HBV突变与HBV相关慢加急性肝衰竭相关性研究

目的探讨HBV突变与慢加急性肝衰竭(acute-on-chronic liver failure,ACLF)之间的相关性。方法以87例HBV相关ACLF患者为研究对象,另选取52例慢性乙型肝炎(chronic hepatitis B,CHB)患者和51例HBV肝硬化(liver cirrhosis,LC)患者为对照,应用聚合酶链反应及基因测序对患者血清标本进行HBV基本核心启动子区、前C区和C区13个位点的突变检测,分析HBV突变与ACLF发生的相关性。结果 CHB、LC、ACLF组人均核苷酸变异检出位点分别为2.31、3.55和3.86个。nt1762、nt1846、nt1899和nt1913 4个突变位点的核苷酸突变率比较,ACLF组高于CHB组;nt1764位点突变率组间比较LC组高于ACLF组,而nt1846和nt1913位点突变率比较LC组低于ACLF组;nt1753、nt1762、nt1764和nt1899 4个突变位点的核苷酸突变率比较,LC组高于CHB组。多因素分析提示A1846T和C1913(A/G)核苷酸突变是预测ACLF的有效指标。结论 CHB或LC患者HBV基因序列的A1846T和C1913(A/G)核苷酸突变可能与其发生ACLF密切相关。     

慢性HBV感染者不同病程阶段HBV前S基因缺失突变的比较分析

目的 探讨慢性HBV感染者不同病程阶段前S(pre-S)基因缺失的临床流行特点及其临床意义. 方法 采用巢式PCR方法扩增测序146例慢性乙型肝炎(chronic hepatitis B, CHB)患者(CHB组)、111例HBV相关肝硬化(liver cirrhosis, LC)患者(LC组)、146例慢加急性肝衰竭(acute-on-chronic liver failure, ACLF)患者(ACLF组)和136例HBV相关肝细胞癌(hepatocellular carcinoma, HCC)患者(HCC组)的HBV pre-S基因(nt 2848-154).分析比较不同组pre-S基因缺失发生率、缺失热点区域和缺失片段长度. 结果 LC组和HCC组HBV pre-S基因缺失率显著高于CHB组(26.1%vs. 15.8%,P=0.040;34.6%vs. 15.8%,P<0.001);LC组pre-S1基因单独缺失率显著高于CHB组(17.1%vs. 4.8%,P=0.001);HCC组pre-S2基因单独缺失率显著高于CHB组(19.1%vs. 4.8%,P<0.001). 不同病程阶段患者发生缺失的热点区域也不相同,CHB组发生缺失的热点区域为nt 3031-3215(30.4%)和nt 24-57(30.4%);LC组为nt 2849-2866(55.2%)和nt 5-55(31.0%);ACLF组为nt 2849-2866(28.6%)和nt 1-54(25.7%);HCC组为nt 5-55(57.4%)和nt 2849-2866(12.8%).不同病程阶段患者发生pre-S基因缺失的片段长度差异无统计学意义. 结论 慢性HBV感染者中,随着疾病进展HBV pre-S基因缺失率呈上升趋势,其中pre-S1基因缺失突变在LC患者中显著增高,pre-S2基因缺失突变在HCC患者中显著增高.pre-S基因缺失可能参与驱动HBV慢性感染的疾病进展.     

HBV感染所致慢加急性肝功能衰竭患者HBV变异的纵向研究

目的明确乙型肝炎病毒（HBV）变异与慢加急性肝功能衰竭（ACLF）发病之间的关系。方法采用纵向研究的方法 ,选取6例HBV感染者为研究对象,其中ACLF4例（P1～P4）,肝硬化（LC）2例（P5、P6）。分别留取每例患者不同疾病阶段血清2份,进行HBVDNA提取、扩增、克隆和测序,测序结果采用VectorNTISuite9.0软件分析,纵向对比HBV核苷酸序列变异情况。结果 6例患者均为基因C型。ACLF患者（P1～P4）在病程中HBV的核苷酸突变位点多于发生肝硬化患者（P5、P6）。ACLF患者中,nt53（C→T或T→C）、nt1846（A→T）、nt1896（G→A）的纵向突变见于2例或2例以上患者。C1913A或T突变可见于患者P1、P2,同时患者P2可见纵向突变。结论 nt53、nt1846、nt1896和nt1913四个核苷酸的突变可能与ACLF的发病相关。     

