的消化道肿瘤术后患者接受化疗时存在HBV再激活并导致肝功能损害,化疗前HBV-DNA高者更易发生HBV再激活。

中药安体维康治疗慢性乙型肝炎临床和实验研究

目的:本研究以中药安体维康(Antivirus Compound,ATVC)进行抗乙型肝炎病毒(HBV)的体外实验和治疗慢性乙型肝炎临床研究。方法:体外实验以HBV-DNA转染细胞系(2.2.15细胞株)为模型。临床治疗慢性乙型肝炎60例,对照组57例,观察治疗3个月和6个月时HBV的复制指标阴转率。结果:体外实验发现ATVC对乙型肝炎表面抗原(HBsAg)和乙型肝炎e抗原(HBeAg)的治疗指数分别为31.75和36.27;对HBV-DNA及其复制中间体成剂量依赖性抑制。临床研究发现ATVC对慢性乙型肝炎治疗组HBeAg和HBV-DNA的转阴率分别为47.8%和54.9%;均高于对照组(P<0.05)。治疗3个月和6个月时的疗效相似。结论:研究表明,ATVC对HBV不论在体内或体外均有明显抑制作用。临床治疗慢性乙型肝炎疗效较好。

恩替卡韦用于干扰素治疗失败的慢性乙型肝炎26例疗效观察

目的观察干扰素治疗失败的慢性乙型肝炎26例患者使用恩替卡韦进行抗病毒治疗疗效。方法选择的26例患者ALT均高于正常,小于正常值上限10倍;乙肝血清标志物(HBsAg、HBeAg、HBcAb)阳性,HBV-DNA阳性,在1年前接受过半年以上干扰素治疗而疗效不佳,本次治疗前1年内未接受过抗病毒药物和免疫调节剂的治疗;均仅用恩替卡韦分散片0.5mg,1次/d,观察疗程1年;治疗前及治疗后的第2、4、8、12、24、36、48周进行肝肾功能、乙肝血清标志物的监测以及HBV-DNA定量检测,同时观察不良反应。结果发现恩替卡韦对干扰素治疗失败的慢性乙型肝炎患者的治疗效果与未经干扰素治疗者应用恩替卡韦治疗达到同样的效果。结论建议对于此类患者可选择恩替卡韦治疗。

汕尾市高考学生乙型肝炎表面抗原携带状况调查研究

目的调查汕尾市应届高考学生乙型肝炎表面抗原(HBsAg)携带状况。方法采用酶联免疫吸附试验对2003—2008年汕尾地区20108名高考学生进行HBsAg检测,对结果进行统计分析。结果高考学生HBsAg平均阳性率为14.36%,与全国普通人群HBsAg阳性率(9.75%)相比,差异有统计学意义(P<0.01);HBsAg阳性率逐年对比差异均无统计学意义(χ2=0.05,0.05,0.17,0.17,0.62,P>0.05);但2008年相对于2003年阳性率对比差异有统计学意义(χ2=5.54,P<0.05),提示HBsAg感染状况有逐年改善的趋势。男女对比差异无统计学意义(χ2=3.79,P>0.05);城乡对比差异有统计学意义(χ2=9.66,P<0.01)。结论汕尾市高考学生HBsAg感染状况不容乐观,但总体有改善的趋附,农村HBsAg感染状况形势严峻,改变现状任重道远。

