阻断乙型肝炎e抗原阳性母亲宫内传播的研究

目的 探讨不同剂量乙肝免疫球蛋白对乙肝表面抗原和乙肝e抗原双阳性孕妇乙型肝炎病毒宫内传播的影响.方法 乙肝表面抗原和乙肝e抗原双阳性孕妇为病例组,乙肝表面抗原阳性但乙肝e抗原阴性的孕妇为对照组,对病例组和对照组均采用3种不同的干预方式:乙肝免疫球蛋白 400U、乙肝免疫球蛋白200U、no-乙肝免疫球蛋白进行干预,观察各组新生儿乙肝表面抗原、乙肝表面抗体、乙肝e抗原、乙肝e抗体、乙肝核心抗体检测情况.采用定性酶联免疫法检测.结果 观察病例乙肝免疫球蛋白400U组新生儿21例,乙肝表面抗原和乙肝e抗原检测均为阴性,未发现宫内感染者,追踪并复查仍未发现乙肝病毒抗原阳性者;观察病例乙肝免疫球蛋白 200U组新生儿22例,乙肝表面抗原均为阴性,1例乙肝e抗原阳性,宫内感染率为4.5%;病例no-乙肝免疫球蛋白组观察新生儿20例,检测乙肝表面抗原均为阴性,5例乙肝e抗原阳性,宫内感染率为25.0%;乙肝免疫球蛋白200U组宫内感染率低于no-乙肝免疫球蛋白组,但无显著性差异(χ2=3.58,P=0.058).结论 孕晚期注射乙肝免疫球蛋白400 U和200 U对阻断乙型肝炎e抗原阳性母亲宫内传播均有效果,乙肝免疫球蛋白400 U干预方案效果更好.     

鸡骨草提取物对体外乙型肝炎病毒的抑制作用

目的 观察鸡骨草提取物对体外HBsAg和HBeAg的抑制作用. 方法 将鸡骨草醇提取液与等容量、滴度为1：64的HBsAg和HBeAg阳性血清混合,使混合后鸡骨草的浓度分别为96,48,24,12和6 g•L^-1,用磷酸缓冲液与1：64的HBsAg和HBeAg阳性血清混合作空白对照. 每组6管, 37 ℃水浴8和16 h后,采用ELISA法测定HBsAg和HBeAg的吸光度（A值）,观察其对HBsAg或HBeAg的抑制作用. 结果 鸡骨草醇提取液对血清中HBsAg和HBeAg均具有明显的抑制作用,其中鸡骨草提取物在浓度为96,48 g•L^-1时对HBsAg、HBeAg抑制作用最为明显（P＜0.05或P＜0.01）. 结论 鸡骨草提取物在体外有较明显的抗HBsAg和HBeAg作用.     

Hepatitis B virus e antigen (HBeAg) may have a negative effect on dendritic cell generation

Hepatitis B virus (HBV) continues to be a serious worldwide health problem despite the use of protective HBV vaccines and therapeutic regimens against chronic HBV infection. Chronic HBV patients cannot induce sufficient immune responses against the virus. HBV and its antigens are believed to suppress immune responses during chronic infection. Hence, studying the role of HBV in immune suppression is very important for the development of alternative therapeutic strategies for HBV infections.     

乙型肝炎病毒DNA与标本物含量及肝功能生化指标间关系探讨

目的探讨血清中乙型肝炎病毒(HBV)DNA含量与HBV标志物[乙型肝炎表面抗原(HBsAg)、乙型肝炎e抗原(HBeAg)]及肝功能指标间的相关性。方法抽取南平市人民医院肝内科住院的200例HBV携带患者血清样本。采用荧光定量聚合酶链反应(PCR)检测血清样本中HBV DNA含量,采用酶联免疫吸附试验(ELISA)测定血清样本中HBV标志物含量,同时检测肝功能。3项检测指标使用同一患者标本,分析3项指标之间的相关性。结果经相关性分析,HBV DNA与肝功能指标的r在±0.2以内,差异无统计学意义(P>0.05);HBsAg、HBeAg含量与肝功能指标的r在±0.2以内,差异无统计学意义(P>0.05);HBsAg、HBeAg含量与HBVDNA含量的r分别为0.457、0.568,差异有统计学意义(P<0.01)。结论 HBsAg、HBeAg含量只反映HBV在血清中的复制情况,不能以HBsAg、HBeAg含量及病毒载量的大小作为判断肝功能受损程度的指标;要综合分析HBV DNA含量与HBV标志物定量检测结果及肝功能,这样才能更有效地对疾病进行评估。     