乙型肝炎病毒前S基因变异与乙型肝炎相关晚期肝病关联性的临床研究

目的研究乙型肝炎病毒(HBV)前S基因变异与乙型肝炎相关晚期肝病及慢加急性肝衰竭(ACLF)的关系。方法收集慢性乙型肝炎(CHB)、乙型肝炎相关性肝硬化(LC)、乙型肝炎相关性肝细胞癌(HCC)患者共71例的血标本及临床资料,提取血清HBV DNA,进行基因分型,PCR扩增HBV DNA前S基因,所得阳性PCR产物进行测序,分析测序结果。结果检测样本中基因型C49例,基因型B21例,混合基因型B+C1例,C为优势基因型。PreS变异率在CHB(不包括ACLF)、LC、HCC三组中分别为18.75%、48.78%、66.67%,CHB组PreS区变异率低于LC组(P0.05)和HCC组(P0.05)。ACLF组PreS区变异率(80%)明显高于CHB组(18.75%)(P0.005)。HBV DNA≥104拷贝/mL在发生PreS区变异组为30例(90.9%),未发生PreS区变异组为19例(50.0%),两组差异有显著统计学意义(P0.001)。Logistic回归分析示HBV DNA≥104拷贝/mL、HBV相关晚期肝病(LC、HCC)与PreS区变异相关(OR值:14.153,95%CI:3.412~58.701;OR值:3.924,95%CI:1.178~13.074)。结论 HBV前S区变异与乙型肝炎相关晚期肝病及肝衰竭的进展有关。     

HBV基因型、变异、X蛋白与HCC相关性的研究

【摘要】 目的  了解HBV病毒基因型、变异位点1762/1764和1896、X蛋白与肝细胞癌的关系,进一步探讨肝癌的发病机理,为肝癌的防治及早期诊断提供理论依据。 方法  采用核酸扩增荧光定量及测序法检测慢性HBV携带者、慢性乙型肝炎、乙肝肝硬化和乙肝合并肝癌患者共159例血清标本的HBV基因型及变异位点,采用免疫组织化学方法检测上述4组肝组织中X蛋白的表达情况,用半定量积分法进行结果判断。 结果  1. 在159例慢性HBV感染者中基因型B型为56例,占35.2％（56/159）,基因型C型为103例,占64.8％（103/159）,基因C型在ASC、CHB、LC、HCC四组中所占比例分别为44.0%（11/25）,63.2%（36/57）,85.7%（36/46）,64.5%（20/31）,差异有统计学意义（χ2 =8.462,P=0.037）。2. 在ASC、CHB、LC、HCC四组中,基因C型HBV感染者发生BCP变异的病例数所占比例分别为36.4%（4/11）,75.0%（27/36）,80.6%（29/36）,75.0%（15/20）,发生PC变异的病例数所占比例分别为45.5%（5/11）,66.7%（24/36）,77.8%（28/36）,70.0%（14/20）,两种变异均多于基因B型感染者,除ASC组外,差异均有统计学意义（P <0.05）。3. X蛋白的表达,以LC组最高（阳性率71.7%）,其次是HCC组（71.0%）、CHB组（59.6%）、ASC组（52.0%）,组间比较差异有统计学意义（P<0.05）。4. ASC组、CHB组、LC组和HCC组基因C型HBV感染者X蛋白的阳性表达率分别为81.8%（9/11）,72.2%（26/36）,86.1%（31/36）和85.0%（17/20）,大于基因B型感染者X蛋白的阳性表达率,即28.6%（4/14）,38.1%（8/21）,20.0%（2/10）,45.5%（5/11）,差异有统计学意义（P<0.05）。ASC、CHB、LC和HCC组中X蛋白阳性表达的HBV感染者BCP变异阳性的比例分别为61.5%,73.5%,69.7%,81.8%,均高于BCP变异阴性感染者所占比例,PC变异阳性的比例分别为46.2%,67.6%,78.8%,63.6%,除了ASC组外均高于PC变异阴性感染者所占比例,但差异无统计学意义。 结论  HBV基因型、变异位点1762/1764和1896、X蛋白之间存在相互关系,与肝细胞癌的发生与发展有关。     

