Is there a need for universal double reflex testing of HBsAg-positive individuals for hepatitis D infection?

Hepatitis D virus(HDV)can infect HBsAg-positive individuals,causing rapid fibrosis progression,early decompensation,increased hepatocellular carcinoma risk,and higher mortality than hepatitis B virus(HBV)mono-infection.Most countries lack high-quality HDV prevalence data,and the collection techniques employed often bias published data.In recent meta-analyses,HDV prevalence in HBsAg-positive patients reaches 5%-15%and is even significantly higher in endemic areas.Since HBV vaccination programs were implemented,HDV prevalence has decreased among younger populations.However,owing to immigrant influx,it has increased in some Western countries.The current practice of HDV screening in HBsAg-positive individuals is stepwise,based on physician’s discretion,and limited to at-risk populations and may require numerous visits.Double reflex testing,which includes anti-HDV testing in all HBsAg-positive individuals and then HDV RNA testing for anti-HDV-positive ones,is uncommon.Reflex testing can identify more HDV infection cases and link identified patients to further care and follow-up.Moreover,laboratory-based double reflex screening is less biased than physician-led testing.Therefore,health-care providers should learn about reflex testing,and federal and provincial hepatitis control programs should implement laboratory-based double reflex testing to obtain reliable HDV prevalence estimates.The test’s cost-effectiveness depends on the number of HBV-positive patients screened to identify one HDV-positive patient.Such testing may be viable in areas with low HBsAg but high HDV prevalence.However,its economic impact on areas with low HDV prevalence needs further study.

Liver histopathological lesions is severe in patients with normal alanine transaminase and low to moderate hepatitis B virus DNA replication

BACKGROUND Chronic hepatitis B virus(HBV)infection remains a major global public health problem.Chronic hepatitis B(CHB)patients can be divided into treatment indication and non-treatment indication individuals according to alanine transaminase(ALT),HBV DNA,serum hepatitis B e antigen status,disease status[liver cirrhosis,hepatocellular carcinoma(HCC),or liver failure],liver necroinflammation or fibrosis,patients’age,and family history of HCC or cirrhosis.For example,normal ALT patients in‘immune-tolerant’phase with HBV DNA higher than 10^(7)or 2×10^(7)IU/mL,and those in‘inactive-carrier’phase with HBV DNA lower than 2×10^(3)IU/mL do not require antiviral therapy.However,is it reasonable to set the defined values of HBV DNA as the fundamental basis to estimate the disease state and to determine whether to start treatment?In fact,we should pay more attention to those who do not match the treatment indications(grayzone patients both in the indeterminate phase and in the‘inactive-carrier’phase).AIM To analyze the correlation of HBV DNA level and liver histopathological severity,and to explore the significance of HBV DNA for CHB with normal ALT.METHODS From January 2017 to December 2021,a retrospective cross-sectional set of 1299 patients with chronic HBV infection(HBV DNA>30 IU/mL)who underwent liver biopsy from four hospitals,including 634 with ALT less than 40 U/L.None of the patients had received anti-HBV treatment.The degrees of liver necroinflammatory activity and liver fibrosis were evaluated according to the Metavir system.On the basis of the HBV DNA level,patients were divided into two groups:Low/moderate replication group,HBV DNA≤10^(7)IU/mL[7.00 Log IU/mL,the European Association for the Study of the Liver(EASL)guidelines]or≤2×10^(7)IU/mL[7.30 Log IU/mL,the Chinese Medical Association(CMA)guidelines];high replication group,HBV DNA>10^(7)IU/mL or>2×10^(7)IU/mL.Relevant factors(demographic characteristics,laboratory parameters and noninvasive models)for liver histopathological severity were analyzed by univariate analysis,logistics analysis and propensity score-matched analysis.RESULTS At entry,there were 21.45%,24.29%,and 30.28%of the patients had liver histopathological severities with≥A2,≥F2,and≥A2 or/and≥F2,respectively.HBV DNA level(negative correlation)and noninvasive model liver fibrosis 5 value(positive correlation)were independent risk factors for liver histopathological severities(liver necroinflammation,liver fibrosis,and treatment indication).The AUROCs of the prediction probabilities(PRE_)of the models mentioned above(<A2 vs≥A2,<F2 vs≥F2,<A2 and<F2 vs≥A2 or/and≥F2)were 0.814(95%CI:0.770-0.859),0.824(95%CI:0.785-0.863),and 0.799(95%CI:0.760-0.838),respectively.HBV DNA level(negative correlation)was still an independent risk factor when diagnostic models were excluded,the P values(<A2 vs≥A2,<F2 vs≥F2,<A2 and<F2 vs≥A2 or/and≥F2)were 0.011,0.000,and 0.000,respectively.For the propensity score-matched pairs,whether based on EASL guidelines or CMA guidelines,the group with significant liver histology damage(≥A2 or/and≥F2)showed much lower HBV DNA level than the group with non-significant liver histology damage(<A2 and<F2).Patients in the moderate replication group(with indeterminate phase)had the most serious liver disease pathologically and hematologically,followed by patients in the low replication group(with‘inactive-carrier’phase)and then the high replication group(with‘immune-tolerant’phase).CONCLUSION HBV DNA level is a negative risk factor for liver disease progression.The phase definition of CHB may be revised by whether the level of HBV DNA exceeds the detection low limit value.Patients who are in the indeterminate phase or‘inactive carriers’should receive antiviral therapy.

