超免疫法

Methods: We divide the rats into 5 groups in random, with Morris method, detect the change of some active oxygen radicals such as super-oxide demitasse, Nitric Oxide Synthase, malondialdehyde etc. Also, we detect the change of cholinesterase, Monoamine Oxidize; using HE,PAS staining, immuno-histochemical technique, detect hippocampus CA1 cerebral pathology slices optical density value of NOS, CHE and lipofuscin of neural cell; detecting apoptosis rate and the expression of P53 , which are neuronal apoptosis related genes.

大鼠随机分为5组，采用"Morris水迷宫"法观察脑缺血性模型大鼠学习记忆功能，检测超氧化物歧化酶、一氧化氮合酶、丙二醛等自由基代谢异常的改变，检测胆碱酯酶、单胺氧化酶等老化相关酶的变化、脑组织神经细胞免疫组织化学技术、检查海马CA1区脑组织的病理切片，检测NOS、CHE光密度值；流式细胞技术：检测细胞凋亡率，及促细胞凋亡基因、P_(53)蛋白表达；病理切片采用HE染色、PAS染色及免疫组化法检查，检测对大鼠海马神经元细胞一般病理变化及脑细胞内脂褐含量。

Wβe measured the levels of MDA in plasma and ultrafiltrate, Scr and 2-MG of 23 ESRD patients undergoing regular maintained on HD before and after CAVH compared with 23 healthy people as control by using TAB chromatometry, radioimmunoassay and base saturation picric acid test.

MDA方法：应用硫代巴比妥酸比色法、放射免疫法、饱和苦味酸法分别检测例维持性血液透析23ESRDHD的患者治疗前后血浆和超滤液中、血清βCAVHMDA2微球蛋白(β-2)和血清肌酐水平，并与例健康人对照。

A series of indexes reflecting free radical changes, anti-oxidations, Immune function, endocrine function was studied by Chemiluminescence method, immunoturbidimetry, ANAE dye method, micro-hemagglutination inhibition test and radio immunoassay in the chicken with encephalomalicia induced by giving the feed being deficient in VE and extra fish oil artificially. The pathological changes including microscopic and ultramicroscopic changes in the relative organs and tissues were observed.

本实验以一日龄150只海兰兰蛋鸡为实验动物，用低维生素E且添加鱼油日粮饲喂雏鸡，复制脑软化症动物模型，采用化学发光法、免疫浊度测定法、酸性α-醋酸萘脂酶染色法、微量血凝抑制试验、放射免疫测定技术等方法系统地检测了脑软化雏鸡的自由基代谢和抗氧化功能、红细胞免疫功能、细胞免疫功能、体液免疫功能及非特异性免疫功能等指标，并对患雏病理组织进行了显微及超微结构观察。

After 24 hours, all rats were anesthetized again and their left common carotid arteries were cannulated to facilitate blood withdrawal for serum sample. The contents of tumor necrosis factor-a and interleukin-6 (IL-6) in serum were determined before and after experiment by ELISA; the activity of superoxide dismutase and content of malondialdehyde were determined by xanthinoxidanse method and modified TBA microdetermination. The content of NO2(superscript -)/NO3(superscript -), and activity of nitric oxide synthase and inducible NOS were determined by nitrate reductase method, and chemical chromogenic reaction, respectively; the immunoglobulin G, IgA, IgM and complement 3 (C3) and C4 were determined by immunoprecipitation method before and after experiment.