慢性HBV基因型C型感染者临床特征分析

目的:在了解延安地区乙型肝炎病毒基因型分布特征的基础上,进一步探讨延安地区HBV基因型C型感染者的临床分型与病毒学和生化学指标的特征.方法:随机选取HBV DNA阳性慢性乙型肝炎患者122例(病史均超过12 mo),在可检出基因型121例患者中,慢性无症状HBV携带者(ASC)58例,慢性乙型肝炎(CHB)46例,肝硬化(LC)17例,采用乙型肝炎病毒核酸扩增荧光定量检测HBV DNA载量;时间分辩荧光免疫分析(TRFIA)技术定量检测乙肝血清标志物,微板核酸杂交-ELISA法测定HBV基因型.结果:在121例可检出基因型的患者中,C型116例(95.87%-95%CI:92.32%-99.42%).C基因型感染者中:ASC组HBV DNA载量、HBeAg定量明显高于CHB组及LC组(t=3.94、4.75,P=0.000,0.000;t=4.82、3.94,P=0.000,0.001):ASC组HBeAb定量、ALT水平明显低于CHB组及LC组(t=3.71、12.09,P=0.000、0.000;t=12.44、7.81.P=0.000、0.000).CHB组与LC组HBV DNA载量、HBeAg定量、ALT水平明没有明显差异(P=0.38,0.58,0.047);ASC组、CHB组、LC组年龄之间比较差异明显(t=5.07、9.72、4.99,P=0.000、0.000、0.000),ASC组年龄最低;CHB组、LC组男性比例明显高于ASC级(X2=9.37,5.18,P=0.002,0.023).结论:HBV基因型C型是延安地区慢性HBV感染者优势株:C型感染者ASC组、CHB组、LC组间分析结果基本符合慢性HBV感染者临床自然史转归特征.     

慢性HBV感染者抗病毒治疗前HBV基本核心启动子及前C区突变的检测及意义

目的研究慢性乙型肝炎病毒（HBV）感染者抗病毒治疗前HBV基本核心启动子（BCP）突变和前C区（PreC）突变与HBeAg、HBV DNA水平和慢性肝病进展的关系。方法收集283例慢性HBV感染者抗病毒治疗前的血清标本,其中慢性乙型肝炎（CHB）185例,肝硬化（LC）98例。采用PCR后直接测序法检测HBV BCP和PreC区突变,同时确定基因型。结果在HBeAg阴性和HBeAg阳性CHB患者中,前C区A1896变异率分别为44.6%（37/83）和21.6%（22/102）（χ2=11.154,P=0.001）,LC患者分别为43.4%（23/53）和17.0%（8/47）（χ2=8.101,P=0.004）。在HBeAg阳性患者中,BCP T1762/A1764双突变率LC组和CHB组分别为89.4%（42/47）和70.6%（72/102）（χ2=6.310,P=0.012）。在单变量分析中,只有年龄（≥45岁）（χ2=27.861,P〈0.001）、BCP T1762/A1764双突变（χ2=8.675,P=0.003）和HBV DNA（≥105拷贝/ml）（χ2=20.499,P〈0.001）与LC进展有关。多因素Logistic回归分析（匹配年龄和性别）发现,BCP T1762/A1764双突变（OR=3.260,95%CI：1.401～7.586;wald=7.517,P=0.006）和HBV DNA（≥105拷贝/ml）（OR=4.640,95%CI：2.331～9.237;wald=19.089,P〈0.001）是LC进展的危险因素。结论前C区A1896突变与HBeAg的消失有关;年龄（≥45岁）、BCP T1762/A1764双突变和HBV DNA高载量（≥105拷贝/ml）与肝硬化进展有关。     

慢性HBV感染前C区变异与病毒复制水平关系

探讨HBV前C基因变异与病毒复制水平的关系在慢性HBV感染者中的意义。应用错配聚合酶链反应（PCR）－限制性片段长度多态性（RFLP）分析与荧光定量聚合酶链反应检测HBVDNA相结合,对30例HBsAg（＋）、抗－HBe( )及抗－HBc( )慢性HBV感染者,其中无症状携带者（AsC)9例、慢性乙型肝炎（CHB）12例及慢性重症肝炎（CHF）9例进行前C区基因变异与HBVDNA水平关系进行分析。AsC组3例（33．33％）,CHB组9例（75％）及CHF组8例（88．89％）有A83(nt1896)变异。荧光定量PCR结果表明HBVDNA含量在CHF组中最高。HBV前C变异与HBV不同感染状态中都可见,其病毒复制水平与肝病活动相关。     

