The cholinergic innervation of the rat fascia dentata: identification of target structures on granule cells by combining choline acetyltransferase immunocytochemistry and Golgi impregnation

A monoclonal antibody against choline acetyltransferase (ChAT), the acetylcholine-synthesizing enzyme, was used to study cholinergic synapses on identified (Golgi stained) granule cells in the rat fascia dentata. Choline acetyltransferase immunocytochemistry was applied to 40-microns Vibratome sections cut perpendicular to the longitudinal axis of the hippocampus. Light microscopy revealed fine varicose ChAT-immunoreactive axons in all layers of the fascia dentata, i.e., in the stratum moleculare, the stratum granulosum, and the subgranular polymorph zone. Most fibers were observed in the vicinity of granule cell bodies where they ran mainly parallel to the granular layer. Next, the immunostained Vibratome sections were sandwiched between small pieces of Parafilm and piled to form a block that was covered with agar and Golgi stained. After that, the sections were separated by cutting away the agar and removing the Parafilm. Sections containing well-impregnated granule cells were gold-toned (Fairén et al., '77), embedded in Araldite, and subjected to ultrathin sectioning for electron microscopy. A total of 14 gold-toned granule cells were examined in the electron microscope for synaptic contacts with cholinergic afferents. Choline acetyltransferase-immunoreactive axon terminals were observed that established symmetric synaptic contacts with the cell bodies and dendritic shafts of the gold-toned identified granule cells. Two types of contact were observed on spines arising from gold-toned granule cell dendrites. Immunoreactive terminals established asymmetric synaptic contacts with the head of small spines and symmetric contacts with the stalk of large, complex spines. The boutons forming asymmetric synaptic contacts with the cup-shaped spine head of the complex spines were not found to be immunoreactive. Our results demonstrate that cholinergic fibers to the rat fascia dentata establish characteristic types of synaptic contact with different postsynaptic elements of granule cells, suggesting a complex function of this afferent system.     

免疫吸附治疗在肾移植致敏受者中的应用

目的：探讨蛋白A免疫吸附（immunoads Orption，IA）治疗对清除肾移植致敏受者体内特异性抗HLA抗体的疗效和安全性。方法：10例肾移植致敏（PRA〉50％）受者用彤新蛋白A免疫吸附柱行IA治疗，测定治疗前后血免疫球蛋白及PRA水平。结果：10例患者IA治疗次数为4～15次（中位数9）。所有患者IA治疗后血清总IgG水平都明显下降（P〈0．001），IgA和IgM也较治疗前显著降低（P〈0．01）。PRA8例转阴，1例〈30％，1例仍为100％。结论：对于肾移植致敏受者，IA是一种特异性高、安全有效的治疗措施。     

抗地高辛人源小分子抗体的获取

目的 从大容量噬菌体抗体库中筛选人源性抗地高辛抗体,并构建双体抗体.方法 以固相化的地高辛对构建的大容量噬菌体抗体库进行筛选,3～4轮后挑取克隆用酶联免疫吸附测定方法鉴定其特异性,对抗地高辛阳性抗体克隆进行DNA指纹分析及测序分析.选取活性好的克隆进行改造,构建双体抗体.结果 在抗体库的筛选过程中可见到明显的富集现象,获得了4株可与地高辛特异性结合的人源抗体,经DNA指纹分析及测序分析证明为不同克隆基因,阳性克隆的可变区基因轻链均属于λ第一亚群,重链分别属于第三和第四亚群.选取4号克隆进行基因改造,构建双体抗体表达载体,获得了活性较好的双体抗体.结论 利用噬菌体抗体技术获得了人源性抗地高辛抗体,并改造成应用前景较好的双体抗体,可能为临床诊断及治疗洋地黄类药物中毒提供具有实用价值的人源抗体.     

