慢性乙型肝炎患者肝组织乙型肝炎病毒cccDNA和血清病毒学标志物相关性的研究

目的探讨CHB患者血清病毒学标志物及肝组织、血清HBV DNA与肝组织HBVcccDNA的关系。方法应用实时荧光定量法进行20例CHB患者肝组织HBVcccDNA、肝组织及血清HBV DNA定量的测定,时间分辨荧光免疫分析法进行HBsAg、HBeAg、HBcAb等血清学标志物的测定,分析肝组织HBVcccDNA与肝组织HBV DNA、血清HBV DNA、HBsAg及HBeAg定量水平之间的关系。相关性分析采用Spearman相关性检验。结果所有患者肝组织中均能检测到HBVcccDNA,含量在7.63×10~4拷贝数/mg~9.46×10~8拷贝数/mg(对数值:7.55±1.08)之间;肝组织HBVcccDNA与肝组织HBV DNA呈显著正相关关系(r=0.859,P0.01),与血清HBV DNA呈显著正相关关系(r=0.640,P0.01);肝组织HBVcccDNA与HBeAg呈显著正相关(r=0.676,P0.01),与血清HBsAg无相关性(r=0.195,P0.05),与HBcAb呈显著负相关(r=-0.624,P0.01)。结论 CHB患者肝组织内HBVcccDNA呈稳定的中等水平复制;血清HBsAg定量尚不能作为反映肝组织中HBVcccDNA水平的指标,结合肝组织HBV DNA、血清HBV DNA等定量检测可以更全面反映HBV复制水平及患者病情评估。     

慢性乙型肝炎、肝硬化患者肝组织病毒载量与肝组织损伤程度的关系

目的了解慢性乙型肝炎（CHB）、肝硬化患者肝内HBV载量的变化与肝组织损伤程度的关系及其对该类患者的预后意义。方法经血清学及肝穿刺病理组织学证实为CHB 46例,经血清学及影像学确诊、手术获得的肝硬化标本17例。采用实时荧光载量系统检测血清及肝组织HBV DNA载量,并观察肝组织炎症及纤维化程度。结果 （1）肝硬化组血清HBV DNA载量低于HBeAg阳性CHB组（P〈0.01）、高于HBeAg阴性CHB组（P〈0.001）;肝硬化组肝组织HBV DNA载量低于HBeAg阳性及HBeAg阴性CHB组（P〈0.001、P〈0.01）;HBeAg阳性及阴性CHB组的肝组织HBV DNA载量均高于血清HBV DNA载量（P〈0.05、P〈0.01）,肝硬化组肝组织HBV DNA载量低于血清（P〈0.001）。（2）CHB、肝硬化血清HBV DNA载量与肝组织的炎症及纤维化程度呈正相关（P〈0.05）,肝组织HBV DNA载量则与肝组织的炎症及纤维化程度呈明显负相关（P〈0.05）。结论肝组织HBV DNA较血清HBV DNA载量更能准确反映肝内HBV复制水平、肝组织损伤程度以及CHB、CHB肝硬化的进展程度。     

HBeAg阳性和阴性慢性乙型肝炎患者肝组织学与血清学关系的比较

目的 研究慢性乙型肝炎(CHB)患者(1×ULN≤ALT≤2×ULN)肝组织学与血清学的关系.方法 对全国多中心516例CHB患者(1×ULN≤ALT≤2×ULN)行血清学及肝穿刺病理检查,依据血清HBeAg表达水平将患者分为HBeAg阳性组和HBeAg阴性组,分析两组肝组织病理学与血清学指标的相关性.结果 HBeAg阳性患者肝组织炎症分级和肝纤维化分期与血清HBsAg和HBeAg表达水平呈负相关(P＜0.01),与血清透明质酸(HA)和α2-巨球蛋白水平呈正相关(P＜0.01),与血清HBVDNA水平无明显相关性;HBeAg阴性患者肝组织炎症分级和肝纤维化分期与血清HBV DNA水平呈正相关(P＜0.01),与HBsAg、HA和α2-巨球蛋白等无明显相关性.结论 血清HBV DNA可做为HBeAg阴性CHB患者肝脏病变程度的有效预测指标,HBsAg、HBeAg、HA和α2-巨球蛋白均可做为HBeAg阳性CHB患者肝组织损伤的预测指标.     

