Factors affecting effectiveness of vaccination against hepatitis B virus in hemodialysis patients

Hepatitis B virus(HBV)is a major global health problem.Despite the success of the general measures against blood transmitted infections in hemodialysis(HD)units,the prevalence of HBV infection among the HD patients is still high.Thus vaccination against HBV is indicating in this population.However,compared with the general population the seroprotection achieved in HD patients remains relatively low,at about 70%.In this review patient,HD procedure and vaccine-associated factors that affect the efficacy of HBV vaccination are analyzed.Also alternative routes of HBV vaccine administration as well as new and more immunogenic vaccine formulations are discussed.However,besides scientific progress,vigilance of HD physicians and staff regarding the general measures against the transmission of blood borne infections and the vaccination against HBV is also required for reducing the prevalence of this viral infection.

Pathogenesis of cholangiocarcinoma in the porta hepatis and infection of hepatitis virus

OBJECTIVE: To study the correlation between human cholangiocarcinoma in the porta hepatis and the infection of hepatitis virus. METHODS: Immunohistochemistry was used to detect HBxAg and HCV-C protein in formalin-fixed and paraffin-embedded samples taken from 68 patients with cholangiocarcinoma in the porta hepatis. The findings were reviewed against the clinical records of the patients. RESULTS: Six patients (8.8%) were positive for HBxAg and 24 (35%) for HCV-C protein, respectively. One patient was positive for both HBxAg and HCV-C protein. There were statistically differences in the extent of differentiation, invasion, lymph-node metastasis, and treatment between the patients with cholangiocarcinomas in the porta hepatis with HB(C) V infection and those without infection. CONCLUSIONS: HB(C) V infection is correlated to the development of cholangiocarcinoma in the porta hepatis. The tumor with HB(C) V infection may have a higher malignancy biologically and poorer prognosis. HBxAg and HCV-C protein may play an important role in the pathogenesis of cholangiocarcinoma in the porta hepatis.

Evaluation of HBsAg Quantification as Surrogate to HBV DNA Viral Load in Hepatitis B Infected Patients in Anambra State, Nigeria

Hepatitis B is an infectious disease of great public health importance. Nigeria is one of the countries with the highest incidence of Hepatitis B Virus (HBV) infection worldwide. However, the accessibility and affordability of HBV DNA quantification (viral load) assay is the key laboratory test for therapy initiation, and monitoring is a challenge to HBV management. This study aimed at determining the relationship between HBV DNA quantification and routine haemato-serological parameters in order to develop a more cost-effective diagnostic algorithm for Hepatitis B management. Cross sectional study design was used with a total of 264 subjects comprising of 88 HBsAg seropositive treatment naïve subjects, 88 HBsAg seropositive subjects on antiviral therapy as case subjects and 88 age-matched apparently healthy HBsAg seronegative individuals were recruited as control subjects. Hepatitis B Virus DNA assay was performed using real time PCR technique while ELISA technique was used for Hepatitis B surface antigen quantification. HBsAg quantification showed strong positive correlation with HBV DNA viral load both in treatment and non-treatment groups (r = 0.673;p < 0.001). However, the Receiver Operation Characteristics curve indicated a very poor performance characteristics (AUC = 0.537, p = 0.002). The non-treatment group has higher viral load (M = 805.50 IU/ml) compared with treatment group (M = 65.50 IU/ml) (p < 0.001). There was a significant difference in HBV DNA levels among the four serological patterns observed in the study (p < 0.001). This study has revealed that HBsAg quantification has strong correlation with HBV viral load but might not be efficient in clinical practice as a predictor of serum HBV viral load due to its poor performance characteristics in identifying high positive viral load.