Correlation between HBeAg and Hepatitis B Virus DNA and RNA Levels in Diverse Liver Disease Cohorts

Objective:To investigate the disparities and associations between HBV DNA and HBV RNA in various liver disease groups with respect to HBeAg status.Methods:Between September 2020 and September 2023,90 patients diagnosed with chronic hepatitis B(CHB),74 patients diagnosed with liver cirrhosis(LC),and 102 patients diagnosed with hepatocellular carcinoma(HCC)from the Department of Gastroenterology or Infection at the First Affiliated Hospital of Xi’an Jiaotong University were selected.HBV DNA,HBV RNA,and HBeAg quantitative tests were conducted using serum samples from the same patients.Results:In the three groups of cases,the HBV RNA load was higher when HBeAg was positive than when HBeAg was negative,and this difference was statistically significant.Only in the HCC group was the HBV DNA load significantly higher when HBeAg was positive than when HBeAg was negative.Additionally,there was a positive correlation between HBV DNA and HBV RNA regardless of HBeAg status.Conclusion:During HBeAg conversion,HBV RNA demonstrates a more sensitive response than HBV DNA.As CHB progresses to LC or HCC,HBV RNA exhibits better diagnostic value than HBV DNA.

不同核酸提取方法对HBV-DNA检测性能验证情况分析

目的评估2种乙型肝炎病毒(HBV)-DNA提取方法及2家检测试剂的性能,有助于选择优化提取试剂和检测试剂。方法2023年4月采用达安全自动核酸提取仪提取法(磁珠法)和手工提取法(一步法),并用达安和圣湘2种HBV-DNA试剂检测,对其进行精密度、正确度、线性范围、检出限及抗干扰能力等性能进行验证和评价。结果达安全自动核酸提取仪提取达安试剂检测、手工提取达安试剂检测和手工提取圣湘试剂检测在精密度、正确度、线性范围、检出限方面验证结果均达标;达安全自动核酸提取仪提取圣湘试剂检测在低值检测中变异系数大于5%,最低检测限验证不合格;抗干扰能力方面,全自动核酸提取仪提取的2.0 g/dL血红蛋白浓度的样本用达安和圣湘试剂检测结果均不受影响。手工提取甘油三酯浓度达3000 mg/dL的样本用达安和圣湘试剂检测的结果均不受影响。结论不同厂家的提取和检测试剂避免混用,达安和圣湘试剂对HBV-DNA定量检测的结果均符合要求。

HBV C基因型有关的HBsAg阴性HBV DNA阳性患者S区突变对HBsAg的影响

目的通过构建HBV C基因型突变质粒研究HBsAg阴性HBV DNA阳性患者HBV S区突变对HBsAg水平的影响。方法收集2022年8月至2023年4月首都医科大学附属北京佑安医院107例HBsAg-/HBV DNA+患者血液样本,对成功提取扩增的HBV DNA S区进行测序,通过构建HBV C基因型突变质粒对HBV S区突变位点进行细胞功能验证,探讨OBI可能发生的分子机制。结果对成功提取扩增的68例患者进行测序,发现HBV S区存在大量突变,包括免疫逃逸突变(如sG145R、sK122R、sS114T、sT131P等)和跨膜结构域(transmembrane domain,TMD)突变(如sT5A、sG10D、sF20S等)。通过构建HBV C基因型突变质粒,进行细胞转染和细胞免疫荧光实验发现sG145R突变会明显降低HBsAg的表达,但是sK122R、sI26N、sQ29N、sR169H、sS114T、sT131P这6个突变位点并未影响细胞内外HBsAg的表达。结论通过测序发现HBsAg-/HBV DNA+患者HBV S区存在大量突变位点,通过构建sG145R、sK122R、sI26N、sQ29N、sS114T和ST131P等突变质粒发现sG145R突变会明显降低细胞内外HBsAg的表达,但是sK122R、sI26N、sQ29N、sR169H、sS114T、sT131P并未明显降低细胞内外HBsAg的表达。

