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Acellular allogeneic dermal matrix is the dermal replacement of heterogenous human skin in which the cellular elements have been removed by special technique and basement membrane complex have been remained. It becomes a kind of 3D porous acellular dermal scaffold. It supports fibroblast infiltration, neovascularization and epithelialization in the absence of an inflammatory response.

异体脱细胞真皮基质(acellular allogeneic dermal matrix, ADM)来源于正常的异体皮肤组织,经过特殊的理化处理去除真皮中的细胞成分,保留原有的胶原三维结构和完整的基底膜,成为一种多孔的三维膜性支架,其空间网状结构有利于细胞的增殖、毛细血管增生,加速组织的愈合过程,移植后可以迅速被血管化和自体细胞重建。

In recent years,masenchymal stem cells have been applied in cartilage tissue engineering and they have shown significant potenital for carriage repair in animal models,and are being used as an ideal cell source in cartilage tissue engineering.

:间充质干细胞具有高度自我新能力,能够分化各类型的细胞近20年间充质细胞在软骨组织工程中得到应用,动物实验显示了良好的软骨修复潜能,成为软骨组织工程的理想的细胞来源。

To sum up, we first conclude that we can isolate mesenchymal stem cells from human fetal articular cartilage and they can be induced to differentiae into several cells under appropriate induction conditions. Anymore, they have their original tissue speciality. Therefore, we first report that mesenchymal stem cells of articular cartilage origin are superior to mesenchymal stem cells of bone marrow origin as seeding cells of cartilage tissue engineering and we provide a kind of new seeding cells of cartilage tissue engineering for the future clinic application.

综上所述,我们首次报道不仅可从人胚胎的关节软骨中获得间充质干细胞,而且软骨来源的间充质干细胞除具有骨髓来源的间充质干细胞的多向分化能力外,还具有其来源的软骨组织特异性,那么,其作为软骨组织工程的种子细胞要优于骨髓来源的间充质干细胞,从而首次提出了一种新型的软骨组织工程的种子细胞,对于未来的软骨组织工程的临床应用提供理论依据。

Results一. Culturing and identification of human conjunctiva cells1. morphous of conjunctiva cellsCells were harvested by tissue cultivation after digestion. Ponderosus cells adhered after 48 hours. Cells were shown in shape of round, ellipse and polygon. Cell body was loose and lucency with nucleus in center, cell membrane was also clearly seen. Cells were arranged in inlay and fuse in film after 12-14 days.2. Immunochemistry testNomogeneous immunochernical positive granules against CK13 were observed in endochylema of human conjunctiva cells.3. growth curve of conjunctival cellsSubcultured cells were aged and feeble after 5th passage.

结果一、正常人结膜细胞的培养与鉴定1、活体细胞的形态消化后组织块培养法:48小时后见大量上皮细胞贴壁,细胞不规则呈圆形、椭圆形、多边形,细胞体肥大透亮,细胞核位于中央,胞膜清楚,约12-14天后细胞呈镶嵌状排列,融合成膜状。2、免疫组化鉴定培养的人结膜上皮细胞CK13免疫组化染色见阳性颗粒均匀分布于胞浆中。3、培养结膜细胞生长曲线传代培养多数样本于P5代以后开始出现衰老征象。

Nano- and non-nano-Al2 O3 could induce the activities of Kupffer cells in rat livers. However, apoptosis induced by NAOs increased significantly than that induced by nNAOs, which may be related to the metabolism characteristics of the two Al2 O3 particles in rat livers.

NAOs及nNAOs均能引起大鼠肝组织中枯否细胞活化,但NAOs引起肝组织细胞凋亡的程度高于nNAOs,可能与两种不同粒径的氧化铝在肝组织的代谢特征不同有关。

Results: the positive cells mostly included ependymal cells of myelocoele, glial cells, anterior horn motor neurons, and epithelial cells of spinal pia mater and their expressions were mainly located on cell membrane. interestingly, aqp4 distribution in glial cells and ependymal cells showed certain directivity, but the former was mainly in the membrane side contacting with capillary endothelial cells and the latter mostly in the basolateral membrane of ependymal cells.

结果: 免疫组织化学染色和原位杂交结果显示胶质细胞、前角运动神经元、中央管室管膜细胞、软脊膜上皮细胞均表达阳性,aqp4主要分布于细胞膜,其中,胶质细胞和中央管室管膜细胞表达存在极性,前者主要分布在与毛细血管内皮细胞接触的一侧,后者主要分布在中央管室管膜细胞的基底侧膜。

Abjective We detected the quantity and ability of CD4+T and CD8+ T cells in peripheral blood and liver tissues ,and investigated the change of cytokines in the liver to study the probably mechanism about HBV affect cellular immunity system in human body.

目的 通过检测乙型肝炎病毒感染后外周血及感染肝组织内CD4+T和CD8+T细胞的数量及功能,以及检测HBV感染者肝组织内细胞因子TNF-α及TGF-β2表达的变化,探讨HBV对人体T细胞免疫的影响及其可能机制。

As shown,the antigen is a glycoprotein present on the surface of cells. Extracts from Hep G2 hepatocellular carcinoma cells, HT-29 colon cancer cells, SKHep-1 liver adenocarcinoma cells, BT-20 breast cancer cells as well as U937 histiocytic lymphoma cells were positive for the precipitation of Ag-Ab complex, which were later reactive with the anti-3A5 McAb staining. While extracts from normal human endothelial cells showed no precipitation in Western blot.

