江苏省启东市乙肝疫苗干预队列研究：40年随访结果及进展

摘 要：乙型肝炎病毒（hepatitis B virus,HBV）感染显著增加肝细胞癌（简称肝癌）的发病风险。婴幼儿及儿童接种乙肝疫苗是阻断HBV母婴传播、降低HBV流行率、预防慢性乙型肝炎的有效措施。乙肝疫苗在我国不同历史时期的可及性存在一定差异,同时近几十年来的生活环境因素发生了显著改变,有必要确认新生儿期乙肝疫苗接种预防肝癌的效果。全文回顾了于1983年起始的在江苏省启东市现场开展的一项乙肝疫苗免疫干预的随机临床对照的队列研究,包括新生儿乙肝疫苗接种队列的建立、维护与发展全过程,并对该人群队列5年、10年期及成年期随访工作方法以及安全性和HBsAg、抗HBs阳性情况进行了总结。队列研究结果证明,在新生儿期按“0-1-6”程序接种3剂5 μg低剂量乙肝疫苗及对HBV高危儿童加强免疫,可预防慢性HBV感染和降低肝癌发病和终末期肝病死亡风险,不仅对儿童慢性HBV感染起到早期保护作用,且对易感人群加强免疫后可降低HBV感染风险。然而,母亲HBV感染的婴儿出现慢性HBV感染风险依然存在,为进一步降低HBsAg阳性率,需加强HBV易感人群的免疫防护。     

乙肝疫苗母-婴阻断失败者病毒全基因变异分析

Objective To determine the factors responsible for failed postnatal immunoprophylaxis for hepatitis B virus(HBV) in Qidong, China. Methods Eleven children who developed into chronic HBV infection after receiving HBIG and HBV recombinant vaccines were recruited into the study. Eleven paired mothers with chronic hepatitis and other 6 mothers whose children successfully generated anti-HBs after im-munoprophylaxis were included as the control in the study. Full-length HBV DNA was amplified through ser-um sample by PCR method and underwent cloning and sequencing. HBV DNA level was quantified by real-time PCR. Results The mean levels of HBV DNA in mothers who had HBV DNA positive children and healthy children were ( 1.2 ×107± 3.1 × 106 ) copies/ml and ( 1.6× 107±8.8×106 ) copies/ml, respec-tively. There was no significant difference between the groups (P >0.05). Meanwhile, viral load in chil-dren was unrelated to that in their mothers (r2 =0.2429). In 11 HBV DNA positive children, 4(36.4% ) demonstrated amino acid substitutions in HBsAg "a" determinant region with 6 different types, I.e. T125A, I126T, Q129H, M133V, D144V and G145A. All of the mothers showed the wild-type sequence in "a" epitope, indicating surface escape mutants were not acquired from the initial infection, but developed under the immune pressure. The mutation rates after immunoprophylaxis for preS1, preS2, S, X, preC/C and P genes were 0.38%, 0. 22%, 0.27%, 0.17%, 0.11%, and 0.11%, respectively, nt2999-3157 in preS1, nt529-677 in S, nt1955-2016 in C, nt923-1001 and nt2489-2602 in P genes were among the hottest muta-tional spots throughout the HBV genome. Conclusion HBV mutation may occur in all the open readingframes after passive and active immunoprophylaxis. In addition to S gene, HBV preS and P genes could alsoassociate with the escape mutants.     

乙肝随访队列中空腹高血糖发病率及相关因素分析

对427例随访者分为FBG升高组（185例）和FBG正常组（242例）。结果：1FBG升高组的ALT（t=-2.170,P=0.031）、GGT（t=-3.500,P=0.001）、TG（t=-3.099,P=0.002）、TC（t=-2.033,P=0.043）、UA（t=-3.009,P=0.003）、BMI（t=-4.165,P=0.001）、收缩压（t=-4.407,P=0.001）、舒张压（t=-2.249,P=0.025）、抗-HBc阳性（P=3.879,P=0.049）和脂肪肝发病率（t=11.158,P=0.001）分别高于FBG正常组;2两组平均年龄;CREA,BUM;AFP、HBsAg、抗-HBs、HBeAg、抗-HBe阳性率;肝癌、肝硬化和慢肝发病率均相似,P〉0.05;3多因素logistic回归分析显示GGT、UA、BMI和收缩压为FBG的危险因素。结论：乙型肝炎患者的空腹高血糖发病率高于健康人群。GGT、UA、BMI和收缩压为乙肝患者发生FBG的危险因素。     