220例血清HBV DNA≤2000 IU/mL的代偿期肝硬化患者肝功能评价指标、并发症分析

目的了解血清乙型肝炎病毒(HBV)DNA≤2000 IU/mL的乙型肝炎病毒(HBV)相关代偿期肝硬化患者的肝功能及并发症发生情况,为后续临床诊疗提供依据。方法220例血清HBV DNA≤2000IU/mL的HBV相关代偿期肝硬化患者,收集其肝功能评价指标资料和并发症资料。结果220例患者肝功能生化指标血清甲胎蛋白为4.80(2.12,8.35)ng/mL、总胆红素为18.90(14.10,28.76)μmol/L,白蛋白为39.35(34.60,43.90)g/L、肌酐为64(56,73.52)μmol/L、丙氨酸转氨酶为31.73(21.12,52.17)U/L、天冬氨酸转氨酶为30.35(23.85,48.85)U/L、γ-谷氨酰转肽酶42.70(24,66.08)U/L、碱性磷酸酶为82.05(63.38,113)U/L、血小板为105(57.25,158)×109/L、凝血酶原活动度为81.03%(66%,93.97%)、国际标准化比值为1.09(1.02,1.21);肝功能CTP分级以A级为主,终末期肝病模型评分(8.91±3.78)分、天冬氨酸转移酶及血小板比率指数(1.63±2.83)分、基于4因子的肝纤维化指数评分(4.84±5.28)分。220例患者中HBV DNA阳性(血清HBV DNA 100~2000 IU/mL)49例、乙肝表面抗原阳性183例、乙肝e抗原阳性151例。220例患者并发症发生情况为脾大103例、脾功能亢进110例、食管胃底静脉曲张74例、门静脉血栓6例、HCC 6例。与其他年龄比较,年龄<40岁的HBV相关代偿期肝硬化患者脾大发生率高(χ^(2)=6.864,P<0.05)。结论虽然血清HBV DNA≤2000 IU/mL的代偿期HBV相关肝硬化患者的肝功能基本正常,但部分指标处于正常范围的低值。血清HBV DNA≤2000 IU/mL的代偿期HBV相关肝硬化患者脾大、脾功能亢进等并发症发生率较高。

乙型肝炎病毒cccDNA与HBx蛋白在肝细胞性肝癌中表达的关系及意义

目的:探讨肝细胞性肝癌中乙型肝炎病毒(HBV)cccDNA与HBx蛋白表达的关系及其意义.方法:取42例肝细胞性肝癌患者的癌和癌旁组织,采用SABC法检测组织中的HBx蛋白;采用RT-PCR法检测组织中的HBV cccDNA水平.结果:HBx蛋白在癌及癌旁组织中的阳性例数分别为31例(73.8%)和35例(83.3%),无显著性差异;癌旁组织中cccDNA水平较癌组织中的高,但是统计学检验无显著性差异;癌组织和癌旁组织中HBx蛋白(+)者的cccDNA水平均明显高于HBx蛋白(-)者(P<0.05).HBx蛋白表达与cccDNA水平呈正相关(r=0.778,P<0.01).结论:HBx蛋白的表达与cccDNA水平明显相关,他们相互作用、相互影响并在肝癌的发生发展中起重要作用.

HBsAg阳性产妇分娩婴儿乙肝免疫接种低和无应答的影响因素

目的探讨可能导致HBsAg阳性产妇分娩的婴儿乙肝免疫接种后免疫低和无应答的因素。方法回顾性分析2013-2014年在首都医科大学附属北京佑安医院收治的HBsAg阳性产妇分娩的832例新生儿的临床资料,新生儿出生后6 h内及21 d接种乙型肝炎免疫球蛋白200 IU,出生当天和出生后1、6个月注射乙型肝炎疫苗10μg。出生后6 h内、7月龄抽取股静脉血,检查HBV标志物和HBV DNA定量。按照产妇孕期是否接受抗病毒治疗、母乳喂养、父亲HBsAg情况、母亲HBeAg情况分组,评估上述因素对婴儿乙肝免疫接种应答情况的影响。结果产妇孕期接受抗病毒治疗组的婴儿低和无应答率为8.6%（25/290）,未行抗病毒治疗组的婴儿低和无应答率为7.0%（38/542）,差异无统计学意义（χ2=0.60,P=0.44）。母乳喂养组婴儿的低和无应答率为8.8%（18/204）,人工喂养组低和无应答率为7.2%（45/628）,差异无统计学意义（χ2=0.52,P=0.47）。父亲HBsA g阳性组的婴儿低和无应答率为20.0%（5/25）,父亲HBsAg阴性组的婴儿低和无应答率为27.2%（58/807）,差异无统计学意义（χ2=3.00,P=0.08）。母亲HBeAg阳性组的婴儿低和无应答率为9.9%（33/335）,母亲HBeAg阴性的婴儿低和无应答率为6.0%（30/497）,差异无统计学意义（χ2=3.55,P=0.06）。结论 HBsAg阳性产妇分娩的婴儿中,母亲孕期采用抗病毒治疗、母乳喂养、父亲HBsAg阳性和母亲HBeAg阳性对婴儿乙肝免疫应答无明显影响。