实验评估：手术创伤后24h进行采用酶联免疫吸附法测定实验前后血清肿瘤坏死因子α、白细胞介素6含量；应用黄嘌呤氧化酶法测定超氧化物歧化酶活性、改良TBA微量法测定丙二醛水平；以硝酸还原酶法、化学显色法分别检测NO2/NO3浓度及一氧化氮合酶、诱导型一氧化氮合酶活胜；以免疫比浊法经全自动生化分析仪检测血清免疫球蛋白IgG、IgM、IgA及补体C3、C4含量。

In the study, we established the animal model of diabetic ED successfully. Modern medical methods such as routine lightscopy, pathology, transmission electroscopy, scanning electroscopy, fluorospectrophotometer and radio-immunity were employed. From the aspects of pathomorphism, cellular supermicrostucture and neurocrine, we observed the mixture's effect on status of the whole animal model and penial erection, the fast plasma glucose, protein non-enzyme gycosylation, nitric oxide synthase , testosterone. In contrast with control groups, the mixture could apparently slow down the corpus cavernosum penial, vascular, neural pathologic progression of the diabetic rats and had a better result than diabetic pill, invigorating yang and restoring normal capsule and amino guanidine.

通过实验研究，成功建立了糖尿病性阴茎勃起功能障碍动物模型，利用常规光镜病理、透射电镜、扫描电镜、荧光分光光度计、放射免疫法等现代医学研究方法，从病理形态学、细胞超微结构、神经内分泌学等方面，观察了降糖起萎合剂对模型动物整体状态和阴茎勃起情况、空腹血糖水平、蛋白非酶糖基化、一氧化氮合酶、睾酮等实验指标的影响，与对照组比较提示降糖起萎合剂能明显延缓大鼠基于DM基础上的阴茎海绵体及血管、神经病变的病理进程，其疗效优于消渴丸、壮阳复春灵胶囊及氨基胍。

①Rat MSC and VSMC were cultured and identified, respectively. MSC were labeled with DAPI firstly, and then co-cultivated with VSMC. The changes of morphology and ultrastructure of co-cultured cells were observed. Immunfluorescence analysis was performed by using monoclonal antibodies against specific antigen.②We established the regulatable system in two steps: a stable MSC line expressing rtTA has been constructed and characterized firstly by transfected with pUHD 17-1hyg and then selected by hygromycin B; in a second step, this line was used for trandfer the AT2R gene to MSC to get the well establishing double stable MSC lines;③The expression of AT2R regulated by doxycycline was evaluated by western blot;④The MSCs were transduced into rat carotid arteries with regulatable AT2R gene after the establishment of rat carotid balloon injury restenosis model. The intimal/medial area ratio were measured by digital analysis system.

研究方法：(1)密度梯度离心法及胶原酶消化法分别培养原代大鼠MSC及VSMC，细胞共培养并行免疫荧光化学染色和透射电镜观察超微结构；(2)组成受Dox调控的哺乳动物表达系统的四种成分的转化、扩增及提纯并酶切鉴定；(3)采用常规分子生物学方法连续两个回合转染体外培养的MSC，并分别采用发光计检测不同细胞克隆萤光素酶活性改变以及RT-PCR方法检测AT2R目的基因mRNA表达情况，根据各个细胞克隆受Dox调控表达的程度，选择低背景、高诱导表达AT2R的细胞系，作为双重稳定MSC细胞系；蛋白免疫印迹法观察该细胞系在Dox调控下AT2R表达的时相性、持续性及在不同浓度Dox调控下的表达情况；(4)建立大鼠颈动脉球囊损伤动物模型，将双重稳定MSC在术中导入血管，分别于14 d、28 d进行病理切片，检测可调控AT2R对新生内膜增生的影响；采用RT-PCR免疫组织化学免疫荧光等技术观察AT2R基因在新生内膜中的表达以及细胞外基质成分表达的改变，TUNEL法检测血管组织中细胞凋亡的变化情况。

We observed the effect of chronic hypoxia on intracelular levels of CGRP in pulmonary endocrine cells of SD rats by means of immunocytochemistry and supraoptimal dilution method.

