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胶质细胞

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Small intestinal biopsies showed subepithelial collagen deposition with varying degrees of villous atrophy and varying numbers of intraepithelial lymphocytes. Four patients had previous biopsies showing enteropathic changes without collagen deposition. Seven cases were associated with collagenous colitis and 1 also had features of lymphocytic colitis. Three patients also had collagen deposition in gastric biopsies. One case was associated with lymphocytic gastritis. Celiac disease (CD, gluten-sensitive enteropathy) was documented in 4 patients. Five patients made a clinical improvement with combinations of a gluten-free diet and immunosuppressive therapy. Two patients died of complications of malnutrition and 1 of another illness. Clonal T-cell populations were identified in 5 of 6 cases tested. Four of these patients improved clinically after treatment but 1 has died. Collagenous sprue evolved on a background of CD in 4 cases. There was no history of CD in others and these cases may be the result of a biologic insult other than gluten sensitivity.

小肠活检表现为上皮下胶原沉积,绒毛萎缩程度不一,上皮内淋巴细胞浸润数量不等。4例患者以往曾做过活检,表现为其他肠病改变,并没有胶原沉积。7例患者合并胶原性结肠炎,且1例还有淋巴细胞性结肠炎的特征。3例患者胃活检也发现有胶原沉积。1例患者合并淋巴细胞性胃炎。4例患者有乳糜泻(CD,谷蛋白敏感性肠病)病史。5例患者经无麸质饮食和免疫抑制治疗后,临床症状有所改善。2例患者因营养不良合并另一种疾病而死亡。6例患者做了T细胞受体基因重排,其中5例发现有克隆性T细胞群,这5例中有4例治疗后临床症状加重,且1例死亡。4例患者在CD的基础上病发胶原性口炎性腹泻;其余患者无CD病史,他们的发病也许是生物源性损害,而非谷蛋白敏感。

RESULTS A eukaryotic expression system for high expression humanmutantCD59 were successfully set up : The recombinant PALTER-MAX plasmid containing human mutantCD59 cDNA and PCDNA plasmid were co-transfected into CHO cell by cation lipoid mediating method ;and the cells were grown in F12 medium containing 400ug/ml G418 for 14 days, positive clones were grown in RPMI1640 medium to get stable expressing cell lines . Highly expressing clones were selected by flow cytometry ,and were named PALTER-CD59-CHO1PALTER-CD59-CHO2 . Flow cytometry indicated that expression rates of PALTER-CD59-CHO1 and PALTER-CD59-CHO2 were 53.7%and 54.5%. Further more, Stable highly expressing CHO cell lines were more detected by immunocytochemistry and immunofluorescence technology . PALTER-CD59 -CHO1 and PALTER-CD59-CHO2 were grown in RPMI1640 to get a large of cells . CD59 protein were obtained by spalling PALTER- CD59- CHO1 and PALTER - CD59 - CHO2 cells . Stable highly expressing cells were further validated by SDS-PAGE, immunoblot analysis and solid enzyme immunoassay . PALTER - CD59 - CHO1 and PALTER - CD59 - CHO2 were glycated in RPMI1640 of 50mM ribose for 72 hours .BCECF releasing test indicted that the releasing rate of PALTER-CD59-CHO1 or PALTER-CD59-CHO2 was less high than PALTER-CHO ,and the releasing rate of glycated PALTER-CD59 -CHOI or PALTER-CD59-CHO2 was higher than unglycated ones . PALTER -CD59-CHO1 and PALTER -CD59 -CHO2 were glycated in RPMI1640 of 50mM ribose for 72 hours .BCECF releasing test indicted that the releasing rate of PALTER - CD59 - CHOI or PALTER - CD59 - CHO2 was less high than PALTER-CHO ,and the releasing rate of glycated PALTER-CD59-CHO1 or PALTER - CD5 9-CHO2 was higher than unglycated ones .

结果 成功构建突变人CD59的真核细胞表达系统:运用阳离子脂质体介导法将含有突变人CD59的PALTER—MAX重组质粒与PCDNA共转染入CHO细胞:用含有400ug/mlG418的F12培养基培养14天,筛选出稳定阳性表达克隆,RPMI1640培养基扩增获得稳定表达细胞株,并用流式细胞术进一步筛选出高效表达细胞株分别命名为PALTER—CD59—CH01、PALTER—CD59—CH02,表达率分别为53.7%、54.5%;应用免疫组化方法、免疫荧光技术进一步鉴定阳性细胞株;RPMI1640培养基大量扩增PALTER—CD59—CH01、PALTER—CD59—CH02细胞株,裂解细胞得到CD59蛋白质;通过SDS—PAGE凝胶电泳技术、免疫印迹技术、固相酶联免疫吸附试验验证了这两中文摘要个阳性细胞株CO59蛋白的高效表达;50mM核糖培养72小时,获得突变人CD59糖化细胞株,BCECF染料释放试验结果显示,PALTER一CD59一CHOI、pALTER一CD59一CHOZ细胞较PALTER一CHO细胞染料释放率低,未糖化PALTER一CD59一CHOI、PALTER一CD59一CHOZ细胞比较糖化后细胞染料释放率低。

FN and LN (the non-collagenous index) out of ECM were measured by immuno-histochemistry mean, and the accumulation of ECM was measured by typeⅠand Ⅲ collagen ,α-SMA was chosen as a symbol of the activated reposit fat cell. TGF-β〓 was chosen as a symbol of the cellular factor made the reposit fat cell activated. We observed the mechanism of 286 through the variety of the upper indexes during the experiment.

