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Methods The immobilizing conditions of neutrophil alkaline phosphatase staining were detected by the stationary liquid of 40%, 60% and 80% acetone-citric acid, and at the same time, the staining conditions were detected at 10, 15 and 20 minutes in the matrix liquid prepared respectively with 2-amino-2-methyl-1.3-propylene glycol (PH9.4-9.6) buffer, barbital (PH9.2) buffer and Tris (PH9.2) buffer.

用40%、60%、80%的丙酮-枸缘酸固定液对中性粒细胞碱性磷酸酶染色固定条件进行测定。同时用2-氨基-2-甲基-1.3-丙二醇(pH9.4-9.6)缓冲液、巴比妥(pH9.2)缓冲液、Tris(pH9.2)缓冲液配制的基质液分别在10、15、20分钟的染色条件进行测定。

In addition to glial cells, CNTF immunoreactivity are widely localized at neurons in the olfactory bulb, cortex, hippocampus, cerebellum, diencephalon and brainstem.

结果表明,CNTF免疫反应物质除广泛存在于中枢神经系统的胶质细胞外,亦有神经元的广泛染色,而且分布广泛,在嗅脑、大脑皮层、海马、小脑、间脑、脑干中均有明显的染色。

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。

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: High-purity ADSCs were acquired successfully. The ADSCs were induced into osteoplastic differentiation, and displayed typical adipocytes changes 7 days after induction. the cells turned to slender fusiform and mixed together, which showed positive PPARγ immunohistochemical staining and the lipid droplet was stained orange-red by oil red staining.

结果:①成功获得纯度较高的脂肪基质细胞,成功诱导其脂向分化,诱导培养后7 d观察到细胞形态学上表现出典型的脂肪细胞改变:细胞相互融合,成片生长,变为细长纺锤形;PPARγ免疫细胞化学染色呈阳性;油红O染色脂滴被染成橙红色。

After the cell growth curves was recorded, RPE cells of the 3-5th passages were utilized. 2、Three different siRNA (siRNAl,siRNA2,siRNA3) targeting against human cx43 gene and one negative control siRNA were designed and transfected into cultured human RPE cells via liposome reagent. The most effective siRNA can be determined by semi-quantitative reverse transcription PCRRT-PCR. 3、To the most effective siRNA, after transfected into human RPEs with different concentration, the cellular proliferate activities were messured by MTT colorimetry ; the percentages of RPE in different cell circle phase was assayed by FCM; the changes of phenotypical properities were observed with SCM; the protein expression of cx43 was studied through immunocytochemistry stain and Weston blot; the communication intercellular was calculated with FRAP; and the ability of recovery was assessed by using an in vitro wound healing model.4、The total proteins of siRNA1 and RPE were seperated by two-dimensional gel electrophoresis and visualized by silver staining. Proteins with significant expression alterations were selected and their peptide mass fingerprints (PMFs were obtained by matrix-assisted laser desorption/ionization time of flying mass spectrometry (MALDI-TOF-MS).The PMFs were used to search NCBInr database by Auto MS-Fit software.

实验方法:1、培养原代的人RPE细胞,经过细胞角蛋白、S-100和神经胶质原纤维酸性蛋白免疫细胞化学鉴定后,通过AO/PI染色技术确定培养细胞的存活率,描记其生长曲线,第3-5代用于以下细胞实验2、生物合成针对人cx43基因的三条小干扰RNA和一条阴性RNA通过脂质体转染RPE细胞后,通过RT-PCR的方法确定抑制效率最高的干扰片断3、将该片段以不同浓度通过阳离子脂质体转染培养的人RPE细胞后,采用MTT法观察其对细胞的增殖力的作用;通过流式细胞仪观察其对细胞周期的影响;通过扫描电镜观察其对细胞形态的影响;通过免疫细胞化学和Weston blot观察其对cx43蛋白表达的作用;采用激光共聚焦和荧光淬灭恢复技术观察荧光恢复速率平均百分率,评价其对细胞间通讯功能的影响;通过制作RPE细胞损伤模型,观察其对损伤修复能力的作用4、分离纯化转染siRNA的RPE组和正常对照组RPE细胞的全部蛋白质,应用等电聚焦电泳和SDS-PAGE双向电泳技术,银染显示分离出的蛋白质斑点,经凝胶图像分析软件对两个样本进行胶图分析,寻找差异蛋白点。

And Sirius Red in a supersaturated picric acid solution viewed by polarization microscopy. The changes of mRNA expression of collagen type I were analyzed by RT-PCR. The results were compared with those after intervention with Losartan.

染色及苦味酸天狼猩红胶原特异染色偏振光显像进行心脏间质胶原的组织化学观察,RT-PCR观察心脏I型胶原mRNA表达的变化,并与氯沙坦干预后比较。

Methods We reported a case of PXA located in the cerebellum of a young adult in view of its extremely unusual location. Morphologic observation, immunohistochemical staining using DAKO EnVision system , histochemical stains including hematoxylin eosin and Gordon Sweet's reticulin methods were used.

采用免疫组织化学DAKOEnVison法、组织化学网质纤维染色和HE染色及形态学观察的方法,报道 1例极罕见于年轻人小脑的多形性黄色星形细胞瘤,探讨其临床病理特点。

Results The expression of Vimentin declined gradually during passaging process from the primary generation to the fifth generation, while the highest level of V...

结果 Vimentin在原贴壁基质细胞系由原代传至 5代的细胞中,其表达逐渐下降,而在再贴壁基质细胞传至第 3代中表达最高;Desmin、α-SmoothMuscleActin单克隆抗体和VonWillebrandFactor抗体染色呈阴性。

It was observed by Nissl and enzyme histochemistry staining that exogenous GDNF decreased lesion-induced motor neuron death in lateral nucleus of spinal anterior horn and the changes in activity of cholinesterase and acid phosphatase in spinal cord and sensory ganglions.

采用硅管套接大鼠切断的坐骨神经模型,局部给予胶质细胞源性神经营养因子,应用尼氏染色、酶组织化学染色方法,观察到外源性GDNF能减少脊髓修复侧前角运动神经元死亡的数目,降低脊髓前角运动神经元及脊神经节感觉神经元中胆碱酯酶及酸性磷酸酶变化的幅度。

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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。

There is no differences of cell proliferation vitality between labeled and unlabeled NSCs.

双标记神经干细胞的增殖、分化活力与未标记神经干细胞相比无改变。