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Rat glioma cell C6 was infected by the deficiency recombinant retrovirus (named C6TK). The sensitivity of C6TK cells to GCV or ACV increased 450 or 10 times than that of C6 cells respectively. Rat C6 glioma model was successfully built, and SD rats inoculated intracerebrally C6TK cells had normal gliomas. The average survival periods of the rats were about 15 days. However, after treated with ACV, the growth of C6TK tumors obviously regressed, and the survival periods extended to 57.8±8.1 days, especially in rats injected with mixed PA317TK and C6 cells or in situ PA317TK cells, which the tumors nearly disappeared after ACV administration and the survival periods extended to over 120 or 71.4±36.1 days respectively, P<0.001.
结果:构建了带有HSV-tk基因的反转录病毒载体GINaTK,应用脂质体转移方法将GINaTK导入反转录病毒包装细胞PA317,成为产病毒细胞PA317TK,用带有HSV-tk基因的复制缺陷型反转录病毒感染C6细胞,命名为C6TK细胞,对GCV和ACV的敏感性分别高于亲本450倍和10倍;成功地建立了大鼠脑胶质瘤模型,并证实转染细胞C6TK的成瘤效应未改变,存活期约为15天;而经ACV治疗后,含有C6TK细胞的肿瘤生长明显被抑制,大鼠生存期延长为57.8±8.07天,尤其是采用PA317TK细胞混和治疗组和原位治疗组,肿瘤基本消失,大鼠生存期显著延长,混和治疗组存活120天以上,原位治疗组存活至71.4±36.1天。t检验,P值均小于0.001。
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Results: GINaTK retroviral vector containing HSV-tk was constructed and transduced into PA317 retrovirus packaging cell by lipofectin (named PA317TK). Rat glioma cell C6 was infected by the deficiency recombinant retrovirus (named C6TK). The sensitivity of C6TK cells to GCV or ACV increased 450 or 10 times than that of C6 cells respectively. Rat C6 glioma model was successfully built, and SD rats inoculated intracerebrally C6TK cells had normal gliomas. The average survival periods of the rats were about 15 days. However, after treated with ACV, the growth of C6TK tumors obviously regressed, and the survival periods extended to 57.8±8.1 days, especially in rats injected with mixed PA317TK and C6 cells or in situ PA317TK cells, which the tumors nearly disappeared after ACV administration and the survival periods extended to over 120 or 71.4±36.1 days respectively, P <0.001.
结果:构建了带有HSV-tk基因的反转录病毒载体GINaTK,应用脂质体转移方法将GINaTK导入反转录病毒包装细胞PA317,成为产病毒细胞PA317TK,用带有HSV-tk基因的复制缺陷型反转录病毒感染C6细胞,命名为C6TK细胞,对GCV和ACV的敏感性分别高于亲本450倍和10倍;成功地建立了大鼠脑胶质瘤模型,并证实转染细胞C6TK的成瘤效应未改变,存活期约为15天;而经ACV治疗后,含有C6TK细胞的肿瘤生长明显被抑制,大鼠生存期延长为57.8±8.07天,尤其是采用PA317TK细胞混和治疗组和原位治疗组,肿瘤基本消失,大鼠生存期显著延长,混和治疗组存活120天以上,原位治疗组存活至71.4±36.1天。t检验,P值均小于0.001。
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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。
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One protein band, which was named as abalone triploid special expression protein, was found to be distinguishingly existed in one to six years old triploid abalone by PAGE analysis. While in Pacific oyster and Zhikong scallop, there is no special expressed protein could be observed among diploid, triploid or tetraploid individuals.
用常规聚丙稀酰胺凝胶电泳技术研究发现皱纹盘鲍的二倍体和三倍体个体中存在稳定的差异表达蛋白质,命名为a-TSP;在长牡蛎的2n、3n、4n之间,及栉孔扇贝的2n和3n之间未获得类似的差异表达蛋白。
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Leptotyphlops carlae is the world's smallest species of snake, with adults averaging just under four inches in length.
这张未标明拍摄日期的照片显示,被命名为&Leptotyphlops carlae&的蛇在一枚0.25美元硬币上休息。
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As she looked at Warrington's manly face, and dark, melancholy eyes, she had settled in her mind that he must have been the victim of an unhappy attachment.
每逢看到沃林顿那刚毅的脸,那乌黑、忧郁的眼睛,她便会相信,他一定作过不幸的爱情的受害者。
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Maybe they'll disappear into a pothole.
也许他们将在壶穴里消失
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But because of its youthful corporate culture—most people are hustled out of the door in their mid-40s—it had no one to send.
但是因为该公司年轻的企业文化——大多数员工在40来岁的时候都被请出公司——一时间没有好的人选。