PTEN基因转染的胶质瘤细胞对γ刀照射敏感性的变化

目的:观察转入正常PTEN基因的胶质瘤细胞经γ刀照射后生长能力及黏附力的改变,探索PTEN基因对胶质瘤细胞放射敏感性的影响。方法:以RT-PCR方法体外扩增PTEN全长基因,与质粒pcDNA3.1连接,构建重组真核载体pcD-NA3.1-PTEN,并用之转染PTEN失活的U251细胞(人胶质母细胞瘤细胞系),用48 h的半数致死剂量(中心剂量和周边剂量分别为9.00和4.50 Gy)进行γ刀等中心照射,并采用MTT法和纤维粘连蛋白黏附实验观察照射前后细胞的生长及黏附力改变。结果:空载体转染组与未转染组照射前后细胞存活率和黏附率均无显著性差异,而PTEN基因转染的细胞存活率及黏附率在照射前后均显著低于空载体转染组及未转染组(P<0.01),且照射后下降程度更加明显。结论:PTEN基因转染的胶质瘤细胞对于γ刀照射的敏感性提高。

应用RNA干扰下调多药耐药蛋白4表达对结直肠癌细胞照射敏感性的影响

目的探讨多药耐药蛋白4（MRP4）表达对结直肠癌细胞照射敏感性的影响。方法应用慢病毒感染RNA干扰技术（RNAi）稳定下调人结直肠癌细胞株HCT116中MRP4的表达。将HCT116细胞分为未感染任何病毒的细胞株（CON）、加阴性对照病毒感染的细胞株（NC）和加RNAi靶点病毒感染的细胞株（KD）3组,应用实时定量RT-PCR和Western blot分别从RNA和蛋白水平检测MRP4表达变化,以验证RNAi的有效性。4Gy剂量照射后24h，流式细胞术检测细胞凋亡,MTT检测细胞增殖,比较RNAi后HCT116细胞株照射敏感性的差异。结果成功构建并转染慢病毒质粒,获得稳定沉默MRP4表达的HCT116-KD细胞株,HCT116-KD细胞株MRP4mRNA和蛋白表达水平较对照均明显下调（P〈0．05）。照射后24h，KD细胞株凋亡率为（71．7±0．8）％，明显高于CON组[（56．1±0．9）％]和NC组[（59．8±0．8）％]（P〈0．05）。照射后48h和72h，KD细胞增殖能力较对照组明显下降（火0．05）。结论MRP4在结直肠癌细胞中的表达水平与照射耐受显著相关，应用慢病毒感染RNA干扰下调MRP4表达可以增强结直肠癌细胞照射敏感性。MRP4有可能成为预测放疗敏感性的分子标志物。

Efficacy of Chaihu Jisang decoction on post-infection cough and its effect on cough reflex sensitivity

OBJECTIVE:To investigate the efficacy of Chaihu Ji Sang Decoction(CJSD),(a modification of Xiao Chaihu Tang(Minor Decoction of Bupleurum) was evaluated in patients with post-infection cough(PIC).Its effect on cough reflex sensitivity(CRS) was also described.METHODS:One hundred and eighteen patients with PIC were enrolled and randomly divided into an experimental group and control group(54 patients in each group).The experimental group was given CJSD orally,and the control group was given cetirizine hydrochloride for 7 d.Cough symptom scores and TCM symptom scores were compared before and after treatment.Efficacy,cough onset time,recurrence,and adverse drug reactions were additionally evaluated.Finally,CRS was evaluated before and after treatment of PIC patients in the experimental group.RESULTS:Fifty-two patients in the experimental group and fifty control patients completed the study.Cough symptom scores and the TCM symptom scores of the two groups were significantly improved after treatment,however cough symptoms in the experimental group 1(0,2) were significantly lower than the control group 2(1,2) after treatment(P < 0.05).Similarly,TCM symptom scores 4(0,7) in the experimental group were significantly lower than the control group 6(3,12)(P < 0.01).Overall,the effective rate of the experimental treatment(84.60%) was significantly higher than that of the control treatment(68.00%)(P < 0.01).The onset time of efficacy of the experimental treatment was one day faster than the control treatment [2(2,3) d vs 3(2,3) d,P < 0.01].The relapse rate in the experimental group(9.09%) was lower than that of the control group(29.41%)(P < 0.05).39 PIC patients in the experimental group completed a capsaicin cough challenge test before treatment.The lgC2 and lg C5 of the experimental group prior to treatment was 1.19(0.59,1.49) and 1.80(1.49,2.10),respectively.The lgC2,lg C5 of the healthy control group was 1.49(0.89,2.40) and 2.70(2.40,3.00),respectively.The differences in lgC2 and lg C5 between experimental group and healthy groups were significant at this time point(both P < 0.01).After treatment,34 patients in the experimental group received re-examination.lg C5 increased from 1.80(1.49,2.10) to 2.10(1.49,3.00).CRS after treatment/re-examination was significantly lower than those of baseline in the experimental group(P < 0.05).CONCLUSION:CJSD could effectively improve the clinical symptoms of PIC patients,with demonstrable low recurrence and safety.CJSD also significantly decreased the CRS in patients with PIC.