抗病毒药物对肝组织乙型肝炎病毒共价闭合环状DNA的影响

目的 探讨抗病毒药物对肝组织HBV共价闭合环状DNA(cccDNA)的影响.方法 71例HBeAg阳性的慢性乙型肝炎患者分别接受48周的拉米夫定-干扰素序贯治疗、单用拉米夫定治疗和24周的干扰素治疗,随访24周.检测治疗前、后肝组织HBV DNA和cccDNA水平;检测治疗前、后及停药24周时的血清HBV DNA和ALT水平.比较C、B基因型HBV感染患者肝组织HBV DNA和cccDNA水平.结果 治疗结束时,序贯治疗、拉米夫定治疗和干扰素治疗组患者肝组织HBV DNA分别为(4.7±1.1)log10、(4.6±1.5)log10和(5.6±1.5)log10,均低于治疗前水平(P＜0.05);cccDNA分别为(3.4±1.3)log10、(3.8±1.1)log10和(5.0±1.5)log10,均低于治疗前水平(P＜0.05).17例患者出现了HBeAg血清学转换,其肝组织cccDNA下降幅度明显大于HBeAg阳性患者(3.0 log10比1.6 log10,P＜0.05).停药24周,18例患者获得持续病毒学应答,其cccDNA基线值明显低于停药后出现病毒反跳患者(P＜0.05).肝组织cccDNA的变化与肝组织HBV DNA的改变正相关(P＜0.05);治疗结束时,肝组织cccDNA水平与血清HBeAg滴度正相关(P＜0.01).C基因型与B基因型HBV感染患者治疗前、后肝组织HBV DNA和cccDNA的变化无统计学意义(P＞0.05).结论 48周的拉米夫定-干扰素序贯治疗和拉米夫定治疗对肝组织cccDNA抑制作用强于24周的干扰素治疗.肝组织cccDNA低水平患者易获得较好的抗病毒疗效.HBV基因型对肝组织cccDNA含量无明显影响.     

慢性乙型肝炎患者血清HBV DNA、HBeAg与肝组织HBcAg及炎症分级的关系

目的 探讨慢性乙型肝炎（CHB）患者血清HBV DNA、HBeAg与肝组织HBcAg及炎症分级的关系.方法 回顾性分析250例CHB肝穿刺患者,按血清HBV DNA水平,分为A组（＜3 log10 IU/ml）45例,B组（3～6 log10 IU/ml）138例,C组（＞6 log10 IU/ml）67例.按HBeAg阴阳性不同分为阳性组142例,阴性组108例.分别与肝组织中HBcAg水平、肝组织炎症分级进行比较,分析彼此的相关性,率的比较用卡方检验及确切概率法,相关性分析采用直线相关分析.结果 血清HBV DNA不同水平与肝组织HBcAg免疫组化比较差异有统计学意义（χ2=11.1,P=0.05）,两者之间呈正相关（r=0.75,P=0.001）;与肝组织炎症分级（G）比较差异无统计学意义（χ2=13.3,P=0.075）,两者之间也无相关性（r=0.04,P=0.325）.HBeAg阳性和阴性分组与肝组织中HBcAg水平比较差异有统计学意义（χ2=6.64,P=0.01）,两者之间呈正相关（r=0.56,P=0.001）;与肝组织炎症分级（G）比较差异无统计学意义（χ2=8.43,P=0 065）,两者之间也无相关性（r=0.06,P=0.415）.结论 CHB患者肝组织HBcAg表达更能反映出肝内HBV复制状态,患者血清中测不出病毒标志物时,可以考虑肝穿刺以观察肝组织中HBcAg表达来判断HBV复制状态具有重要意义.     

血清HBsAg和HBeAg水平变化与CHB患者抗病毒疗效预测

血清HBsAg或HBeAg定量检测在CHB患者对治疗应答有预测作用,因而可以指导和调整治疗方案。本文阐述了CHB患者血清HBsAg和HBeAg变化与HBV复制指标如血清HBV DNA、肝内HBV cccDNA和HBV DNA的关系,以便对CHB患者血清HBsAg或HBeAg水平在判定抗病毒疗效方面有一个相对清晰的认识。     

