苯胺蓝

Methods: A 5ml bone marrow was extracted from the lilac of human volunteers. By Percoll fluid and density gradient centrifugation, the MSC was obtained; after the cells filled the bottom of vessel, subcultured them, when they subculture in third generation, redigested them, 500 R/min centrifugate, alter the completed medium to chemical definition medium, examined the form change and prolifration of cells by invert microscope, toluidine blue stain、immunocytochemical stain and RT-PCR to test the type Ⅱ collagen mRNA and proteoglycan.

取健康成人髂后上棘处骨髓5ml，经percoll液分离后密度梯度离心，〓／ml密度接种培养，观察原代细胞的贴壁、增殖状况，细胞长满瓶底后进行传代培养；传至第三代细胞，重新消化后以500转／分钟轻度离心5分钟，〓／ml接种，改用化学限定培养基代替完全培养基培养，倒置显微镜观察细胞生长情况及形态变化，甲苯胺蓝染色观察诱导细胞合成细胞外基质中的蛋白多糖，免疫细胞化学染色检测ECM中Ⅱ胶原的蛋白合成，RT-PCR鉴定诱导细胞Ⅱ胶原mRNA的表达。

Part one Isolating, culturing, chondroblast induction and evaluation of human MSC in vitroObjective:To examine the property and proliferation ability of primary culturing MSC ,mvestgate the effect of lightly centrifugation and chemical definition medium to induce MSC differentiate into chondroblast.

结果：原代骨髓基质干细胞易于体外培养，增殖旺盛；经轻度离心和化学限定培养基诱导后细胞呈高密度生长，仍保持较旺盛的增殖能力，细胞转化成圆形肥大细胞，甲苯胺蓝染色显示软骨细胞外基质 v的特异性异染，11胶原蛋白免疫细胞化学染色呈强阳性，BT干CR鉴定显示＊胶原mRNA丰富的表达。

The aim are:lTo examine the proliferation ability and potential chondroblast differentiation ;2To find an ideal condition stimulated BMSC differentiate into chondroblast;3To examine the chondroblast proliferation in porous scaffolds and to explore the interaction between cells and materials;4To examine the release of cytokine in vitro.

取健康成人略后上棘处骨髓 5ml，经 percoll液分离后密度梯度离心，10'砌l密度接种培养，观察原代细胞的贴壁、增殖状况，细胞长满瓶底后进行传代培养；传至第三代细胞，重新消化后以500转／分钟轻度离心 5分钟，10V加 l接种，改用化学限定培养基代替完全培养基培养，倒置显微镜观察细胞生长情况及形态变化，甲苯胺蓝染色观察诱导细胞合成细胞外基质QCM）中的蛋白多糖，免疫细胞化学染色检测ECM中＊胶原的蛋白合成，RT干CR鉴定诱导细胞＊胶原mRNA的表达。

RESULTS: At 16 weeks after the compound of after-induced chondrogenetic BMSCs and SIS were transplanted, the materials had been completely absorbed and there were new cambium in the experimental group, which proved to be chondrocytes after hematoxylin-eosin stain, Massan stain and Toluidine blue dyeing.

结果：诱导软骨细胞-小肠黏膜下层复合物在植入体内16周时材料明显被吸收完全，且有软骨生成，与正常组织未见明显区别，但修复层软骨较薄。经苏木精-伊红染色，Massan染色，甲苯胺蓝染色检查证实为软骨组织。

The tissue structure, elastic fiber, collagenic fiber and smooth muscle were stained by HE, Weigert, Aniline blue and Orange G respectively.

用HE法、Weigert法、苯胺蓝法及桔黄G法分别染组织结构、弹性纤维胶原纤维和平滑肌。

The uranine staining can be used to observe the whole growing phases of Sphaerotheca fuliginea, while aniline blue staining is suitable for conidial observation.

荧光素钠可以观察到菌体整个发育阶段。苯胺蓝只适合于前期菌体入侵过程的观察。？

Following studies had been performed:(1)repetitive WLT and real-time ultrasonography had been performed to 18 HS and 40 FD toassess the reproducibility and influencing factors.(2) correlations of WLT with symptomscore and electrogastrography had been analysed.(3) differences ofWLT between HS and FD had been compared.(4) methophenolane andimmunohistochemistry stain of gastric fundus mucosa biopsy from 40 FD patients hadbeen performed to analyse the correlations of symptom score and WLT with count andactivity of mast cells, 5-HT level, and H.Pylori infectious status.(5) the effect of oralsumatriptan to proximal stomach function in 10 HS and 10 FD had been assessed, so as toestablish the possible regulating mechanism of fundic relaxation, explore the therapeuticpotential and feasibility of sumatriptan on FD.Results:I. Evaluation of WLT and its application in FD management.

我们主要进行以下几个方面的研究：(1)对18名健康人及40名FD患者重复进行水负荷试验并结合B超声对近端胃的实时监测，评价水负荷试验结果的可靠性并分析其影响因素；(2)观察水负荷试验结果与症状积分及体表胃电结果的相关性；(3)比较FD患者和健康成人水负荷试验结果的差别；(4)对40名FD患者胃底粘膜活检组织进行免疫组化染色和甲苯胺蓝染色，观察FD患者的症状及水负荷试验结果与胃底粘膜肥大细胞、5-HT表达情况及H.pylori感染情况的相关性：(5)观察口服舒马曲坦对10名健康人及10名FD患者近端胃功能的影响，并由此推测胃底舒张运动调节的生理机制，探讨口服舒马曲坦治疗FD的可行性。

Staining observation on complex mesochondrium of marrow-derived mesenchymal stem cells-TCP: It was found with TB staining that the complex of marrow-derived mesenchymal stem cells-TCP inside the joint cavity obviously had positive staining, which indicated that there were glycosidoprotein base formed.

骨髓基质干细胞-磷酸三钙复合体软骨基质特染观察结果：甲苯胺蓝染色发现，植入关节腔的骨髓基质干细胞-磷酸三钙复合体有明显阳性染色，表明有糖蛋白基质形成。

The central part was composed of hypertrophic chondrocytes, for which the number increased with time. The cartilaginous tissue at the time of 2 weeks' cultivation appeared strongly positive staining of collagen type Ⅱ immunohistochemistry and safranine "O", as well as strongly metachromatic to toluidine blue.

其中心为肥大软骨细胞，且随培养时间的延长而增多。2周时，软骨组织甲苯胺蓝染色呈强的红色异染反应，Ⅱ型胶原免疫组织化学染色呈强阳性，番红&O&染色呈强阳性。

Then the bFGF was replaced by transforming growth factor-β1 (TGF-β1 10 μg·L-1) for next three weeks. Results Some spindle MSCs turned into round or elliptic shape and secreted metachromatic matrix with Toluidine blue.

结果：细胞形态由纺锤形变为圆形或椭圆形，细胞分泌了大量的甲苯胺蓝异染的基质，形成了类软骨陷窝。

This one mode pays close attention to network credence foundation of the businessman very much.

这一模式非常关注商人的网络信用基础。

Cell morphology of bacterial ghost of Pasteurella multocida was observed by scanning electron microscopy and inactivation ratio was estimated by CFU analysi.

扫描电镜观察多杀性巴氏杆菌细菌幽灵和菌落形成单位评价遗传灭活率。