HBXIP基因对乙肝病毒X蛋白诱导细胞凋亡的影响

探讨乙型肝炎病毒X蛋白结合蛋白(hepatitisBXinteractingprotein ,HBXIP)基因在乙型肝炎病毒X蛋白(HBX)诱导肝癌细胞凋亡时对细胞周期的影响.构建HBXIP基因真核表达载体pcDNA3 hbxip ,进行瞬时基因转染,将克隆有HBx基因的pCMV X (分别为1μg、2 μg和3μg)和pcDNA3 hbxip质粒分别和共转染至人H74 0 2肝癌细胞中(总体积分别为5 0 μl) .发现瞬时转染3μgpCMV X质粒后,肝癌细胞凋亡发生率为34 4 % ,肝癌细胞的细胞周期相关蛋白p2 7表达水平发生明显上调;与对照组相比,瞬时转染1μg、2 μg和3μg时,细胞周期蛋白D和细胞周期蛋白E的表达水平均发生明显上调,但随着HBX水平的增加细胞周期蛋白D和细胞周期蛋白E的表达水平发生明显下降;在稳定转染pCMV X质粒的H74 0 2 X肝癌细胞中无明显的细胞凋亡发生,研究发现p2 7的表达水平发生了明显下调,而细胞周期蛋白D和细胞周期蛋白E的表达水平发生了明显上调;当pcDNA3 hbxip质粒与pCMV X质粒进行共瞬时转染时,细胞凋亡发生率由pcDNA3质粒与pCMV X质粒共转染时的2 9 2 %下降为13 3% ,p2 7的表达水平发生了下调,但细胞周期蛋白D和细胞周期蛋白E的表达水平无明显变化.研究结果表明,瞬时转染一定剂量的x基因可导致肝癌细胞发生凋亡,细胞周期相关蛋白p2 7。

血清IL-34水平评估非病毒性肝细胞癌患者预后的临床价值

目的评估血清IL-34预测非病毒性肝细胞癌患者(HCC)预后的临床价值。方法选择2013年3月至2014年3月我院收治的100例非病毒性肝细胞癌患者(非病毒性HCC组)和100例健康体检者(对照组)。通过ELISA评估血清IL-34水平。结果非病毒性HCC组血清IL-34水平[(15.89±6.21)pg/mL]显著高于对照组[(3.03±0.83)pg/mL,t=22.122,P<0.001]。PLT、ALT、AST、ALP、总胆红素、AFP、Child-pugh分级、肿瘤大小、肿瘤分期与IL-34呈正相关。ROC分析血清IL-34对非病毒性HCC的AUC=0.889,截断值8.87 pg/mL,P<0.001,95%CI 0.820~0.958,敏感性82.98%,特异性92.45%。高IL-34(≥8.87 pg/mL,n=41)和低IL-34(<8.87 pg/mL,n=59)生存率具有统计学差异(χ2=14.360,P=0.002)。Cox比例风险模型分析AFP、肿瘤大小、肿瘤分期、IL-34是非病毒性HCC预后危险因素。结论IL-34是非病毒性肝细胞癌患者生存预后的独立因素,IL-34可能与非病毒性肝细胞癌预后有关。

血清白细胞介素⁃6水平对非病毒性肝细胞癌患者预后的影响

目的研究血清白细胞介素⁃6(IL⁃6)对非病毒性肝细胞癌(HCC)患者预后的影响。方法选择2009年1月1日至2015年12月31日在九江学院附属医院消化内科和肝胆外科被诊断为非病毒性HCC的患者共107名,本研究的随访终点是患者死亡,观察时间不少于3个月。通过随访终点将患者分为2组,分别为存活组和死亡组,分析影响患者生存的因素。采用酶联免疫吸附测定法定量检测血清IL⁃6水平。根据血清IL⁃6的中位水平对患者进行分层,以分析其对生存的影响。同时采用多变量分析评估IL⁃6对非病毒性HCC患者预后的影响。结果血清IL⁃6水平[风险比(HR):1.96,95%CI:1.06~3􀆰13,P<0.01]、肿瘤分期(HR:2.14,95%CI:1.13~3.59,P<0.01)及血清谷丙转氨酶水平(HR:1.21,95%CI:0.84~1.93,P<0.01)是非病毒性HCC患者生存的独立预测因素。高IL⁃6组的5年生存率(25.42%)显著低于低IL⁃6组(43􀆰75%),差异有统计学意义(P<0.05)。结论高血清IL⁃6水平是非病毒性HCC患者预后不良的独立因素。