不同病情乙型肝炎患者HBV-DNA定量、两对半检测结果差异

目的分析乙型肝炎患者乙型肝炎病毒(HBV)-DNA定量检测与两对半检测结果的相关性,并观察不同病情HBV-DNA阳性率和两对半检测结果差异。方法选取江苏省泰兴市人民医院2021年4月至2023年4月收治的375例乙型肝炎患者病例为研究资料。比较不同乙肝两对半检测[乙肝表面抗原(HBsAg)、乙肝表面抗体(HBsAb)、乙肝e抗原(HBeAg)、乙肝e抗体(HBeAb)、乙肝核心抗体(HBcAb)]结果患者HBV-DNA定量检测结果差异,将患者根据HBV-DNA定量检测结果分为阳性组和阴性组,比较乙肝两对半结果差异,使用Pearson相关性分析;将患者根据疾病类型分为肝炎组、肝硬化组和肝癌组,比较三组患者乙肝两对半和HBV-DNA定量检测结果差异。结果不同乙肝两对半类型患者HBV-DNA阳性率比较,差异有统计学意义(P<0.05);HBV-DNA阳性组患者HBsAg、HBeAg水平高于HBV-DNA阴性组,HBsAb、HBeAb、HBcAb水平低于HBV-DNA阴性组,差异有统计学意义(P<0.05);HBsAg、HBeAg水平与HBV-DNA呈正相关(P<0.05),HBsAb、HBeAb、HBcAb水平与HBV-DNA呈负相关(P<0.05);不同疾病类型患者乙肝两对半类型HBV-DNA阳性率比较,差异有统计学意义(P<0.05),不同疾病类型患者HBV-DNA定量检测结果比较,差异有统计学意义(P<0.05)。结论乙型肝炎患者HBV-DNA定量检测与两对半检测结果有相关性,不同乙肝两对半类型间及不同疾病类型间HBV-DNA均有差异,HBV-DNA定量检测联合两对半检测可较好地反映病毒复制和疾病进展情况,为临床治疗提供指导。

血清ALT串联HBsAg和串联HBV DNA识别HBeAg阳性慢性HBV感染非活动性肝炎的性能评价

乙型肝炎病毒(hepatitis B virus,HBV)e抗原(hepatitis B e antigen,HBeAg)阳性的慢性HBV感染依次经历非活动性肝炎(non-aggressive hepatitis,NAH)和活动性肝炎(aggressive hepatitis,AH)2个分期,但仍缺乏界定HBeAg阳性NAH与AH的可靠标准。本文根据179例患者的长期随访队列,以自发性HBeAg血清转换作为终点事件,采用Kaplan-Meier生存分析,指定了丙氨酸转氨酶(alanine transaminase,ALT)、HBV表面抗原(hepatitis B surface antigen,HBsAg)和HBV DNA识别HBeAg阳性NAH的功能截断值;在此基础上,评价了ALT串联HBsAg和串联HBV DNA识别HBeAg阳性NAH的性能。结果显示,ALT≤60 IU/L、HBsAg>4.602 log10IU/mL和HBV DNA>7.477 log10IU/mL为识别HBeAg阳性NAH的功能截断值。基于功能截断值,ALT串联HBsAg的患者中,病理学分级≤G1和“分级≤G1且分期≤S2”的构成比均为100%,病理学分期≤S1和“分级≤G2且分期≤S1”的构成比均为68.2%;ALT串联HBV DNA的患者中,病理学分级≤G1和“分级≤G1且分期≤S2”的构成比均为86.2%,病理学分期≤S1和“分级≤G2且分期≤S1”的构成比均为69.0%;ALT串联HBsAg识别病理学分级≤G1和“分级≤G1且分期≤S2”的阳性似然比均为+∞,识别病理学分期≤S1和“分级≤G2且分期≤S1”的阳性似然比均为2.034;ALT串联HBV DNA识别病理学分级≤G1和“分级≤G1且分期≤S2”的阳性似然比分别为3.000和3.068,识别病理学分期≤S1和“分级≤G2且分期≤S1”的阳性似然比均为2.106。以上结果提示,ALT串联HBsAg和串联HBV DNA均可有效识别HBeAg阳性NAH;且ALT串联HBsAg识别HBeAg阳性NAH的性能优于ALT串联HBV DNA。

