慢性乙型肝炎病毒感染S区基因缺失及相关因素研究

HBV为环状、部分双股的DNA病毒，慢性HBV感染可引起严重的肝脏疾病，临床表现为慢性肝炎、肝硬化、肝细胞肝癌及肝功能衰竭等。HBV—DNA的负链上有S、P、C及X4个开放的编码区，S区基因全长1167bp，由S基因、前S基因（Pre—S）组成，Pre-S包括Pre-S1／Pre-S2基因，各有其起始密码ATG。Pre-S还包含T、B淋巴细胞表位，它在HBV复制、宿主免疫反应及病毒进入肝细胞中具有重要作用。本文研究慢性HBV感染的不同肝脏病患者血清HBV全序列，分析S区基因序列缺失的模式、频率、临床相关因素。     

乙型肝炎病毒S基因变异与临床

变异是生物界普遍存在的现象，是物种适应生存压力的结果。乙型肝炎病毒(hepatitis B virus，HBV)与其它DNA病毒相比具有很高的变异率。随着研究的深入，表明乙型肝炎病毒S基因变异与乙肝疫苗预防接种失败、HBIG预防移植后肝脏再感染、HBsAg(-)的HBV感染及疾病严重性等均有密切关系。本文就HBV S基因变异及其在临床中的意义作一综述。     

乙型肝炎病毒DNA基因分型与相关慢性肝病关系的临床研究

人类感染不同基因型乙型肝炎病毒(HBV)及受宿主因素的不同影响，可引起不同的慢性肝脏疾病，包括慢性无症状乙型肝炎表面抗原携带者(ASC)、慢性乙型肝炎(CHB)、肝硬化(LC)及肝细胞癌(HCC)等。我们分析了我院264例慢性HBV感染者的HBV基因型，旨在探讨HBV基因型与HBV感染相关肝病的相互关系及临床意义。     

HBeAg阴性慢性乙型肝炎患者B、C基因型HBVS蛋白变异比较

目的 探讨乙型肝炎病毒(hepatitis B virus,HBV)基因型与S蛋白变异的关联,为分析病毒基因型在疾病预后转归中的作用提供理论基础.方法 选择上海中医药大学附属曙光医院东院2019年1月至2019年10月门诊或住院的HBeAg阴性慢性乙型肝炎患者99例,其中,B基因型50例,C基因型49例.基因测序法用于...     

慢性乙型肝炎患者HBV基因型和哑型分布调查

目的研究慢性乙型肝炎患者HBV基因型和亚型流行情况。方法应用HBV基因型和亚型特异性引物PCR法对北京、长春、大连、西安、石家庄、郑州和合肥7个城市660份HBV DNA阳性慢性乙型肝炎患者血清进行基因型和亚型分析。结果在660份HBV DNA阳性血清中,B基因型、C基因型和B／C混合感染分别为16．67％（110／660）、74．54％（492／660）和8．79％（58／660）;在C基因型中,C1亚型6例（1．22％）、C2亚型473例（96．14％）、C1／C2混合基因亚型13例（2．64％）;B基因型均为Ba亚型,B基因型和C基因型混合感染者均为Ba与C2亚型混合感染,未发现其他基因型和基因亚型;不同基因型感染患者HBeAg阳性率差异无统计学意义（P=0．153）;B基因型和C基因型患者之间血清HBV DNA水平差异无统计学意义（6．37±1．62lg copies／ml对6．29±1．76lg copies／ml）,但均高于B和C基因型混合感染患者（5．25±1．65lg copies／ml）。结论这7个城市慢性乙型肝炎患者以B基因型和C基因型感染为主,有部分B／C基因型混合感染。HBV亚型以Ba和C2亚型占优势。     

