血液病患者治疗后乙型肝炎病毒再激活的临床研究

目的观察血液病患者经过化疗及免疫抑制治疗后,乙型肝炎病毒（HBV）再激活情况及拉米夫定对其治疗作用。方法收集2005年1月至2010年1月在本院进行治疗的携带HBV的血液系统疾病患者62例,统计HBV再激活以及再激活后肝炎发生情况,并根据拉米夫定使用情况将其分为未预防组（n=50）和拉米夫定预防组（n=12）,检测两组化疗前后天冬氨酸转氨酶（AST）、丙氨酸转氨酶（ALT）、总胆红素、白蛋白（ALB）、HBV DNA水平。结果 62例患者中,治疗后HBV激活并发肝损害的有21例（33.9%）。治疗前两组的AST、ALT、总胆红素、ALB比较差异均无统计学意义,HBV DNA水平未预防组显著低于拉米夫定预防组;治疗后拉米夫定预防组的患者8.3%出现肝功能异常,而未预防组为40.0%（P〈0.01）,同时未预防组的AST、ALT和HBV DNA水平明显增高（P〈0.01）,而总胆红素和ALB无明显变化。结论化疗和免疫抑制治疗可使HBV再激活,拉米夫定可预防和治疗HBV感染的血液病患者化疗后病毒的重新激活,且不良反应较小。     

不同基因型乙型肝炎病毒患者拉米夫定治疗期间YMDD突变株的变化分析

目的研究不同基因型乙型肝炎病毒（HBV）患者拉米夫定治疗期间酪氨酸-蛋氨酸-天冬氨酸-天冬氨酸（YMDD）突变株的变化。方法采用实时荧光聚合酶链反应（PCR）技术和基因测序法分别对115例接受拉米夫定治疗的乙型肝炎患者血清进行乙型肝炎病毒基因（HBV DNA）水平、YMDD突变株及基因型检测。结果拉米夫定治疗时间〈12个月、12~24个月及〉24个月患者的HBV DNA阳性率分别为75%、35%、16%,YMDD变异率分别为22%、54%、100%。对115例乙型肝炎患者进行HBV基因分型发现,B型占17%,C型占75%,B、C混合型占8%;其中,B型、C型患者中YMDD变异率分别为21%、19.5%,B、C混合型患者中未发现YMDD变异。HBV DNA与HBV的YMDD变异率无明显相关性（P〉0.05）。结论拉米夫定治疗期间,HBV的YMDD变异不受病毒基因型的影响。     

拉米夫定治疗乙型肝炎病毒相关性肾炎的临床效果观察

将收治的50例乙型肝炎病毒相关性肾炎患者作为研究对象,对照组采用血管紧张素阻断剂治疗,观察组采用口服拉米夫定治疗,观察两组的治疗效果。治疗后,观察组总有效率、尿蛋白量及内生肌酐清除率均优于对照组,差异具有统计学意义(P0.05)。拉米夫定治疗乙型肝炎病毒相关性肾炎的临床效果显著,安全性高,无不良反应,值得推广应用。     

拉米夫定预防淋巴瘤化疗后乙型肝炎病毒再激活的临床观察

目的 探讨拉米夫定预防淋巴瘤化疗后乙型肝炎病毒(HBV)再激活的临床疗效及方法.方法 36例HBV感染的淋巴瘤患者(拉米夫定组)化疗同时及化疗期间口服拉米夫定100 mg/d,并持续至化疗结束后半年;对照组42例HBV感染的淋巴瘤患者未接受预防性拉米夫定治疗.对2组患者HBV病毒激活发生率及危险因素进行分析.结果 拉米夫定组36例,病毒再激活5例(13.9%),无一例死亡;对照组42例,病毒再激活19例(45.2%),死亡2例(4.8%),2组病毒再激活率差异有统计学意义(x2=8.943,P=0.003),病死率差异无统计学意义(x2=1.759,P=0.497).对照组使用CEOP方案24例,7例(29.2%)病毒再激活;使用R-CEOP方案18例,12例(66.7%)病毒再激活,CEOP和R-CEOP方案病毒再激活率差异有统计学意义(x2=5.893,P=0.016).Logistic多因素分析显示利妥昔单抗与对照组HBV再激活相关(P=0.042,OR=4.139,95%CI为1.052～15.207).结论 对于HBV感染的淋巴瘤患者,化疗同时应使用拉米夫定预防HBV再激活,特别是使用含利妥昔单抗方案的患者.     

