The significance of antiendothelial cell antibodies in patients with lupus n ephritis and immunoblotting analysis of the target components

Objective To investigate the prevalence of antiendothelial cell antib odies and its possible role in the pathogenesis in lupus nephritis Methods Sera from 58 patients with lupus nephritis were studied Cellu l ar enzyme linked immunoabsaborbent assay was used to detect antiendothelia l cell a ntibodies, and immunoblotting technique was performed to determine its specific endothelial target antigens Results 362% of the patients with lupus nephritis had elevated IgG is o type antiendothelial cell antibodies The association was revealed between th e presence of antiendothelial cell antibodies (AECA) and some laboratory and re nal biopsy features, such as thrombocytopenia, higher serum IgG level, lower ser um C3 level, higher erythrocyte sedimentation rate (ESR), presence of antin uc lear antibody (ANA), antidsDNA antibodies and glomerular thrombosis Antien d othelial cell antibodies were consisted of a group of heterogeneous antibodies, and reacted with different endothelial cell antigens which ranged from 12-90?kD in molecular size in immunoblot A correlation was demonstrated between heavy p roteinuria (>35?g/24?hr) and the existence of antibodies against both 27 and 29 ?kD endothelial cell antigens, an association between the presence of glomerula r capillary thrombi and antibody against the 29?kD endothelial cell antigen was also found Conclusions Lupus nephritis patients with antiendothelial cell antibo dies showed a complex autoimmune disorder and clinical disease activity, and the results also provided an indirect evidence that antiendothelial cell antibodi es might contribute to the pathogenesis of lupus nephritis

中华硬蜱唾液腺抗原成分的分析研究

中华硬蜱唾液腺提取物（ＳＧＥ）经ＳＤＳ聚丙烯酰胺凝胶电泳显示出２４条蛋白带，其中主带有６条，分子量分别为１４２、１０５、９４、６６、６４和５６ＫＤ。用中华硬蜱叮刺家兔后兔血清做免疫印渍，特异性抗原带为１０５和９４ＫＤ。说明中华硬蜱１０５和９４ＫＤ的蜱唾液腺蛋白抗原能够有效地诱导宿生产生抗体。

2008~2012年杭州地区招聘人员梅毒血清学检测结果分析

目的 了解2008～2012年杭州地区招聘人员梅毒螺旋体感染情况.方法 采用梅毒快速血浆反应素试验（RPR）和明胶凝集法梅毒螺旋体抗体试验（TPPA）对2008～2012年20 876例招聘人员进行梅毒血清学筛查,阳性结果采用免疫印迹试验（WB）进行确认,并按年度、年龄、拟聘职业及性别分组进行统计分析.结果 20 876例招聘人员的梅毒阳性检出率为0.44%（91/20 876）,呈缓慢的逐年增长（χ2=8.12,P〈0.01）,男性梅毒总阳性例数、TPPA单项阳性数及RPR/TPPA双阳性数均高于女性（χ2=8.34～19.23,P均〈0.01）;三个年龄组之间（〈30岁,31～40岁,〉41岁）的梅毒检出率差异有统计学意义（χ2=39.03～135.53,P均〈0.01）,且随年龄增加而上升（χ2=131.18,P〈0.01）;在五个职业类别中,服务业员工的梅毒阳性检出率最高（1.85%,22/1 186）,国家公务员、企事业员工最低,分别为0.04%,0.05%.结论 招聘人员的梅毒感染率处于较低水平,但呈逐年缓慢上升趋势,应将梅毒血清学筛查试验作为招聘人员体检的必检项目,对有争议或可疑的结果应采用免疫印迹试验（WB）进行确认,从而有效控制梅毒流行.

家蚕微孢子虫蓖麻毒蛋白B链基因RBL463-4的克隆及在毕赤酵母中的表达

基于家蚕微孢子虫(Nosema bombycis,Nb)基因组中发现的蓖麻毒素B链(ricin B chain)序列数据设计引物,提取感染Nb第8天的家蚕幼虫中肠组织总RNA,利用RT-PCR方法扩增出RBL463-4基因,并将其克隆进毕赤酵母表达载体pPICZα-A中,构建重组酵母表达载体pPIC-R4。重组质粒pPIC-R4经SacⅠ线性化后,电击导入毕赤酵母X33中,利用博莱霉素(zeocin)筛选获得重组酵母X33/RBL463-4。以2%甲醇诱导表达后,经SDS-PAGE、Western blot检测表达产物,结果出现25kD和20kD2种蛋白,其中25kD蛋白与预测融合蛋白大小一致,表明获得了RBL463-4的融合蛋白。研究结果为进一步探究RBL463-4蛋白在家蚕微孢子虫侵染宿主过程中的功能作用奠定了基础。

大鼠脑缺血再灌注损伤后脑组织MMP-9的表达及神经调节素的干预作用

目的观察大鼠脑缺血再灌注损伤基质金属蛋白酶(MMP-9)表达及神经调节素-1β(NRG-1β)的干预作用。方法取成年健康雄性Wister大鼠100只,应用线拴法经颈外-颈内动脉插线建立大脑中动脉闭塞再灌注(MCAO/R)动物模型,经颈内动脉单剂量注射1.5%NRG-1β5μL干预治疗。采用免疫组化、免疫荧光、免疫印迹法,观察脑组织MMP-9表达。结果脑缺血再灌注损伤可诱导脑组织MMP-9表达。随着缺血缺氧时间的延长,对照组MMP-9蛋白的表达逐渐增加,阳性细胞形态大小不一,分布于脑组织各区。NRG-1β治疗可降低MMP-9的表达水平,与对照组相应脑区相比,差异显著(P<0.01)。结论MMP-9参与脑缺血再灌注损伤后的炎症反应过程。NRG-1β可能通过调节脑组织中MMP-9的活性,干扰脑缺血再灌注损伤后炎症的发生发展过程,改善神经元的生存环境,延迟神经元损伤的时间和程度,进而对缺血性脑损伤具有积极的保护作用。