血清HBsAg和HBeAg水平变化与慢性乙型肝炎患者抗病毒疗效预测

血清HBsAg或HBeAg定量水平的变化在慢性乙型肝炎（CHB）患者抗病毒治疗的应答中有重要预测价值,可以用于指导和调整治疗方案。本文综述了CHB患者血清HBsAg和HBeAg水平变化与HBV复制指标如血清HBV DNA和肝内HBV cccDNA间的关系,以阐明CHB患者血清HBsAg或HBeAg水平在判定抗病毒疗效方面的新认识。     

干扰素治疗慢性乙型肝炎时e抗原血清学转换的相关因素

目的探讨HBeAg阳性慢性乙型肝炎在聚乙二醇干扰素α-2a（PEG-IFN α-2a）治疗过程中HBeAg血清学转换和病毒学应答的相关因素，HBeAg血清学转换与HBV DNA应答的相关性。方法患者采用PEG-IFNα-2a每次180μg，皮下注射，每周1次，共治疗48周，治疗结束后随访24周。用Abbott公司生产的第三代HBV血清学检测试剂和AXSYM自动酶标检测仪检测血清HBeAg、抗-HBe，实时荧光定量PCR检测HBVDNA载量，分析不同治疗阶段和随访结束的病毒学应答率（HBV DNA〈1．0×10^5拷贝／ml），HBeAg血清转换率及变化规律和影响病毒学应答和HBeAg血清转换的因素。结果治疗12周和随访结束时HBeAg血清转换组和非转换组的ALT水平比较，差异有统计学意义。病毒学应答无论在治疗期还是随访结束时，应答组与非应答组之间的ALT水平差异均有统计学意义。HBeAg血清学转换与非转换组之间的HBV DNA载量之间在治疗12周、治疗结束和随访结束时，差异无统计学意义。治疗期间病毒学应答与非应答组的HBV DNA载量之间的差异有统计学意义，但持续病毒学应答与HBV DNA载量无显著相关性。治疗12、24周和48周获得病毒学应答组的HBeAg血清转换率分别为43．8％、21．4％和18．9％。治疗12、24周和48周时病毒学应答组，在随访结束时的HBeAg的血清转换率分别为42．9％、33．3％和27．6％。多因素分析显示，治疗72周的HBeAg血清转换与治疗结束时的HBV DNA阴转显著相关（OR=2．15，95．0％CI=1．744-2．664，P〈0．01）。结论治疗12周和持续HBeAg血清学转换以及病毒学应答均与ALT基线水平相关，HBeAg血清学转换与基础HBV DNA载量无关，但与治疗过程中病毒学应答相关。     

HBV感染恢复期血清标志物与肝细胞HBV cccDNA检测对照

目的:了解肝组织乙肝病毒共价闭合环状脱氧核糖核酸(HBV cccDNA)与血清HBV复制指标的关系,以探索判定抗HBV的疗程终结点的观察方法。方法:既往HBV感染者、乙型肝炎肝硬化肝癌及慢性乙型肝炎(CHB)患者各13例,将血清HBV DNA、HBV-M及肝组织HBV cccDNA检测结果进行对照。结果:肝硬化肝癌及CHB患者抗-HBe阳性分别为12例和9例,血HBV DNA阳性分别为6例和8例,肝细胞HBV cccDNA均阳性;既往HBV感染13例中,HBsAg阴性,抗-HBs和/或抗HBc阳性各7和6例,有3例肝细胞中HBV cccDNA阳性。抗-HBs阳性的7例肝细胞HBV cccDNA阴性,抗-HBe阳性及抗HBs合并抗-HBe阳性者,肝细胞HBV cccDNA分别为105拷贝/ml和103拷贝/ml,与抗-HBs阳性组对照,t=4.5、4.0(P均<0.01),差异有非常显著性意义。CHB患者核苷类治疗后全应答,肝组织HBV cccDNA仍均为阳性。结论:抗-HBs阳性HBV感染后恢复期血清,肝组织HBV cccDNA阳性比率明显低于CHB抗HBe阳性HBV DNA阴性病例。抗病毒治疗CHB的终点,应是血清抗-HBs阳性的出现。     