用免疫组化方法测评肝细胞外基质中的非胶原指标FN、LN,胶原指标Ⅰ、Ⅲ型胶原等在实验过程中的变化作为判定ECM沉积增多的事实指标;用a-平滑肌肌动蛋白作为贮脂细胞被激活的标志物指标;用转化生长因子-β〓作为激活贮脂细胞的细胞因子指标;通过观测上述指标在实验中的变化,来探讨"268方"的作用机理。

FN and LN(the non-collagenous index) out of ECM were measured by immuno-histochemistry mean, and the accumulation of ECM was measured by type I amd III coHagen, a -SMA was chosen as a symbol ef .the activated reposit fat cell.TGF- P1 was chosen as a symbol of th ieccellular factor made the reposit fat cell activated.We observed the mi sclnanism of 286 through the variety of the upper indexes during the experiment.

用免疫组化方法测评肝细胞外基质中的非胶原指标FN、LN,胶原指标Ⅰ、Ⅲ型胶原等在实验过程中的变化作为判定ECM沉积增多的事实指标;用a-平滑肌肌动蛋白作为贮脂细胞被激活的标志物指标;用转化生长因子-β_1(TGF-β_1)作为激活贮脂细胞的细胞因子指标;通过观测上述指标在实验中的变化,来探讨"268方"的作用机理。

Result:After removal of the bilateral lower molars, the remodeling and degenerative activities in different layers of the condyles increased, such as vesiculation of endoplasmic reticulum, distortion of mitochondria and increase of lysosome. Furthermore, the sequential arrangement of collagen fibrils was disturbed, the collagen network broke down and the proteoglycan granules decreased.

结果:细胞发生了不同程度的改建及退行性反应,如粗面内质网的扩张,线粒体肿胀,溶酶体增多等;基质中胶原原纤维的有序排列结构消失,胶原网受到破坏,同时蛋白多糖颗粒减少;基质中脂质小体及基质小囊等成分增多;负重区较非负重区的改变更显著。

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。

This pattern is characterized by the combination of pro-MMP-2 and membrane type 1 (MT1)-MMP expression, which drie pericellular generation of actie MMP-2 and local degradation of normal lier matrix. In addition there is a marked increase in expression of TIMP-1 leading to a more global inhibition of degradation of fibrillar lier collagens by interstitial collagenases (MMP-1/MMP-13). These pathways play a significant role in the progression of lier fibrosis.

其降解肝正常基质,同时抑制引起肝纤维化的纤维胶原降解,这种情况的特点是前明胶酶A合成和膜型1-MMP表达,使得小叶内细胞激活MMP-2和降解当处的正常肝基质,此外血浆1型组织基质金属蛋白酶抑制剂(TIMP-1)明显升高引起广泛的抑制间隙胶原酶(MMP-1/MMP-13)对纤维肝脏胶原的降解。

Immunoelectron microscopy showed that the distinct ultrastructure of fibroblast and the group, spot gold particle specificity distribution shape. The positive markers of CTGF were mainly located in the RER,while there are expression in RER ribosome, Cf, ECM, desmosome connection, euchromatin,et al.

免疫电镜标记显示成纤维细胞细胞超微结构清晰,金颗粒呈团状、点灶状分布,特异性强,CTGF蛋白阳性标记主要位于粗面内质网,粗面内质网核糖体、胶原纤维、细胞外基质、桥粒连接、常染色质等部位也有表达。

Some cell dropped into the cavity and became free. Thrombosis or part organization could be seen. The internal elastic layer became thin, disappear or broken. In internal and middle layer existed fibroblasts, fibrocytes and collagen. Some of the wall indicated hyaline change, soomth muscle cell decreased greatly. The massive inflammatory cells invaded the middle and external layer. There were many foam cells in the capsule tissue. Cytoplasm was filled with fatty tissue and cholesterol. some cavities were full of thrombosis. Some thrombosis was fibrosis, the bottom was organization. The surface of the thrombosis existed red blood cell and librae.(2)Elatic fibrila staining: the internal elastic menbrane almost completely disappeared, the intact internal elastic menbran could be seen in the new small vessels.

动脉瘤管壁厚薄明显不均,全层或局部区域显著变薄向外膨出,内皮细胞空泡变性或坏死脱落,部分内皮细胞剥离并突入管腔成游离状,可见血栓形成及部分血栓机化;内弹力板变薄、消失或突然中断;在内膜及中膜部位主要为纤维母细胞、纤维细胞和大片胶原;部分动脉瘤壁呈均质状玻璃样变,平滑肌细胞明显减少;中膜和外膜可见大量的炎性细胞浸润;瘤壁组织有纤维母细胞、纤维细胞、大片胶原成分及较多泡沫细胞,胞浆内充满脂类物质及胆固醇结晶;部分动脉瘤腔内充满血栓,有的血拴已经纤维化,血栓基部机化,血栓表面有红细胞和纤维素。

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