慢性乙型肝炎病毒感染者病毒前C区和基本核心启动子区变异检测及意义

目的 探讨乙型肝炎病毒（HBV）前C区和基本核心启动子（BCP）区变异与基因型及疾病进展间的关系。方法 收集HBV携带者（ASC）、慢性乙型肝炎（CHB）、肝炎肝硬化（LC）、肝细胞肝癌（HCC）患者血清148份，用半巢式聚合酶链反应扩增HBV前C／C基因部分片段，产物纯化后直接测序，检测前C区A1896及BCP区T1762／A1764变异。用S基因聚合酶链反应-限制性片段长度多态性分析（PCR-RFLP）方法确定HBV基因型。结果 有128份血清能够成功分型和测序，其中B基因型60份，C基因型68份。在B基因型感染者中前C区A1896变异检出率（48．33％）明显高于C基因型感染者（29．41％，X^2=4．83，P〈0．05）；而BCP区T1762／A1764变异检出率却明显低于C基因型感染者，差异亦有统计学意义（30．00％：73．54%，X^2=24．25。P〈0．05）。前C区A1896变异在CHB、LC、HCC中的阳性检出率分别为46．88％（15/32）、39．39％（13／33）、51．52％（17／33）。与ASC的13．33％（4／30）相比，P分别〈0．05，差异有统计学意义。BCP区T1762／A1764变异检出率在HCC、LC组分别为87．88％（29／33）和72．73％（24／33）．明显高于CHB组的37．50％（12／32）及ASC组10.00％（3／30）（P〈0．05）。结论 前C区A1896变异常见于B基因型感染者，而BCP区T1762/A1764变异C基因型感染者多见。除ASC外．前C区A1896变异与疾病进展关系不大．而BCP区T1762/A1764变异与乙型肝炎进展及顶后相关。     

肝细胞癌患者血清乙型肝炎病毒X区及前S/S区基因变异特点

H CC是常见的恶性肿瘤之一。众多流行病学资料显示，慢性HBV感染是HCC的主要病因。HBV感染致HCC的确切机制尚未彻底阐明，有学者认为HBV基因变异与肝癌发生发展密切相关!‘]。HBVC区变异与HBeAg血清转换及肝炎活动相关，与HCC发生关系不大121。而P基因区变异与抗病毒药物耐药有     

乙型肝炎病毒S区准种与疾病活动性的关系

目的　通过对慢性乙型肝炎不同病型患者S区准种复杂性差异的分析 ,探讨S区准种与疾病活动性的关系。方法　选择HBVDNA阳性患者共 112例 ,其中慢性重型肝炎 (FHF) 6 0例 (同时有肝硬化者 38例 ) ,慢性轻、中型肝炎 (CH) 30例 ,病毒携带者 (ASC) 2 2例 ,采用单链构像多态性(SSCP)和DNA序列分析方法检测HBVS区准种。结果　不同疾病期SSCP条带数不同 ,分别为ASC( 1.4 5± 0 .13) ,CH( 3.70± 0 .2 2 ) ,FHF( 5 .93± 0 .2 4 )。准种复杂性随疾病进展而增加 ,组间比较差异有显著性 ,F =72 .73,P <0 .0 1。HBeAg阳性与HBeAg阴性患者SSCP条带数分别为 3.4 1± 0 .2 7和 5 .2 7± 0 .30 ,两组差异有显著性 (t =4 .5 3,P <0 .0 1)。HBV基因型B、C患者SSCP条带数分别为 4 .71± 2 .36和 3.0 6± 1.76 ,差异有显著性 (t =2 .6 6 ,P <0 .0 1)。 2例重型肝炎患者随访 1年后丙氨酸转氨酶 (ALT)分别由 5 6 7.2U/L、4 6 2 .3U/L降至正常和接近正常 ( 4 2 .8U/L) ,SSCP条带数分别由 9、7条减至 5、2条 ;1例ASC随访 1年SSCP方式仍维持 1条带不变 ,ALT水平高低与SS CP条带数多少相一致。经DNA序列分析 ,证实了SSCP显示的HBV准种性。结论　HBVS基因准种随疾病病情加重而复杂性增加 ,其增加程度与HBeAg阴性、病情轻重及基因型     

