【作者】 汤小湄；

【导师】 肖小敏；

【作者基本信息】 暨南大学 ， 妇产科学， 2007， 硕士

【摘要】 目的：①探讨HBV母婴垂直传播的危险因素；②初步探索hIL-6和hIL-6R在胎盘组织的分布及其与胎盘感染HBV的关系。方法：选取2004年10月至2006年10月在暨南大学附属第一医院妇产科产检和住院分娩的血清HBsAg阳性的孕产妇及其分娩的婴儿各318例为研究对象，收集孕期及产后的流行病学资料，采集孕产妇和婴儿的静脉血及产妇产后24小时内的乳汁检测乙肝病毒标志物(即乙肝病毒两对半和HBV-DNA)，同时收集分娩时胎盘组织进行研究。①根据结果分感染组和非感染组，分析临床资料，筛选HBV母婴垂直传播的危险因素。②采用酶联免疫吸附试验(ELISA)方法，检测血清HBsAg阳性的孕妇及其分娩的婴儿0、1、6月龄的静脉血，根据孕妇产前是否应用HBIG分为HBIG组和非HBIG组，分别比较两组婴儿HBV感染率。③采用酶联免疫吸附试验(ELISA)方法，检测血清HBsAg阳性的孕妇及其分娩的婴儿的静脉血，根据分娩方式的不同分为顺产组和剖宫产组，比较两组婴儿HBV感染率。④采用酶联免疫吸附试验(ELISA)方法和实时荧光定量PCR技术，检测血清HBsAg阳性的孕产妇血清、乳汁及所分娩的婴儿静脉血的乙肝病毒标志物，比较母亲血清HBV-M状态与母亲血清、乳汁的HBV-DNA阳性率的关系，并选取其中母亲乳汁HBsAg阳性，婴儿出生时HBsAg阴性的病例91例为研究对象，根据喂养方式的不同分母乳喂养组和人工喂养组，比较两组婴儿HBV感染率。⑤免疫组化PV-9000法检测胎盘组织中HBsAg分布，分析胎盘组织中各层细胞的感染状况。⑥免疫荧光双标记技术检测HBsAg阳性胎盘组织中HBsAg和hIL-6，激光共聚焦显微镜下观察两者的分布情况。⑦免疫荧光双标记技术检测HBsAg阳性胎盘组织中HBsAg和hIL-6R，激光共聚焦显微镜下观察两者的分布情况。结果：①根据随访的结果，以六月龄时HBsAg阳性作为垂直感染的指标，感染组17例，非感染组301例，六月龄婴儿HBV母婴垂直传播率为5.35％，母血HBV-DNA、HBeAg阳性是HBV母婴垂直传播的危险因素。②176例HBIG组(其中有25例应用HBIG＜3次)，142例非HBIG组，两组婴儿在1、6月龄时的HBV感染率差别有统计学意义(p＜0.05)，HBIG组低于非HBIG组。③158例顺产组，132例剖宫产组，两组婴儿在0、6月龄时的HBV感染率差别具有统计学意义(p＜0.05)，剖宫产组低于顺产组。④41例母乳喂养组，50例人工喂养组，两组婴儿各月龄HBV感染率差别无统计学意义(p＞0.05)。⑤检测132例血清HBsAg阳性孕妇的胎盘，其中67例HBsAg阳性，胎盘HBV感染率为50.77％，免疫组化结果显示阳性信号位于胎盘组织中的各层细胞的胞浆和胞膜。⑥免疫荧光双标记法检测胎盘组织中HBsAg和hIL-6，结果显示HBsAg和hIL-6存在于同一位置。⑦胎盘组织从滋养层细胞、间质细胞至绒毛毛细血管内皮细胞有hIL-6R的表达。⑧免疫荧光双标记法检测胎盘组织中HBsAg和hIL-6R，结果显示HBsAg和hIL-6R存在于同一位置。结论：①母血HBV-DNA、HBeAg阳性可能是HBV母婴垂直传播的危险因素。②产前应用HBIG可有效降低HBV母婴垂直传播率。③婴儿垂直感染HBV可能与分娩方式有关，与阴道顺产相比剖宫产可以降低HBV的母婴垂直传播率。④婴儿感染HBV可能与喂养方式无关，在婴儿采取适当的免疫措施后，HBV携带产妇母乳喂养是安全的。⑤HBV宫内感染可能存在经胎盘细胞表面hIL-6R介导的感染途径。更多还原