HBV DNA与乙肝五项定量在不同年龄段乙肝病毒感染患者病情进展中的意义

目的探讨乙肝病毒(HBV)DNA与乙肝五项定量[包括乙肝表面抗原(HBsAg)、乙肝表面抗体(HBsAb)、乙肝e抗原(HBeAg)、乙肝e抗体(HBeAb)、乙肝核心抗体(HBcAb)]在HBV感染进程中各个阶段的差异,从而为慢性乙型肝炎、乙肝肝硬化、肝癌不同阶段的诊治提供参考依据。方法选择2020年1月至2021年12月收治的295例HBV感染患者为研究对象,根据年龄将其分为青年组(≤45岁,105例)、中年组(46~59岁,128例)和老年组(≥60岁,62例)。比较三组慢性乙型肝炎、乙肝肝硬化、肝癌患者的HBV DNA及乙肝五项定量。结果三组慢性乙型肝炎、乙肝肝硬化和肝癌患者的年龄比较,差异无统计学意义(P>0.05)。青年组和中年组中,慢性乙型肝炎、乙肝肝硬化和肝癌患者的HBV DNA、HBsAg、HBeAg水平比较,差异具有统计学意义(P<0.05);慢性乙型肝炎患者的HBV DNA、HBsAg、HBeAg水平明显高于乙肝肝硬化和肝癌患者(P<0.05)。老年组中,慢性乙型肝炎、乙肝肝硬化和肝癌患者的HBV DNA、HBsAg、HBsAb、HBeAg、HBeAb、HBcAb水平比较,差异无统计学意义(P>0.05)。结论青中年患者中,随着慢性乙型肝炎向乙肝肝硬化、肝癌的病程进展,HBV DNA、HBsAg、HBeAg水平均呈现下降趋势,监测相关指标趋势对于提示疾病发展具有一定意义。

阿拉尔市某医院HBsAg阳性孕妇血清标志物模式分布与HBV-DNA载量分析

目的分析阿拉尔市某医院的乙型肝炎病毒表面抗原(HBsAg)阳性孕妇乙型肝炎病毒(HBV)感染血清标志物模式与HBV-DNA载量之间的关系,为监控孕期HBV感染和预防阻断HBV母婴传播提供理论依据。方法选取2021年10月至2023年10月期间在新疆生产建设兵团第一师阿拉尔医院产科建档及生产的HBsAg阳性孕妇为研究对象,通过磁微粒化学发光法检测孕妇五项HBV血清标志物包括测HBsAg和乙肝病毒表面抗体(HBsAb)、乙肝病毒e抗原(HBeAg)、乙肝病毒e抗体(HBeAg)以及乙肝病毒核心抗体(HBcAb),并利用荧光定量PCR检测HBV-DNA载量,分析HBV-DNA载量以及孕妇HBV感染模式对新生儿的影响。结果共纳入145例HBsAg阳性孕妇,HBV感染模式以感染模式Ⅱ[HbsAg(+)、HBeAb(+)和HBcAb(+)]为主,占62.76%。HBV-DNA阳性(>0.5×10^(3)IU/mL)阳性率为63.45%(92/145),不同HBV感染模式的HBsAg阳性孕妇HBV-DNA阳性率及载量分布情况比较差异有统计学意义(χ^(2)=16.532、64.367,P<0.05),主要表现为感染模式Ⅰ比例随HBV-DNA载量升高而增高。同时,HBV-DNA阳性组随HBeAg浓度升高而比例增高,而HBV-DNA阴性组正好相反,Spearman相关分析表明HBsAg阳性孕妇的HBeAg浓度与HBV DNA载量呈现明显正相关性(r=0.327,P<0.05)。不同HBV感染模式的HBsAg阳性孕妇的新生儿脐带血HBV阳性率比较差异有统计学意义(χ^(2)=16.851,P=0.002),同时HBsAg阳性孕妇的新生儿脐带血HBV阳性率随着HBV-DNA载量增高而明显增高,差异有统计学意义(χ^(2)=15.062,P=0.005)。结论HBsAg阳性孕妇血清标志物模式与HBV-DNA病毒载量密切相关,产前筛查对孕产妇及新生儿的乙肝预防及干预有重要意义,是母婴传播防控的重点。

妊娠合并慢性乙型肝炎患者乙肝标志物、前S1抗原与HBV-DNA的关系研究

目的 研究妊娠合并慢性乙型肝炎(CHB)患者乙肝标志物、前S1抗原(Pre-S1Ag)与乙型肝炎病毒核酸(HBV-DNA)的关系。方法 选取2022年5月至2023年12月我院收治的100例妊娠合并CHB患者为研究对象,依据HBV-DNA定量测定结果将其分为阳性组与阴性组,每组50例。比较两组的乙肝标志物、Pre-S1Ag水平、肝功能指标及不良妊娠结局;分析妊娠合并CHB患者乙肝标志物、Pre-S1Ag以及各项肝功能指标与HBV-DNA的相关性。结果 阳性组的乙型肝炎表面抗原(HBsAg)、乙型肝炎e抗原(HBe Ag)、Pre-S1Ag水平均高于阴性组,乙型肝炎表面抗体(HBsAb)、乙型肝炎e抗体(HBeAb)、乙型肝炎核心抗体(HBcAb)水平低于阴性组(P<0.05)。阳性组的各项肝功能指标水平均高于阴性组(P<0.05)。阳性组的剖宫产、胎膜早破、产后出血及羊水粪染发生率均高于阴性组(P<0.05)。Pearson相关性分析结果显示,妊娠合并CHB患者HBsAg、HBe Ag、Pre-S1Ag及各项肝功能指标均与HBV-DNA呈正相关,HBsAb、HBeAb及HBcAb与HBV-DNA呈负相关(P<0.05)。结论 妊娠合并CHB患者的乙肝标志物、Pre-S1Ag以及肝功能指标均与HBV-DNA密切相关。