慢性HBV感染者肝脏HBV cccDNA含量相关因素分析

目的探讨慢性HBV感染者肝组织中HBV cccDNA含量与血清病毒标志物、HBV DNA及肝脏病理分级的关系,为临床评价抗病毒治疗效果及疗程确定提供理论依据。方法以2007年5月-2008年2月住院的30例慢性HBV感染者为研究对象,应用实时荧光定量聚合酶链反应（RT-PCR）方法检测患者肝组织中HBV cccDNA、肝组织总HBV DNA（HBV tDNA）和血清HBVDNA,同时用化学发光免疫分析法检测HBsAg、HBeAg定量,分析感染者肝组织内HBV cccDNA与肝组织内HBV DNA、血清HBVDNA、HBsAg及HBeAg定量水平之间的关系,并比较肝组织中HBV cccDNA含量与肝脏病理炎症和纤维化分级的关系。采用Pear-son简单相关和Spearman等级相关法进行相关性分析。结果 30例慢性HBV感染者肝组织中均可检出HBV cccDNA,范围在3.15×103～1.06×107拷贝/mg;肝组织cccDNA定量与肝组织总HBV DNA定量呈正相关（r=0.375,P〈0.05）,与血清HBV DNA无相关性（r=0.174,P〉0.05）;肝组织中HBV cccDNA水平与血清HBsAg定量呈高度正相关（r=0.562,P〈0.001）,而与血清HBeAg定量无相关性（r=0.152,P〉0.05）。肝组织cccDNA定量与肝组织炎症活动度（G）及纤维化程度（S）无相关性（r=0.082,P〉0.05）。结论慢性HBV感染者肝组织内HBV cccDNA成稳定的中等水平复制;血清HBV DNA载量不能直接代表其肝组织中的HBV cccDNA水平;血清HBsAg定量可作为反映肝组织中HBV cccDNA水平的指标。     

慢性乙型肝炎患者肝组织HBV cccDNA、总HBV DNA与血清HBV DNA的相关性及其与临床的关系

目的探讨慢性乙型肝炎患者肝组织HBV共价闭合环状DNA（cccDNA）、肝组织总HBV DNA（HBV tDNA）与血清HBV DNA之间的相关性及其与临床的关系。方法 78例慢性乙型肝炎患者入选本研究。肝组织β- globinDNA、HBV cccDNA和HBV tDNA采用实时荧光定量PCR方法检测,平均每个肝细胞HBV cccDNA和HBV tDNA含量（拷贝/cell）=HBV cccDNA（实测值）/β-globin DNA（实测值）和HBV tDNA（实测值）/β3-globin DNA（实测值）,肝组织HBV cccDNA和HBV tDNA含量的计算单位定义为log₁₀拷贝/10⁶cell;采用实时荧光定量PCR、ELISA法检测血清HBVDNA和HBV标志物;采用免疫组织化学方法检测肝细胞中HBsAg和HBcAg的表达。统计分析采用pearson相关分析及t检验。结果 （1）肝组织HBV cccDNA与HBV tDNA定量呈正相关（r=0.696,P<0.001）;肝组织HBV cccDNA与血清HBV DNA定量呈正相关（r=0.304,P<0.01）;肝组织HBV tDNA与血清HBV DNA定量呈正相关（r=0.341,P<0.01）;（2）肝细胞内HBcAg定性检测阳性患者的血清HBV DNA定量明显高于阴性患者,且差异有统计学意义（P<0.05）;肝细胞内HBcAg定性检测阳性患者与阴性患者的肝组织HBV cccDNA和HBV tDNA定量差异均无统计学意义和（P均>0.05）;（3）肝细胞内HBsAg定性检测阳性患者的血清HBV DNA定量明显高于阴性患者,且差异有统计学意义（P<0.05）;肝细胞内HBsAg定性检测阳性患者与阴性患者的肝组织HBV cccDNA和HBV tDNA定量差异均无统计学意义（P>0.05）;（4）HBeAg（+）/抗-HBe（-）患者血清HBV DNA定量明显高于HBeAg（-）/抗-HBe（+）患者,且差异有统计学意义（P<0.05）;HBeAg（+）/抗-HBe（-）患者肝组织HBV cccDNA和HBV tDNA定量与HBeAg（-）/抗-HBe（+）患者比较差异均无统计学意义（均P>0.05）;（5）肝组织HBV cccDNA、HBV tDNA以及血清HBV DNA三者与肝脏炎症活动度及纤维化程度均无显著相关性（P>0.05）。结论 （1）血清HBV DNA定量结果并不一定能完全反映患者肝组织中HBV cccDNA和HBV tDNA含量,尤其在血清HBV DNA<500拷贝/mL时,肝组织中仍存在HBV cccDNA和HBV tDNA,且含量大小不等。（2）肝细胞内HBcAg定性检测阳性或者HBsAg定性检测阳性患者的血清HBV DNA定量均明显高于阴性患者;而两者的肝组织HBV cccDNA和HBV tDNA定量均没有显著差异;（3）HBeAg（+）/抗-HBe（-）患者血清HBV DNA定量明显高于HBeAg（-）/抗-HBe（+）患者,而两者的肝组织HBV cccDNA和HBV tDNA均没有显著差异;（4）肝组织HBV cccDNA、HBV tDNA及血清HBV DNA与肝脏炎症活动度和纤维化程度均无显著相关性。     