慢性重型肝炎患者乙型肝炎病毒S基因区变异对其RNA二级结构的影响

重型肝炎患者由于肝功能衰竭和机体免疫功能低下，病死率较高，但目前的相关研究主要集中在前C区变异及e抗原的存在状态对肝炎进展的影响上，关于RNA方面的研究多见于ε环变异对病毒包装的影响。我们选取慢性重型肝炎患者的血清佯本，通过扩增HBVs基因及测序，应用软件模拟构建其RNA二级结构并考察其变化，以探讨慢性重型肝炎患者血清中HBV变异的存在情况及其对疾病的影响。     

乙型肝炎病毒相关慢性肝病前C区/BCP区基因突变的临床意义

实验采用PCR技术及芯片杂交原理，对65例乙型肝炎病毒(HBV)相关性慢性肝病患者进行前C区及BCP区A1896、A1899及nt1762／nt1764联合突变检测，探讨HBV相关性慢性肝病前C区／BCP区基因突变的临床意义。     

慢性HBV感染者肝脏HBV cccDNA含量相关因素分析

目的探讨慢性HBV感染者肝组织中HBV cccDNA含量与血清病毒标志物、HBV DNA及肝脏病理分级的关系,为临床评价抗病毒治疗效果及疗程确定提供理论依据。方法以2007年5月-2008年2月住院的30例慢性HBV感染者为研究对象,应用实时荧光定量聚合酶链反应（RT-PCR）方法检测患者肝组织中HBV cccDNA、肝组织总HBV DNA（HBV tDNA）和血清HBVDNA,同时用化学发光免疫分析法检测HBsAg、HBeAg定量,分析感染者肝组织内HBV cccDNA与肝组织内HBV DNA、血清HBVDNA、HBsAg及HBeAg定量水平之间的关系,并比较肝组织中HBV cccDNA含量与肝脏病理炎症和纤维化分级的关系。采用Pear-son简单相关和Spearman等级相关法进行相关性分析。结果 30例慢性HBV感染者肝组织中均可检出HBV cccDNA,范围在3.15×103～1.06×107拷贝/mg;肝组织cccDNA定量与肝组织总HBV DNA定量呈正相关（r=0.375,P〈0.05）,与血清HBV DNA无相关性（r=0.174,P〉0.05）;肝组织中HBV cccDNA水平与血清HBsAg定量呈高度正相关（r=0.562,P〈0.001）,而与血清HBeAg定量无相关性（r=0.152,P〉0.05）。肝组织cccDNA定量与肝组织炎症活动度（G）及纤维化程度（S）无相关性（r=0.082,P〉0.05）。结论慢性HBV感染者肝组织内HBV cccDNA成稳定的中等水平复制;血清HBV DNA载量不能直接代表其肝组织中的HBV cccDNA水平;血清HBsAg定量可作为反映肝组织中HBV cccDNA水平的指标。     

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.     

Double point mutation in the core promoter region of hepatitis B virus (HBV) genotype C may be related to liver deterioration in patients with chronic HBV infection

BACKGROUND AND AIM: Hepatitis B virus (HBV) genotype C has a more severe pathogenesis than genotype B in Japan. We retrospectively investigated the relationship between HBV genotype and the core promoter (CP) (nt 1762 and 1764) and precore (PreC) (nt 1896) mutations of the HBV genome. METHODS: A total of 129 Japanese patients (42 genotype B and 87 genotype C) with chronic HBV infection, living in two different geographical areas in Japan, were evaluated (mean follow-up period 10.1 +/- 3.8 years). In 2000, CP and PreC HBV mutations were analyzed by direct sequencing from sera. Hepatitis B e antigen (HBeAg), HBV DNA and serial alanine aminotransferase (ALT) changes were followed and determined using serological methods. RESULTS: Genotype C patients had significantly higher rates of HBeAg (40.2%vs 2.4%), HBV DNA positivity (75.9%vs 7.1%) and ALT abnormality (71.3%vs 11.9%) than genotype B patients (all P < 0.05). Among genotype B patients, CP wild type (92.9%) was predominant and PreC mutation (88.1%) was predominant. However, among genotype C patients, CP mutation (75.9%) was predominant and PreC mutation (66.7%) was predominant. The CP mutation was found significantly more in genotype C than in genotype B (P < 0.05). Of the 67 patients with ALT abnormality, five (7.5%) genotype B and 62 (92.5%) genotype C patients (31 HBeAg positive and 31 negative) were found. Among the 31 genotype C patients who were HBeAg positive, the combination of CP mutation and PreC wild (54.8%) was predominant, while among the remaining 31 genotype C patients who were HBeAg negative, the combination of CP mutation and PreC mutant (71.0%) was predominant. CONCLUSION: Genotype C might be one of the worse prognostic markers in patients with chronic HBV infection, possibly because of mutation in the CP region.     

