拉米夫定相关性HBV变异对乙型肝炎预后的影响

目的 进一步探讨拉米夫定相关性ＨＢＶ变异对患者临床经过的影响。方法 将接受拉米夫定治疗１００ｍｇ／ｄ的８２例患者在第１０４周按其发生变异程序分为完全变异、部分变异和无变异３组,分别进行肝功能和血清学指标比较。结果 在第１０４周,Ｆ和Ｍ组无１例ＨＢｃＡｇ阴国专,而Ｎ组有１８例ＨＢｅＡｇ阴转 ,１１例ｃＡｇ／抗－ＨＢｅ血清转换;拉米夫定相关性ＨＢＶ变异发生后ＡＬＴ可增高,完全变异组Ｈ ＢＶＤＮＡ水平明     

拉米夫定加IFN-α联合治疗慢性乙型肝炎疗效分析

为提高慢性乙型肝炎的疗效 ,采用拉米夫定加ＩＦＮ α联合治疗慢性乙型肝炎和单用拉米夫定作同期对照。对临床确诊为慢性乙型肝炎血清ＨＢｅＡｇ、ＨＢＶＤＮＡ同时阳性 ,ＡＬＴ异常者 4 0例 ,随机分为两组 :联合治疗组拉米夫定 10 0～ 150ｍｇ/ｄ ,疗程 4 8周 ,ＩＦＮ α30 0万Ｕ ,肌注每周 3次 ,疗程 2 4周 ;单用拉米夫定组 (对照组 ) 10 0～ 150ｍｇ/ｄ ,疗程 4 8周。动态观察血清ＨＢＶ复制指标和肝功能的变化。结果表明 ,用药第 2 4周内两组病例血清ＨＢＶＤＮＡ均阴转 ,ＨＢｅＡｇ阴转率联合治疗组为 60 % ( 12 / 2 0 ) ,对照组为 15%…     

拉米夫定治疗2年时乙肝病毒的YMDD变异情况

观察核苷类似物拉米夫定治疗慢性乙肝病人２年时ＹＭＤＤ变异情况及其与血清ＨＢＶＤＮＡ，ＡＬＴ水平等指标的关系。第一阶段（１－１２周）为随机、双盲、安慰剂对照研究，７２名ＨＢｓＡｇ．ＨＢｅＡｇ阳性至少６个月，ＨＢＶ－ＤＮＡ阳性的慢性乙肝患者分别口服拉米夫定１００ｍｇ／ｄ（ｎ＝５４）或安慰剂（ｎ＝１８）；第二阶段（１３－１０４周）所有患者均服用拉米夫定 １００ｇ／ｄ。５２周和 １０４周检查病毒的 ＹＭＤＤ变异．其总变异率分别为 １３．７％（８／５８）和 ３９．７％（２３／５８）。１０４周时变异组血清 ＨＢＶＤＮＡ，ＡＬＴ水平高于无变异组（３９４．９±７２７．９比 １６．３±５０．９，Ｐ＝０．００４８；６２．７±５７．９比２６ ４±２７．５，Ｐ＝０．００３），ＨＢＶＤＮＡ阴转率低于未变异组（１７．４％比４８．６％，Ｐ＜０．０５）；服用拉米夫定的慢性乙肝患者的ＹＭＤＤ发生率与服药时间长短有关，血清ＨＢＶＤＮＡ及ＡＬＴ水平与ＹＭＤＤ相关。     