乙型肝炎肝硬化和肝癌患者肝组织HBV复制状态分析

目的了解肝硬化和肝癌患者HBV的复制状态。方法在1982例肝癌和1995例乙型肝炎肝硬化患者,常规检测HBV标记物,并在其中12例患者取得肝组织进行HBVcccDNA检测。结果肝硬化和肝癌患者抗HBe阳性所占比例分别为61.7%和74%(P<0.01);将肝硬化和肝癌患者合并后,血清抗HBe阳性者血清HBVDNA≤103拷贝/毫升者占69.1%,高于HBeAg阳性者的21.6%(P<0.01);抗HBe阳性患者HBV DNA≥107拷贝/毫升者占6.4%,低于HBeAg阳性者的42.0%(P<0.01);HBeAg阳性和抗HBe阳性患者HBV DNA载量分别为6.1lg拷贝/毫升和3.8lg拷贝/毫升(P<0.01);在送检的12例肝组织,HBV DNA均阳性,HBVcccDNA阳性8例。结论肝硬化和肝癌患者抗-HBe阳性比例较高,肝组织HBVcccDNA仍可被检出。     

Intracellular levels of hepatitis B virus DNA and pregenomic RNA in peripheral blood mononuclear cells of chronically infected patients

Summary.  It remains uncertain whether hepatitis B virus (HBV) covalently closed circular DNA (cccDNA) and pregenomic RNA (pgRNA) can be detected in the serum or peripheral blood mononuclear cells (PBMC) of patients with chronic hepatitis B (CHB) infection. We examined HBV cccDNA and pgRNA in the serum and PBMC, and investigated the effect of lamivudine therapy on the viral loads in the PBMC of CHB patients. Paired serum and PBMC samples from 50 treatment-naïve CHB patients [25 hepatitis B e antigen (HBeAg) positive and 25 HBeAg negative] were quantified for total HBV DNA, cccDNA and pgRNA by real time polymerase chain reaction. HBV cccDNA and pgRNA were below the lower detection limit in all serum samples, and in 84% of PBMC. HBV DNA ( r  = 0.889, P  <   0.001) and pgRNA ( r  = 0.696, P  <   0.001) in PBMC correlated with the HBV DNA in serum. In the longitudinal study, 30 patients treated with lamivudine therapy for a median duration of 34 weeks (range 12–48 weeks) were examined. The median HBV DNA reduction in PBMC before and after treatment was 1.318 (range −0.471 to 3.846) log units, which was significantly lower than serum HBV DNA reduction [3.371 (range −0.883 to 9.454) log units, P  <   0.05]. HBV cccDNA and pgRNA were undetectable in the serum of CHB patients. HBV viral loads in PBMC correlated with serum HBV DNA. Lamivudine therapy had less effect on the HBV viral loads in PBMC compared with the serum viral loads.     

Peginterferon alpha-2b plus adefovir induce strong cccDNA decline and HBsAg reduction in patients with chronic hepatitis B