抗原分子量为63kDa,是一种存在于细胞膜上的糖蛋白,在肝细胞肝癌Hep G2细胞,肠癌HT-29细胞,肝管癌SK-Hep-1细胞,乳腺癌BT-20和人组织细胞淋巴瘤U937细胞膜提取物中,均有此蛋白带,并能被单抗3A5特异性地识别,而人正常内皮细胞则不见此反应带。

The PCR products were examined by agarose gel electrophoresis. The target gene fragments were purified by gel extraction kit and ligated to cloning vector pMD18-T. The recombinant vectors were transformed into host strain E. coli K802 by lithium chloride method, screened and identified with PCR and restrictive enzymatic digestion. Their sequences were confirmed by DNA sequencing.(2) sTWEAK1 gene was subcloned into expression vector pProEx HTb and transformed into E. coli BL21. sTWEAK2 gene was subcloned into expression vector pMAL-C2x and transformed into E. coli TB1. The recombinant vectors were screened and identified with PCR and restrictive enzymatic digestion. The recombinant fusion proteins were induced to express with IPTG, detected by coomassie brilliant blue-stained SDS-polyacrylamide gel electrophoresis , and confirmed by Western blot analysis.(3) The sTWEAK1 fusion protein was purified with Ni-NTA Spin Kit.(4) The biological activity was assayed on transformed and tumor cells by microplate photometer after crystal violet or sulfur rodamine B staining.(5) The contents of IL-8 in the supernatant of 1990 cell cultures were determined by ELISA.(6) The morphological changes of the sensitive cells were observed by light and transmission electron microscopies.(7) The cell cycle and apoptotic rate were assayed by flow cytometry in 1990 and M85 cells.(8) The effect of fusion proteins on induction of NF-κB in 1990 and LOVO cells was detected with Dual-Luciferase Reporter Assay system.(9) The TWEAK gene was subcloned into Adeno-X Viral DNA with pShuttle vector and transfected into HEK293 cells by lipofectamine method.

(1)本研究用RT-PCR方法,从人组织细胞总RNA中扩增可溶性TWEAK胞外区(sTWEAK1和sTWEAK2)的cDNA序列及全长编码序列,用琼脂糖凝胶电泳分析PCR产物,胶回收目的基因片段,连接到pMD18-T克隆载体中,转化大肠杆菌K802,PCR和酶切筛选阳性克隆,全自动DNA测序验证序列;(2)sTWEAK1和sTWEAK2分别亚克隆到pProEx HTb和pMAL-C2x表达载体中,分别转化大肠杆菌BL21和TB1,PCR筛选和酶切鉴定,阳性克隆用IPTG诱导表达,表达产物用SDS-PAGE分析和Western blot验证融合蛋白;(3)用NTA-Ni Spin试剂盒初步分离纯化sTWEAK1融合蛋白;(4)用体外培养的肿瘤细胞和正常对表达产物进行活性检测,贴壁细胞用结晶紫染色法,悬浮细胞用磺酰罗丹明B染色法,酶标仪检测OD值;(5)敏感细胞用ELISA法检测细胞培养上清中IL-8的含量;(6)用光镜和电镜观察敏感细胞死亡和细胞凋亡情况;(7)用流式细胞仪分析表达产物对敏感细胞凋亡率和细胞周期的影响;(8)用双荧光素酶报告基因检测法,测定表达产物对敏感细胞NF-κB的影响;(9)用pShuttle穿梭质粒将TWEAK重组到腺病毒载体上,用脂质体转染法转染HEK293细胞,PCR鉴定重组质粒。

Second is studying the traumatic condition changes in a choosey pressure at different post-PT phases. Moreover, we detected model PLA_2 activity, express of PLA_2 subtype mRNA by fluorescent quantitative RT-PCR technique, protein express and site by Western blot and IHC at different post-PT phases. At last, we studied the PLA_2 inhibited effects and model protection of Variabilin in vivo.

研究在不同压强的外力作用下,模型指定时相点的一般情况和存活率、大体、光镜及电镜下胰腺组织的病理变化和常规酶学变化,得到稳定建模致伤压强;然后,固定致伤压强,研究胰腺创伤后不同时相点的伤情变化(包括一般情况、胰腺组织的病理变化、AMS和LPS活性变化、血常规、血生化、胰腺细胞凋亡以及胰腺组织细胞生长周期变化情况);再次,采用3H-花生四烯酸标记大肠杆菌膜作为底物的方法检测大鼠胰腺创伤后不同时相点的PLA_2活性,利用荧光定量RT-PCR技术检测其亚型mRNA的表达,利用Western blot和IHC技术检测其蛋白表达量和蛋白组织定位;最后,将Variabilin体内应用于急性胰腺创伤大鼠模型,研究其对模型的保护作用和对PLA_2的抑制效应。

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