乙型肝炎病毒核心区DNA疫苗接种后H-2d小鼠的特异性免疫应答

目的观察乙型肝炎病毒（ＨＢＶ）核心区ＤＮＡ疫苗接种后ＢＡＬＢ／ｃ（－２ｄ）小鼠的特异性免疫应答。方法构建ＨＢＶ核心区ＤＮＡ疫苗（ＰＪＷ４３０３／ＨＢｃ）;用基因枪法和肌肉注射祛将该ＤＮＡ疫苗接种ＢＡＬＢ／ｃ小鼠;ＥＬＩＳＡ法检测小鼠血清抗－ＨＢＣ（ＩｇＧ）及ＩｇＧ亚类（ＩｇＧ１,ＩｇＧ２ａ）;５１铬释放法检测小鼠脾细胞ＨＢｃＡｇ特异性ＣＴＬ活性。结果该ＤＮＡ疫苗在体外转染细胞中可良好表达ＨＢｃＡｇ。血清抗－ＨＢｃ终点滴度在ＤＮＡ疫苗基因枪组和肌肉注射接种组小鼠分别为１：３２８０５０和１：１０９３５０．两组小鼠的抗－ＨＢｃＩｇＧ亚类均以ＩｇＧ２ａ略占优势。两组小鼠的ＨＢＣＡｔｈｅ异性ＣＴＬ杀伤活性分别达到５１．１％和５５．２％。结论ＨＢＶ核心区ＤＮＡ疫苗在小鼠实验中具有良好的细胞免疫和体液免疫原性。     

基因疫苗诱导小鼠抗HBV皮下移植瘤免疫研究

目的　观察 ＨＢＶ ＤＮＡ 疫苗（ｐＣＲ３－１Ｓ） 诱导Ｂａｌｂ／ ｃ 小鼠（ Ｈ２ｄ） 的特异性细胞免疫应答及其对稳定表达ＨＢｓＡｇ 的小鼠肥大细胞瘤Ｐ８１５ 细胞（Ｐ８１５ＨＢＶＳ） （ Ｈ２ｄ） 成瘤性的影响．方法　肌肉注射ＤＮＡ 疫苗,背部皮下接种Ｐ８１５ＨＢＶＳ 细胞,观察成瘤情况,４ ｈ ５１Ｃｒ 释放法检测小鼠脾细胞ＣＴＬ 活性．结果　接种 ＤＮＡ 疫苗后小鼠成瘤率为１２－５ ％ , 对照组为１００ ％ ． 小鼠平均存活期大于３８－２ ｄ ,对照组为２８－４ ｄ ,４０ ｄ 后小鼠存活率为８７－５ ％ ,对照组为０ ％ ． ＣＴＬ 细胞杀伤活性明显增加,ｐＣＲ３－１Ｓ 组５１ ％ ,对照组为２１ ％ （ Ｐ＜ ０－００１） ．结论　ＤＮＡ 疫苗可以诱导细胞免疫应答,对体内ＨＢＶ 感染具有预防及治疗作用．     

拉米夫定治疗慢性乙型肝炎2年临床试验的总结

目的 评估拉米夫定长期治疗慢性乙型肝炎的疗效和安全性,以及治疗过程中产生病毒变异的临床影响。方法 系多中心的双盲,随机、安慰剂对照的临床试验。４２９例ＨＢｓＡｇ和ＨＢｅＡｇ阳性的慢性乙型肝炎,按３：１随机分成拉米夫定治疗组和对照组,分别服用拉米夫定１００ｍｇ／ｄ和安慰剂共１２周,此后所有病人均服用拉米夫定,共观察２年,结果 治疗１２周,拉米夫定组血清ＨＢＶ ＤＮＡ累计阴转率（〈１．６ｐｇ／ｍｌ）为     

HBV转基因小鼠暴露ATB-1后肝脏ATB-1-DNA加成物水平的测定

目的探讨ＨＢＶ与ＡＴＢ１协同致肝癌的机理．方法用ＲＩＡ法测定ＨＢＶ转基因小鼠与正常小鼠暴露ＡＴＢ１后０，０５，１，２，４，８，２４ｈ７个不同时相肝脏ＡＴＢ１ＤＮＡ加成物的含量变化．结果暴露ＡＴＢ１后，ＨＢＶ转基因小鼠肝脏ＡＴＢ１ＤＮＡ加成物含量在各时相均高于正常小鼠，尤以１ｈ高峰时相值（５５９２ｐｍｏｌ／ｍｇ±４１５ｐｍｏｌ／ｍｇ比４１３６ｐｍｏｌ／ｍｇ±２８２ｐｍｏｌ／ｍｇＤＮＡ，Ｐ＜００１）及２４ｈ时相值（２４８７±２０３比９８９±８５，Ｐ＜００１）最显著．２４ｈ后，转基因小鼠肝脏ＡＴＢ１ＤＮＡ加成物仍维持高水平，但正常小鼠已基本恢复至暴露前水平．结论ＨＢＶ转基因小鼠暴露ＡＴＢ１后肝脏ＡＴＢ１ＤＮＡ加成物含量增加可能为ＨＢＶ与ＡＴＢ１协同致肝癌的直接原因．     

