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Methods: Density gradient centrifugalization and absorption technique wererespectively applied to detach peripheral blood mononuclear cells andperipheral blood lymphocytes,T lymphocytes were got by Nylon postabsorption method, and CD4+T and CD8+T lymphocytes divided with anti- CD4 andanti-CD8 antibody by alexin cytotoxic method.

密度梯度离心法分离外周血单个核细胞,玻璃器皿吸附法分离外周血淋巴细胞,尼龙棉柱法分离 T 淋巴细胞,补体介导的细胞毒法分别制备 CD4+T和 CD8+T 淋巴细胞,采用 T 淋巴细胞与角质形成细胞共培养技术研究银屑病患者外周血 T 细胞对角质形成细胞的影响,免疫组化法检测共培养角质形成细胞 Ki67、c-Myc 及 Bcl-xL 蛋白的表达。

Furthermore, from the result of inoculation with fungi in vivo, some animalcula stimulate the formation and accumulation of dragon's blood. 3. According to the results of zymolysis culture of some animalcula, there are a small quantity of loureirins in the zymolysis products, contents of loureirin A range from 0.090 to 0.439mg/g, loureirin B from 0.012 to 0.133mg/g.

从某些代谢产物呈红色或菌体本身呈红色的菌的液体发酵培养结果来看,这些菌的发酵产物中有龙血素成分存在,但量极少,龙血素A、B的含量分别在0.090-0.439mg/g,0.012-0.133mg/g范围之类,并且其含量随菌的培养时间的增长有增加的趋势。

At the same time,the effect of injections such as Traditional Chinese Medicine injection of danshen and huangqi on the HPMC in vitro from patients with high coagulaˉtive state was investigated.Methods HPMCs from these patients and normal persons were sored by density gradient Fieoll-Hypaque and cultured48hours in vitro.PCA and PA from these patients and mormal persoms were examined by assay of chromophorous substrate.

分离高凝状态患者及正常人外周血单核细胞,短期培养后用发色底物法检测其促凝血活性和纤溶活性,检测加丹参注射液及黄芪注射液对体外培养HPMC表达PCA和PA的作用。

Methods Density gradient centrifugalization and absorption technique were respectively applied to detach peripheral blood mononuclear cells and peripheral blood lymphocytes.

密度梯度离心及吸附法分离外周血单个核细胞和外周血淋巴细胞,采用抗CD4和CD8抗体分别制备CD~(8+)和CD~(4+)T细胞进行培养,应用MTT比色法及ELISA法分别检测T细胞增殖活性及培养上清液IL-2水平。

Part II The experiment study on anti-tumour effect of dendritic cells derived from peripheral blood monocyte of patients with oral cancer after pulsed by tumour antigen in different ways. Collected 50ml peripheral blood from each patients and volunteers to induced DC in vitro. We parted the test group and control group into three subgroups respectively, untreated group A, freezed and thawn group B, RNA transfected group C. To group A, after induced DCs for five days , we added 200U/mlTNF-α in each hole and cultured on. To group B, we added protein antigen (DC/ tumor cells = 1 : 1 ,each hole) for 4h at 5d, and then added 200U/mlTNF-α in each hole and cultured on. To group C, transfected immature DC with tumor cell RNA(DC:RNA=1×105/ml: 5ug)at 5d, operated completely with the specification of invitrogen DMRIE-C reagents. After transfection we replaced transfection agent with complete medium, and then added 200U/mlTNF-α in each hole and cultured on.

第二部分口腔癌患者外周血来源的树突状细胞体外负载抗原后抑瘤作用的实验研究无菌采集20例口腔癌患者和20例健康志愿者外周血50ml,分离单个核细胞体外诱导培养DC,并将实验组和对照组细胞各分为三组,A组为未处理组,B组为冻融组,C组为RNA转染组。A组DC在培养5d后,每孔加入TNF-α200U/ml;B组DC培养第5d后,每孔中加入癌细胞蛋白抗原,使DC/肿瘤细胞=1:1,继续培养4h后,每孔加入TNF-α200U/ml;C组将培养至第5d的未成熟DC用肿瘤细胞RNA进行转染(DC:RNA=1×105/ml: 5ug,按照invitrogen DMRIE-C试剂说明书进行操作),转染结束后用完全培基取代转染剂,每孔加入TNF-α200U/ml。