长期核苷酸类似物治疗的慢性乙型肝炎患者血清HBV pgRNA与抗原状态变化的相关性研究

目的研究长期核苷酸类似物治疗的慢性乙型肝炎患者血清乙型肝炎病毒前基因组RNA(HBV pgRNA)与抗原状态变化的相关性,阐明慢性乙型肝炎长期核苷酸类似物抗病毒治疗,达到HBeAg血清转换后停药复发率高的原因和可能的机制。方法选择长期口服核苷酸类似物抗病毒治疗(2年以上)的慢性乙型肝炎患者94例,在其血清HBV DNA低于检测下限(<20 IU/ml)的基础上,根据其HBeAg和HBsAg水平分为5组:HBeAg阳性组(组1)、HBeAg阴性且HBsAg>1500IU/L组(组2)、HBeAg阴性且100IU/L<HBsAg<1500IU/L组(组3)、HBeAg阴性且HBsAg<100IU/L组(组4)及HBeAg阴性同时HBsAg阴性组(组5)。分析比较不同抗原状态下HBVpgRNA的水平及检出率。此外,为了排除其他因素对本研究结果的影响。研究分别按照年龄、性别和服药治疗时间分组。按资料分别用t检验、Mann-Whitney U检验、Kruskal Wallis H检验或线性回归相关性分析进行统计学分析。结果HBeAg阳性患者(组1)血清HBV pgRNA检出率为95.0%;而HBeAg阴性患者则为43.2%,明显低于HBeAg阳性者,差异有统计学意义(P<0.05)。血清HBV pgRNA检出率在HBeAg阴性患者中,组2为75.0%;组3为65.0%;组4为15.0%及组5为0。其中,组1、组2及组3的血清HBVpgRNA检出率明显高于组4和组5,差异有统计学意义(P值均<0.05)。然而,统计学分析发现:组1、组2和组3之间血清HBV pgRNA检出率差异无统计学意义(P>0.05)。组4和组5之间的血清HBV pgRNA检出率比较,差异无统计学意义(P>0.05)。而且,血清HBV pgRNA检出率与患者的年龄、性别及核苷酸类似物治疗时间无相关性(P>0.05)。结论慢性乙型肝炎长期核苷酸类似物抗病毒治疗后的血清学抗原状态可能与血清HBV pgRNA的存在有关。HBV pgRNA的持续存在也可能是HBeAg血清转换率低及HBsAg持续高水平的原因。

Inhibition of hepatitis B virus production by Boehmeria nivea root extract in HepG2 2.2.15 cells

AIM: To explore the anti-hepatitis B virus (HBV) effects of Boehmeria nivea (B. nivea) root extract (BNE) by using the HepG2 2.2.15 cell model system. METHODS: Hepatitis B surface antigen (HBsAg), hepatitis B virus e antigen (HBeAg), and HBV DNA were measured by using ELISA and real-time PCR, respectively. Viral DNA replication and RNA expression were determined by using Southern and Northern blot, respectively. RESULTS: In HepG2 2.2.15 cells, HBeAg (60%, P < 0.01) and particle-associated HBV DNA (> 99%, P < 0.01) secretion into supernatant were significantly inhibited by BNE at a dose of 100 mg/L, whereas the HBsAg was not inhibited. With different doses of BNE, the reduced HBeAg was correlated with the inhibition of HBV DNA. The anti-HBV effect of BNE was not caused by its cytotoxicity to cells or inhibition of viral DNA replication and RNA expression. CONCLUSION: BNE could effectively reduce the HBV production and its anti-HBV machinery might differ from the nucleoside analogues.