中老年乙型肝炎病毒DNA的动态变化

应用聚合酶链反应（ＰＣＲ）和酶联免疫吸附试验（ＥＬＩＳＡ）动态检测了中老年乙型肝炎病人不同时期的血清标志。结果表明，ＨＢｅＡｇ阳性的血清其ＰＣＲ检测ＨＢＶＤＮＡ均阳性，两者呈平行关系，１～４ｗ后有２０％病人血清ＨＢＶＤＮＡ转阴，同时伴有部分病人ＨＢｅＡｇ消失或ＨＢｅＡｂ出现，其他ＨＢＶ血清学标志物阳性的病人血清ＨＢＶＤＮＡ阳性率明显低于ＨＢｅＡｇ阳性血清，且ＨＢＶＤＮＡ暂转阴率高于ＨＢｅＡｇ阳性血清，ＨＢＶＤＮＡ转阴的时间也短（１～４ｗ）。动态检测ＨＢＶＤＮＡ可以动态观察ＨＢＶＤＡＮ的复制，对指导临床治疗具有很大的意义。     

肺结核合并HBV感染患者血清HBV－DNA的检测及其意义

采用分子斑点杂交和酶连免疫吸附试验（ＥＬＩＳＡ）方法检测２６８例肺结核患者血清乙型肝炎病毒ＤＮＡ（ＨＢＶ－ＤＮＡ）及其它乙肝病毒标志物（ＨＢＶ－Ｍ），观察肺结核患者抗结核化疗中肝功能变化与血清ＨＢＶ－ＤＮＡ的关系。结果，２６８例肺结核患者血清ＨＢＶ－Ｍ阳性率为１９．０％，ＨＢＶ－ＤＮＡ阳性２０例（７．５％）；化疗中，肺结核合并ＨＢＶ－ＤＮＡ阳性组、肺结核合并ＨＢＶ－ＤＮＡ阴性其它ＨＢＶ－Ｍ阳性组、单纯肺结核组，肝功能受损率分别为９５％、１０％及５．１％。肺结核合并ＨＢＶ感染组、单纯ＨＢＶ－Ｍ携带者组、单纯肺结核组，肝功能受损率分别为４９％、１０％及５．１％。肺结核合并ＨＢＶ－ＤＮＡ阳性组死亡４例（１５％），且死亡与肝病直接相关（肝衰竭），３．５年后发生肝硬化２例。结果提示，检测肺结核患者血清ＨＢＶ－ＤＮＡ有重要意义。抗乙型肝炎病毒治疗对改善存在ＨＢＶ复制的肺结核患者的预后可能有重要作用，化疗前常规检查并有ＨＢＶ感染的肺结核患者血清ＨＢＶ－ＤＮＡ，可指导用药。     

拉米规定治疗慢性乙型肝炎2年临床疗效及病毒的变异

目的 研究拉米夫定治疗慢性乙型肝炎病人２年的临床疗效。方法 ８８例病人按３：１比例随机分成治疗组和安慰剂组,每日服拉米夫定１００ｍｇ和安慰剂各一片,１２周以后全部服拉米夫定每日１００ｍｇ,持续到１０４周。定期检测ＡＬＴ、ＨＢＶＤＮＡ、ＨＢｅＡｇ和抗－ＨＢｅ。用ＰＣＲ后直接测序方法检测ＹＭＤＤ变异。结果 ７５例病人完成２年试验。１２周时,治疗组和安慰剂组ＨＢＶＤＮＡ阴转分别为４７例（８０％）和４例     

Inhibitory effect of oxymatrine on serum hepatitis B virus DNA in HBV transgenic mice

