目的研究抗Fas锤头状核酶对T细胞Fas表达及其凋亡的影响,探讨增强供者淋巴细胞输注时移植物抗白血病(graft versus leukemia,GVL)效应的新策略。方法构建可有效切割Fas mRNA的锤头状核酶真核质粒,电穿孔法将其导入小鼠细胞毒T淋巴细胞(cytotoxic T lymphocyte,CTL)细胞株CTLL-2中,借助RT-PCR和Western blot检测细胞Fas的表达,同时检测转染前后细胞Caspase-3活性的改变和细胞凋亡(AnnexinⅤ-FITC法)。MTT法检测空白对照组、转染空载体组及转染pU6-RZ596组CTLL-2细胞的增殖情况和体外杀伤粒-单核白血病细胞(WEHI-3)的活性。结果构建的U6嵌合型锤头状核酶RZ596在细胞内能有效切割Fas,RT-PCR电泳检测其与β-actin条带的灰度比值:CTLL-2细胞空白对照组为(1.06±0.12),转染空载体组为(0.98±0.15),转染pU6-RZ596组为(0.43±0.11)(n=3);Fas蛋白的Western blot检测显示:以空白对照组灰度值为1,转染空载体组和转染pU6-RZ596组电泳条带的灰度比值分别为(0.98±0.13)和(0.45±0.08)(n=3),提示抗Fas锤头状核酶可明显降低小鼠活化CTLL-2的Fas水平。与高表达Fas配体(FasL)的WEHI-3细胞共孵育,CTLL-2细胞的存活率和体外杀伤活性明显增加,空白对照组、转染空载体组和转染pU6-RZ596组的细胞凋亡率分别为88%、84%和37%,对WEHI-3细胞杀伤活性分别为32%、31%和67%。结论抗Fas锤头状核酶能显著降低小鼠活化CTLL-2细胞的Fas表达,使其免于Fas途径的凋亡。
目的:分析探讨可吸收棒内固定技术治疗MasonⅡ型桡骨小头骨折的临床疗效。方法:11例桡骨小头骨折患者,其中男7例,女4例;年龄10～16岁,平均11岁。所有病例骨折分型为MasonⅡ骨折,骨折块分离移位。术前X线及CT显示骨折情况,均采用切开复位可吸收棒内固定技术进行治疗。术后常规拍摄X线片了解骨折复位情况,患肢屈肘90°、前臂中立位长臂石膏托固定4周,去石膏后积极进行功能锻炼。通过拍摄X线片及Broberg and Morrey肘关节评分标准进行评价。结果:所有病例均获得随访,随访时间6～22个月,平均15个月,复查X线证实骨折愈合,平均愈合时间3个月。所有病例均未出现骨折移位、关节疼痛、关节不稳及畸形愈合等并发症。患肢功能恢复良好,无关节活动障碍影响生活者。结论:应用可吸收棒内固定技术治疗MasonⅡ型桡骨小头骨折是一种疗效可靠、简单经济的临床治疗方法。
Objective: To investigate the relationship between peroxisomeproliferators-activated receptor-gamma (PPAR-γ) and clinical pathology of lung cancer, and to studythe inhibitory effect of PPAR-γ activators on lung cancer growth and the mechanism of inducingapoptosis of lung cancer. Methods: Expression of PPAR-γ in 15 cases of non-cancerous lung tissuesand 64 cases of lung cancer tissues was detected by using immunohistochemistry. The average A valueswere measured by using image analysis. The expression of PPAR-γ in lung cancer cells was detectedby using RT-PCR. After being treated with PPAR-γ activators, apoptosis was detected by using flowcytometry, and meanwhile, the change in caspase-3 activity was detected by caspase-3 kits. Results:Expression levels of PPAR-γ in cancerous tissues were higher than those in non-cancerous lungtissues. In four types of lung cancer, the sequence of PPAR-γ expression from high to low levelswas small-cell lung cancer, squamous carcinom, large-cell lung cancer, adenocarcinoma in turns. Theexpression of PPAR-γ was correlated with differentiation and postoperative TNM staging of lungcancer tissues, but not with lymph node metastasis. PPAR-γ expressed in two lung cancer cell lines,which could induce apoptosis of lung cancer cells after being treated with PPAR-γ activators, andcaspase-3 activity in cells treated with PPAR-γ activators was significantly increased. Conclusion:PPAR-γ is correlated with clinical pathology and apoptosis of lung cancer closely, and activatedPPAR-γ can increase caspase-3 activity to induce cells apoptosis. PPAR-γ will be able to become anew target for therapy of lung cancer in the future.