Hepatitis B virus (HBV) covalently closed circular DNA (cccDNA) is responsible for persistent infection of hepatocytes. The aim of this study was to determine changes in intrahepatic cccDNA in patients with chronic hepatitis B (CH-B) during 48 weeks of antiviral therapy and its correlation to virological, biochemical, and histological parameters. Twenty-six HBsAg-positive CH-B patients received combination treatment with pegylated interferon alpha-2b (peg-IFN) and adefovir dipivoxil (ADV) for 48 weeks. Paired liver biopsies from before and at the end of treatment were analyzed for intrahepatic HBV-DNA. Median serum HBV-DNA had decreased by -4.9 log10 copies/mL at the end of treatment and was undetectable in 13 individuals (54%). Median intrahepatic total HBV-DNA and cccDNA had decreased by -2.2 and -2.4 log10, respectively. Changes in intracellular HBV-DNA positively correlated with HBsAg serum reduction and were accompanied by a high number of serological responders. Eight of 15 HBeAg-positive patients lost HBeAg, and five developed anti-HBe antibodies during treatment. These eight patients exhibited lower cccDNA levels before and at the end of therapy than did patients without HBeAg loss. Four patients developed anti-HBs antibodies. ALT normalized in 11 patients. The number of HBs-antigen- and HBc-antigen-positive hepatocytes was significantly lower after treatment, suggesting the involvement of cytolytic mechanisms. In conclusion, combination therapy with peg-IFN and ADV led to marked decreases in serum HBV-DNA and intrahepatic cccDNA, which was significantly correlated with reduced HBsAg.     

Intrahepatic hepatitis B virus covalently closed circular DNA can be a predictor of sustained response to therapy

BACKGROUND & AIMS: This study aimed to determine whether intrahepatic hepatitis B virus (HBV) covalently closed circular (ccc) DNA and total HBV DNA levels at the end of therapy would predict sustained response to therapy. METHODS: Hepatitis B e antigen (HBeAg)-positive chronic hepatitis B patients receiving either lamivudine monotherapy or combination of peginterferon and lamivudine had liver biopsy at the end of 1 year therapy and were followed for 52 more weeks after cessation of therapy. Serum HBV DNA, intrahepatic HBV ccc DNA, and total HBV DNA levels were determined. RESULTS: Forty-seven patients, including 34 males and 13 females, were studied. Twenty-seven patients received combination therapy, and 20 patients received lamivudine monotherapy. Twenty-nine patients had end-of-treatment virologic response, and 15 patients had sustained response 52 weeks after therapy. At the end of treatment, log serum HBV DNA levels correlated well with log intrahepatic HBV cccDNA and log intrahepatic total HBV DNA levels. Log intrahepatic cccDNA and log intrahepatic total DNA levels were significantly lower among patients with sustained virologic response. The adjusted odds ratio for log cccDNA was 5.3 (95% CI: 1.5-18.2, P = .009) and, for log intrahepatic HBV DNA, was 4.4 (95% CI: 1.3-14.7, P = .015) to predict sustained virologic response. Using log cccDNA at -0.80 copies/genome equivalent as cutoff, the sensitivity, specificity, and positive and negative predictive values and accuracy of predicting sustained virologic response were 73%, 78%, 56%, 86%, and 77% respectively. CONCLUSIONS: Intrahepatic HBV cccDNA and intrahepatic total HBV DNA levels at the end of therapy are superior to serum HBV DNA as surrogates of sustained virologic response.     

Lamivudine therapy for children with chronic hepatitis B

AIM: To assess the effectiveness and side-effects of lamivudine therapy for children with chronic hepatitis B (CHB) who fail to respond to or have contraindications to interferon-α(IFN-α) therapy. METHODS: Fifty-nine children with CHB were treated with 100 mg lamivudine tablets given orally once daily for 12 mo. Alanine aminotransferase (ALT) activity was evaluated monthly during the therapy and every 3 months after its discontinuation. HBe antigen, anti-HBe antibodies, HBV DNA level in serum were evaluated at baseline and every six months during and after the lamivudine therapy. Sustained viral response (SVR) to lamivudine therapy was defined as permanent (not shorter than 6 mo after the end of the therapy), namely ALT activity normalization, seroconversion of HBeAg to anti-HBe antibodies, and undetectable viral HBV-DNA in serum (lower than 200 copies per mL). The analysis of the side-effects of the lamivudine treatment was based upon interviews with the patients and their parents using a questionnaire concerning subjective and objective symptoms, clinical examinations, and laboratory tests performed during clinical visits monthly during the therapy, and every 3 mo after the therapy. RESULTS: ALT normalisation occurred in 47 (79.7%) patients between the first and 11th mo of treatment (mean 4.4±2.95 mo, median 4.0 mo), and in 18 (30.5%) of them after 2 mo of the therapy. There was no correlation between the time of ALT normalization and the children's age, the age of HBV infection, the duration of HBV infection, inflammation activity score (grading), staging, ALT activity before treatment, serum HBV DNA level, and lamivudnie dose per kg of body weight. HBeAg/anti HBe seroconversion was achieved in 27.1% of cases. The higher rate of seroconversion was connected with lower serum HBV DNA level and longer duration of HBV infection. There was no connection between HBeAg/ anti HBeAb seroconversion and the children's age, age of HBV infection, grading, staging, ALT activity before treatment, and lamivudnie dose per kg of body weight. No complaints or clinical symptoms were observed during lamivudine therapy. Impairment of renal function or myelotoxic effect was noted in none of the patients. CONCLUSION: One year lamivudine therapy for children with chronic hepatitis B is effective and well tolerated. Seroconversion of HBeAg/HBeAb and SVR are connected with lower pre-treatment serum HBV DNA level.     