AIM: To study the inhibitory effect of oxymatrine on serum hepatitis B virus (HBV) DNA in HBV transgenic mice. METHODS: HBV transgenic mice model was established by microinjection, and identified by HBV DNA integration and replication. Transgenic mice with replicating HBV were divided into 3 groups, and injected with normal saline (group A, n=9), 50 mg/kg (group B, n=8) and 100 mg/kg (group C, n=9) oxymatrine intraperitoneally once a day for 30 d, respectively. Quantitation of serum HBV DNA in HBV transgenic mice was performed by competitive polymerase chain reaction (PCR) in combination with DNA hybridization quantitative detection technique before and after treatment. RESULTS: Compared with pre-treatment, the serum HBV DNA in group A (F=1.04, P=0.9612) and group B (F=1.13, P=0.8739) had no changes after treatment. However, in group C serum HBV DNA was significantly decreased (F=13.97, P=0.0012). The serum HBV DNA after treatment was lower in group C than in groups B and A (F=8.65, P=0.0068; F=12.35, P=0.0018; respectively). The serum HBV DNA after treatment was lower in group B than in group A, but there was no statistical significance (F=1.43, P=0.652). CONCLUSION: Oxymatrine has inhibitory effects on serum HBV DNA in HBV transgenic mice.     

乙型肝炎病毒转基因小鼠用于研究治疗乙型肝炎药物初探

目的：建立乙型肝炎病毒（HBV）转基因小鼠模型,探讨该模型能否用于抗HBV药物的验证,筛选,方法：原核显微注射法制备HBV转基因小鼠,用巢式PCR,Southern杂交,免疫组化学,ELISA等检测HBV基因的整合与表达,筛选60只血清HBVDNA,HBsAg阳性的小鼠,随机分为6组,3组为实验组,分别给予拉米夫定灌胃,胸腺肽腹腔注射,HBVS基因DNA疫苗,多点肌肉注射,另3组分别为生理盐水对照组,结果：显微注射产生的HBV转基因小鼠中有HBV的复制和表达,拉米夫定,胸腺肽,DNA疫苗用于转基因小鼠后,均能使血清HBVDNA阴转,以拉米夫定阴转率最高,并可看出作用开始,持续时间及反跳情况,结论：拉米夫定,胸腺肽,DNA疫苗可抑制转基因小鼠HBV的复制,HBV转基因小鼠有可能成为抗HBV药物验证,筛选的有力工具。     

Twenty-four-week clevudine therapy showed potent and sustained antiviral activity in HBeAg-positive chronic hepatitis B

Clevudine is a pyrimidine analogue with potent and sustained antiviral activity against HBV. The present study evaluated the safety and efficacy of 30 mg clevudine once daily for 24 weeks and assessed the durable antiviral response for 24 weeks after cessation of dosing. A total of 243 hepatitis B e antigen (HBeAg)-positive chronic hepatitis B patients were randomized (3:1) to receive clevudine 30 mg once daily (n=182) or placebo (n=61) for 24 weeks. Patients were followed for a further 24 weeks off therapy. Median serum HBV DNA reductions from baseline at week 24 were 5.10 and 0.27 log10 copies/mL in the clevudine and placebo groups, respectively (P<0.0001). Viral suppression in the clevudine group was sustained off therapy, with 3.73 log10 reduction at week 34 and 2.02 log10 reduction at week 48. At week 24, 59.0% of patients in the clevudine group had undetectable serum HBV DNA levels by Amplicor PCR assay (less than 300 copies/mL). The proportion of patients who achieved normalization of alanine aminotransferase (ALT) levels was 68.2% in the clevudine group and 17.5% in the placebo group at week 24 (P<0.0001). ALT normalization in the clevudine group was well maintained during post-treatment follow-up period. The incidence of adverse events (AEs) was similar between the clevudine group and the placebo group. No resistance to clevudine was detected with 24 weeks of administration of drug. CONCLUSION: A 24-week clevudine therapy was well tolerated and showed potent and sustained antiviral effect without evidence of viral resistance during treatment period in HBeAg-positive chronic hepatitis B.     

