沙门氏菌鞭毛蛋白属共同抗原表位的定位

用淋巴细胞杂交瘤技术研制成４株沙门氏菌属特异单抗ＣＢ８，ｄｅ７，ｃｃ６，ＤＤ４，在免疫转印试验中，证实它们所识别的抗原表位位于鞭毛蛋白上，ＥＬＩＳＡ相加试验及竞争试验显示，ＣＢ８和ｃｃ６与ｄｅ７，ＤＤ４识别的抗原表位不同，表明鞭毛蛋白的属共同抗原具有多种表位的特性。用胰蛋白酶消化鞭毛蛋白，ＳＤＳ－ＰＡＧＥ结果出现两条新带，分子量分别为４２０００（Ｆ４２），２７０００（Ｆ２７），长时间消化，最终只剩下Ｆ２７条带，它可抵制该酶的消化。对该酶切样品的免疫转印结果表明，沙门氏菌鞭毛蛋白属共同抗原表位出现于Ｆ４２和Ｆ２７上，而相应的沙门氏菌分型Ｈ抗原单抗或因子血清识别的表位也在Ｆ４２和Ｆ２７上都得到了证实。这为沙门氏菌鞭毛蛋白抗原的多样性及结构和功能研究提供了证据。     

真核表达载体构建表达ＰＲＶ闽Ａ株gE基因表位抗原编码区片段的重组质粒

根据伪狂犬病病毒闽Ａ株gE基因表位抗原编码区的序列与身份种真核表达载体pPICZaA、pAcGP67A序列与特性分别设计了两对ＰＣＲ引物。通过ＰＣＲ方法扩增到了两端具有不同酶切位点的gE基因表位抗原编码片段。将这２个片段分别克隆到pPICZaA与pAcGP67A载体，转化大肠杆菌ＴＯＰ１０菌档及ＸＬ１－Ｂlue菌株，获得了含伪狂犬病病毒闽Ａ株gE基因表位抗原编码区的重组质粒pICZaA-FS与pAcGP67A-FS。序测定结果显示两个重组质粒中插入片段的大小与方向均正确。     

Astrocyte proliferation in the rat hippocampus after middle cerebral artery occlusion

AIM: To study rat astrocyte proliferation in ipsilateral hippocampus following focal cerebral ischemia. METHODS: Ischemia was induced by temporary middle cerebral artery occlusion (MCAO). In hippocampus of rats at 3, 7 and 30 days after MCAO, the numbers and anatomic distribution of glial fibrillary acidic protein (GFAP) were detected by immunohistochemistry. The protein expression of GFAP and proliferating cell nuclear antigen (PCNA) in the ipsilateral hippocampus were analyzed by Western blot analysis. RESULTS: Astrocytes appeared hypertrophic, with increased process thickness and numbers at 7 days after MCAO, and the highest density of astrocytes were seen at 30 days in the CA1, CA2 regions of the ipsilateral hippocampus. Western blot analysis revealed that GFAP levels were normal at 3 days, but increased by 7 days and remained elevation at 30 days. Western blot analysis of PCNA protein also revealed identified upregulation PCNA at 3 days after MCAO and the expression peaked at 7 days. CONCLUSION: This study demonstrates that focal cerebral ischemia in the rat results in a rapid response, a process often referred to as reactive astrogliosis or glial scarring, from resident astrocytes of the ipsilateral hippocampus to the side of ischemia.     

携带猪流行性腹泻病毒S1抗原表位的乙肝核心抗原病毒样颗粒的构建与鉴定

旨在构建以乙肝核心抗原(HBcAg)为载体呈现猪流行性腹泻病毒(PEDV)S1抗原表位的病毒样颗粒.将PEDV S1蛋白中含B细胞表位的270 bp片段插入到HBcAg主要免疫显性区域(MIR),构建重组质粒pET-32a(+)-HBcAg-PEDV S1,转化到感受态细胞BL21(DE3)中,经IPTG诱导表达,SD...     

Effects of swimming on experimental endometriosis in rats

AIM To evaluate the effect of swimming on experimental endometriosis in rats. METHODS 80 female SD rats were divided into 8 groups, including control group, model group and animals performed light exercise （swimming once a week）, moderate exercise （swimming 3 times a week）, and intense exercise （swimming 5 times a week） before or after endometriosis induction,10 rats in each group. The mRNA and protein expressions of fatty acid synthase （FAS）, matrix metalloproteinase 9 （MMP9） and proliferating cell nuclear antigen （PCNA） in endometrium of rats were detected. RESULTS The swimming before the induction of the edometriosis lesions did not prove to have aprophylactic role against endometriosis, whereas the swimming after induction of the lesions had a beneficial effect regardless of frequency, with a greater reduction in the groups practicing moderate and intense activity （P<0.05）, an increase in FAS levels and a decrease in MMP9 and PCNA levels were also observed （P<0.05）. CONCLUSION Swimming after induction of the edometriosis is beneficial for the treatment of endometriosis, the mechanism may be related to the expression of FAS, MMP9 and PCNA protein.     

