结核分枝杆菌Ag85B真核表达质粒的构建及其表达

目的：构建结核分枝杆菌Ag85B真核表达质粒并表达。方法：从结核分枝杆菌（H37Rv）基因组中扩增出Ag85B编码基因，经限制性内切酶消化后，定向克隆入pcDNA3.1( )中。采用脂质体法将pcDNA/Ag85B转染COS－7细胞，采用RT－PCR、ELISA和斑点印迹法检测其表达。结果：扩增出Ag85B基因，经双向DNA序列测定，与Genbank注泵的序列一致；重组质粒转染COS－7细胞后经检测证实，该基因能在真核细胞中表达。结论：成功地构建了pcDNA/Ag85B质粒，其在真核细胞中表达的蛋白具有良好的抗原性。     

Immunohistochemical localization of vascular endothelial cell factor viii/von Willebrand factor antigen in human normal and disordered gastric tissues

Vascular endothelial cell factor VIII/von Willebrand factor antigen (FVIII/vWF Ag) of normal and disordered gastric tissues was studied with staphyloceocus protein A-gold (PAG) labelling followed by photochemical silver reaction. FVIII/vWF Ag was localized clearly in the tissue fixed with various common fixatives and embedded in paraffin without enzyme treatment. The most satisfactory staining and the least nonspecific background were observed in the tissues fixed with Zamboni’s and Bouin’s solutions. The staining reaction could be enhanced, if the sections were preteateed with trypsin and subtilisin. Under the electron microscope, the gold particles were found over the Weibel-Palade bodies of vascular endothelial cells in the tissues fixed either in Zamboni’s solution or in Zamboni’s solution-osmium tetroxide, and embedded either with Lowicryl K₄M or With Epon 812. It has been proved to be a better technique in investigation of FVIII/vWF Ag in vascular endothelial cells.     

HBsAg阳性人群乙肝病毒前S1抗原检测的临床意义

目的：探讨乙型肝炎病毒前S1抗原与血清乙肝病毒两对半、HBVDNA的相互关系，了解乙肝病毒感染者血清Pre-S1Ag的临床意义。方法：双抗体夹心ELISA法测定血清Pre-S1Ag，EIA法检测血清乙肝病毒两对半，荧光定量PCR检测血清乙肝病毒DNA。结果：1122例HBsAg阳性人群中，Pre-S1Ag阳性率为50.1％。114例HBVDNA阳性血清，Pre-S1Ag阳性率为78.1％，HBVDNA阴性人群，Pre-S1Ag阳性率15.7％(8/51)；HBeAg阳性人群Pre-S1Ag阳性率为82.2％，HBeAg阴性人群Pre-S1Ag阳性率为35.7％；抗-HBe阳性人群Pre-S1Ag阳性率为31.6％。172例乙肝“两对半”阴性人群Pre-S1Ag阳性率为1.74％。结论：血清Pre-S1Ag与HBVDNA、HBeAg相互关联，可以作为HBV存在和复制的标志。联合检测Pre-S1Ag和HBeAg可相互补充、印证，对判断乙肝病毒的复制和存在更有价值。血清Pre-s1Ag测定还是乙肝患者疗效评估和预后判断的可靠指标。     

口腔粘膜良恶性病变Ag—NORs形态观察和定量研究

应用细胞核仁组成区嗜银蛋白染色技术，对８２例口腔粘膜急性恶性病变进行研究。结果表明，口腔粘恶性病变Ａｇ－ＮＯＲｓ团块形态不规则，分布离散；Ａｇ－ＮＯＲｓ计数明显高于正常口腔粘膜和良性病变组，但不同组织类型和分级的病性病主Ａｇ－ＮＯＲｓ计数无显著差异，与其生物学行为相一致。由此可见，Ａｇ－ＮＯＲｓ形态观察和定量研究有助于口腔粘膜良恶性病变的鉴别和癌前病变的早期诊断，但对不同组织类型恶性病变的区分和分     

100例HBV血清标志物、HBV—DNA、HBV—PreS1—Ag对比监测与临床分析

目的 探讨临床监测HBV－PreS1-Ag对乙肝患诊断和预后的意义。方法 对100例乙型病毒性肝炎患HBV血清标志物、HBV－DNA及PreS1-Ag进行量、性跟踪监测并加以临床分析。结果 100例患HBsAg阳性91例,PreS1-Ag阳性57例,HBeAg阳性43例,HBV－DNA阳性56例,PreS1-Ag阳性率与HBV－DNA相似且略高于HBsAg。结论 PreS1-Ag比HBsAg更为精确的反映了HBV在人体内感染和复制的情况,因而动态监测PreS1-Ag的变化有助于临床评估急慢性肝炎的治疗效果及预后。     

Safety,tolerability, and immunogenicity of a 4-antigen Staphylococcus aureus vaccine (SA4Ag): Results from a first-in-human randomised,placebo-controlled phase 1/2 study

