穿透支原体脂质相关膜蛋白诱导人单核细胞产生炎症性细胞因子

穿透支原体（M.penetrans，Mpe）是从感染人类免疫缺陷病毒的患者球液中分离出来的，被称为艾滋病相关支原体。Mpe能依靠其特殊的顶端结构黏附和入侵宿主的红细胞、CD4^＋T细胞和巨噬细胞等，但关于其对宿主的致病性及其可能的致病机制还不十分清楚。脂质相关膜蛋白（LAMPs）是暴露在Mpe表面与周围环境各种成分不断相互作用因而影响机体的免疫系统的一种主要蛋白。我们的研究证实，Mpe LAMPs能诱导小鼠巨噬细胞表达诱导性一氧化氮合酶和产生一氧化氮。     

Transforming Growth Factor-β_2 Gene Cloning and Protein Expression in Human Trabecular Meshwork Cells

Primary open- angle glaucoma (POAG) is aleading cause of blindness,which involves optic neu-ropathy accompanied by characteristic visual field de-fects and is often associated with elevated intraocularpressure due to disturbance ofaqueoushumor outflowthrough the trabecularmeshwork (TM) .The patho-physiology of the TM in POAG has been character-ized by an increasein extracellularmatrix componentsand a decrease in the number of TM cells[1,2 ] .It hasbeen found that,compared with the non- P…     

Apoptosis of Human Trabecular Meshwork Cells Induced by Transforming Growth Factor-β2 in vitro

Agraduallossofcellsoccurswithinthehumantrabecularmeshwork (TM )duringnormalagingandappearstobeincreasedinthepatientswithprimaryopenangleglaucoma (POAG) ,whichhasbeensug gestedtobeassociatedwith pathogenesisofPOAG[1,2 ].Ithasbeenfoundthat,comparedwiththeno…     

沙眼衣原体pORF5 蛋白通过激活PI3K / Akt 信号通路抑制细胞凋亡

目的:探讨沙眼衣原体(Chlamydia trachomatis,Ct)pORF5 蛋白对细胞凋亡的影响,并进一步分析其分子机制,为阐明Ct 致病机制提供实验依据。方法:pGEX-6p/ pORF5 重组质粒转化XL1-blue 大肠杆菌,IPTG 诱导表达GST-pORF5融合蛋白,融合蛋白经谷胱甘肽琼脂糖凝胶4B 纯化及蛋白酶切除GST 标签后得到pORF5 蛋白。用不同浓度的pORF5 蛋白刺激HeLa 细胞,Western blot 检测不同时间Bax 和Bcl-2 的表达水平以及PI3K/ Akt 磷酸化水平,Hoechst 33342 及流式细胞技术分析细胞凋亡情况;HeLa 细胞经PI3K/ Akt 特异性抑制剂LY294002 预处理1 h 后,再用pORF5 蛋白刺激24 h,测定细胞凋亡率,并进一步分析凋亡相关蛋白Bax 和Bcl-2 的表达水平及PI3K/ Akt 磷酸化水平。结果:凋亡相关蛋白Bax 和Bcl-2 的表达变化与pORF5 蛋白浓度呈现一定的浓度和时间依赖性,当pORF5 蛋白浓度达10 μg/ ml 时,Bax 表达下调,Bcl-2 的表达上调,当升高至15 μg/ ml 时,Bax 和Bcl-2 的表达量变化最明显;15 μg/ ml 的pORF5 蛋白刺激HeLa 细胞24 h,Bax 和Bcl-2 表达变化最明显;流式细胞检测结果显示:pORF5 蛋白刺激组较TNF鄄琢处理组和未处理组细胞凋亡率分别降低了27.3% (P<0.01)和8.4% (P<0.05);Akt 在pORF5 蛋白刺激15 min 后发生磷酸化,30 min 后磷酸化水平达到峰值,用PI3K 抑制剂LY294002 预处理Hela 细胞后,发现Akt 的磷酸化显著减少,Bax 蛋白表达明显上调,Bcl-2 表达明显下调;LY294002 处理组细胞凋亡率相比于对照组增加了13.0% (P<0.01)。结论:pORF5 蛋白通过激活PI3K/ Akt 信号通路调节Bcl-2 和Bax 蛋白的表达抑制细胞凋亡。     

