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  fo cell
Experiments were carried out with a specially designed FO cell described in Section 2.5.
      
The Reynolds number was found to be around 2100 for the flow conditions in the new FO cell described above.
      
  fo cell
Experiments were carried out with a specially designed FO cell described in Section 2.5.
      
The Reynolds number was found to be around 2100 for the flow conditions in the new FO cell described above.
      


The expression plasmid PCEA-TK or pCEA-CD was constructed by ligating the CEA gene promoter to suicide gene such as HSV-TK (herpes simplex virus thymidine kinase) gene or EC-CD (E. colt cytosine deaminase) gene. A human colorectal carcinoma cell line LoVo or a human uterine cervical cancer cell line HeLa was co-transfected with pSV2-neo and PCEA-TK (or PCEA-CD). After selection with G418, the transgenetic cell clones (hoVo/CEA-TK,LoVo/CEA-CD, HeLa/CEA-TK and HeLa/CEA-CD) were obtained. There was no significant...

The expression plasmid PCEA-TK or pCEA-CD was constructed by ligating the CEA gene promoter to suicide gene such as HSV-TK (herpes simplex virus thymidine kinase) gene or EC-CD (E. colt cytosine deaminase) gene. A human colorectal carcinoma cell line LoVo or a human uterine cervical cancer cell line HeLa was co-transfected with pSV2-neo and PCEA-TK (or PCEA-CD). After selection with G418, the transgenetic cell clones (hoVo/CEA-TK,LoVo/CEA-CD, HeLa/CEA-TK and HeLa/CEA-CD) were obtained. There was no significant difference in either morphology or cell growth ctirve between suicide gene transduced cells and wild type cells. But LoVo/CEA-TK (or LoVo/CEA-CD) cells were 2000 (or 700) times more sensitive to the cytotoxicity of prodrug ganciclovir (or 5-Fluorocytosine ) than parental LoVo cells. However, HeLa/CEA-TK or HeLa/CEA-CD cells were still resistant to Ganciclovir (or 5-Fluorocytosine). These results showed the POssibility of gene therapy for human colorectal carcinoma by cell type-specific expression of suicide genes.

构建了以癌胚抗原(CEA)基因启动子控制的HSV-TK和ECCD的表达质粒PCEA-TK和pCEA-CD.将它们分别与pSV2-neo共转染人结肠癌细胞株LoVo和人宫颈癌细胞株HeLa,G418筛选得到细胞克隆LoVo/CEA-TK、toVo/CEA-CD、HeLa/CEA-TK和HeLa/CEA-CD.与野生型LoVo细胞相比,LoVo/CEA-TK和LoVo/CEA-CD形态无明显改变,生长曲线也相似,但对GCV或5-FC的细胞毒的敏感性分别提高了2000倍或700信而HeLa/CEA-TK(或HeLa/CEA-CD)仍对低浓度GCV(或5-FC)不敏感.以上结果显示了应用组织专一性表达的自杀基因治疗人结肠癌的可能性.

Objective To investigate the cytotoxic effect of fraction C from Naja Naja atra venom (FC) on human glioma cell line (U251). Methods Normal human gliocyte and U251 were incubated with different concentrations of FC, adriamycin (ADM) and nimustine hydrochloride (ACNU). The cytotoxic effects of FC, ADM and ACNU on gliocyte and U251 were assessed according to the inhibition rate of cell growth by MTT rduction assay. Result The inhibitory effect of FC on U251 was obvious, with the inhibitory concentrations (IC50)...

Objective To investigate the cytotoxic effect of fraction C from Naja Naja atra venom (FC) on human glioma cell line (U251). Methods Normal human gliocyte and U251 were incubated with different concentrations of FC, adriamycin (ADM) and nimustine hydrochloride (ACNU). The cytotoxic effects of FC, ADM and ACNU on gliocyte and U251 were assessed according to the inhibition rate of cell growth by MTT rduction assay. Result The inhibitory effect of FC on U251 was obvious, with the inhibitory concentrations (IC50) of 2.95 and 2.21μg/ml at 24, 48 h respectively. Compared with the IC50 of 8.60 μg/ml and 6.61 μg/ml respectively for glial cell, and the difference was significant (P<0.05). Conclusion FC can inhibit the cell growth effectively and induce the apoptosis of U251 and is more effective in inhibiting the growth of glioma cells compared with ADM or ACNU.