Profile, spectrum and significance of HBV genotypes in chronic liver disease patients in the Indian subcontinent

BACKGROUND AND AIM: Certain hepatitis B virus (HBV) genotypes have been alleged to be associated with the development of cirrhosis and hepatocellular carcinoma (HCC), and the response to interferon therapy in Taiwanese patients. We undertook to study the prevalence and significance of HBV genotypes in the Indian subcontinent. METHODS: One hundred and thirty histopathologically proven chronic HBV-infected patients, including 52 incidentally detected asymptomatic hepatitis B surface antigen (HBsAg)-positive subjects (IDAHS) with chronic HBV infection (group I), 48 cirrhotics (group II) and 30 hepatocellular carcinoma (HCC; group III) patients were studied. Hepatitis B virus genotypes were determined by using restriction fragment length polymorphism, and direct sequencing of the s gene including the 'a' determinant region. RESULTS: Only genotypes A (46%) and D (48%) were found in the chronic HBV-infected patients. A mixed infection with genotypes A and D was seen in 6% of patients. Genotype A was found in 42, 48 and 50%, and genotype D in 48, 50 and 47% of group I, II and III patients, respectively (P = NS). The patients who had mixed genotypes were significantly younger (P < 0.05). In group I (IDAHS) patients infected with genotype D, none had a histological activity index (HAI) of < four. Genotype D was significantly more common in group I patients with HAI > 4 as compared to genotype A (53 vs 32%, P < 0.05). Similarly, genotype D was associated with more severe liver diseases (61 vs 30%, P < 0.05). Genotype D was more prevalent in HCC patients of < 40 years of age, as compared to IDAHS (63 vs 44%, P = 0.06). CONCLUSIONS: (i) Hepatitis B virus genotypes A and D are prevalent in chronic liver disease patients of Indian origin; and (ii) HBV genotype D is associated with more severe diseases and may predict the occurrence of HCC in young patients.     

HBV 相关慢性肝衰竭患者白介素-17 及白介素-6检测分析

目的：探讨HBV相关慢性肝衰竭（HBV related chronic liver failure，HBV-CLF）患者细胞因子表达及其与预后的关系。方法：采用流式液相多重蛋白定量技术检测HBV—CLF患者及慢性乙型肝炎（CHB）患者血清IL．17及IL-6水平，并对HBV—CLF患者进行为期24周随访，分为死亡组及存活组。结果：HBV-CLF患者IL-17及IL-6水平高于CHB患者（P〈0．05）；HBV—CLF患者死亡组IL-17水平高于存活组（P〈0．05），两组间IL-6水平之差异无显著性意义（P〉0．05）。结论：IL．17和IL-6可能与肝衰竭的发病机理有关，其中IL．17水平与患者预后有一定相关性。     

Distribution of hepatitis B virus genotypes among patients with chronic infection.