HBV感染相关性肝细胞癌患者血甲胎蛋白、转氨酶水平与HBV-DNA载量和病情进展的关系

目的探讨乙型肝炎病毒(HBV)感染相关性肝细胞癌(HCC)患者血甲胎蛋白(AFP)、转氨酶水平与HBV-DNA载量和病情进展的关系。方法回顾性分析2022年1月至2023年4月本院收治的50例HBV感染相关性HCC患者资料并设为HCC组,另筛选同期HBV相关肝硬化(LC)和慢性乙型肝炎(CHB)患者资料各30例设为LC组和CHB组,比较三组血AFP及转氨酶水平;比较HCC组不同HBV-DNA载量患者AFP及转氨酶水平差异,并分析肝癌组患者AFP及转氨酶水平与病情进展的关系。结果HCC组患者AFP和丙氨酸氨基转移酶(ALT)水平均高于CHB组和LC组(P<0.05);50例HCC患者中,中低分化及Ⅲ~Ⅳ期患者血AFP和ALT水平高于高分化及Ⅰ~Ⅱ期患者(P<0.05);HBV-DNA高载量患者血AFP和ALT水平高于低载量患者(P<0.05),且患者HBV-DNA载量与其血AFP和ALT水平呈正相关(P<0.05)。结论HBV感染相关性HCC患者血AFP和ALT水平显著提升,且血AFP和转氨酶水平与HBV-DNA载量、肿瘤分期分化程度关系密切,二者对临床评估HBV感染相关性HCC病情进展有一定提示作用。

HBV感染患者HBV-DNA载量与乙肝五项指标的相关性研究

目的探讨乙肝病毒(Hepatitis B Virus,HBV)感染患者血清中乙型肝炎病毒脱氧核苷酸(HBV-DNA)载量与乙肝五项指标的相关性。方法选取沛县人民医院于2022年1月—2023年1月收治的100例HBV感染患者作为研究对象,所有患者均接受HBV-DNA载量与乙肝五项指标检测,并按照其HBV-DNA载量进行分组,分别为<50 IU/mL组69例、50~<10^(4) IU/mL组10例、10^(4)~10^(10) IU/mL组21例,对比3组检测结果。结果10^(4)~10^(10) IU/mL组乙型肝炎e抗体(Hepatitis B Virus e Antibody,HBeAb)、乙型肝炎e抗原(Hepatitis B e Antigen,HBeAg)、乙型肝炎表面抗原(Hapatitis B Surface Antigen,HBsAg)指标高于50~<10^(4) IU/mL组、<50 IU/mL组,50~<10^(4) IU/mL组各指标水平高于<50 IU/mL组,差异有统计学意义(P均<0.05)。相关性分析后显示,HBeAb、HBeAg、HBsAg与HBV-DNA载量呈正相关(r=0.512、0.549、0.673,P均<0.05)。结论HBV-DNA载量与HBeAb、HBeAg、HBsAg指标之间有正性关联,可为评估HBV感染程度提供一定参考依据。

血清肝纤维化指标、HBV-DNA病毒载量联合检验在乙型肝炎肝纤维化中的评估价值

目的:探究血清肝纤维化指标、HBV-DNA病毒载量联合检验在乙型肝炎肝纤维化中的评估价值。方法:选取2022年1月至2023年5月本院收治的100例乙型肝炎肝纤维化患者作为观察组,并选择同期100例慢性乙型肝炎患者作为对照组。分别采用化学发光法和荧光定量PCR法检测两组患者肝纤维化指标及HBV-DNA水平,并比较分析其与乙型肝炎肝纤维化的关系。结果:观察组透明质酸(HA)、层粘连蛋白(LN)、Ⅲ型前胶原(PCⅢ)、Ⅳ型胶原(Ⅳ-C)水平均显著高于对照组(P<0.05);HBV-DNA水平显著低于对照组(P<0.05)。结论:HBV在慢性乙型肝炎患者具有高度复制性,在肝纤维化阶段复制水平下降。随着乙肝的进一步发展,肝纤维化指标水平逐渐升高。联合检测肝纤维化指标和HBV-DNA病毒载量对临床诊断乙型肝炎肝纤维化有一定的参考价值。