慢性乙型肝炎肝组织Fas抗原表达及与HBV抗原的关系

用免疫组织化学技术检测了５２例慢性乙型肝炎（ＣＨＢ）肝组织内的Ｆａｓ抗原、ＨＢｓＡｇ和ＨＢｃＡｇ。结果表明，４８例Ｆａｓ抗原阳性表达（９２．３％），主要在肝细胞浆内，其表达程度与肝组织活动性炎症分度密切相关，提示Ｆａｓ抗原介导的细胞凋亡在肝细胞损伤中起一定作用；肝细胞Ｆａｓ抗原的表达程度与ＨＢｓＡｇ相关而与ＨＢｃＡｇ不相关。

急性乙型肝炎患者Pres1抗原和HBVDNA与ALT的动态观察

目的了解前S1蛋白(PreS1)、乙型肝炎病毒(HBVDNA)及丙氨酸氨基转移酶(ALT)与急性乙型肝炎(AHB)病程的关系。方法采用酶联免疫吸附试验(ELISA)、核酸扩增(PCR)荧光定量检测技术和双试剂法分别检测PreS1抗原、HBVDNA和ALT。结果不同病程AHB患者血清PreS1抗原与ALT阳性符合率较高(P>0.05),HBVDNA与ALT相比阳性符合率较低(P<0.01)。结论AHB患者治疗过程中,PreS1抗原与ALT阴转基本一致,先于HBVDNA阴转,提示疾病的转归及预后较好。

血清HBcAg对判断病毒复制的临床价值

目的 探讨血清 HBc Ag对判断病毒复制的临床价值。方法 对 5 82份有一项乙型肝炎病毒标志物阳性的血清以 EL ISA法进行检测 HBc Ag、抗 - HBc Ig M,并以 PCR- EL ISA法检测 HBV DNA定量。结果 结果 HBc Ag阳性组与阴性组的 HBV DNA阳性率分别为 91 .4 % (6 4 / 70 ) ,6 2 .5 % (32 0 / 5 1 2 ) ,两组相比 ,P<0 .0 0 1 ,前者明显大于后者 ;两组的抗 - HBc Ig M阳性率相比未见差异。结论 HBc Ag与 HBVDNA同是

pHBx-EGFP载体构建及其在肝癌Bel7402细胞中的表达

目的:构建乙型肝炎病毒(hepatitis B virus,HBV)-X蛋白(hepatitis B virus x protein,HBx)和增强型绿色荧光蛋白(enhanced green fluorescentprotein,EGFP)真核融合蛋白表达载体(pHBx-EGFP),转染肝癌Bel7402细胞,观察其表达和定位,为进一步研究HBx功能提供工具.方法:以pcDNA3.1-HBx为模板,应用PCR和DNA重组技术构建增强型绿色荧光蛋白HBx-EGFP融合蛋白表达载体pHBx-EGFP,经脂质体转染Bel7402细胞,应用倒置荧光显微镜观察融合蛋白表达和定位;同时提取转染pHBx-E G F P24h后的B e l7402细胞总蛋白,应用Western blot技术,鉴定HBx在细胞的表达情况.结果:成功扩增HBx片段插入pEGFP载体,经酶切验证后测序正确;pHBx-EGFP转染Bel7402细胞24h后,荧光显微镜下观察显示HBx-EGFP存在于细胞核周区域,而EGFP则弥散分布于整个细胞;Western blot得到HBx-EGFP、EGFP和HBx目的条带.结论:成功构建pHBx-EGFP真核重组蛋白表达载体,通过检测绿色荧光蛋白标记显示其在Bel7402细胞中表达及定位,并证实pHBx-EGFP载体能在Bel7402细胞正确表达.