Of CBMC can be purified by Mini-MACS as CD34〓 stem cells. B. The number of CD34〓 stem cells can expand to 40.24±9. 86 fold after 14 days. C. No matter in the expression of CD1a, CD80, CD86, and HLA-DR, or in the function of stimulating xenogenous lymphocyte proliferation, there was no difference between CD34+ stem cell DCs or monocyte DCs. D. The percentage of CD3〓CD56〓 cells is the same in CIK cells co-culture with DCs transfected with SKOV3 RNA, CIK cells co-culture with DCs, and CIK cells. E. The expansion rate of CIK cells can be accelerated by co-culturing with loaded or unloaded DCs. However, the expansion rate between loaded or unloaded group is the same. F. The strongest cytotoxicity against SKOV3 cell line was achieved by CIK cells co-cultured with DCs loaded with SKOV3 RNA.

结果:1、Mini-MACS分选系统可自CBMC中提取0.78±0.31%的CD34〓细胞;2、体外培养14天后可获得原始CD34〓细胞量40.24±9.86倍的细胞;3、不论在CD1a、CD80、CD86及HLA-DR的表达上,或是刺激异体淋巴细胞增生的功能上,脐带血CD34〓细胞与单核细胞来源的DC都没有差别;4、CIK细胞中CD3〓CD56〓双阳性的表达率在与RNA转染DC共培养的CIK细胞组、与DC共培养的CIK细胞组及单纯CIK细胞组3组间比较无差异;5、脐带血CIK细胞增殖显著,培养14天时可扩增18.18±5.59倍,培养21天时可扩增35.02±6.30倍;5、与未转染或转染DC共培养的CIK细胞在培养第14天后增殖速率大于单纯CIK细胞。

D. The function of stimulating xenogenous lymphocyte proliferation was the same between peripheral DCs and ascites PCs. E. The percentage of CD3〓CD56〓 cells was the same in CIK cells co-culture with DCs transfected with SKOV3 RNA, CIK cells co-culture with DCs, and CIK cells. F. The expansion rate of CIK cells can be accelerated by co-culturing with loaded or unloaded DCs. However, the expansion rate between loaded or unloaded group is the same. F. The strongest cytotoxicity against SKOV3 cell line was achieved by CIK cells co-cultured with DCs loaded with SKOV3 lysate.

结果:1、腹水可获得0.83±0.24×10〓个AMC/ml,单核细胞有0.74±0.25×10〓个/ml;2、卵巢癌患者外周血可获得0.87±0.20×10〓个AMC/ml,单核细胞有0.92±0.17×10〓个/ml;3、除CD86外周血单核细胞来源DC表达较高以外,其他表面分子在不同来源DC间没有统计学差异;4、不同来源DC的异基因刺激能力没有差异;5、与负载或未负载卵巢癌抗原的DC共培养并不能提高CIK细胞群中CD3〓CD56〓细胞的数量;6、CIK细胞增殖显著,培养14天时可扩增19.18±4.70倍,培养21天时可扩增35.82±4.36倍;7、与未负载或负载DC共培养的CIK细胞在培养第14天后增殖速率大于单纯CIK细胞。

Chromosome analysis of metaphases from bone marrow cells or cul-tured peripheral blood lymphocytes of 15 patients with blood disorderswas carried out.

对15例血液病患者进行了骨髓或外周血培养细胞的染色体观察。15例中12例临床确诊为慢性粒细胞白血病,2例诊断为急性粒细胞白血病,1例为继发性红细胞增生症。

Objective To evaluate function and Analyze the factor for false positive for BacT/Alert-blood culture system.

目的 评价全自动血培养系统BacT/Alert系统的功能及影响因素。

Objective To evaluate the value of automated blood culture system (BacT/Alert3D).

目的 评价全自动血培养仪BacT/Aler3D的临床应用情况。

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