替比夫定治疗慢性乙型肝炎108周的临床疗效

目的 探讨替比夫定治疗慢性乙型肝炎(CHB)患者108周的临床疗效.方法 随机选择就诊于吉林大学中日联谊医院未经过抗病毒治疗的72例CHB患者,其中包括35例肝硬化患者,给予口服替比夫定600 mg,1次/d,连续治疗108周.观察患者治疗前、后血清HBV DNA水平,肝功能及HBV标志物.根据12周和24周时的HBV DNA水平将患者分为＜3 log_(10)拷贝/ml和≥3 log_(10)拷贝/ml两组,比较其在治疗108周时的疗效.疗效比较采用χ~2检验.结果 替比夫定治疗4周时,HBV DNA不可测率及ALT复常率分别为37.5%和33.3%,至108周时达到87.5%和91.7%.46例HBeAg阳性患者治疗108周时HBeAg消失率及血清学转换率分别为39.1%和23.9%.12周时HBV DNA水平＜3 log_(10)拷贝/ml的患者,108周时的HBV DNA不可测率及HBeAg血清学转换率明显高于HBV DNA≥3 log_(10)拷贝/ml的患者(χ~2值分别为7.96和3.94,P值均＜0.05).24周时HBV DNA＜3log_(10)拷贝/ml的患者,108周时的HBV DNA不可测率高于HBVDNA≥3 log_(10)拷贝/ml的患者(χ~2=10.13,P＜0.05),耐药发生率低于HBV DNA≥3 log_(10)拷贝/ml的患者(χ~2=4.82,P＜0.05).替比夫定抗病毒治疗108周时肝硬化患者Child-Pugh分级较治疗前明显改善,其中Child A级患者比例明显增加(χ~2=5.83,P＜0.05).结论 替比夫定可强效、快速抑制HBV DNA复制,HBeAg血清学转换率高,长期治疗可明显改善肝硬化患者Child-Pugh评分,获得早期病毒学应答者的耐药发生率低.     

三种免疫抑制剂对HBV质粒感染小鼠HBV复制的影响*

目的 应用小鼠模型评价3种临床常用的免疫抑制剂对乙型肝炎病毒（HBV）复制的影响。方法 利用高压水注射法将HBV质粒pAAV/HBV1.2导入Balb/C小鼠肝内,建立小鼠HBV复制模型;将Balb/C小鼠随机分成4组,每组10只,分别给予生理盐水（Saline）、FK506、DEX和CYP处理。1w后,将供体C57BL/6小鼠皮瓣移植至上述经免疫抑制剂或生理盐水处理的Balb/C小鼠,皮肤移植后继续使用上述免疫抑制剂或生理盐水至观察期结束。采用电化学免疫发光法检测血清HBsAg、HBsAb和HBcAb水平,采用Real-time PCR法检测血清病毒载量。结果 FK506、地塞米松和环磷酰胺处理小鼠移植皮瓣的存活时间分别为（21.50±3.09） d、（21.50±3.09） d和（21.40±1.52） d,分别较生理盐水处理组（8.67±1.86） d明显延长[P=0.037、P=0.000和P=0.000];FK506处理组小鼠HBV复制及其抗体产生与对照组无显著性差异,地塞米松处理组在药物处理期间能维持病毒复制,但停药后病毒DNA和HBsAg迅速下降至检测水平以下,至观察终点仍未检出HBsAb和HBcAb阳性,环磷酰胺能明显延长小鼠HBV复制的持续时间,即使在停药后16 w血清HBV DNA和HBsAg均维持在较高水平,HBcAb和HBsAb持续阴性。结论 环磷酰胺而非FK506处理可在Balb/C小鼠模型中建立HBV持续复制模型。     

Antiviral effect of Chinese medicine jiaweisinisan in hepatitis B virus transgenic mice