精浆抗精子抗体阳性不育患者顶体酶、一氧化氮合酶及超氧化物歧化酶活力变化的研究

目的:研究精浆抗精子抗体(AsAb)阳性对精子顶体酶、精浆一氧化氮合酶(NOS)及超氧化物歧化酶(SOD)活力的影响。方法:精浆AsAb阳性不育者40例,对照组为40例正常生育男性。通过吸光度变化分别计算顶体酶活力(BAEE/ADH联合法)、NOS活力(氧化还原反应)、SOD活力(黄嘌呤氧化酶法)。结果:精浆As-Ab阳性组与正常生育组比较,精子顶体酶活力明显降低(P<0.01),NOS活力明显升高(P<0.01),精浆中SOD活力明显降低(P<0.01)。结论:精浆AsAb阳性引起不育可能与精子顶体酶、精浆中SOD及NOS活力改变有关。     

PCR-SSP法HLA-A、B基因分型与血清学分型的比较

目的比较聚合酶链反应-序列特异性引物(PCR-SSP)进行HLA-Ⅰ类A、B抗原位点分型的准确性,并探讨血清学分型错误发生的原因。方法用PCR-SSP以及单克隆抗体血清学分型技术对HLA-A、B分型并比较。结果34例样本PCR-SSP基因分型无假阳性和假阴性出现。PCR-SSP法与血清学比较,血清学检出错误或漏检率分别为HLA-A位点23.5%,B位点26.5%。血清学发生错误或易混淆的抗原有:A2和A68、A32和A33,B5、B60和61。结论PCR-SSP法进行HLA-A、B抗原等位基因分型具有分辨率高、特异性强、重复性好、实验过程简捷快速、分型结果较血清学更加准确可靠的优点。     

特异性神经元抗体对神经系统副肿瘤综合征筛检价值的研究

目的　探讨特异性神经元抗体 (尤其是抗 Hu抗体 )对国内人群神经系统副肿瘤综合征 (paraneoplasticsyndromesofnervoussystem ,PNSNS)筛检诊断的特异性和灵敏性。方法　采用免疫组织化学ABC法和Westernblot对 2 0例PNSNS和 12 0例非PNSNS患者的血清进行检测 ,并采用流行病学筛检公式计算出特异度和灵敏度。结果　PNSNS组和非PNSNS组ABC法阳性结果差异有统计学意义 (χ2 =33 4 5 ,P <0 0 1) ,特异性神经元抗体筛检诊断PNSNS的灵敏度为 95 % ,特异度为 72 5 % ;PNSNS组和非PNSNS组Westernblot法阳性率差异有统计学意义 (χ2 =117 12 ,P <0 0 1) ,抗 Hu抗体筛检诊断的PNSNS的灵敏度为 95 % ,特异度为 98 3%。结论　血清特异性神经元抗体 (尤其是抗 Hu抗体 )是国内人群中PNSNS患者较敏感和特异性的血清标志物 ,在神经元的免疫损伤中具有重要作用     

Fumonisins as a possible contributory risk factor for primary liver cancer: A 3-year study of corn harvested in Haimen, China, by HPLC and ELISA

Employing HPLC fluorometry, gas-liquid chromatography (GLC) and a novel enzymelinked immunosorbent assay (ELISA) based on a monoclonal antibody, 40 corn samples, each collected in 1993 from agricultural stocks for human consumption in Haimen (Jiangsu County) and Penlai (Shandong Province), high- and low-risk areas for primary liver cancer (PLC) in China, respectively, were analysed for fumonisins (FBs), aflatoxins (AFs) and trichothecenes. Levels and positive rates of FBs and deoxynivalenol (DON) were significantly higher in Haimen than in Penlai. ELISA of the 40 corn samples harvested in the two areas in 1994 revealed that FB contamination levels and rates in these areas were comparable to those observed in 1993 in Haimen. ELISA analysis of 1993 and 1994 products revealed a wide occurrence of AFB₁ but the positive rates as well as levels were not significantly different between these areas. ELISA of the same sample number of corn harvested in 1995 revealed that FB contamination in Haimen was significantly higher than in Penlai. These 3-yearly surveys of corn samples (240 in total) demonstrated that corn harvested in Haimen was highly contaminated with FBs and that the contamination level, as well as positive rate in 1993 and 1995, were 10–50-fold higher than those in Penlai, suggesting FBs as a risk factor for promotion of PLC in endemic areas, along with the trichothecene DON. Co-contamination with AFs, potent hepatocarcinogens, was assumed to play an important role in the initiation of hepatocarcinogenesis.     

多种底物在免疫金银染色检测抗核抗体试验中的联合应用

将多种底物联合应用于免疫金银染色检测ANA试验,与各底物单独应用比较,可明显改善检测的敏感性与核型显示。其中,两种鼠脏器印片联合应用的结果可与Hep-2细胞的结果相当,三种及四种鼠脏器印片联合应用的结果可与Hep-2细胞和一种鼠脏器印片联合应用的结果相当,且均不使正常人ANA阳性率明显升高。