化学发光免疫测定法检测HBV、抗HCV、抗HIV和TP-Ab结果分析

目的：对患者输血前或手术前的血液传染性疾病标志物进行检测分析。方法采用化学发光免疫测定（CLIA）法和ELISA法对湘雅萍矿合作医院3542例血清标本的HBsAg、抗HBs、HBeAg、抗HBe、抗HBc、抗丙型肝炎病毒（抗HCV）、抗人类免疫缺陷病毒（抗 HIV）、梅毒螺旋体抗体（TP-Ab）进行了检测。结果利用CLIA法检测3542例标本中乙型肝炎阳性704例，检出率为19．8％；3032例标本中丙型肝炎阳性52例，检出率为1．72％；3049例标本中抗 H IV阳性3例，检出率为0．09％；3153例标本中TP-Ab阳性200例，检出率为6．34％；不同性别乙型肝炎病毒（HBV）、抗 HCV、抗 HIV和 TP-Ab感染率有所不同，其中HBV、抗HCV和TP-Ab 3项，男性检出率明显高于女性（P＜0．05）；不同年龄段 HBV、抗 HCV、抗 HIV和TP-Ab感染率有所不同，HBV和抗 HCV以21～40岁检出率最高，分别为31．9％和2．33％；TP-Ab检出率随着年龄增大有增加的趋势，60岁以上检出率最高，达7．55％；经上述两种方法检测，HBsAg测值不一致的有51例，占1．44％，抗 HCV测值不一致的有1例，占0．03％，TP-Ab测值不一致的有3例，占0．1％，抗 HIV测值完全符合，均有3例阳性，不一致的标本经第三方试剂验证，结果CLIA法测值较为准确。结论 CLIA法是一种高灵敏度的检测方法，可有效地避免阳性标本漏检，应用CLIA法在输血或手术前对患者进行血液传染病性标志物检测具有重要意义。

Development of a universal immunoenzyme quantitative assay for detecting amplified products of nucleic acid and its preliminary application in hepatitis C virus

Objective To develop a universal quantitative immunoenzyme assay (EIA) for detecting amplified products of nucleic acid and its application in hepatit is C virus (HCV). Methods The appropriate cycle number of amplification was selected to s top polymerase chain reaction (PCR) before the “plateau stage”. At the same ti me, primers HCV(3) of the second PCR were modified with biotin so that the ampli fied products were labeled. The products were diluted and subsequently added t o the streptavidin coated wells, and the biotinylated products were captured, f ollowed by denaturation of NaOH, and non biotinylated strands were removed. Hy bridization was performed by adding the specific probe labeled with fluorescein. Finally anti fluorescein horse radish peroxidase (HRP) conjugates were added, after washing, 3,3',5,5', tetramethylbenzidine (TMB) was added to the well s and then measured on a microplate reader. Results EIA detection of amplified products of HCV showed that this ass ay was rapid, sensitive, specific and accurate. Correlation between the initial number of viral template and the EIA of amplified products was good. We also pro spectively investigated the response to interferon in five patients with HCV coi nfection. Results showed that this assay could be used as a guidance to the clin ical therapy in directing the use of antiviral drugs. Conclusions This assay could be widely used as a universal technique fo r the quantitative detection of amplified products of all nucleic acid (such as virus, bacterium) and other human genes (such as HLA B 27 ), it has vast vistas.

Efficacy of interferon alpha-2b and lamivudine therapy for chronic hepatitis B in children

To evaluate the efficacy of interferon (IFN) alpha-2b and lamivudine therapy in children with chronic hepatitis B virus (HBV) infection Method Ten children who developed chronic hepatitis B infection received IFN alpha-2b 10 million international units (IU)/m 2 body surface area, subcutaneously three times a week for six months IFN+lamivudine therapy began to be used in the cases who were unresponsive to IFN treatment Results Among 27 HBsAg (+) subjects in this study, interferon treatment was given to 11 subjects who developed chronic hepatitis One case was excluded from the study due to detection of Herpes type 1 encephalitis At the end of six months of follow-up, complete response was obtained in three (30%) patients and partial response in four (40%) patients, whereas no response was detected in three (30%) patients Fifty percent of the cases experienced serological response, 70% biochemical response, and all (100%) had histological response In three cases started concomitant IFN+lamivudine therapy, HBV-DNA became negative at the second month of treatment Conclusions IFN-alpha and lamivudine can be used for the treatment of chronic hepatitis B infection in children