AIM: To study the antiviral effect of Chinese medicine jiaweisinisan (JWSNS) on hepatitis B virus (HBV) infection in transgenic mice (TGM). METHODS: Twenty two 6-8 wk old HBV TGM in the third generation were divided into TGM control group and TGM treated group randomly. The normal control group included ten normal BC 57L/6 mice at the same age. The mice in treated group were administrated with JWSNS at the concentration of 4 g/mL and the dosage of 50 g/kg per d for 30 d, while the mice in TGM control group and normal control group were administrated with normal saline at the same dosage and the same time. Polymerase chain reaction (PCR) was used to assess the contents of HBV DNA in serum of HBV TGM before and after treatments, whereas blot hybridization was utilized to measure the contents of HBV DNA in the liver of both HBV TGM and normal BC 57L/6 mice. RESULTS: The levels of serum HBV DNA in TGM treated group were remarkably decreased after the treatment of JWSNS (7.662±0.78 vs 5.22±3.14, P<0.05), while there was no obvious change after administration of normal saline in TGM control group (7.125±4.26 vs 8.932±5.12, P>0.05). The OD values of HBV DNA in the livers of the mice in TGM treated group were significantly lower than those of TGM control group (0.274±0.096 vs 0.432±0.119,P<0.01). CONCLUSION: JWSNS exerts suppressive effects on HBV DNA in the serum and liver of TGM.     

Perioperative reactivation of hepatitis B virus replication in patients undergoing partial hepatectomy for hepatocellular carcinoma

Background and Aim: Reactivation of hepatitis B virus (HBV) replication happens in patients who receive transarterial chemoembolization or systemic chemotherapy for hepatocellular carcinoma (HCC). The incidence and risk factors of HBV reactivation during the perioperative period in HCC patients receiving hepatic resection is unknown. Methods: Between May 2009 and November 2010, 164 consecutive patients with HBV‐related HCC who underwent hepatic resection were prospectively enrolled in the study. Among these, 126 patients received antiviral treatment before the operation (the antiviral group) and 38 patients did not receive any antiviral treatment (the non‐antiviral group). Results: Ten patients (6.1%) developed HBV reactivation perioperatively (within 1 month after hepatectomy). The incidence of HBV reactivation in the antiviral group and non‐antiviral group were 1.6% (2/126) and 21.1% (8/38), respectively (P < 0.001). On univariate analysis, preoperative HBV DNA < 1.0 × 10³ copies/mL and non‐antiviral therapy were significantly correlated with the occurrence of HBV reactivation (P = 0.044 and P < 0.001, respectively). Only non‐antiviral therapy remained as a predictive factor on multivariate analysis (odds ratio, 15.46; 95% confidence interval, 2.80–85.46, P = 0.002). The recovery of liver function (defined as a decrease of alanine aminotransferase back to normal) was achieved in 86.8% (132/152) patients without HBV reactivation and in 37.5% (3/8) patients with HBV reactivation when evaluated on day 30 after hepatectomy (P < 0.001). Conclusion: Hepatectomy could reactivate HBV replication during the perioperative period, especially in patients who did not receive any antiviral therapy. A close monitoring of HBV DNA during the perioperative period was necessary irrespective of the preoperative HBV DNA level. Once HBV was reactivated, antiviral therapy should be given.     

Structure-Based Discovery of N-Sulfonylpiperidine-3-Carboxamides as Novel Capsid Assembly Modulators for Potent Inhibition of HBV Replication

As a key element during HBV replication, a nucleocapsid is considered a promising target for the treatment of chronic hepatitis B. The present study aimed to identify small molecules as novel capsid assembly modulators with antiviral activity. Structure-based virtual screening of an integrated compound library led to the identification of several types of HBV inhibitors. Among these inhibitors, N-sulfonylpiperidine-3-carboxamides (SPCs) potently reduced the amount of secreted HBV DNA. Through structure–activity relationship studies, we identified an SPC derivative, namely, C-39, which exhibited the highest antiviral activity without causing cytotoxicity. Mechanism studies showed that C-39 dose-dependently inhibited the formation of HBV capsid, synthesis of cccDNA, e antigen (HBeAg), viral pregenomic RNA (pgRNA), and HBV DNA levels, thereby restraining HBV replication. In summary, SPCs represent a new class of capsid assembly modulators. Further optimization of SPCs is expected to obtain new antiviral drugs against HBV infection.