Mechanism of T cell hyporesponsiveness to HBcAg is associated with regulatory T cells in chronic hepatitis B

AIM： To study the mechanisms of hyporesponsiveness of HBV-specific CD4^＋ T cells by testing TH1 and TH2 commitment and regulatory T cells. METHODS： Nine patients with chronic hepatitis B were enrolled. Peripheral blood mononuclear cells were stimulated with HBcAg or HBsAg to evaluate their potential to commit to TH1 and TH2 differentiation. HBcAg-specific activity of regulatory T cells was evaluated by staining with antibodies to CD4, CD25, CTLA-4 and interleukin-10. The role of regulatory T cells was further assessed by treatment with anti-interleukin-10 antibody and depletion of CD4^＋CD25^＋ cells. RESULTS： Level of mRNAs for T-bet, IL-12R β2 and IL-4 was significantly lower in the patients than in healthy subjects with HBcAg stimulation. Although populations of CD4^＋CD25^highCTLA-4^＋ T cells were not different between the patients and healthy subjects, IL-10 secreting cells were found in CD4^＋ cells and CD4^＋CD25^＋ cells in the patients in response to HBcAg, and they were not found in cells which were stimulated with HBsAg. Addition of anti-IL-10 antibody recovered the amount of HBcAgspecific TH1 antibody compared with control antibody （P 〈 0.01, 0.34% ± 0.12% vs 0.15% ± 0.04%）. Deletion of CD4^＋CD25^＋ T cells increased the amount of HBcAgspecific TH1 antibody when compared with lymphoo/tes reconstituted using regulatory T cells （P 〈 0.01, 0.03% ± 0.02% vs 0.18% ± 0.05%）.CONCLUSION： The results indicate that the mechanism of T cell hyporesponsiveness to HBcAg includes activation of HBcAg-induced regulatory T cells in contrast to an increase in TH2-committed cells in response to HBsAg.

A combination treatment of entecavir and early-phase corticosteroid in severe exacerbation of chronic hepatitis B

Of patients with severe exacerbation of chronic hepatitis B accompanied by jaundice and coagulopathy,20%-30%have a fatal outcome.In this report,we describe 2 cases of severe exacerbation of chronic hepatitis B with jaundice and coagulopathy who were successfully treated with a combination of entecavir and corticosteroid.In both cases,rapid reductions in serum hepatitis B virus(HBV)-DNA levels were observed,and corticosteroid was stopped after serum HBV-DNA levels became undetectable.Entecavir treatment was continued.Generally,entecavir treatment reduced serum HBV-DNA levels rapidly,although the improvement in liver function was delayed by a few weeks.During this time lag,liver cell injury continued and the disease progressed.Corticosteroid suppressed the excessive host immune response and was useful for stopping progressive deterioration.A combination of entecavir and early-phase corticosteroid may be a useful treatment in severe exacerbation of chronic hepatitis B.

A comparative review of HLA associations with hepatitis Band C viral infections across global populations

Hepatitis B （HBV） and hepatitis C （HCV） viral infection or co-infection leads to risk of development of chronic infection, cirrhosis and hepatocellular carcinoma （HCC）. Immigration and globalization have added to the challenges of public health concerns regarding chronic HBV and HCV infections worldwide. The aim of this study is to review existing global literature across ethnic populations on HBV and HCV related human leukocyte antigen （HLA） associations in relation to susceptibility, viral persistence and treatment. Extensive literature search was conducted to explore the HLA associations in HBV and HCV infections reported across global populations over the past decade to understand the knowledge status, weaknesses and strengths of this information in different ethnic populations. HLA DR13 is consistently associated with HBV clearance globally. HLADRB1＊11/＊12 alleles and DQB1＊0301 are associated with HBV persistence but with HCV clearance worldwide. Consistent association of DRB1＊03 and ＊07 is observed with HCV susceptibility and non-responsiveness to HBV vaccination across the population. HLA DR13 is protective for vertical HBV and HCV transmission in Chinese and Italian neonates, but different alleles are associated with their susceptibility in these populations. HLA class I molecule interactions with Killer cell immunoglobulin like receptors （KIR） of natural killer （NK） cells modulate HCV infection outcome via regulating immune regulatory cells and molecules. HLA associations with HBV vaccination, interferon therapy in HBV and HCV, and with extra hepatic manifestations of viral hepatitis are also discussed. Systematic studies in compliance with global regulatory standards are required to identify the HLA specific viral epitope, stage specific T cell populations interacting with different HLA alleles during disease progression and viral clearance of chronic HBV or HCV infections among different ethnic populations. These studies would facilitate stage specific therapeutic strategies for clearance of HBV and HCV infections or co-infections across global populations and aid in identification of HBV-HCV combined vaccine. HLA associations of chronic HBV or HCV development with confounding host factors including alcohol, drug abuse, insulin resistance, age and gender are lacking and warrant detailed investigation across global populations.