黏膜佐剂大肠埃希菌不耐热肠毒素B亚单位优化的淋病奈瑟菌孔洞蛋白B核酸疫苗的构建及其诱导小鼠的免疫应答

Objective To investigate the specific humoral immune response and cellular immune response induced by DNA vaccine with Neisseria gonorrhoeae porin B (PorB) fused with B subunit of Escherichia coli heat-labile enterotoxin B (LTB) in mice. Methods Target genes of porB, ltB and ltB-porB were amplified by polymerase chain reaction (PCR) and cloned into eukaryotic vector pcDNA3.1(-). The recombinants were identified by PCR, enzyme digestion and DNA sequencing.The vectors were transfected into Hela cells, and expressed proteins were checked by cytoimmunofluorescence. Female BALB/c mice were intranasally immunized with recombination vectors. The humoral immune response and cellular immune response were detected by enzyme linked immunosorbent assay (ELISA) and methyl thiazolyl tetrazolium (MTT) colorimetric assay. The expressions of recombination vectors in intranasal mucosal tissues of the immunized mice were detected by immunohistochemistry. The means between groups were compared by analysis of variance. Results All the three recombinants were expressed in Hela cells and intranasal mucosal tissues. The PorB specific IgG in serum and sIgA in vaginal secretions in DNA vaccine immunized mice were significantly higher than those in controls (P＜0.01 ; P＜0.05). Moreover, the sIgA level in pcDNA3.1 (-)/ltB-porB group was higher than that in peDNA3, 1(-)/porB group (P＝0. 002). The levels of interferon-gamma (IFN-γ) and interleukin-4 (IL-4) in the supernatants and stimulation index (SI) of spleen lymphocyte culture in pcDNA3, 1(-)/porB group were (170.04±23.89) pg/mL, (114.68±14.27) pg/mL and 1. 68±0.19, respectively; and those in pcDNA3, 1(-)/ltB-porB group were (161.42±27.50) pg/mL, (124.16±19.04) pg/mL and 1.73±0.28, respectively; which were both higher than those in pcDNA3.1(-)/ phosphate buffered saliae (PBS) group (P＜0. 01; P＜0.05) and pcDNA3.1 (-)/ltB group (all P＜0.05), while there was no significant difference between pcDNA3.1 (-)/ltB-porB group and pcDNA3. 1 (-)/porB group (0. 998, 0. 696, 0. 994; all P＞0.05). Conclusions The constructed DNA vaccines are all successfully expressed in Hela cells and murine intranasal mucosal tissues. The mucosal immunization of the vaccines [pcDNA3. 1 (- )/porB and pcDNA3.1 ( -)/ltBporB] could induce humoral immune response and cellular immune response, especially mucosal immune response. It is confirmed that mucosal adjuvant LTB could promote PorB to induce higher level of mucosal immune response in mice.     

沙眼衣原体D型生殖道感染小鼠动物模型的建立及评价

目的 建立沙眼衣原体(Chlamydia trachomatis,Ct)D型感染小鼠生殖道动物模型,为研究人类生殖道Ct感染的病理变化和致病机制提供实验基础.方法 分离20个临床株,经阴道接种C3H/HeJ小鼠,得到清除时间明显延长的临床株(UT0603).应用该临床株建立Ct D型感染模型,免疫荧光检测阴道脱落上皮细胞中衣原体的含量;原位组织免疫荧光检测上生殖道的衣原体感染定植情况;分离生殖道组织,肉眼观察并进行病理学评分.结果 Ct 临床株感染小鼠下生殖道其排菌量及排菌周期明显延长,可见衣原体有上行感染和定植,并能引发类似人类生殖道感染的病理变化.结论 成功建立了Ct D型生殖道感染动物模型.     

吡喹酮治疗297例少年及儿童慢性血吸虫病的临床观察

吡喹酮是一种治疗血吸虫病的新药,可以治疗各型血吸虫病。根据1980年7月全国血研会吡喹酮协作组所拟订的临床研究实施方案,于1980年10月在浙江常山县阁底公社治疗297例少年期及儿童期慢性血吸虫病人。现将治疗情况与疗效考核的结果总结于后。     

植物血凝素(PHA)治疗血吸虫病的免疫学和临床观察

植物血凝素(Phytohemagglutinin,PHA)作为免疫刺激剂已广泛应用于肿瘤和白血病等患者,在传染病领域中,亦已有用于流行性出血热及病毒性肝炎等病的报道,在血吸虫病方面除徐兆骥等曾有初步报道外,国内外尚未见类似报告。为探讨PHA对血吸虫病患者的免疫功能、