Hepatitis B virus (HBV) can be classified into at least eight genotypes, A-H. We evaluated the distribution HBV genotypes among patients with chronic infection. METHODS: We consecutively evaluated adult patients with chronic HBV infection from Salvador, Brazil. Patients were classified according to HBV infection chronic phases based on HBV-DNA levels and presence of serum HBV markers. HBV-DNA was qualitatively and quantitatively detected in serum by polymerised chain reaction (PCR). Isolates were genotyped by comparison of amino acid mutations and phylogenetic analysis. RESULTS: One-hundred and fourteen patients were evaluated. HBV-DNA was positive in 96 samples. HBV genotype was done in 76. Mean age was 36 +/- 11.3. In 61 of 76 cases subjects were classified as inactive HBsAg carriers. Their mean HBV serum level was 1760 copies/ml and 53 of 61 were infected with HBV genotype A, seven with HBV genotype F and one with genotype B. Twelve of the 76 patients had detectable hepatitis B e-antigen (HBeAg) in serum. Ten were infected with HBV genotype A and two with genotype F; most had increased alanine aminotransferase and high HBV-DNA levels. Three patients were in the immunotolerant phase, two were infected with HBV genotype A and one with genotype F. HBV subtyping showed subtypes adw2 and adw4. CONCLUSIONS: HBV genotype A adw2 and genotype F adw4 were the most prevalent isolates found. We could not find differences in genotype distribution according to HBV clinical phases and DNA levels. We did not detect HBV genotype D in contrast to a previous study in our center with acute hepatitis B. All inactive HBsAg carriers had low HBV-DNA levels.     

Sequence analysis of the S gene region in HBV DNA from patients positive for both HBsAg and HBsAb tests

Aims: To study the characteristics of mutation in the amino acids coded by the S gene region in the HBV DNA sequence and to comprehensively explore and analyze the cause of the double positive result phenomena in both HBsAg and HBsAb tests. Methods: Specimens collected from 43 cases of chronic hepatitis B patients with positive results for both HBsAg and HBsAb tests were used as the experimental group; specimens collected from 43 cases randomly picked from all patients with chronic hepatitis B with a single positive result for HBsAg test were used as the control group. In HBV DNA, the S gene region was amplified and sequenced. Amino acid sequences were grouped, and mutations were analyzed based on the sequencing results. Results: The patients were infected with HBV of the genotype B and C and those who with genotype C show more mutations than genotype B carriers. Compared with the control group, the experimental group had a marked increase in S gene amino acid mutations; a higher amino acid mutation rate was observed in the first loop (aa124–137) of the a‐determinant (aa124–147) and there was a statistical difference (genotype B: 2.68% vs. 0.00%, P = 0.041; genotype C: 7.14% vs. 2.01%, P < 0.001). Conclusion: The first loop in a‐determinant of S gene sequence possesses a large numbers of mutated amino acids, leading to changes of antigenicity and simultaneous positive results in both HBsAg and HBsAb tests finally.     

97例慢性乙型肝炎病毒感染者临床诊断和肝组织病理特征的关系

目的总结分析慢性HBV感染者临床诊断与肝组织病理特征之间的关系,以期对临床诊断及治疗提供依据。方法对97例慢性HBV感染者（慢性HBV携带、慢性乙型肝炎）进行临床诊断分析及肝穿刺活检病理诊断、免疫组化检查。结果 55例慢性HBV携带者中S0～4分别为31、21、2、1、0例,分别占56.3%、38.1%、3.6%、1.8%、0.0%;G0～4分别为28、23、3、1、0例,分别占50.9%、41.8%、5.5%、1.8%、0.0%;30～40岁年龄段肝组织炎症及纤维化程度均高于其他两组。97例临床血清检测诊断为HBV感染者肝组织活检免疫组化结果有6例HBsAg阴性,7例HBcAg阴性。结论临床诊断为慢性HBV携带者及慢性肝炎轻度患者常常存在不同程度的肝脏炎症和纤维化,需要积极的干预治疗。肝组织活检病理诊断为临床诊疗提供巨大帮助,但也有一定局限性     

Hepatitis A in patients with chronic liver disease - severity of illness and prevention with vaccination

Acute hepatitis A in patients with pre-existing chronic liver disease (CLD) may theoretically predispose such patients to a more severe outcome of infection. The results of reports covering epidemics and case series suggest but do not universally support this possibility. An analysis of the larger case series support the suggestion that acute hepatitis A superimposed on chronic hepatitis B infection is associated with higher peak laboratory abnormalities, more severe disease and a higher fatality rate. The safety and immunogenicity of an inactivated hepatitis A vaccination was evaluated in patients with CLD and found to be satisfactory for the protection of this group of patients.