乙肝表面抗原阳性人群糖代谢结果分析

调查分析随访10年的乙肝表抗阳性和阴性队列的体检结果。结果：HBsAg（＋）与HBsAg（-）组比较：DM、IFG、IGT、脂肪肝的发生率无统计学差异。而脂肪肝组的糖尿病发病率41.8%高于非脂肪肝组的糖尿病发病率20.5%。     

小切口硬核白内障囊外摘出术的临床观察

目的观察小切口手法碎核在硬核白内障囊外摘出术中的临床效果。方法对80例(80眼)硬核白内障行小切口手法碎核。结果术后1天矫正视力0.3以上41眼(51.25%),术后1周矫正视力0.3以上65眼(81.25%)。术后1月矫正视力0.3以上71眼(88.75%),术中主要并发症为后囊膜破裂,术后为角膜内皮水肿。结论切口手法碎核硬核白内障囊外摘出术是一种经济、有效、安全、简便的方法。     

乙型肝炎病毒e抗原与肝癌关系的17年前瞻性队列研究

目的:探讨启东肝癌高发区乙型肝炎病毒e抗原(hepatitis B e antigen,HBeAg)与肝癌发生之间的关系。方法:利用1992年在启东建立的由807例乙型肝炎病毒表面抗原(hepatitis B surface antigen,HBsAg)携带者和761例性别、年龄匹配的HBsAg阴性者组成的肝癌前瞻性队列,分析1992年5月-2010年3月肝癌发生与HBeAg之间的关系。结果:队列观察总人数为24715人年。HBsAg阳性组中156例发生肝癌,发生率为1288.83/10万人年;HBsAg阴性组中9例发生肝癌,发生率为71.37/10万人年。肝癌发病在HBsAg(+)/HBeAg(-)和HBsAg(+)/HBeAg(+)组中的相对危险度分别为13.25[95%可信区间(con?dence interval,CI)为6.67～26.33,P<0.001]和28.05(95%CI为13.87～56.73,P<0.001)。在HBsAg阳性者中,HBeAg效价<24、1:24～1:27、1:28～1:212和>212者其罹患肝癌的风险分别是HBeAg阴性者的2.55(95%CI为1.54～4.22,P<0.001)、5.02(95%CI为2.89～8.73,P<0.001)、1.71(95%CI为0.91～3.18,P>0.05)和1.19(95%CI为0.64～2.23,P>0.05)倍。结论:HBeAg是预测肝癌高危的一个重要指标,且低效价者的危险度更高。     

肝癌患者发病前血清中高尔基体糖蛋白73水平的回顾性动态观察

目的 探讨肝癌发病前后高尔基体糖蛋白73 (GP73)的动态水平及相关因素.方法 2007至2012年对一组乙型肝炎表面抗原(HBsAg)阳性者人群开展一年两次的周期性的筛查,统一保留血标本于生物样本库,直至肝癌发生.将肝癌发病时及发病前30个月内6个时点中,至少有3次血标本并有B超临床检查结果的39例肝癌患者列为血清GP73检测分析对象,最终获得162个标本.采用双抗体夹心ELISA法检测GP73.用stata软件作时序间分析并进行不同分组间差异的统计学检验.结果 肝癌发病时的39例患者GP73检测值为(126.77±73.73)μg/L,发病前5次GP73检测的平均值分别为(128.32±81.18)、(129.97±83.62)、(127.38±80.10)、(135.52±97.88)及(138.24 ±93.58) μg/L,差异无统计学意义(F=0.07,P=0.997).39例肝癌发病前后的GP73水平未见明显的趋势变化.162个检测样本按B超检查结果划分为不伴有肝硬化(63例次)和伴有肝硬化(99例次)两组,GP73平均水平分别为(97.16 ±51.39)、(151.20 ±91.68) μg/L,差异有统计学意义(F=18.22,P＜0.01).进一步以GP73均值(130.19 μg/L)将调查对象分为两组,不伴有肝硬化者中只有1/14的患者GP73水平高于平均值,而伴有肝硬化者(25例)中有12例的GP73水平高于平均值,差异有统计学意义(P =0.013).拟合回归模型也显示GP73与时间序列不相关(t=0.75,P=0.455),而与肝硬化相关(t=4.30,P＜0.01).结论 肝癌患者发病时及发病前30个月内的GP73水平动态变化不大.肝癌患者GP73水平的高低是由肝病背景所决定的;肝硬化可能是一个主要的影响因素或混杂因素.     