慢性乙型肝炎和肝硬化患者血浆miR-181a水平变化*

目的 探讨慢性HBV感染者血浆微小RNA(miRNA)-181a的变化。方法 采用实时荧光定量PCR法检测20例乙型肝炎病毒携带者（ASC）、20例慢性乙型肝炎（CHB）患者、20乙型肝炎肝硬化（LC）患者和20例健康人血浆miR-181a水平,常规检测血清HBV DNA。结果 NC组、ASC组和LC组血浆miR-181a水平分别为（1.01±0.03）、（0.97±0.02）、（0.91±0.02）,而CHB组患者为（0.74±0.03）,显著低于前三组（P<0.001）;ASC组和LC组血清HBV DNA（3.86±2.89）lg copies/ml和（2.47±3.26）lg copies/ml,而CHB组患者为（8.96±1.63）lg copies/ml（P<0.001）;CHB患者血浆miR-181a水平与HBV DNA水平呈负相关（r=0.76,P<0.001）,ASC组和LC组血浆miR-181a水平与HBV DNA水平无显著相关性。结论 CHB患者血浆miR-181a水平降低,其意义还需要进一步探讨。     

慢性乙型肝炎、乙型肝炎肝硬化及原发性肝癌患者HBV-X基因突变分析

目的研究慢性HBV感染者,如慢性乙型肝炎(CHB)、乙型肝炎肝硬化(LC)、原发性肝癌(PLC)患者血清中HBV-X基因序列的突变与肝癌发生的关系。方法收集2011-2013年间于重庆医科大学附属第二医院就诊的慢性HBV感染者血清共89例,从血清中提取HBV DNA,扩增全长HBV-X基因序列,经测序后与已知HBV-X基因相应序列比较该患者体内HBV-X基因变异位点以及变异形式,并用卡方检验、单因素方差分析处理数据,NCBI的genotype工具测定基因型。结果所有患者均属于B/C基因型,HBeAg阳性患者中B基因型占46.2%,C基因型占53.8%;HBeAg阴性患性中B基因型占81.2%,C基因型占18.8%(P=0.001)。在PLC组中,启动子(BCP)区的突变显著高于CHB、LC(69.2%vs34.4%和61.3%,P0.05),且nt1821位点存在明显的T碱基的缺失(88.5%vs 53.1%和71%,P=0.014)。在CHB、LC中,C基因型BCP的双突变率显著高于B基因型(61.5%vs 15.8%,P=0.007;83.3%vs 47.4%,P=0.045),HBV DNA低病毒载量(≤106拷贝/ml)中BCP的突变率较高病毒载量(106拷贝/ml)更显著(81.3%vs 47.9%,P=0.015)。结论 BCP区的双突变及nt1821位点的缺失可能与PLC的发生密切相关。     

Association of Hepatitis B Virus Mutations of A1846T and C1913A/G With Acute-on-Chronic Liver Failure Development From Different Underlying Chronic Liver Diseases

Background

As most HBV-related acute-on-chronic liver failure (ACLF) have concurrent cirrhosis, it is important to clarify the association of viral factors with ACLF with or without cirrhosis.

Objectives

The aim of this study was to analyze the association of HBV genotypes and mutations with ACLF development underlying different chronic liver diseases.

Patients and Methods

Eighty-seven ACLF patients including 29 patients with chronic hepatitis (ACLF-CHB) and 58 patients with liver cirrhosis (ACLF-LC) were enrolled. Age and sex matched patients with chronic hepatitis (CHB) and liver cirrhosis (LC) were enrolled as controls. The genotypes and mutations at HBV basic core promoter (BCP), precore (PC), and partial C regions were determined by nested PCR and direct sequencing.

Results

Our results revealed significantly higher incidences (P < 0.05) of genotype B with C1913A/G or A1846T in patients with ACLF-CHB than those with CHB; genotype C with C1913A/G or A1846T in patients with ACLF-CHB and ACLF-LC than those with CHB and LC, respectively. Multivariable analysis indicated that A1846T and C1913A/G mutations were independent factors for ACLF (OR = 2.86 and 5.93, respectively), suggesting an association between the mutations and development of ACLF. In addition, there were no significant differences in mutations at T1753V, A1762T, G1764A, G1896A, and G1899A which were found between either CHB and ACLF-CHB or LC and ACLF-LC patients， suggesting no associations of these mutations with ACLF development.