Prevalence of hepatitis B and C markers among refugees in Athens

AIM: To assess the prevalence of hepatitis B and C serological markers in a population of refugees living in Athens.METHODS: One hundred and thirty refugees (81 males and 49 females, mean age ±SD: 31.7±8 years) were included in the study. The hepatitis B virus surface antigen (HBsAg),the hepatitis B virus core antibody (anti-HBc) and the hepatitis C virus antibody (anti-HCV) were detected using a third-generation immunoassay.RESULTS: Twenty individuals (15.4%) were HBsAg positive and 69 (53.1%) were anti-HBc positive. The prevalence of HBsAg and anti-HBc was higher among refugees from Albania and Asia (statistical significant difference, P＜0.008 and P＜0.001 respectively). The prevalence of these markers was found irrelevant to age or sex. Anti-HCV was detected in the serum of 3 individuals (2.3 %). No differences among age, sex or ethnicity regarding anti-HCV prevalence were found.CONCLUSION: It can be concluded that refugees living in Athens are an immigrant population characterized by a high incidence of HBV infection. The prevalence of HBV markers is higher among refugees from Albania and Asia. It is therefore believed that the adherence to general precautions and the initiation of HBV vaccination programs will be necessary in the future, especially in these communities.Although the prevalence of HCV infection seems to be relatively low, extended epidemiological surveys are needed to provide valid results.

Existence and significance of hepatitis B virus DNA in kidneys of IgA nephropathy

AIM: To investigate the existence and significance of hepatitis B virus (HBV) DNA in the pathogenesis of IgA nephropathy(IgAN).METHODS: Fifty cases of IgAN with HBV antigenaemia and/or hepatitis B virus antigens (HBAg, or HBsAg, HBcAg)detected by immunohistochemistry in renal tissues were enrolled in our study. The distribution and localization of HBV DNA were observed using in situ hybridization.Southern blot analysis was performed to reveal the state of renal HBV DNA.RESULTS: Among the 50 patients with IgAN, HBs antigenemia was detected in 17 patients (34%). HBAg in renal tissues was detected in 48 patients (96%), the positive rate of HBAg, HBsAg, and HBcAg was 82% (41/50), 58% (29/50),and 42% (21/50) in glomeruli, respectively; and was 94%(47/50), 56% (28/50) and 78% (39/50) in tubular epithelia,respectively. Positive HBV DNA was detected in 72% (36/50)and 82% (41/50) cases in tubular epithelia and glomeruli respectively by in Situ hybridization, and the positive signals were localized in the nuclei of tubular epithelial cells and glomerular mesangial cells as well as infiltrated interstitial lymphocytes. Moreover, 68% (34/50) cases were proved to be HBV DNA positive by Southern blot analysis, and all were the integrated form.CONCLUSION: HBV infection might play an important role in occurrence and progress of IgAN. In addition to humoral immune damages mediated by HBAg-HBAb immune complex,renal tissues of some IgAN are directly infected with HBV and express HBAg in situ, and the cellular mechanism mediated by HBV originating from renal cells in situ may also be involved in the pathogenesis of IgAN.