青海地区野生狐狸棘球绦虫感染情况调查研究

野生狐狸是棘球绦虫的主要终末宿主,在棘球蚴病的传播中有重要作用.为了解青海省狐狸中棘球绦虫感染状况,我们用棘球绦虫粪抗原检测试剂盒,对收集于青海省称多县、贵南县、祁连县的野生狐狸粪便进行了检测.结果表明,称多县收集的129份狐狸粪便阳性15份,阳性率11.6％;祁连县收集的84份狐狸粪便阳性3份,阳性率3.6％;贵南县...     

旋毛虫p53糖蛋白抗原研究进展

旋毛虫p53糖蛋白是旋毛虫肌幼虫排泄分泌抗原的主要抗原组分之一,具有肌幼虫期特异表达特性,主要定位在肌幼虫β-杆细胞.不同旋毛虫种p53糖蛋白氨基酸序列相对保守,但抗原性有差异,在同一种内变异极少,具有种及属特异性抗原表位.论文就旋毛虫p53糖蛋白抗原的分子结构、不同发育时期的特异性表达、抗原性等方面的研究进展进行综述...     

检测狂犬病病毒抗原的夹心间接Dot—ELISA的建立和应用研究

本研究建立了检测狂犬病病毒抗原的夹心间接斑点酶联免疫吸附试验(SI-Dot-ELISA).本法检出狂犬病病毒抗原的最低浓度为:0.01IU/mL的标准抗原,1:100000 (相当于20 LD_(50))的狂犬病病毒CVS株和鹿8202株的鼠脑悬液,1:400(相当于7906TCID_(50)/mL)的狂犬病病毒SAG株的细胞培养物.对多种健康动物的组织、健康细胞培养物以及犬瘟热病毒、犬腺病毒等检测均为阴性.用SI—Dot—ELISA、夹心间接ELISA(SI—ELISA)和小鼠脑内接种3种方法,检测了388份材料,检出的阳性份数分别为173、171和179,经统计学处理,3种方法在狂犬病病毒抗原的检测上,可以相互代替.甲醛灭活病毒不影响本方法的检测,而加入氢氧化铝胶后的病毒悬液则不能用本法检测.本法适用于狂犬病的诊断、流行病学调查、疫苗效价的测定和实验研究中病毒抗原的测定.     

捻转血矛线虫HLJ株15 ES抗原基因的原核表达

为了对羊源捻转血矛线虫(H.contortus)15 ES抗原基因进行原核表达,本研究利用RT-PCR技术从黑龙江省羊源H.contortus成虫总RNA中扩增得到相对分子质量15Ku ES蛋白基因(H15 ES),序列分析表明,其核苷酸序列与国外已发表的15 ku排泄分泌抗原基因的同源性为99.78%.将H15 ES克隆至pET-32a(+)中,构建重组表达质粒pET-32a-H15 ES,转化大肠杆菌,IPTG诱导表达,经SDS-PAGE电泳和western blot分析表明,表达蛋白相对分子质量约为35.2 ku.本研究为H.contortus病诊断抗原和保护性抗原的大量制备及H.contortus核酸疫苗的研究奠定了基础.     

金黄色葡萄球菌FnbpA、ClfA、Ebps抗原表位的串联表达及多克隆抗体的制备

为获得金黄色葡萄球菌特异性抗原蛋白并以此制备多克隆抗体,应用DNAStar软件筛选金黄色葡萄球菌FnbpA、ClfA、Ebps的抗原表位,以柔性氨基酸序列连接后进行全基因合成,将目的基因克隆到pGEX-4T-1原核表达载体,转化至大肠杆菌感受态细胞,诱导表达,采用Sepharose 4B柱亲和层析纯化表达蛋白,制备抗原后对家兔进行皮下多点注射,于第42天采集外周血,获得血清。结果表明,本试验成功构建了金黄色葡萄球菌抗原基因表达载体,并获得了高免血清,经ELISA的方法检测血清效价可达到1∶10000以上,成功制备多克隆抗体,为以后采用免疫学方法快速检测金黄色葡萄球菌奠定了基础。