隐匿性乙型肝炎病毒感染者血清表面抗体性质鉴定

目的 了解隐匿性HBV感染者抗-HBs的特性及其与HBsAg的结合能力.方法 对2例抗-HBs阳性的隐匿性HBV感染者进行长期随访,使用多种试剂盒对患者血清进行多次HBsAg检测,利用不同血清型HBsAg对患者血清进行中和反应,了解血清中抗体亚型情况.PCR扩增S基因构建真核表达质粒,分析S基因变异情况,并将质粒转染HepG2细胞,取培养上清液及转染细胞分别混合进行HBsAg检测.使用患者血清及抗-HBs阳性血清(对照组)对部分HBsAg阳性克隆上清液进行中和反应.不同组间数据比较采用t检验.结果 多种试剂盒进行的多次检测结果均表明患者血清HBsAg为阴性;HBsAg的3种不同血清型(adr、adw、ay)均能够中和患者血清中大部分抗-HBs(82.1%～100.0%).S基因序列分析表明核苷酸同源性和氨基酸同源性分别为95.13%～97.79%和92.04%～95.58%;培养上清液和转染细胞裂解液中HBsAg定量分别为对照组的48.1%和59.3%,上清液/细胞裂解液比值分别为0.85和0.38.中和试验结果显示转染上清液中HBsAg定量较混合前有所降低,但是仍然可以检测到,而对照组检测不到HBsAg,差异均有统计学意义(F值分别为353.6和645.2,P值均<0.01).结论 抗-HBs阳性隐匿性HBV感染者体内HBsAg的抗原性及分泌能力有所下降,抗HBs主要针对不同血清型HBsAg的共同表位,但可能与疫苗注射产生的抗-HBs有所不同.

Abstract：

Objective To investigate the properties of HBsAb in occult hepatitis B virus infection and its affinity to different serotypes of hepatitis B virus surface antigen (HBsAg). Methods Long-term follow-up was conducted in 2 HBsAb positive patients with occult hepatitis B virus infection. HBsAg was detected using multiple diagnostic kits and the HBsAb subtype was determined by performing neutralization experiments with different serotypes of HBsAg. The viral S gene was PCR-amplified and mutation analysis was conducted. Plasmids expressing HBsAgs were constructed by inserting these PCR products into an eukaryotic expression vector and were then transfected into HepG2 cells. The cell culture supernatant and cellular extracts were detected for HBsAg respectively. Neutralization experiments were carried out in the cell culture supernatant from HBsAg plasmids transfected HepG2 cells and serum samples from these patients and others who had been confirmed to be positive for HBsAb. Results Multiple tests using various diagnostic kits showed that the 2 patients were negative for HBsAg and the three different serotypes of HBsAg (adr, adw, ay) could neutralize 82.1%-100% of HBsAb existed in the 2 patients. Sequence analysis of S gene cloned from these patients revealed that the homology to reference strain were 95.13%-97.79% and 92.04%-95.58% respectively at the nucleotide and amino acid levels. Quantitation of HBsAg showed that the expression levels of HBsAg from the two patients were 41.1% and 22.6% respectively of that of control HBsAg in cell culture supernatant and 48.1% and 59.3% respectively in cellular extract, and the supernatant/cell lysate ratios were 0.85 and 0.38 respectively. In neutralization experiments, HBsAg could be totally absorbed by control serum, whereas could only be partially neutralized by HBsAbs from the two patients (F = 353.6 and 645.2, P < 0.01). Conclusion Both the antigenicity and the ability of HBsAg secreted outside of the cells are decresed in these HBsAb-positive patients with occult HBV infection. The HBsAbs are mainly specific for common epitopes among different serotypes of HBsAg and are probably different as compared with those produced by vaccine inoculation.     