Durability of viral response after off-treatment in HBeAg positive chronic hepatitis B

AIM:To evaluate the durability in hepatitis B e antigen (HBeAg) positive chronic hepatitis B patients who discontinued antiviral treatment. METHODS:A total of 48 HBeAg positive chronic hepatitis B patients who were administered nucleoside analogues and maintained virological response for ≥ 6 mo [hepatitis B virus (HBV) DNA < 300 copies/mL and HBeAg seroconversion] before cessation of treatment were enrolled between February 2007 and January 2010. The criteria for the cessation of the antiviral treatment were defined as follows:(1) achievement of virological response; and (2) duration of consolidation therapy (≥ 6 mo). After treatment cessation, the patients were followed up at 3-6 mo intervals. The primary endpoint was serologic and virologic recurrence rates after withdrawal of antiviral treatment. Serologic recurrence was defined as reappearance of HBeAg positivity after HBeAg seroconversion. Virologic recurrence was defined as an increase in HBV-DNA level > 104 copies/mL after HBeAg seroconversion with previously undetectable HBV-DNA level. RESULTS:During the median follow-up period of 18.2 mo (range:5.1-47.5 mo) after cessation of antiviral treatment, the cumulative serological recurrence rate was 15 % at 12 mo. The median duration between the cessation of antiviral treatment and serologic recurrence was 7.2 mo (range:1.2-10.9 mo). Of the 48 patients with HBeAg positive chronic hepatitis, 20 (41.6%) showed virological recurrence. The cumulative virologic recurrence rates at 12 mo after discontinuing the antiviral agent were 41%. The median duration between off-treatment and virologic recurrence was 7.6 mo (range:4.3-27.1 mo). The mean age of the virological recurrence group was older than that of the non-recurrence group (46.7 ± 12.1 years vs 38.8 ± 12.7 years, respectively; P = 0.022). Age (> 40 years) and the duration of consolidation treatment (≥ 15 mo) were significant predictive factors for offtreatment durability in the multivariate analysis [P = 0.049, relative risk (RR) 0.31, 95% CI (0.096-0.998) and P = 0.005, RR 11.29, 95% CI (2.054-65.12), respectively]. Patients with age (≤ 40 years) who received consolidation treatment (≥ 15 mo) significantly showed durability in HBeAg positive chronic hepatitis B patients (P = 0.014). These results suggest that additional treatment for more than 15 mo after HBeAg seroconversion in patients who are ≤ 40 years old may be beneficial in providing a sustained virological response. CONCLUSION:Our data suggest that HBeAg seroconversion is an imperfect end point in antiviral treatment. Long-term consolidation treatment (≥ 15 mo) in younger patients is important for producing better prognosis in HBeAg positive chronic hepatitis B.

Generation of the regulatory protein rtTA transgenic mice

AIM: To translate Tet-on system into a conditional mouse model, in which hepatitis B or C virus (HBV or HCV) gene could be spatiotemporally expressed to overcome 'immune tolerance' formed during the embryonic development and 'immune escape' against hepatitis virus antigen(s), an effector mouse, carrying the reverse tetracycline-responsive transcriptional activator (rtTA) gene under the tight control of liver-specific human apoE promoter, is required to be generated. METHODS: To address this end, rtTA fragment amplified by PCR was effectively inserted into the vector of pLiv.7 containing apoE promoter to create the rtTA expressing vector, I.e., pApoE-rtTA. ApoE-rtTA transgenic fragment (-6.9 kb) released from pApoE-rtTA was transferred into mice by pronucleus injection, followed by obtaining one transgene (+) founder animal from microinjection through PCR and Southern blot analysis.RESULTS: rtTA transgene which could be transmitted to subsequent generation (F1) derived from founder was expressed in a liver-specific fashion. CONCLUSION: Taken together, these findings demonstrate that rtTA transgenic mice, in which rtTA expression is appropriately targeted to the murine liver, are successfully produced, which lays a solid foundation to 'off-on-off' regulate expression of target gene (s) (e.g., HBV and/or HCV) in transgenic mice mediated by Tet-on system.