乙型肝炎病毒e抗原在肝细胞内作用蛋白AK026018亚细胞定位的研究

目的：确定肝细胞内乙型肝炎病毒e抗原(HBeAg)作用蛋白AK026018的亚细胞定位，初步研究该蛋白的生物学功能。方法：应用逆转录聚合酶链反应(RT-PCR)技术，从HepG2细胞中扩增编码AK026018蛋白的全基因，构建真核表达载体pEGFP-AK，转染HepG2、293T细胞系，在激光共聚焦荧光显微镜下与转染空载体pEGFP-N1的细胞比较观察。结果：经限制性内切酶分析和DNA序列测定鉴定构建的重组表达载体正确。通过激光共聚焦荧光显微镜观察，转染了重组栽体pEGFP-AK的细胞绿色荧光信号较集中分布于胞浆中。而空载体pEGFP-N1转染的细胞绿色荧光信号在整个细胞中均匀分布。结论：成功分离了AK026018全基因序列并构建了真核表达载体，该基因表达产物亚细胞定位于细胞浆中。     

Relationship of clinical and virological factors with hepatitis activity in hepatitis B e antigen-negative chronic hepatitis B virus-infected patients

To investigate the factors associated with active disease among hepatitis B surface antigen (HBsAg) positive/hepatitis B e antigen (HBeAg)-negative chronic hepatitis B virus (HBV) infection we studied chronic HBV infected patients who had undetectable HBeAg at the first visit. HBV DNA was determined by the cross-linking assay (NAXCOR) and polymerase chain reaction (PCR). Mutations in the core promoter and precore regions and viral genotypes were studied. Clinical outcome of these patients were followed and categorized as: (i) relapse (ALT > 200 IU/L or three times the previous levels); (ii) active hepatitis (elevated ALT < 200 IU/L with concomitant detectable HBV DNA); or (iii) remission. A total of eighty-five patients were followed up for 5.5 ± 1.0 years. At first visit, 31 (36.5%) patients had elevated ALT levels, 12 (14.1%) had measurable HBV DNA by the cross-linking assay and 26 (30.6%) by PCR. Sixteen (18.8%) patients had hepatitis relapse, 13 (15.3%) had active hepatitis, and 56 (65.9%) remained in remission. Core promoter and precore stop codon mutants were found in 27 and 12 patients, respectively. Eleven and 20 had genotype B and C HBV, respectively. Initial elevated ALT and detectable HBV DNA were associated with active liver disease. Patient demographics, viral mutants or genotypes failed to predict disease activity. Hence, serum ALT and HBV DNA levels offer the best prediction of natural course of HBeAg-negative chronic HBV infection.     

Lamivudine vs lamivudine and interferon combination treatment of HBeAg(-) chronic hepatitis B

To determine whether combination treatment of HBeAg(-) chronic hepatitis B is beneficial we studied 78 patients with HBeAg(-), HBV DNA-positive chronic hepatitis B who were randomized to lamivudine, 100 mg, qd, for 12 months or lamivudine-interferon (9 MU, t.i.w.) in combination. In the combination arm, 2 months of lamivudine treatment preceded 10 months of combination treatment. Biochemical, virologic and histologic responses were assessed at the end of treatment, after six and a median 27 months of drug-free follow-up (short- and long-term follow-up, respectively). Virologic response was defined as undetectable HBV DNA with a hybridization assay and biochemical response as normal alanine aminotransferase (ALT). Change in HBV DNA was also assessed by real-time polymerase chain reaction (PCR). Presence of YMDD mutants at the end of treatment was investigated with a line probe assay. Both treatment regimes led to a median 2 log decline in HBV DNA levels. Virologic end of treatment responses were 90 and 92% with mono- and combination treatment, respectively. Corresponding virologic responses at short- and long-term follow-up were 59 and 54%, and 27 and 25%, respectively. Patients having a baseline HBV DNA value > or =200 pg/mL were more likely to relapse within 6 months off therapy than those patients with a baseline HBV DNA level <200 pg/mL (P = 0.041). YMDD mutants were observed in 53% of patients receiving lamivudine compared with 24% of patients receiving the combination regime (P = 0.017). In conclusion, efficacy of combination treatment is similar to lamivudine monotherapy. However, combination treatment decreases the development of YMDD mutant strains compared with lamivudine monotherapy.     