NS5ATP9与HBx相互作用促进HBV cccDNA的形成与转录

目的探讨丙型肝炎病毒NS5A反式调节蛋白9(hepatitis C virus NS5Atransactivated protein 9,NS5ATP9)在乙型肝炎病毒(hepatitis B virus,HBV)共价闭合环状DNA(covalently closed circular DNA,cccDNA)形成与转录中的作用机制。方法利用1.3拷贝HBV表达质粒转染Huh7和HepG2细胞、整合有4拷贝HBV基因组的HepG2.2.15细胞、在诱导型四环素启动子控制下表达HBV的HepAD38细胞构建NS5ATP9过表达或干扰的HBV细胞模型,收集样品和细胞上清液,提取RNA、HBV核心DNA(coreDNA)、cccDNA和蛋白,利用酶联免疫吸附试验、实时荧光定量聚合酶链反应(polymerase chain reaction,PCR)、Southern blot和Western blot技术检测HBV总RNA、前基因组RNA(pregenomic RNA,pgRNA)、乙型肝炎病毒s抗原(hepatitis B virus s antigene,HBsAg)、乙型肝炎病毒e抗原(hepatitis B virus e antigene,HBeAg)、松弛环状DNA(relax circular DNA,rcDNA)以及cccDNA水平。在HepG2细胞中转染乙型肝炎病毒x蛋白(hepatitis B virus x protein,HBx),通过免疫荧光成像及免疫共沉淀方法检测NS5ATP9与HBx的结合情况。双荧光素酶报告基因实验检测NS5ATP9对HBx启动子活性的影响。利用Huh7细胞转染HBV1.3及HBV稳定表达细胞株HepG2.2.15和HepAD38转染NS5ATP9过表达/干扰质粒,通过Western blot技术检测DDB1和SMC6的蛋白水平。结果在HBV病毒活跃的细胞中,NS5ATP9 mRNA水平[HepG2.2.15细胞:1.891±0.567比1.00±0.034,t=2.87,P=0.0351;HepAD38 tet+细胞:1.978±0.399比1.00±0.034,t=4.131,P=0.0091;HepAD38 tet-细胞:2.642±0.672比1.00±0.034,t=4.127,P=0.0091]和蛋白水平均显著增加。过表达NS5ATP9后可显著增加HBeAg[(5.402±0.327)S/COV比(2.68±0.552)S/COV,t=7.35,P=0.0018]、HBsAg[(2.846±0.185)S/COV比(1.512±0.221)S/COV,t=8.02,P=0.0013]、HBV pgRNA及rcDNA的表达水平,而干扰NS5ATP9后此增加作用消失[HBeAg:(2.029±0.09)S/COV比(3.733±0.445)S/COV,t=6.501,P=0.0029;HBsAg:(1.501±0.105)S/COV比(1.878±0.174)S/COV,t=3.216,P=0.0324)]。机制研究显示,NS5ATP9和HBx蛋白主要位于细胞核核仁内,并具有共定位信号,且NS5ATP9可显著提高HBx启动子(1071.06±79.44比488.47±40.12,t=13.09,P=0.00012)的转录活性。另外,过表达NS5ATP9可显著降低DDB1和SMC6的蛋白水平,而沉默NS5ATP9则可显著提高DDB1和SMC6的蛋白水平。结论HBV上调NS5ATP9的表达,形成HBV-NS5ATP9-HBV cccDNA-HBV的正反馈环路,NS5ATP9通过与HBx相互作用上调肝细胞中HBV cccDNA的形成与转录,进而促进慢性乙型肝炎的发生发展。

慢性乙型肝炎患者HBV DNA载量与血清miR-122、miR-223水平的关系

目的分析慢性乙型肝炎(CHB)患者乙型肝炎病毒脱氧核糖酸(HBV DNA)载量与血清微小RNA-122(miR-122)、微小RNA-223(miR-223)水平的关系。方法回顾性分析郑州市第七人民医院2022年6月—2023年3月收治的540例CHB患者的临床资料。依据血清HBV DNA载量分为低载量(<10^(5)拷贝/mL)组(n=230)、中载量(10^(5)~10^(7)拷贝/mL)组(n=180)、高载量(>10^(7)拷贝/mL)组(n=130),另选取同期于我院体检中心体检的健康者300名设为对照组。比较不同HBV DNA载量组血清miR-122、miR-223水平,绘制ROC曲线评价血清miR-122、miR-223诊断CHB的效能,采用多因素Logistic回归分析CHB的危险因素,采用Pearson相关性分析HBV DNA载量与血清miR-122、miR-223水平的关系。结果三组血清miR-122(1.45±0.37、2.84±0.72、4.11±0.90)、miR-223(1.34±0.33、1.69±0.37、1.91±0.45)比较差异有统计学意义(F=716.128、104.744,P<0.05);CHB组患者的血清miR-122(2.56±0.49)、miR-223(1.79±0.42)及HBV DNA载量>10^(5)拷贝/mL的人数占比(310/540)显著高于对照组(1.07±0.21、1.05±0.30、75/300)(t=51.844、26.935,χ^(2)=81.585,P<0.05);经ROC分析证实,血清miR-122、miR-223水平均可用于预测CHB,曲线下面积分别为miR-122(AUC=0.794,95%CI:0.690~0.898)、miR-223(AUC=0.813,95%CI:0.720~0.906),均有P<0.05;多因素logistic回归分析显示,miR-122≥1.528、miR-223≥1.210及HBV DNA载量>105拷贝/mL是CHB的危险因素,OR值分别为2.011(95%CI:1.211~3.339)、1.696(95%CI:1.026~2.804)、2.117(95%CI:1.974~3.987)均有P<0.05;相关性分析显示,血清miR-122、miR-223水平与HBV DNA载量呈正相关(r=0.753、0.712,P<0.05)。结论血清miR-122、miR-223水平与HBV DNA载量呈正相关,且miR-122≥1.528、miR-223≥1.210及HBV DNA载量>10^(5)拷贝/mL是CHB的危险因素,可将以上指标作为诊断CHB的生物学标志物,从而为临床病情评估和治疗提供参考。