乙型肝炎病毒基因突变与耐药株监测进展

拉米夫定(Lamivudine)为核苷酸类似物,主要用于抗乙型肝炎病毒治疗,能有效地抑制HBV的复制。但长期使用可引起乙型肝炎病毒基因突变,其突变部位主要在 HBV基因组 P区中高度保守的聚合酶基因YMDD模序,导致耐药病毒株的出现,影响临床治疗。目前临床检测突变的手段是建立在PCR技术基础上的方法。     

拉米夫定治疗临床表现为肾炎综合征的乙型肝炎病毒相关性肾炎的疗效观察

目的观察拉米夫定治疗临床表现为肾炎综合征的乙型肝炎病毒相关性肾炎(HBV-GN)的疗效。方法32例符合条件的HBV-GN患者,随机分成治疗组和对照组,治疗组口服拉米夫定100mg/d,对照组服用血管紧张素转换酶抑制剂(ACEI)或血管紧张素受体拮抗剂(ARB),治疗12个月,随访6个月。结果拉米夫定治疗组蛋白尿较对照组明显减少,肾功能稳定,仅1例患者因使用拉米夫定出现YVDD变异,无明显不良反应发生。结论拉米夫定治疗临床表现为肾炎综合征的乙型肝炎病毒相关性肾炎安全有效。     

阿德福韦酯与拉米夫定抗乙型肝炎病毒作用比较

阿德福韦酯(adefovir dipivoxil)是新一代抗乙型肝炎病毒新药,欧美等国已用于乙型肝炎治疗,我国目前也已进入临床观察阶段.拉米夫定是迄今我国用于乙型肝炎治疗的主要首选药物之一,临床已有其成熟的治疗方案,现将上述两药在作用机制、临床疗效、安全性及耐药性等方面的特点进行比较,供读者参考.     

拉米夫定联合化疗药物对乙型肝炎病毒感染合并肿瘤患者乙型肝炎病毒再激活的预防作用

目的 探讨拉米夫定联合化疗药物对乙型肝炎病毒（hepatitis B virus,HBV）感染合并肿瘤患者乙肝病毒再激活的预防作用.方法 2014年7月至2017年2月在南充市中心医院住院治疗的HBV感染合并肿瘤患者共120例作为研究对象,据治疗方法不同分为观察组与对照组各60例,对照组给予常规化疗与保肝治疗措施,观察组在对照组治疗的基础上给予拉米夫定（100 mg/d,1次/d）预防性治疗,疗程均为8周.结果 观察组与对照组的HBV再激活率分别为5.0％（3/60）和33.3％（20/60）,两组比较差异有统计学意义（χ^2=15.692,P〈0.05）.治疗前、后观察组患者血清谷氨酸转移酶、天冬氨酸转移酶比较差异均无统计学意义（P均〉0.05）,对照组呈现上升趋势（P均〈0.05）;治疗后观察组的血清谷氨酸转移酶与天冬氨酸转移酶[（31.98±6.33）U/L和（26.38±4.98）U/L）]均低于对照组[（43.89±6.73）U/L和（51.78±5.99）U/L],两组比较差异均有统计学意义（t值分别为8.294、11.842,P均〈0.05）.治疗后观察组与对照组的HBV DNA[（0.16±0.04）×10^3 copies/ml和（5.02±1.72）×10^3 copies/ml]均低于治疗前[（14.55±2.14）×10^3 copies/ml和（14.09±1.98）copies/ml],差异均有统计学意义（t值分别为25.498、8.142,P均〈0.05）;治疗后观察组患者较对照组低,两组比较差异有统计学意义（t=24.292,P〈0.05）.结论 拉米夫定联合化疗药物应用于HBV感染合并肿瘤患者具有预防乙肝病毒再激活作用,不影响患者的肝功能,有利于抑制乙肝病毒复制,有很好的应用价值.     