乙型肝炎病毒cccDNA定量与乙型肝炎临床及病理关系

目的探讨慢性乙型肝炎（CHB）肝组织HBVcccDNA定量与乙型肝炎的关系。方法分别采用荧光定量PCR、酶联免疫吸附分析法（ELISA）检测48例CHB肝组织HBVcccDNA定量、肝组织和血清HB VDNA定量、乙型肝炎病毒标志物。同时用链霉菌抗生素蛋白-过氧化物酶连接法（SP）检测肝细胞中HBcAg表达。分析肝组织HBVcccDNA与组织和血清HBV DNA、HBeAg、肝细胞内HBcAg水平及肝脏炎症活动度的关系.结果1．肝组织HBVcccDNA定量与组织和血清HBV DNA定量呈正相关（r=0．837，P〈0．001；r=0．627，P〈0．005）；2．肝组织HBV cccDNA定量与肝细胞内HBcAg半定量呈正相关（r=0．618，P〈0．005）；3．肝组织HBV cccDNA定量与肝脏炎症活动度尤明显相关（P〉0．05）：4．HBeAg阳性较抗-HBe阳性患者肝组织HBV cccDNA定量、肝组织和血清HBV DNA定量高（P〈0．05）。结论荧光定量PCR法检测肝组织HBV cccDNA定量是评价HBV复制最直接可靠的指标，在CHB的诊断和抗病毒治疗中有重要意义。但与肝组织炎症无明显相关。     

Comparison between quantitative hepatitis B surface antigen,hepatitis B e‐antigen and hepatitis B virus DNA levels for predicting virological response to pegylated interferon‐α‐2b therapy in hepatitis B e‐antigen‐positive chronic hepatitis B

Aim: The aim of this study was to compare the clinical applicability of quantitative serum hepatitis B surface antigen (HBsAg), hepatitis B e‐antigen (HBeAg) and hepatitis B virus (HBV) DNA for predicting virological response (VR) to pegylated interferon (PEG‐IFN) therapy. Methods: Thirty HBeAg‐positive chronic hepatitis B patients who received PEG‐IFN‐α‐2b for 48 weeks were enrolled. Quantitative HBsAg, HBeAg and HBV DNA were measured before, during and after the therapy. Paired liver biopsies were performed before and after treatment for covalently closed circular (ccc)DNA and intrahepatic HBV DNA analysis. Results: VR at 48 weeks post‐treatment, defined as HBeAg seroconversion and HBV DNA less than 10 000 copies/mL was achieved in 10 (33.3%) patients. Responders had significantly lower baseline HBsAg, HBeAg, cccDNA and intrahepatic HBV DNA levels than non‐responders. Baseline and reduced levels of log₁₀ HBsAg and log₁₀ HBeAg correlated well with those of log₁₀ cccDNA and log₁₀ total intrahepatic HBV DNA. Responders showed consistent decrease in serum HBsAg, HBeAg and HBV DNA levels during therapy. HBeAg level of 2.0 log₁₀ sample to cut‐off ratio at week 24 on therapy provided the best prediction of sustained virological response, with sensitivity and negative predictive values of 85% and 92%, respectively. One patient (3.3%) who cleared HBsAg at follow up exhibited a more rapid decline in serum HBsAg during therapy than those who developed VR without HBsAg clearance. Conclusion: Quantitative measurement of serum HBeAg during therapy may be superior to serum HBsAg and HBV DNA as a prediction of HBeAg seroconversion. Kinetics of HBsAg levels on therapy may help predict HBsAg clearance after treatment.