血清HMGB1、GP73、IL-37与HBV-ACLF患者HBV-DNA载量的关系及对预后的预测价值

目的探究乙型肝炎病毒相关慢加急性肝衰竭(HBV-ACLF)患者血清高迁移率族蛋白B1(HMGB1)、高尔基体蛋白73(GP73)、白细胞介素-37(IL-37)与HBV-DNA载量的关系及对预后的预测价值。方法选取2018年7月至2023年7月中国人民解放军联勤保障部队第九八〇医院收治的112例HBV-ACLF患者的临床资料进行回顾性分析,根据HBV-CALF患者入院90 d内临床结局分为死亡组(42例)与生存组(70例)。比较两组临床资料及血清HMGB1、GP73、IL-37水平,分析HBV-CALF患者预后的影响因素,分析血清HMGB1、GP73、IL-37与HBV-DNA载量的关系,评价血清HMGB1、GP73、IL-37对HBV-CALF患者预后的预测价值。结果死亡组在病情分期、腹膜炎、肝性脑病、肺部感染、HBV-DNA载量、终末期肝病评分模型(MELD)评分方面与生存组比较,差异有统计学意义(P<0.05);死亡组血清HMGB1、GP73、IL-37水平均较生存组高(P<0.05)。病情分期、腹膜炎、肝性脑病、肺部感染、HBV-DNA载量、MELD评分及血清HMGB1、GP73、IL-37水平均为HBV-ACLF患者预后的独立影响因素(P<0.05)。HBV-ACLF患者血清HMGB1、GP73、IL-37水平均与HBV-DNA载量呈负相关(P<0.05)。血清HMGB1、GP73、IL-37预测HBV-ACLF患者预后的曲线下面积(AUC)分别为0.781、0.790、0.782,三者联合预测的AUC最大,为0.944,灵敏度、特异度分别为86.33%、91.43%。结论血清HMGB1、GP73、IL-37水平与HBV-CALF患者HBV-DNA载量及预后密切相关,3项指标联合检测有助于提高对预后的预测效能。

血清PIVKA-Ⅱ、AFP与HBV-DNA联合检测对HBV所致肝癌的诊断及预后预测价值

目的探究血清异常凝血酶原Ⅱ(PIVKA-Ⅱ)、甲胎蛋白(AFP)与乙肝病毒脱氧核糖核酸(HBV-DNA)联合检测对HBV所致肝癌(HCC)的诊断及预后预测价值。方法选取2018年8月至2020年7月在本院接受治疗的98例HCC患者作为研究对象(肝癌组),另取同期95例体检健康人群作为健康组,95例肝硬化患者作为肝硬化组,观察3组受试者血清PIVKA-Ⅱ、AFP与HBV-DNA表达水平和一般资料差异。根据肝癌组3年内预后情况将患者分为生存组(57例)和死亡组(41例)。比较肝癌组一般资料和血清PIVKA-Ⅱ、AFP与HBV-DNA水平关系;多因素Logistic和COX回归分析分别分析影响受试者患肝癌和患者预后不良的影响因素;四格表法计算血清PIVKA-Ⅱ、AFP与HBV-DNA水平对HCC的预测价值;ROC曲线分析评估血清PIVKA-Ⅱ、AFP与HBV-DNA水平对HCC患者预后的预测价值。结果肝癌组患者血清PIVKA-Ⅱ、AFP与HBV-DNA水平显著高于健康组和肝硬化组(P<0.05);HCC发病与血清PIVKA-Ⅱ、AFP与HBV-DNA水平有关,且是危险因素(P<0.05)。HCC患者预后不良与血清PIVKA-Ⅱ、AFP与HBV-DNA水平以及肿瘤数量有关(P<0.05),且是危险因素。血清PIVKA-Ⅱ、AFP与HBV-DNA水平以及3项联合诊断HCC发病的准确度分别为77.43%、72.57%、77.43%和84.72%。血清PIVKA-Ⅱ、AFP与HBV-DNA水平以及3项联合诊断HCC预后不良的AUC分别为0.823、0.841、0.824和0.958,3项联合诊断效能优于单一诊断(P<0.05)。结论血清PIVKA-Ⅱ、AFP与HBV-DNA水平在HCC患者和预后不良患者中呈高表达,且3项联合可有效预测HCC发病和HCC患者预后情况。