不同基因型乙型肝炎病毒患者拉米夫定治疗期间YMDD突变株的变化分析

目的研究不同基因型乙型肝炎病毒(HBV)患者拉米夫定治疗期间酪氨酸-蛋氨酸-天冬氨酸-天冬氨酸(YMDD)突变株的变化。方法采用实时荧光聚合酶链反应(PCR)技术和基因测序法分别对115例接受拉米夫定治疗的乙型肝炎患者血清进行乙型肝炎病毒基因(HBV DNA)水平、YMDD突变株及基因型检测。结果拉米夫定治疗时间<12个月、12~24个月及>24个月患者的HBV DNA阳性率分别为75%、35%、16%,YMDD变异率分别为22%、54%、100%。对115例乙型肝炎患者进行HBV基因分型发现,B型占17%,C型占75%,B、C混合型占8%;其中,B型、C型患者中YMDD变异率分别为21%、19.5%,B、C混合型患者中未发现YMDD变异。HBV DNA与HBV的YMDD变异率无明显相关性(P>0.05)。结论拉米夫定治疗期间,HBV的YMDD变异不受病毒基因型的影响。     

拉米夫定对激素治疗的肾病综合征患者乙型肝炎病毒复制的防治

目的:观察拉米夫定对肾上腺皮质激素(激素)治疗的肾病综合征患者乙型肝炎病毒(hepatitisBvirus,HBV)复制的防治效果。方法:8例肾病综合征伴HBV标志物阳性患者口服足量泼尼松或甲泼尼龙,同时口服拉米夫定治疗3个月以上,观察血清HBVDNA滴度、HBV标志物及ALT、AST的变化。结果:拉米夫定治疗1个月后血清HBVDNA滴度由(6.9±2.8)×108copy/mL下降至(3.0±1.9)×108copy/mL(P<0.05),治疗3个月后下降至(1.1±0.6)×108copy/mL(P<0.01),8例中2例病人治疗后血清HBVDNA滴度阴转,其中1例HBsAg、抗-HBe及抗-HBc阳性患者治疗2个月后血HBVDNA滴度阴转,ALT、AST正常,但无1例HBeAg阴转。结论:拉米夫定对激素治疗的肾病综合征患者的HBV复制有防治作用。     

Rapid detection of lamivudine-resistant hepatitis B virus mutations by PCR-based methods

Hepatitis B virus (HBV) is one of the major causes of liver disease worldwide, and chronic HBV infection may progress to cirrhosis and hepatocellular carcinoma. Mutations at the active site of DNA polymerase of HBV, tyrosine-methionine-aspartate-aspartate (YMDD) motif, render infected patients resistant to antiviral drug (Lamivudine) therapy. Hence, sensitive and specific methods aimed at detecting the mutants are essential. The purpose of this study was to develop methods for detecting the mutations at YMDD by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and real-time PCR using locked nucleic acid (LNA)-mediated TaqMan probes. The results obtained by these methods were compared with those examined by conventional direct sequencing on serum samples of 77 patients treated with lamivudine. Our results show that both PCR-RFLP and real-time PCR could detect wild type, YMDD, and its mutants, tyrosine-isoleucine-aspartate-aspartate and tyrosine-valine-aspartate-aspartate. In addition, the mixtures of the wild-type virus and its mutants in the serum sample were detected. Importantly, real-time PCR is less time-consuming, and more sensitive for the detection of mixed populations than PCR-RFLP. The real-time PCR with LNA-mediated TaqMan probes is a sensitive, specific and rapid detection method for mutations at the YMDD motif, which will be essential for monitoring patients undergoing lamivudine antiviral therapy.     

