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Collagen Ⅱ immunohistochemistry and toluidine blue staining had a weak expression on primary and subcultured cells.

原代和传代后细胞Ⅱ型胶原免疫组织化学和甲苯胺蓝异染反应均较弱。

The other cartilage specimens were stained with hematoxylin and eorin、toluidine blue to examine the histological structure under light microscopy.

另3 只动物的软骨标本分别行HE 染色和甲苯胺蓝染色,光镜下观察修复软骨的组织学结构。

Carbon nanotube ; toluidine blue ; synergistic effect ; glucose oxidase

碳纳米管;甲苯胺蓝;协同作用;葡萄糖氧化酶

RESULTS: The AF cells cultured in different substrates were morphologically undistinguishable. Toluidine blue staining showed that there was also no difference between AF cells cultured on flexible silicone membranes and in plastic plates. The cells growing in different substrates expressed the same levels of collagen typeⅠ,Ⅱ and aggrecan mRNA and integrin β1. AF cells grew well on silicone membranes.

结果:种植于不同基质的纤维环细胞形态学观察无明显差别;在两种基质上的纤维环细胞甲苯胺蓝染色无差异;Ⅰ,Ⅱ型胶原及聚集蛋白聚糖mRNA及细胞膜整合素β1的表达亦无明显差异;纤维环细胞可以黏附于硅橡胶膜上生长。

Methods Rats were exposed to S band HPM which average power densities were respectively 2~90 mW/cm^2. HE staining, Toluidine blue staining and electron telescope were used to study the change of the structure of cortex, and high performance liquid chromatography was used to study the changes of 4 amino acids (glutaminic acid, asparagic acid, GABA and glycine) of cortex in 6 h and 1d after exposure to 10 and 50mW/cm^2 HPM.

采用2~90mW/平方公分S波段高功率微波辐照Wistar大鼠,通过苏素素-伊红染色、甲苯胺蓝染色和电镜观察大脑皮层的形态结构改变;采用高效液相色谱仪检测10和50mW/平方公分照后6h和ld大脑皮层中4种氨基酸(谷氨酸、天门冬氨酸、γ-氨基丁酸和甘氨酸)含量的变化。

The results have indicated that the introduction of electron-donating methoxy or electron-withdrawing cyano-group at the 7-position of 3-phenylcoumarins caused a bathochromic shift for both absorption and fluorescence spectra. The phenyl group at the 4-position derivative has a hypsochromic shift for both absorption and fluorescence spectra.

结果发现,在3-苯基香豆素的7-位引入给电子基团甲氧基或吸电子基团氰基均使它们的吸收光谱和荧光光谱产生红移,4-苯基香豆素的衍生物的吸收光谱和荧光光谱均产生蓝移。3-苯基香豆素衍生物与4-苯基香豆素的衍生物的基态和激发态的电子转移方向相反。

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。

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