【Abstract】 Objective:1. To investigate the risk factors of hepatitis B virus (HBV) materno-fetal vertical transmission.2. To investigate the distribution of hIL-6 and hIL-6R in placental tissue and the correlation between hIL-6、hIL-6R and placental HBV infection.Methods:318 cases of HBsAg positive pregnant women and their babies respectively, who were examined and laboured at the first affiliated Hospital of Ji Nan University, were studied. The epidemiologic informations of mothers and babies during pregnant period and after-born were collected. The peripheral blood hepatitis B virus markers (HBV-M) of the pregnant woman and their newborns, as well as the HBV markers detected in maternal milk within 24 hours after parturition, were collected. And the placenta tissues during parturition were collected for research.1. According to the follow-up results, all cases were divided into either infection group or non-infection groups, the clinical datas were analyzed, and the risk factors of placental HBV infection were selected.2. The HBV-M of peripheral blood in the pregnant woman and their newborns (0, 1, 6 months old) were detected by ELISA. The vertical HBV infection rates of the 2 newborn groups which applied and didn’t applied HBIG were compared.3. The peripheral blood in the pregnant woman and their newborns were detected by ELISA. According to the parturition manner, all cases were divided into either normal labour group or Caesarean birth group. The vertical HBV infection rates of the 2 newborn groups were compared.4. The peripheral blood and milk of the HBsAg positive pregnant woman and the HBV-M of their newborns were detected by ELISA and Fluorescence Quantitative PCR (FQ-PCR). The correlation of maternal in the HBV-M situation and HBV-M positive rate of milk were analyzed. 91 HBsAg negative newborns, whose mothers’ milk were HBsAg positive, were selected. According to the different feeding manner, these 91 cases were divided into either mother-milk feed group or manpower feed group, the neonatal infection rates of these 2 groups were compared.5. Immunohistochemical staining PV-9000 was applied to detect HBsAg in placentas and analyze the HBsAg positive rate of cells in every layer of them.6. HBsAg and hIL-6 in the HBsAg positive placenta tissue were detected by double-label immunofluorescence assays and their distributions were observed by the confocal laser scanning technique.7. HBsAg and hIL-6R in the HBsAg positive placenta tissue were detected by double-label immunofluorescence assays and their distributions were observed by the confocal laser scanning technique.Results:1. The follow-up results of infants showed that, using the HBsAg positive at 6-month infants as a diagnostic criteria for intrauterine HBV infection, infection group had 17 cases, non-infection group had 301 cases, and the rate of placental HBV infection was 5.35%. Maternal serum HBV-DNA positive, HBeAg positive were the risk factors of placental HBV infection.2. 176 cases that applied HBIG (among them which 25 cases applied HBIG＜3 times), 142 cases that didn’t apply HBIG. There is a significant difference between these 2 groups in HBsAg positive rate of one-month and six-month old infant (P＜0.05).3. There are 158 cases of normal labour group and 132 cases of Caesarean birth group. There is a significant difference between these 2 groups in HBsAg positive rate of zero-month and six-month old infant (P＜0.05).4. There are 41 cases of mother-milk feed group and 50 cases of manpower feed group. No significant difference existed between these 2 infant groups in the HBsAg positive rate of zero-month、one-month and six-month old infant P＞0.05).5. There were 67 HBsAg positive placentas in 132 HBsAg positive pregnant women (the positive rate was 50.77%). The immunohistochemical result showed that there is a positive signal in the cell plasm and membrane of every layer of the placental tissue.6. Double-label immunofluorescence assays and the confocal laser scanning technique showed that HBsAg and hIL-6 existed in the same position of placental tissue.7. The positive signal of hIL-6R was indicated by trophoblastic cells, villous mesenchymal cells and vascular endothelial cells of placental tissue.8. Double-label immunofluorescence assays and the confocal laser scanning technique showed that HBsAg and hIL-6R existed in the same position of placental tissue.Conclusion:1. Maternal serum HBV-DNA、positive HBeAg and parturition manners are high risk factors of HBV materno-fetal vertical transmission.2. Applying HBIG takes effects lower the rate of blocking the HBV vertical transmission.3. The parturition manners are related with the placental HBV infection of infants. Compared with normal labour, Caesarean birth may reduce the the rate of HBV materno-fetal vertical transmission.4. The HBV infection of infants could be nothing to do with the feeding manners. Feeding by HBV positive lying-in women will be safe if proper immunity protection measures are used.5. The route of infection mediated by hIL-6R on placental cells maybe exists in placental HBV infection.更多还原