Conclusions

Our findings suggest that CHB or LC patients infected with HBV A1846T and C1913A/G mutants are more susceptible to develop ACLF.     

Hepatitis B virus genotype B and mutations in basal core promoter and pre-core/core genes associated with acute-on-chronic liver failure: a multicenter cross-sectional study in China

Background and aim

In China, acute-on-chronic liver failure (ACLF) is mostly caused by hepatitis B virus (HBV). However, the mechanism remains unclear. This study aims to investigate the association between both HBV genotype and mutations in basal core promoter (BCP) and pre-core/core (pre-C/C) regions with the development of HB-ACLF.

Methods

A multicenter cross-sectional study was performed in China. Serum samples from 522 patients were analyzed, including 231 patients with mild-chronic hepatitis B (CHB-M), 84 with severe-chronic hepatitis B (CHB-S) and 207 with HB-ACLF. HBV genotype and related mutations in the BCP and pre-C/C regions were determined by direct sequencing.

Results

A significantly higher ratio of HBV genotype B to C was detected in HB-ACLF patients than in CHB-M or CHB-S patients. The A1762T/G1764A, A1846T and G1896A mutations were significantly more common in HB-ACLF patients infected with either genotype B or C as compared with CHB-M, whereas the C1913A/G and A2159G mutations were more associated with HB-ACLF in genotype C patients. Comparing with CHB-S, the A1762T/G1764A mutation in genotype B and the A2159G mutation in genotype C were significantly more common in HB-ACLF patients. A multivariate analysis showed that factors such as HBV genotype B, age ≥40 years and A1762T/G1764A, A1846T and G1896A mutations were independently associated with the development of HB-ACLF.

Conclusion

Chronic HBV infection with genotype B, A1762T/G1764A, A1846T and G1896A mutations has a higher possibility to develop HB-ACLF. These virological factors could serve as possible molecular markers for prediction of the clinical outcomes of chronic HBV infection.     

慢性乙型肝炎、乙肝肝硬化、乙肝肝癌患者Th1/Th2型细胞因子水平变化研究

目的分析慢性乙型肝炎(CHB)、乙肝肝硬化(LC)、乙肝肝癌(HCC)患者血清中Th1/Th2型细胞因子水平变化,为慢性乙型肝炎至肝癌的发生发展过程中的免疫变化研究提供线索,并为患者的临床治疗研究提供免疫学指标。方法选取2010年2月-2011年11月于首都医科大学附属北京佑安医院就诊的40例CHB患者、40例LC患者、53例HCC患者,用Luminex技术检测血清中Th1类细胞因子[白细胞介素(interleukin,IL)-12、干扰素(interferon,IFN)-γ和肿瘤坏死因子(tumor necrosis factor,TNF)-α]水平及Th2类细胞因子(IL-4、IL-6和IL-10)水平,并将25例健康志愿者作为正常对照组进行比较。结果除TNF-α外,CHB组、LC组和HCC组Th1类IL-12、IFN-γ和Th2类IL-4、IL-6、IL-10细胞因子水平均低于正常对照组;CHB组中大部分Th1类和Th2类细胞因子都高于LC组和HCC组;HCC组中TNF-α细胞因子水平要高于CHB组、LC组。结论 CHB、LC和HCC患者体内Th1/Th2细胞因子分泌水平受到抑制,Th1/Th2平衡发生漂移,对HBV病毒的清除作用受到抑制。TNF-α在肝癌发生过程中发挥着重要作用。     

Clinical and virological implications of A1846T and C1913A/G mutations of hepatitis B virus genome in severe liver diseases

Objective: Mutations occurring within different genes of hepatitis B virus (HBV) genome may have different clinical implications. This study aimed to observe the clinical and virological implications of the A1846T and C1913A/G mutations of HBV genome in the development and treatment outcome of severe liver diseases, which has not been previously determined.

Materials and methods: A total of 438 cases of patients with liver diseases were retrospectively reviewed, including 146 with mild chronic hepatitis B infection (CHB-M), 146 with severe chronic hepatitis B infection (CHB-S), and 146 with acute-on-chronic liver failure (ACLF). Partial or full-length HBV genome was directly sequenced. Replicons containing A1846T, C1913A or other mutant sequences, or the wild-type counterparts were constructed respectively, and then transfected into HepG2 cells for phenotype analysis.