兔脂肪源性间充质干细胞的扩增及其基本生物学特性

目的探讨兔脂肪间充质干细胞在体外的基本生物学特性。方法选取清洁级新西兰大白兔4只，采用Ⅱ型胶原酶消化兔腹股沟区皮下脂肪组织，贴壁法培养脂肪间充质干细胞。检测细胞生长动力学，测定生长因子EGF对细胞生长的影响，采用免疫细胞荧光法检测基质细胞表面抗原CD44、CD45。结果所培养细胞由P0传代至P8，每代倍增数目未发现有下降趋势；EGF具有显著的促进ADSCs增殖的作用，所培养细胞表达CD44抗原，荧光显微镜下呈红色荧光，CD45近似于阴性表达。结论由兔脂肪组织分离扩增脂肪间充质干细胞，不仅具备干细胞的基本生物学特性，而且增殖稳定，可获得充足的细胞数量，有望成为组织工程的种子细胞来源。     

乙型肝炎免疫球蛋白聚氰基丙烯酸正丁酯纳米粒抑制HBsAg、HBV DNA分泌的体外实验

Objective To investigate the inhibitive activities of hepatitis B immunoglobulin(HBIG)poly(butylcynaoacrylate)nanoparticles(HBIG-PBCA-NP)to hepatitis B surface antigen(HBsAg)and hepatitis B virus(HBV)DNA secretions using HBV infected cell model in vitro.Methods HepG 2.2.15 cells were cultured with media containing HBIG-PBCA-NP or HBIG for several days,or cultured with HBIG-PBC-NP and HBIG for 2 days and without HBIG-PBCA-NP and HBIG from day 3.The supernatants at different time points were collected for quantitative detection of HBsAg and HBV DNA.The comparisons between groups were done by variance analysis.Resalts Secretions of HBsAg and HBV DNA in supernatants of HepG2.2.15 cultured with 0.1-10.0 IU/mL of HBIG-PBCA-NP and HBIG were inhibited significantly compared with control group.HBsAg titers and HBV DNA levels in supernatants of HBIG-PBCA-NP group and HBIG group cultured with media without HBIG-PBCA-NP and HBIG kept decreasing at day 5 and 7,then rebounded at day 9 and 11.HBsAg titera in supernatants of 0.1,1.0,5.0 IU/mL HBIG-PBCA-NP group were all significantly different from those in HBIG group at day 9[(31.31±1.98)μg/L vs(40.62±2.99)μg/L,(23.79±1.31)μg/L vs(36.51±2.12)μg/L,(19.91±1.74)μg/L vs(33.03±1.65)μg/L;F=412.24,P＜0.01].Couclusion HBIG-PBCA-NP can inhibit secretions of HgsAg and HBV DNA in vitro,which is more effective than HBIG.     

恩替卡韦长疗程后联合或序贯IFNα治疗慢性乙型肝炎的效果与安全性分析

目的探讨经恩替卡韦(ETV)长期治疗,有效抑制HBV DNA后,采取序贯或联合IFNα治疗与单用ETV治疗的疗效与安全性比较。方法收集2013年1月-2015年12月南通大学附属吴江医院门诊和住院的慢性乙型肝炎患者27例,所有患者已经ETV抗病毒治疗3年或以上,HBV DNA阴性,无IFN治疗禁忌证,将纳入患者随机分为3组,每组9例。对照组:继续ETV治疗;序贯组:更换为IFNα序贯治疗;联合组:联合IFNα治疗。检测治疗0、12、24、36、48周时血清HBV DNA、HBV血清标志物、血常规、肝功能,观察治疗情况,记录结果并进行统计分析。计量资料多组间比较采用单因素方差分析,进一步两两比较采用Scheffe检验。结果无退出研究病例,无严重不良事件发生;序贯组有1例24周后出现了病毒反弹,但无临床症状,肝功能各项生化指标正常。治疗结束时,序贯组和联合组HBsAg下降的绝对值均显著高于对照组(P值均<0.001);序贯组和联合组各有2例出现了HBsAg的阴转,阴转率22%;对照组未出现HBsAg的阴转病例。结论 ETV有效抑制病毒复制后的IFNα序贯或联合治疗可能是安全的,HBsAg下降水平优于ETV单药治疗,少数患者出现临床治愈。     