启东市非新生儿期接种乙肝疫苗儿童远期效果观察

目的:了解非新生儿期接种乙肝疫苗儿童的远期效果。方法:上述对象(非新生儿组)在12岁时全程接种乙肝疫苗(10μg/支),同龄的对照组(新生儿组)0岁起全程接种乙肝疫苗(10μg/支)。18岁时两组对象静脉采血,检测HBsAg、抗-HBs、抗-HBc和丙氨酸氨基转移酶(ALT)。结果:非新生儿组与新生儿组相比:前者HBsAg(8.0%)、抗-HBs(62.3%)和抗-HBc(23.1%)阳性率显著高于后者HBsAg(2.0%)、抗-HBs(37.2%)和抗-HBc(4.5%)(P<0.001)。HBsAg阳性者中抗-HBc阳性率前者为100%,后者71.4%,P=0.035;抗-HBs阳性者中的抗-HBc阳性率前者为19.7%,后者6.0%,P=0.000。ALT异常率:前者(5.4%)和后者(6.4%)无显著差异(P=0.575)。将HB-sAg分为阳性者和HBsAg阴性者且进行组内比较时,两组HBsAg阴性者的ALT异常率(前者5.1%,后者6.4%)均显著高于HBsAg阳性者(前者0.6%,后者0.0%)(P=0.000)。结论:非新生儿期接种乙肝疫苗儿童在远期(18岁)仍具有较高抗-HBs阳性率,对HBV感染具有一定保护性;但HBV的感染标志HBsAg和抗-HBc高于新生儿期接种儿童。     

乙肝疫苗母-婴阻断失败者病毒全基因变异分析

目的 探讨乙肝疫苗母-婴阻断失败者中乙肝病毒(HBV)全基因变异状况.方法 用PCR方法对启东地区11对乙肝疫苗免疫失败儿童-母亲配对血清及6例疫苗接种成功儿童的"大三阳"母亲血清扩增HBV全长基因,克隆测序后以Clustal X软件进行序列比对.病毒载量采用实时荧光定量PCR方法测定.结果 疫苗阻断失败和成功儿童的母亲HBV平均滴度分别为(1.2×107±3.1×1010)copies/ml和(1.6×107±8.8×1010)copies/ml,两组之间差异无统计学意义(P>0.05).在11例HBV DNA阳性的儿童中,4例(36.4%)有HBsAg"a"决定簇的氨基酸改变,表现为T125A、I126T、Q129H、M133V、D144V和G145A等6种不同突变形式,但母亲中HBsAg"a"决定簇均为野生型.疫苗接种失败的儿童中HBV preS1、preS2、S、X、preC/C和P基因的平均突变率分别为0.38%、0.22%、0.27%、0.17%、0.11%和0.11%.preS1区nt2999-3157、S区nt529-677、C区nt1955-2016和P区nt923-1001、nt2489-2602为病毒基因组中最易发生突变的区域.结论 经主动和被动免疫后,儿童体内HBV突变可发生于所有开放阅读框架内.除s基因外,preS和P基因的突变可能也与免疫逃逸有关.     

乙肝疫苗母-婴阻断失败者病毒全基因变异分析

Objective To determine the factors responsible for failed postnatal immunoprophylaxis for hepatitis B virus(HBV) in Qidong, China. Methods Eleven children who developed into chronic HBV infection after receiving HBIG and HBV recombinant vaccines were recruited into the study. Eleven paired mothers with chronic hepatitis and other 6 mothers whose children successfully generated anti-HBs after im-munoprophylaxis were included as the control in the study. Full-length HBV DNA was amplified through ser-um sample by PCR method and underwent cloning and sequencing. HBV DNA level was quantified by real-time PCR. Results The mean levels of HBV DNA in mothers who had HBV DNA positive children and healthy children were ( 1.2 ×107± 3.1 × 106 ) copies/ml and ( 1.6× 107±8.8×106 ) copies/ml, respec-tively. There was no significant difference between the groups (P >0.05). Meanwhile, viral load in chil-dren was unrelated to that in their mothers (r2 =0.2429). In 11 HBV DNA positive children, 4(36.4% ) demonstrated amino acid substitutions in HBsAg "a" determinant region with 6 different types, I.e. T125A, I126T, Q129H, M133V, D144V and G145A. All of the mothers showed the wild-type sequence in "a" epitope, indicating surface escape mutants were not acquired from the initial infection, but developed under the immune pressure. The mutation rates after immunoprophylaxis for preS1, preS2, S, X, preC/C and P genes were 0.38%, 0. 22%, 0.27%, 0.17%, 0.11%, and 0.11%, respectively, nt2999-3157 in preS1, nt529-677 in S, nt1955-2016 in C, nt923-1001 and nt2489-2602 in P genes were among the hottest muta-tional spots throughout the HBV genome. Conclusion HBV mutation may occur in all the open readingframes after passive and active immunoprophylaxis. In addition to S gene, HBV preS and P genes could alsoassociate with the escape mutants.