In vitro susceptibility of lamivudine-resistant hepatitis B virus to adefovir and tenofovir

Emergence of lamivudine-resistant hepatitis B virus (HBV) is a major concern in human immunodeficiency virus (HIV) and HBV coinfected patients. Following selection of resistant mutants, hepatitis flare or rapid progression to cirrhosis may occur. Treatment of patients with new nucleotide analogues such as adefovir dipivoxil (ADV) or tenofovir disoproxil fumarate (TDF) has shown good efficacy in controlling wild-type or lamivudine-resistant HBV replication. The purpose of this study was to assess the in vitro efficacy of new nucleotide analogues on HBV strains isolated from lamivudine-treated patients. After purification of HBV DNA from patient sera, the whole HBV genome was PCR-amplified and cloned. Drug sensitivity was measured after transfection of the isolated full genomes into HepG2 cells and measurement of HBeAg, HBsAg and viral replication in the culture media under increasing drug concentrations. A wild-type strain isolated from an untreated patient served as control. In a clinical study of ADV (Gilead 460i study), seven of the 35 patients carried HBV strains with the triple lamivudine resistance-associated amino-acid changes rtV173L/L180M/M204V at baseline. Although all patients responded to ADV in this clinical study, the serum HBV reduction was lower in the seven patients with the triple mutation (median -3.3 log copies/ml) compared to the patients who had only the rtL180M/M204V mutations (median -4.1 log copies/ml) at week 48 (P=0.04, Mann-Whitney test). In our in vitro system, lamivudine IC50 on lamivudine-resistant HBV carrying amino-acid substitutions rtL180M and rtM204V within the polymerase encoding region increased by more than 16,000-fold (from 6 nM to over 100 microM) when compared to wild-type HBV. For ADV and TDF, comparison of wild-type and lamivudine-resistant HBV IC50 (rtL180M-M204V) showed, respectively, 2.85-fold (from 0.07 to 0.2 microM) and 3.3-fold (from 0.06 to 0.2 microM) increases, indicating a mild decrease of both drug activities, in vitro. At the ADV concentration of 0.1 microM, presence of the V173L mutation reduced the inhibition of HBsAg production from 50 to 30% (P<0.01) and the viral replication from 45 to 32% (P<0.01, Mann-Whitney). Conversely, tenofovir had similar potency on both HBV mutation profiles with 60% inhibition of HBsAg production and 45% inhibition of viral replication at 0.1 microM. Our study supports the high efficacy of ADV and TDF seen in patients after lamivudine breakthrough. The excellent activity of TDF on lamivudine-resistant virus independently of the resistance mutation profile offers an interesting treatment alternative to HIV-HBV coinfected patients.     

Adefovir dipivoxil for treatment of breakthrough hepatitis caused by lamivudine-resistant mutants of hepatitis B virus

OBJECTIVE: Adefovir dipivoxil (ADV) is a nucleoside analogue that inhibits wild-type hepatitis B virus (HBV) and lamivudine-resistant HBV mutants in vitro and in vivo. The aim of this study was to evaluate the efficacy of ADV against lamivudine-resistant mutants and of adefovir and interferon (IFN) add-on to lamivudine for patients with severe acute exacerbation of hepatitis caused by lamivudine-resistant mutants. METHODS: Fourteen patients with breakthrough hepatitis were treated with ADV. Four of the 14 patients also received IFN as combined treatment for severe acute exacerbation of hepatitis. RESULTS: At week 24, serum HBV DNA levels had significantly decreased by a median of over 4.8 log copies/ml in the ADV group and over 5.9 log copies/ml in the ADV + IFN group compared to baseline. The median decrease in alanine aminotransferase (ALT) levels from baseline to week 24 was -1.05 times the upper limit of normal (ULN) in the ADV group [significant at week 24 compared with baseline (p = 0.012)] and -22.3 times the ULN in the ADV + IFN group. CONCLUSIONS: Administration of ADV add-on to lamivudine for patients with breakthrough hepatitis reduced HBV DNA and ALT levels. ADV and IFN add-on to lamivudine could prevent a fatal course in patients with severe acute exacerbation of hepatitis.     