Hepatitis B virus-DNA in the serum of patients followed-up longitudinally with acute and chronic hepatitis B

Sera from 79 patients with acute self-limiting hepatitis, 17 patients with acute hepatitis B evolving into chronic HBsAg carriership, and 43 chronic HBsAg carriers without a history of acute hepatitis were analyzed for presence of hepatitis B virus (HBV)-DNA by a molecular hybridization technique. In acute self-limiting hepatitis, HBV-DNA was cleared within a few weeks after the onset of clinical symptoms. The longest period of DNA positivity observed in this group was 42 days. In 29 of 52 patients HBV-DNA was cleared before HBeAg disappeared. Among 17 patients who became chronic HBsAg carriers, HBV-DNA was present for more than 6 months in all but one. Most of the HBsAg carriers eventually cleared HBV-DNA. The DNA clearance frequently preceeded the conversion of HBeAg to anti-HBe. Thus, in many patients there was a transitional period with HBeAg but without HBV-DNA. HBV-DNA was found to be a better index of impending chronicity than HBeAg since persistence of HBeAg for more than 42 days was noted in 10% of the patients who nevertheless cleared HBsAg within 6 months. By that time all those patients had turned negative for HBV-DNA. On the other hand, in 16 of the 17 patients who became chronic carriers of HBsAg, HBV-DNA as well as HBeAg persisted for more than 6 months. The present results also suggest that infectivity in acute hepatitis B is a feature mainly of the presymptomatic and early symptomatic period.     

前S1抗原、HBV—DNA和乙肝病毒血清标志物联合检测的临床意义

[目的]了解乙肝两对半（主要是HBeAg）、乙肝病毒DNA（HBV—DNA）与乙肝前S1抗原（Pre—S1）检测三者之间的关系以及它们的临床应用价值。[方法]采用酶联免疫吸附实验（ELISA）方法对273例乙肝患者血清进行乙肝两对半、Pre—S1抗原的测定，用荧光PCR技术对HBV—DNA进行测定，并分析3者之间的关系。[结果]HbeAg阳性组Pre—S1抗原、HBV—DNA的阳性率分别为85．09％、91．23％。说明Pre—S1抗原与HBV—DNA一样可较好的反映乙肝病毒在体内的活动及复制情况；HbeAg阴性组中Pre—S1抗原与HBVDNA的阳性率分别为37．11％、37．11％，说明在HbeAg阴性组患者仍有病毒复制，不能完全以HbeAg来判断病毒复制与否；与HBV—DNA的检测结果相比较，Pre—S1抗原检测的检出率为88．95％，特异性为87．32％，总符合率为88．28％，HBVDNA与Pre—S1抗原的阳性检出率没有差别（P〉0．05）。[结论]Pre—S1抗原能较好的反映HBV病毒感染及复制情况，三者联合检测互相补充，更有助于临床疾病的诊治。     

137例慢性乙型肝炎患者肝功能与乙型肝炎病毒DNA及乙型肝炎e抗原相关性分析

目的探讨慢性乙型肝炎患者治疗过程中血清丙氨酸氨基转移酶（ALT）、天门冬氨酸氨基转移酶（AST）水平变化与乙型肝炎e抗原（HBeAg）和慢性乙型肝炎病毒（HBV）DNA的相关性。方法 137例血清ALT、AST水平正常的慢性乙型肝炎患者中HBeAg和HBV DNA同时阳性46例,同时阴性48例,单项HBVDNA阳性35例,单项HBeAg阳性8例。3～6个月后分别对各组患者复发前后血清HBV DNA、ALT、AST水平及ALT、AST异常水平的例数进行比较。结果复发前HBeAg和HBV DNA同时阳性组、HBV DNA阳性组、HBeAg阳性组的ALT、AST水平分别为（22.93±18.45）U/L和（24.12±15.67）U/L、（21.74±16.48）U/L和（17.87±18.05）U/L、（20.55±17.33）U/L和（19.72±16.91）U/L,复发后分别为（529.65±252.42）U/L和（434.18±218.53）U/L、（419.06±204.26）U/L和（476.63±189.43）U/L、（326.48±187.95）U/L和（347.27±198.15）U/L,差异均有统计学意义（P〈0.05）。HBeAg和HBV DNA同时阴性组ALT、AST水平复发前后差异无统计学意义（P〉0.05）。结论血清HBeAg和HBV DNA的检测对慢性乙型肝炎患者诊断、治疗、预后判断有重要的作用,提示慢性乙型肝炎治疗HBeAg的阴转和HBV DNA不能检出能维持较低的复发率。