Results: There was significant difference in the detection rates of A1846T (30.82%, 40.41% and 55.48%, respectively) and C1913A/G (15.52%, 28.77%, and 35.62%, respectively) among patients with CHB-M, those with CHB-S, and those with ACLF (p?p?=?.041). In vitro experiment revealed that A1846T mutant resulted in 3.20-fold and 1.85-fold increase of replication capacity and promoter activity, respectively compared with wild type counterpart (p?p?Conclusion: Occurrence of A1846T and C1913A is positively associated with clinical presentations of severe liver disease. A1846T mutation is significantly associated with poor prognosis of ACLF.     

Virological analysis, genotypes and mutational patterns of the HBV precore/core gene in HBV/HCV-related hepatocellular carcinoma

We investigated the replicative profile of hepatitis B (HBV) and hepatitis C (HCV) viruses and the mutational pattern of the HBV precore/core (pre-C/C) domain in hepatocellular carcinoma (HCC). Thirty-eight consecutive patients with HCC were included in the study - 18 of them with HBV/HCV co-infection and 20 with HBV single infection. Twenty-three additional patients with co-infection, without HCC were recruited as the control group. Replication activity was evaluated by detecting and quantitating both HBV and HCV genomes. The HBV pre-C/C region, encompassing the pregenome encapsidation signal involved in viral replication, was analysed by direct sequencing. HBV viraemia levels were significantly lower (P = 0.04) in patients with co-infection in comparison with single-infected HCC, whereas two different HBV viraemia profiles were detected in co-infection with or without circulating HCV. HBV genotype D was prevalent in the three groups and HCV genotype 1b was found to be the infecting strain in all patients. Lower variability in the pre-C/C region was found in co-infection in comparison with HBV single infection (P = 0.0004). A synonymous T1936C mutation was found in all co-infected HCC cases not related to the presence or absence of circulating HCV, and a hypermutated pre-C strain, characterized by the same mutational pattern, was identified in three HCC cases. The mutational pattern of the pre-C/C region was closely related to HBV replication efficiency, and specific HBV mutations selectively associated with HCV co-infection could be linked with accelerated HBV/HCV-related disease progression.     

广州地区424例乙型肝炎患者病毒基因亚型分布与乙型肝炎疾病谱的关系探讨*

目的调查广州地区乙型肝炎病毒（HBV）基因亚型分布情况, 并探讨其与HBV感染疾病谱的关系。方法采用直接测序法测定S基因序列并构建基于S基因进化树,对424例HBV感染者进行研究,随机选择慢性乙型肝炎（CHB）137例,HBV相关慢加急性肝衰竭（HB-ACLF）90例,肝硬化（LC）90例和肝细胞癌（HCC）107例,应用SPSS 13.0软件进行x²检验、方差分析和多元Logistic回归分析。结果在424例HBV感染者中,B2亚型 269例（63.44%）,C1亚型117例（27.59%）,C2亚型36例（8.49%）,C5亚型 2例（0.47%）,未发现其他亚型;CHB患者感染HBV B2亚型、C1亚型、C2亚型和C5亚型分别为70.1%、24.09%、5.11%和0.73%;HB-ACLF患者感染HBV B2亚型、C1亚型和C2亚型分别为87.78%、7.78%、4.44%,其中HBV B2基因亚型显著高于CHB（x²=9.641,P=0.002）、LC（x²=19.565,P=0.000）、HCC患者（x²=26.789,P=0.000）;LC患者感染HBV B2亚型、C1亚型和C2亚型分别为50.00%、36.67%、13.33%,其中HBV C1和C2亚型显著高于CHB患者（x²=6.262,P=0.012;x²=4.790,P=0.029）或HB-ACLF患者（x²=25.894,P=0.000;x²=4.390,P=0.036）;HCC患者感染HBV B2亚型、C1亚型、C2亚型和C5亚型分别为45.79%、41.12%、12.15%、0.93%,其中HBV C1和C2亚型也显著高于CHB患者（x²=11.264,P=0.001;x²=3.957,P=0.047）或HB-ACLF患者（x²=28.327,P=0.000;x²=3.904,P=0.048）;LC和HCC患者HBV C1和C2基因亚型无明显差异（x²=0.429,P=0.512;x²=0.804,P=0.833）;多因素Logistic回归分析提示B2亚型是HB-ACLF发生的危险因素（OR=2.597,95%CI=1.145~5.891,P=0.022）,C型是HCC发生的危险因素（OR=3.257,95%CI=1.49~7.194,P=0.003）。结论广州地区HBV基因亚型主要由B2、C1和C2亚型构成,同时也存在C5亚型;广州地区B2亚型感染者可能更易发生HBV相关慢加急性肝衰竭,而C1和C2亚型感染者可能更易进展为肝硬化和肝细胞癌。     