肝移植后乙肝再感染的因素分析及其防治

肝移植患者由于使用免疫抑制剂以及过多种广谱抗生素,使得肝移植比任何其他腹部手术更易发生感染,国内以肝移植后发生乙肝再感染为主.随着乙型肝炎病毒相关性肝病患者行肝移植后长期存活数目的增加,移植学家们把研究的重点延伸到如何优化肝移植后预防乙型肝炎病毒再感染的治疗方案以降低因乙型肝炎病毒变异导致再感染和减少长期用药、降低治疗费用上来.文章分析了肝移植术后乙肝再感染的因素,介绍了肝移植后用于防治乙肝再感染的药物种类及临床应用,展望了肝移植后药物防治乙肝再感染的新的研究方向.     

Efficacies of entecavir against lamivudine-resistant hepatitis B virus replication and recombinant polymerases in vitro

Entecavir (ETV) is a potent and selective inhibitor of hepatitis B virus (HBV) replication in vitro and in vivo that is currently in clinical trials for the treatment of chronic HBV infections. A major limitation of the current HBV antiviral therapy, lamivudine (3TC), is the emergence of drug-resistant HBV in a majority of treated patients due to specific mutations in the nucleotide binding site of HBV DNA polymerase (HBV Pol). To determine the effects of 3TC resistance mutations on inhibition by ETV triphosphate (ETV-TP), a series of in vitro studies were performed. The inhibition of wild-type and 3TC-resistant HBV Pol by ETV-TP was measured using recombinant HBV nucleocapsids, and compared to that of 3TC-TP. These enzyme inhibition studies demonstrated that ETV-TP is a highly potent inhibitor of wild-type HBV Pol and is 100- to 300-fold more potent than 3TC-TP against 3TC-resistant HBV Pol. Cell culture assays were used to gauge the potential for antiviral cross-resistance of 3TC-resistant mutants to ETV. Results demonstrated that ETV inhibited the replication of 3TC-resistant HBV, but 20- to 30-fold higher concentrations were required. To gain further perspective regarding the potential therapeutic use of ETV, its phosphorylation was examined in hepatoma cells treated with extracellular concentrations representative of drug levels in plasma in ETV-treated patients. At these concentrations, intracellular ETV-TP accumulated to levels expected to inhibit the enzyme activity of both wild-type and 3TC-resistant HBV Pol. These findings are predictive of potent antiviral activity of ETV against both wild-type and 3TC-resistant HBV.     

Low-dose hepatitis B immune globulin and higher-dose lamivudine combination to prevent hepatitis B virus recurrence after liver transplantation

Post-transplant prevention of hepatitis B virus (HBV) infection is based on treatment with lamivudine and/or hepatitis B immune globulin (HBIG). However, optimum doses and duration for these drugs are not yet clear. We tested high doses of lamivudine (300 mg/day) in combination with low doses of HBIG (200-400 IU/2-4 weeks). Eighty patients who had post-transplant prophylaxis of lamivudine and HBIG were included in the study. Of those, 20 had hepatitis D virus co-infection and eight were HBV DNA-positive at the time of transplantation. Ten HBV DNA-positive patients were treated with lamivudine (150 mg/day) before transplantation; all were HBV DNA-negative after lamivudine treatment. All patients in the anhepatic phase were given 4000 IU of HBIG. Following this, 400 or 800 IU HBIG was administered intramuscularly daily for 5-10 days post-transplantation and 2-4 times weekly thereafter, according to serum titre of antibodies to hepatitis B surface antigen (anti-HBs). Lamivudine was maintained or initiated at the time of transplantation and was continued indefinitely. Median follow-up was 21 months (range 3-73 months). Recurrence of hepatitis B surface antigen (HBsAg)-positivity occurred in only three out of 78 (4%) patients; two of these three were HBV DNA-positive. Median anti-HBs titre at the final follow-up was 68 IU. Patient and graft survival was 85% at 1 year. In conclusion, a combination of lamivudine 300 mg/day and low-dose HBIG prevents post-transplantation recurrence of hepatitis B, even in the presence of viral replication in the pre-transplant period.     