Impact of comorbidities on the severity of chronic hepatitis B at presentation

AIM:To evaluate the clinical relevance of each cofactor on clinical presentation of chronic hepatitis B.METHODS:Out of 1366 hepatitis B surface antigen(HBsAg) positive subjects consecutively observed in 79 Italian hospitals,53(4.3%) showed as the only cofactor hepatitis D virus(HDV) infection [hepatitis B virus(HBV)/HDV group],130(9.5%) hepatitis C virus(HCV)(group HBV/HCV),6(0.4%) human immunodeficiencyvirus(HIV)(group HBV/HIV),138(10.2%) alcohol abuse(group HBV/alcohol);109(8.0%) subjects had at least two cofactors and 924 were in the cofactor-free(CF) group.RESULTS:Compared with patients in group CF those in group HBV/alcohol were older and more frequently had cirrhosis(P < 0.001),those in group HBV/HDV were younger(P < 0.001),more frequently resided in the south of the country and had cirrhosis(P <0.001),those in group HBV/HCV were older(P < 0.001) and more frequently had cirrhosis(P < 0.001).These cofactors were all independent predictors of liver cirrhosis in HBsAg positive patients.Multivariate analysis showed that an older age [odds ratio(OR) 1.06,95% CI:1.05-1.08],alcohol abuse with more than 8 drinks daily(OR 2.89,95% CI:1.81-4.62) and anti-HDV positivity(OR 3.48,95% CI:2.16-5.58) are all independently associated with liver cirrhosis.This association was found also for anti-HCV positivity in univariate analysis,but it was no longer associated(OR 1.23,95% CI:0.84-1.80) at multivariate analysis.CONCLUSION:Older age,HDV infection and alcohol abuse are the major determinants of severe liver disease in chronic HBV infection,while HCV replication plays a lesser role in the severity of hepatic damage.

Distinct expression patterns in hepatitis B virus-and hepatitis C virus-infected hepatocellular carcinoma

AIM： To identify biomarkers indicating virus-specific hepatocarcinogenic process, differential mRNA expression in 32 patients with hepatitis B virus （HBV）-/hepatitis C virus （HCV）-associated hepatocellular carcinoma （HCC） were investigated by means of cDNA microarrays comprising of 886 genes. METHODS： Thirty two HCC patients were divided into two groups based on viral markers： hepatitis B virus positive and HCV positive. The expression profiles of 32 pairs of specimens （tumorous and surrounding nontumorous liver tissues）, consisting of 886 genes were analyzed. RESULTS： Seven up-regulated genes in HBV-associated HCC comprised genes involved in protein synthesis （RPSS）, cytoskeletal organization （KRTS）, apoptosis related genes （CFLAR）, transport （ATPSF1）, cell membrane receptor related genes （IGFBP2）, signal transduction or transcription related genes （MAP3KS）, and metastasis-related genes （MMP9）. The up-regulated genes in HCV-infected group included 4 genes： V/M （cell structure）, ACTB （cell structure）, GAPD （glycolysis） and CD58 （cell adhesion）. The expression patterns of the 11 genes, identified by cDNA microarray, were confirmed by quantitative RT-PCR in 32 specimens.CONCLUSION： The patterns of all identified genes were classified based on the viral factor involved in HBV- and HCV-associated HCC. Our results strongly suggest that the pattern of gene expression in HCC is closely associated with the etiologic factor. The present study indicates that HBV and HCV cause hepatocarcinogenesis by different mechanisms, and provide novel tools for the diagnosis and treatment of HBV- and HCV-associated HCC.