目的研究中华眼镜蛇毒组分C(FC)体外对人胶质瘤细胞株U251的抑制作用。方法培养箱孵育U251与正常人胶质细胞;加入不同浓度的FC、阿霉素(ADM)和尼莫斯汀(ACNU)。采用细胞毒实验(MTT法),通过细胞生长抑制率评价FC、ACNU和ADM的细胞毒作用。结果FC对胶质瘤细胞的抑制作用突出,24h和48h的IC50分别为2.95μg/ml和2.21μg/ml,而ADM与ACNU远未达到此抑制率。FC对正常胶质细胞的24h、48hIC50分别为8.60μg/ml和6.61μg/ml,两细胞株间存在显著性差异(P<0.05)。结论人胶质瘤细胞对FC的细胞毒性作用是敏感的。FC对胶质瘤细胞的抑制作用优于ADM和ACNU。

Objective:To study the feasibility of using ~ 19 F nuclear magnetic resonance (NMR) spectroscopy to monitor CD transgene expression in human colon cancer cell line. Methods:The cell line SW1116 was transfected with a gene coding for cytosine deaminase (CD from E.coli) by retroviral mediated method. The assessment of expression of CD gene in colon cancer cell line was carried out by ~ 19 F-NMR spectroscopy drug sensitivity to 5-FC in vitro and in vivo. Results: ~ 19 F -NMR spectroscopy showed that incubation...

Objective:To study the feasibility of using ~ 19 F nuclear magnetic resonance (NMR) spectroscopy to monitor CD transgene expression in human colon cancer cell line. Methods:The cell line SW1116 was transfected with a gene coding for cytosine deaminase (CD from E.coli) by retroviral mediated method. The assessment of expression of CD gene in colon cancer cell line was carried out by ~ 19 F-NMR spectroscopy drug sensitivity to 5-FC in vitro and in vivo. Results: ~ 19 F -NMR spectroscopy showed that incubation of cultured CD+cells with 774 μmol/L 5FC for 24 h , the spectrum of intact cells shows two broad resonances corresponding to 5-FC (chemical shift relative to trifluoroacetate δ-91.7) taken up into cell, 5-FU(δ-93.3) produced by the action of cytosine deaminase. The resulting SWCD_ 2 cell line was more sensitive to the prodrug 5-fluorocytosine (5-FC); IC_ 50 =66 μmol/L, compared to parent cancer cells SW1116;IC_ 50 =16 000 μmol/L. Conclusion:The feasibility of using 19NMR spectroscopy to noninvasively monitor CD gene expression in human colon cancer was demonstrated. This capability provides an new approach for measuring gene expression that will be useful in clinical gene therapy trials.

目的:研究CD基因在结肠癌细胞表达的分子成像分析。方法:通过逆转录病毒介导方法,将编码大肠杆菌的CD基因转染到人结肠癌细胞系SW1116。通过19F-NMR波谱分析CD基因在人结肠癌细胞中的表达状况,MTT方法检测5-FC的细胞毒作用。结果:19F-NMR波谱分析显示,转导CD基因细胞在774μmol/L5-FC浓度培养24h后的样品,在δ-91.7和δ-93.3显示了2个双峰,分别是5-FC和5-FU的19F-NMR的信号,提示5-FU是SWCD2细胞内的CD酶催化5-FC的代谢产物。转基因的肿瘤细胞SWCD2中表达了CD基因,对前药5-FC的敏感性较亲本肿瘤细胞SW1116明显增高,50%细胞生长抑制率(IC50)为66μmol/L,而SW1116细胞IC50为16000μmol/L。结论:CD基因表达的代谢产物不但对结肠癌细胞恶性生长具有抑制作用,而且CD基因具有分子成像标记基因的功能。

 
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