乙型肝炎病毒感染者肾移植前肝穿刺活检与预后分析

背景：乙肝病毒感染者肾移植手术国内有较多报道,但乙肝病毒感染者肾移植前肝穿刺活检观察有限。目的：对慢性肾功能衰竭合并不同程度慢性乙型病毒性肝炎患者进行肾移植前肝穿刺活检,移植后2年随访观察转归情况。方法：对接受肾移植的21例乙型肝炎病毒感染的尿毒症患者进行肝穿刺活检。根据肝活检组织病理学改变,分为轻度（n=9）、中度（n=7）、重度（n=5）3组。肾移植后随访观察2年。3组中各有2例进行重复肝活检组织病理学检查。结果与结论：轻度慢性乙型肝炎组在随访2年中各项观察指标均无明显变化。中度慢性乙型肝炎组从移植后3个月开始谷氨酰转肽酶活性明显高于正常水平,随访至终点时,2例重复肝活检病理显示已处于重度病变。重度慢性乙型肝炎组从移植后3个月开始谷氨酰转肽酶活性持续高于正常水平;18个月开始,血清白蛋白水平低于正常值,球蛋白水平高于正常值;随访至终点时,有4例呈肝硬化改变。提示不同程度的慢性乙型病毒性肝炎患者肾移植后预后不同,肝活检是评价肝脏病变程度的重要手段,具有指导肾移植选择的作用。     

乙型肝炎病毒感染者肾移植前肝穿刺活检与预后分析

背景:乙肝病毒感染者肾移植手术国内有较多报道,但乙肝病毒感染者肾移植前肝穿刺活检观察有限.目的:对慢性肾功能衰竭合并不同程度慢性乙型病毒性肝炎患者进行肾移植前肝穿刺活检,移植后2年随访观察转归情况.方法:对接受肾移植的21例乙型肝炎病毒感染的尿毒症患者进行肝穿刺活检.根据肝活检组织病理学改变,分为轻度(n=9)、中度(n=7)、重度(n=5)3组.肾移植后随访观察2年.3组中各有2例进行重复肝活检组织病理学检查.结果与结论:轻度慢性乙型肝炎组在随访2年中各项观察指标均无明显变化.中度慢性乙型肝炎组从移植后3个月开始谷氨酰转肽酶活性明显高于正常水平,随访至终点时,2例重复肝活检病理显示已处于重度病变.重度慢性乙型肝炎组从移植后3个月开始谷氨酰转肽酶活性持续高于正常水平;18个月开始,血清白蛋白水平低于正常值,球蛋白水平高于正常值;随访至终点时,有4例呈肝硬化改变.提示不同程度的慢性乙型病毒性肝炎患者肾移植后预后不同,肝活检是评价肝脏病变程度的重要手段,具有指导肾移植选择的作用.     

Prediction of breakthrough hepatitis due to lamivudine-resistant hepatitis B virus by a sensitive semiquantitative assay using peptide nucleic acids

OBJECTIVE: The aim of this study was to predict breakthrough hepatitis and analyze the dynamics of lamivudine-resistant hepatitis B virus in patients treated with lamivudine. METHODS: Fifty-five chronic hepatitis B patients treated with lamivudine were included. The emergence of YMDD motif mutants was detected by peptide nucleic acid (PNA) mediated PCR clamping with a detection limit of 10(1) YMDD mutants. We then performed a semiquantitative PCR assay of subjects in whom YMDD mutants were detected. This assay detects 10(2.7)-10(7.7) copies of mutant virus per 1 ml of serum. RESULTS: YMDD mutants were detected in 28 (51%) of the 55 patients. Eight patients stopped medication before viral breakthrough. YMDD mutants appeared transiently despite the continuance of lamivudine therapy in 12 patients. In all 8 patients with breakthrough hepatitis, the quantities of YMDD mutants ranged from 10(2.7)-10(4.7) copies/ml in the two to three months before clinical breakthrough. In contrast, in 12 patients without viral breakthrough, there were always less than 10(2.7) copies/ml YMDD mutants. CONCLUSIONS: Lamivudine-resistant viruses sometimes disappear even during lamivudine administration. Our sensitive quantitative assay proved useful for early detection of YMDD mutants and a threshold of 10(2.7) copies/ml is suggested for predicting viral breakthrough.