乙肝大小三阳状态、不同HBVDNA载量孕妇肝功能指标水平分析

目的研究探讨乙肝孕妇大三阳和小三阳状态中乙型肝炎病毒(Hepatitis B virus,HBV)DNA载量、肝功能指标、肝纤维化标志物含量及不同HBVDNA载量孕妇的肝功能水平。方法选取临清市人民医院于2020年1月—2023年1月收治的120例乙肝孕妇作为研究对象,根据乙肝两对半检测结果将其分为两组,即大三阳组(n=60)和小三阳组(n=60)。比较两组HBVDNA载量、肝功能指标、肝纤维化标志物含量及不同HBVDNA载量孕妇的肝功能指标、肝纤维化标志物含量。结果大三阳组丙氨酸转移酶水平(29.00±7.62)U/L、HBVDNA水平(5.18±1.16)×10^(4)U/mL均高于小三阳组的(26.00±2.85)U/L、(2.08±1.11)×10^(4)U/mL,差异有统计学意义(t=2.856、14.956,P均<0.05)。大三阳组的重组人几丁质酶3样蛋白1水平低于小三阳组,差异有统计学意义(P<0.05)。伴随着HBVDNA载量上升,两组患者甲胎蛋白水平随之增加,但是在同等HBADNA载量下,两组甲胎蛋白水平比较,差异无统计学意义(P>0.05)。结论开展孕前抗乙肝病毒诊疗、孕期风险系数评估,采取科学有效的干预措施,均能够有效抑制乙肝在母婴中的传播。

Evolution of HBV Viral Load during Clinical and Biological Follow-Up of Chronic Hepatitis B Patients at the Saint Camille Hospital in Ouagadougou

Hepatitis B virus (HBV) infection is a major public health problem worldwide. The aim of this study was to document the dynamics of HBV viral load during the follow-up of chronic hepatitis B patients at the Saint Camille Hospital in Ouagadougou (HOSCO) from 2017 to 2021. This descriptive retrospective study was carried out in the Hepato-Gastro-Enterology Department of HOSCO and focused on patients who were undergoing treatment for chronic viral hepatitis B. A total of 260 cases of chronic hepatitis B were included in the study. The most affected age group was 21 to 30 years, accounting for 48.08% of the cases. Lifestyle factors included alcohol consumption (3.08%) and tobacco use (2.69%). Major risk factors for transmission included lack of vaccination (98.46%), family history of HBV infection (68.00%) and engagement in high-risk activities (28.00%). Patients requiring treatment were prescribed Tenofovir 300 mg tablets. FibroScan® showed the presence of stage F3-F4 fibrosis (2.14%) and S3 steatosis (13.33%). After one year of follow-up, 6.92% of patients achieved an undetectable viral load with normalized transaminase levels. The majority of other patients had a detectable viral load but below 20,000 IU/mL. The prevalence of viral hepatitis B remains significant worldwide. Although effective and well-monitored treatment can lead to undetectable viremia, prevention remains the most effective strategy for successful management of this disease.

DNA-based vaccination induces humoral and cellular immune responses against hepatitis B virus surface antigen in mice without activation of Cmyc

AIM To develop a safe and effective DNAvaccine for inducing humoral and cellularimmunological responses against hepatitis Bvirus surface antigen(HBsAg).METHODS BALB/c mice were inoculated withNV-HB/s,a recombinant plasmid that had beeninserted S gene of hepatitis B virus genome andcould express HBsAg in eukaryotes.HBsAgexpression was measured by ABC immunohis-tochemical assay,generation of anti-HBs byELISA and cytotoxic T lymphocyte(CTL),byMTT method,existence of vaccine DNA bySouthern blot hybridization and activation ofoncogene C-myc by in situ hybridization,RESULTS With NV-HB/s vaccination byintramuscular injection,anti-HBs was initiallypositive 2 weeks after inoculation while all micetested were HBsAg positive in the muscles.Thetiters and seroconversion rate of anti-HBs weresteadily increasing as time went on and weredose-dependent.All the mice inoculated with100 μg NV-HB/s were anti-HBs positive onemonth after inoculation,the titer was 1:1024 ormore.The humoral immune response was similarinduced by either intramuscular or intradermalinjection.CTL activities were much stronger(45.26%)in NV-HB/s DNA immunized mice as compared with those(only 6%)in plasma-derived HBsAg vaccine immunized mice.Twomonths after inoculation,all muscle sampleswere positive by Southern-blot hybridization forNV.HB/s DNA detection,but decreased to 25%and all were undetectable by in situhybridization after 6 months.No oncogene C-myc activation was found in the muscle ofinoculation site.CONCLUSION NV-HB/s could generatehumoral and cellular immunological responsesagainst HBsAg that had been safely expressed insitu by NV-HB/s vaccination.