单采血浆和输血途径感染HIV的患者中HBV和HCV感染状况

目的 了解单采血浆和输血途径感染艾滋病病毒(HIV)的人群中,乙型肝炎(乙肝)病毒(HBV)、丙型肝炎(丙肝)病毒(HCV)的感染状况. 方法 应用酶联免疫吸附试验(ELISA),对825 名HIV阳性的单采血浆献血员和受血者,检测乙肝病毒表面抗原(HBsAg)和丙肝病毒抗体(抗-HCV). 结果 单采血浆和输血途径感染HIV的人群中,HBV、HCV的感染率分别为3.76%、72.24%.其中单采血浆献血员中,HCV感染率(81.85%)高于输血人群(56.03%),而HBV感染率(1.54%)低于输血人群(7.49%)(P均＜0.01). 结论 在单采血浆和输血途径感染HIV的患者中,有很高的HCV感染率,而HBV感染率较低.     

旋毛虫成虫抗原的免疫保护性研究

目的 比较旋毛虫成虫虫体抗原、排泄分泌抗原和表面抗原对小鼠产生的免疫保护作用。 方法 检查免疫鼠和对照鼠肠道成虫、肌幼虫和血液中的嗜酸性粒细胞数;用ELISA测血清中抗旋毛虫肌幼虫IgG抗体滴度。 结果旋毛虫成虫虫体抗原、排泄分泌抗原和表面抗原免疫组的成虫减虫率分别为84.48％、89.98％和85.16％;肌幼虫减虫率分别为69.82％、78.80％和73.94％。3种抗原免疫组小鼠血中的嗜酸性粒细胞(EOS)数明显增多,血清中IgG抗体滴度明显升高,IgG抗体的几何平均倒数滴度(GMRT)分别是未免疫组的6.96、7.99和6.06倍。 结论 旋毛虫成虫虫体抗原、排泄分泌抗原和表面抗原均能诱导宿主产生较强的抗攻击感染保护力,且可激发特异性体液免疫和细胞免疫。成虫的排泄分泌抗原显示出更强的免疫原性。     

HBV感染者134例的HBsAg·Re研究

近年采用ＲＩＡ夹心法测定１３４例ＨＢＶ感染者及３６例非患者的血清ＨＢｓＡｇ；Ｒｅ并以同期测得的血清ＨＢｃＡｇ等作对照。结果在１１０例ＨＢｓＡｇ阳性者中，检出６９例ＨＢｓＡｇ．Ｒｅ阳性，３３例ＨＢｅＡｇ阳性者中，检出２６例ＨＢｓＡｇ．Ｒｅ阳性。     

慢性乙型肝炎处理的现代观点和实践

HBV感染的定义为乙肝病毒表面抗原(HBsAg)阳性超过6个月。HBV感染是发展为肝硬化、肝功能失代偿和肝细胞癌(HCC)的高危因素。有15％～40％的HBV感染在生命过程中可发生严重的肝脏并发症。全世界约有3．5亿慢性HBV感染(包括无症状HBV携带(ASC)和乙型肝炎病毒)。     

北海市2006—2011年不同生源地学生HBsAg及HBsAb检测结果

目的了解不同生源地学生乙肝表面抗原(HBsAg)及乙肝表面抗体(HBsAb)检测结果,为乙型肝炎防治提供科学依据。方法用ELISA法检测HBsAg及HBsAb,并对结果进行分析。结果 2006—2009年3所学校学生HBsAg阳性率在6.0%~13.7%,生源来自市、区内的学生HBsAg阳性率高于来自区外的学生,男生阳性率高于女生;3类生源地学生HBsAb阳性率43.9%~52.9%。结论 2006—2009年学生HBsAg阳性率处于较高水平,来自市、区内的学生HBsAg阳性率较高,应进一步加强健康教育及血清学检测,加强乙肝在学校中的预防和控制。