我们用免疫细胞化学方法及超稀释法观察了慢性缺氧对SD大鼠肺中神经内分泌细胞内降钙素基因相关肽的含量的影响。

In Part 1 of our research, BGSCs were sorted through immunomagnetic beads marking by CD133 and cultured in vitro, and character as a stem cell was identified by stem cell markers (CD133 and Nestin) and differentiated cell markers [ microtubule-associated protein 2(MAP2), glial acidic fibrillary protein and myelin basic protein] , ultrastructure observing with electron microscopeand engrafting to severe combined immunodeficiency mice for tumorigenesis test. The results were as following: Only a small subset of CD133+ glioma cells in glioma cell lines and fresh specimens from various pathologic grade could express stem cell markers CD133 and Nestin, view ultrastructure of a stem cell and be capacity of serial passage in culture. These CD133+ cells possese a marked capacity for multipotent differentiation and could differentiate into tumor cells expressing MAP2,β-TubulinⅢ, GFAP and MBP; When engrafted into SCID mice, they can generate and form tumors that phenotypically resembl the tumor from the patient.

在本课题中，在第一部分实验中采用以CD133为标志的免疫磁珠法从人脑胶质瘤组织和细胞株中分离脑胶质瘤干细胞并进行体外培养，通过免疫荧光技术检测干细胞标志物CD133、Nestin，诱导分化后检查分化细胞标志物MAP2、GFAP、MBP以及电镜超微结构观察和移植SCID鼠致瘤实验，对其干细胞特性加以鉴定，得到如下结果：不同病理分级的新鲜胶质瘤标本和胶质瘤细胞株中存在一小部分CD133+的胶质瘤细胞，能表达干细胞的标志物CD133和Nestin，符合干细胞的超微结构特点，体外培养能连续传代；具有多向分化潜能：诱导分化后能产生MAP2、β-TubulinⅢ、GFAP、MBP染色阳性的细胞；移植SCID鼠后能形成与亲本肿瘤表型一致的移植瘤。

Six and twelve boors after sepsis, leptin levels in liver homogenate were detected by radioimmunoassay serum alanine aminotransferase and 4 enzymes in liver homogenate. myeloperoxidase, glutathin-S-transferase, xanthine oxidase and superoxide dismutase, all related with synthesis of free radicals, detoxication and purine metabolism, were detected by 96 well spectrophotometry. H-E staining was used to examine the histopathologic changes of liver.

于脓毒症后6 h和12 h，采用放射免疫法测量肝组织匀浆液中Leptin水平，采用96孔分光光度法检测血清丙氨酸氨基转移酶及髓过氧化物酶、谷胱甘肽－S转移酶、黄嘌呤氧化酶、超氧化物歧化酶等4种与自由基合成、解毒和嘌呤生成代谢相关的酶的水平，同时以H-E染色法观察肝组织病理学改变。

We killed some mice at the day 6th , the others were continue to expose to formaldehyde and observe their estrous cycle. HE staining was used to study the pathological changes happened in ovarian tissue. Uranium acetate and lead acetate double staining was used to study the ultrastructure of oocyte. E2 and LH level in blood serum was measured by radioimrnunoassay reagent kit.

第6d处死一批小鼠，用HE染色观察小鼠卵巢组织的病理改变，用醋酸铀—醋酸铅双重染色电镜下观察小鼠卵母细胞超微结构的损伤；用GSH测定试剂盒测定小鼠卵巢组织中的GSH含量；用E_2和LH放射免疫分析试剂盒分别测定小鼠血清中E_2及LH水平；采用超排卵法，计数超排卵数并收集卵母细胞，于油镜下观察生殖细胞遗传物质的损伤。

However, as the name（read－only memory）implies, CD disks cannot be written onorchanged in any way.

然而，正如其名字所指出的那样，CD盘不能写，也不能用任何方式改变其内容。

Galvanizes steel pallet is mainly export which suits standard packing of European Union, the North America. galvanizes steel pallet is suitable to heavy rack. Pallet surface can design plate type, corrugated and the gap form, satisfies the different requirements.

镀锌钢托盘多用于出口，替代木托盘，免薰蒸，符合欧盟、北美各国对出口货物包装材料的法令要求；喷涂钢托盘适用于重载上货架之用，托盘表面根据需要制作成平板状、波纹状及间隔形式，满足不同的使用要求。