聚合酶链反应在732份临床标本检测中的应用

利用聚合酶链反应（ＰＣＲ）检测７３２份痰、血清、尿、分泌物、胸腹水、脑脊液等临床标本中之结核杆菌、乙型肝炎病毒和淋球菌的ＤＮＡ。结果表明，活动性肺结核病人痰标本阳性率最高，达７４．２３％；乙型肝炎病毒阳性率为３２，１４～６９．２３％，淋球菌阳性率为４４．１２％，内一例病人分泌物涂片阴性而ＰＣＲ阳性。本技术具有灵敏、特异、简便、快速和不需培养等优点，适合于临床应用。

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。

反义寡核苷酸靶向肝细胞体外抗乙型肝炎病毒的实验研究

作者在对２．２．１５细胞中乙型肝炎病毒（ＨＢＶ）基因测序的基础上，设计合成了１段针对Ｕ５样序列区的１６－聚硫代磷酸反义寡核苷酸（ＡＳＯＮ），并将其与肝导向性载体——半乳糖化白蛋白和半乳糖化多聚赖氨酸连接，在２．２．１５细胞中观察了它们的抗病毒作用。在相同实验条件下，非导向组ＡＳＯＮ对表面抗原和ｅ抗原的抑制仅为７０％和５８％，而导向组ＡＳＯＮ可高达９０％～９６％和７９％～８２％，两者差异有显著意义。可见肝导向性载体能明显提高ＡＳＯＮ的抗病毒活性。ＡＳＯＮ对ＨＢＶ的抑制作用是特异性的，呈剂量依赖性关系；而对肝细胞本身的甲胎蛋白合成无影响，亦未见细胞毒性作用。无关序列寡核苷酸无明显抑制作用，反义抑制机理可能是作用在病毒逆转录和翻译水平上。

成人肾膜性病变的临床与病理特点分析

目的分析膜性肾病(MN)中,特发性膜性肾病(IMN)、Ⅴ型狼疮性肾炎(V-LN)及乙肝相关性膜性肾病(HBV-MN)的临床与病理特点。方法回顾性分析肾活检病理类型为MN的221例患者的临床与病理特点。结果 IMN、V-LN和HBV-MN分别占MN的66.52%、22.17%和11.31%。三种MN临床表现以肾病综合征和浮肿较常见。V-LN患者血清C3、C4显著减少,并伴有肾外表现。三种MN病理分期以Ⅱ、Ⅲ期为主,免疫荧光检测均有IgG和C3的大量沉积,电子致密物多部位沉积均较常见。V-LN中,C3和C1q的沉积多于IMN和HBV-MN。结论肾膜性病变各有其临床与病理特点,确诊需综合进行分析。

本院2013年-2015年乙型肝炎两对半检测结果分析及表面抗原钩状效应的处理

目的对2013年-2015年5 115例就诊者HBV血清标志物(HBs Ag、抗-HBs、HBe Ag、抗-HBe和抗-HBc)结果进行比较分析,为HBV感染者早发现早治疗提供依据。方法采用全自动化学发光分析仪对5 115例标本及其中320例阳性和5例HBs Ag低值而HBe Ag阳性标本进行检测。结果 3年来5 115例就诊者HBs Ag阳性率男性分别为7.12%、6.58%、6.47%,女性分别为5.75%、5.67%、5.59%;HBV感染与年龄有关,男性阳性率从50.27%降至2.70%,女性从51.11%降至0.74%;320例HBV感染者不论男女血清HBs Ag、HBe Ag、抗-HBc水平较正常对照组显著增高,抗-HBs和抗-HBe水平基本正常;1号~5号样倍比稀释后计算结果逐渐增加,呈现钩状效应。结论对患者乙型肝炎病毒检验,不仅能准确判断患者有无HBV感染,还能对乙型肝炎病毒的传染性进行判断,对临床治疗及预防HBV感染带来重要的指导意义。