HBV基因型和S基因序列特点与慢性乙型肝炎患者HBsAg/抗-HBs共存关系

目的探讨HBV基因型及S基因序列特点与慢性乙型肝炎（CHB）患者HBsAg和抗-HBs共存的内在相关性以及HBsAg＋/抗-HBs＋的发生机制及意义。方法共收集49例HBsAg＋/抗-HBs＋CHB患者的血清HBVDNA样本,采用PCR-RFLP及测序法鉴定其基因型,并与267例HBsAg＋/抗-HBs-CHB患者的HBV基因型分布进行比较。对6例HBsAg＋/抗-HBs＋CHB患者（Ⅰ组）的HBVS基因进行克隆测序,并与HBsAg＋/抗-HBs-CHB患者（Ⅱ组）进行对比,分析变异种类及频率等的差异。结果 HBsAg＋/抗-HBs＋CHB患者HBV基因型B、C及B/C混合感染的构成比分别为63.3%（31/49）、26.5%（13/49）、10.2%（5/49）,而对照组上述构成比分别为52.8%（141/267）、46.1%（123/267）、1.1%（3/267）,两组构成比差异有统计学意义（P〈0.01）。Ⅰ组HBsAg各区段,特别是主要亲水区（MHR）的变异位点明显多于Ⅱ组;发现了若干新变异、少见的W196和C69终止突变,1例患者检测到G145R变异。某些患者HBsAgMHR-2（包含a决定簇）并未发现变异。结论 HBV基因型的差异及HBsAg变异增多可能是部分CHB患者出现HBsAg＋/抗-HBs＋的原因之一;基因型B或B/C混合感染相对更易出现HBsAg＋/抗-HBs＋。HBsAg变异增多是CHB患者出现HBsAg＋/抗-HBs＋的重要机制之一。某些患者HBsAgMHR-2并无变异,提示HBsAg/抗-HBs共存还存在其他机制。     

慢性乙型肝炎肝硬化结节样变与病毒变异的关系

目的：分析慢性乙型肝炎肝硬化结节样变与 HBV 变异的关系以及 HBV 变异在慢性肝硬化、肝癌进展中的作用。方法收集临床诊断慢性乙型肝炎肝硬化患者104例,其中不典型结节样增生病例43例,单纯肝硬化患者41例,肝癌患者20例。采用荧光探针实时定量 PCR 试剂盒提取 HBV 基因组,选取 PCR 强阳性产物用 Sanger 双脱氧链末端终止法对 HBV 前 C 区变异基因 G1896A ,BCP 区 G1764A、A1762T 位点在全自动核苷酸分析仪上测序,测序结果与 Gene Bank 中 pADR 标准株序列作对比分析。结果结节性增生组、单纯肝硬化组、肝癌组患者 HBV 前 C Gl896A 变异检出率分别是52．3％、40．1％和37．4％,差异无统计学意义（χ2＝0．547,P ＝0．05）;结节性增生组 BCP A l762T 变异率68．4％,与单纯硬化组36．3％比较,差异有统计学意义（P ＝0．038）,与肝癌组 G1764A 变异检出率分别是73．0％,与对照组比较具有显著性差异（P ＝0．0411）。单纯肝硬化组、结节性增生组 BCP 基因变异（＋）组,HBV DNA 载量2．18×107、1．2×106高于基因变异（-）组1．18×104、2．95×103,差别有统计学意义（P ＝0．0451,P ＝0．0412）;结节性增生组、肝癌组 BCP 基因变异（＋）组HBeAg 定量750．00 IU／mL,1300．00 IU／mL 高于基因变异（-）组416．13 IU／mL,927．60 IU／mL 差别有统计学意义（P ＝0．0451,P ＝0．0073）。结论前 C 区变异与乙肝肝硬化结节样变临床进展为肝癌无关,而 BCP 区变异与乙肝结节样变临床进展为肝癌有关。