BackgroundA prophylactic Staphylococcus aureus four-antigen vaccine (SA4Ag) is under development for prevention of invasive S. aureus disease. A preliminary S. aureus three-antigen vaccine (SA3Ag) was reformulated to include a novel manganese transporter protein (MntC or rP305A). This study describes the first-in-human dose-finding, safety, and immunogenicity results for SA4Ag.MethodsIn this double-blind, sponsor-unblind, placebo-controlled, phase 1/2 study, 454 healthy adults aged 18–64 years were randomised to receive a single dose of one of three formulations of SA4Ag with escalating dose levels of rP305A or placebo. Functional immune responses were measured using opsonophagocytic activity (OPA) killing and fibrinogen-binding inhibition (FBI) assays; antigen-specific immunogenicity was assessed using a four-plex competitive Luminex® immunoassay (cLIA).ResultsA high proportion of SA4Ag recipients met the pre-defined antibody thresholds for each antigen at Day 29. A substantial and dose-level dependent immune response was observed for rP305A, with up to 18-fold rises in cLIA titres at Day 29. Robust functional responses were demonstrated, with >80-fold and >20-fold rises in OPA assay titres at Day 29 using S. aureus strains expressing capsular polysaccharide serotypes 5 and 8, respectively. Durable antibody responses were observed through month 12, gradually waning from peak levels achieved by days 11–15. SA4Ag was well tolerated, and no vaccine-related serious adverse events were reported.ConclusionsSingle-dose vaccination of SA4Ag in healthy adults aged 18–64 years safely induced rapid and robust functional immune responses that were durable through month 12, supporting further development of this vaccine. Trial registration number: NCT01364571     

Prevalence and significance of intrahepatic hepatitis delta virus antigen expression in HBeAg-negative patients with chronic type-B hepatitis in Taiwan

A consecutive series of 97 HB_eAg-negative patients with chronic type-B hepatitis, who were not drug addicts or hemophiliacs, were examined for the prevalence of intrahepatic expression of hepatitis delta virus (HDV) antigen (HDAg) by direct immunofluorescence. The clinical and histological features were compared between those with and without intrahepatic HDAg expression. Intrahepatic HDAg expression was detected in 15 (15.5%) of the 97 patients. All of the 15 were males between 18 and 71 years of age. Each had elevated SGPT levels, with 46.8% being greater than 10 times normal and 73.4% greater than five times normal. Histological study revealed chronic lobular hepatitis in nine and chronic active hepatitis in six, including two with cirrhosis. Compared to the 82 patients without intrahepatic HDAg expression, no difference in the age (34.1±14.3 vs 36.7±10.8) and sex (150 vs 7210) distribution was evident, but greater biochemical and histological activity was present in HDV-positive patients.This work was supported by Chang Gung Medical Research grant MRP 197 and a grant from the Prosperous Foundation, Taipei.     

多发性骨髓瘤细胞系中ANGPT1/2基因的表达

目的 检测ANGPT1/2基因在多发性骨髓瘤细胞系中的表达.方法 应用实时荧光定量(RQ)-PCR方法检测两种多发性骨髓瘤细胞株U266和P3X63Ag653 (653)中ANGPT1/2的表达.结果 以人脐静脉内皮细胞为对照,ANGPT1在U266和653中的表达升高,与人脐静脉内皮细胞相比差异无统计学意义.ANGPT2在U266中表达降低,与人脐静脉内皮细胞相比差异无统计学意义;而ANGPT2在653中明显高表达,与人脐静脉内皮细胞相比差异有统计学意义.结论 应用RQ-PCR检测ANGPT1/2在2株多发性骨髓瘤细胞系中表达,为下一步在体内外研究ANGPT-TIE在多发性骨髓瘤中的作用机制奠定基础.     

探讨血清preS1-Ag抗HBc-IgM检测在急、慢性乙肝患者中的临床价值

目的:探讨急、慢性乙肝患者病变及治疗过程中Pre-S1Ag、抗HBc-IgM的变化及其临床价值.方法:应用酶联免疫法、荧光定量PCR技术及化学法对40例急、慢性乙肝患者病变及治疗过程中血清Pre-S1Ag、抗HBc-IgM、HBV-DNA及肝功能等指标进行检测并观察其与预后的关系.结果:Pre-S1Ag、抗HBc-IgM在急性乙肝中的检出率达92.9%和96.4%,与HBV-DNA检出率差异无显著性(P＞.05);急性肝炎患者血清Pre-S1Ag、抗HBc-IgM检出率明显下降(P＜0.05)但肝功能恢复不佳的急肝患者,其血清Pre-S1Ag、抗HBc-IgM检出率下降不明显(P＞0.05);慢性乙肝中,活动性乙肝患者血清Pre-S1Ag、抗HBc-IgM、HBV-DNA检出率均显著高于非活动性患者(P＜0.05).结论:检测Pre-S1Ag和抗HBc-IgM对急性乙肝的早期诊断、监测复制及预后评估及慢性乙肝的分类均具有较大价值.     

Formation of ZnSe on Ag(111) by electrochemical atomic layer epitaxy

ZnSe is an ideal candidate for optoelectronic devices. However, modern technology is quite demanding as to the quality of materials employed. The electrochemical atomic layer epitaxy (ECALE) method ensures good atomic-level control of thin film formation, as it is based on the alternated deposition of atomic layers of the elements making up the compound. The electrochemical conditions necessary to form ZnSe deposits of up to 35 Zn layers and 35 Se layers on Ag(111) by ECALE are described here. The 1:1 stoichiometric ratio of Zn to Se suggests that ZnSe is formed. A miniaturized electrochemical cell is also described.