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广东省自然科学基金(06021352)

作品数:3 被引量:22H指数:2
相关作者:李华王欣璐姜楠杨扬汪根树更多>>
相关机构:广州军区广州总医院中山大学附属第三医院南方医科大学南方医院更多>>
发文基金:国家自然科学基金广东省自然科学基金国家重点基础研究发展计划更多>>
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靶向人端粒酶逆转录酶进行RNAi的逆转录病毒载体的构建被引量:2
2006年
目的构建靶向性干扰人端粒酶逆转录酶表达的特异性小干扰RNA的真核表达载体。方法采用PCR法分别扩增增强型绿色荧光蛋白的DNA序列、U6启动子序列和靶向性干扰人端粒酶逆转录酶的特异性小干扰RNA对应的DNA序列,随后将与之相应的DNA序列片断依次克隆入真核表达载体pLXSN中,并通过限制性内切酶和测序对该重组表达载体进行鉴定。以荧光显微镜和利用流式细胞仪分析重组病毒表达EGFP蛋白的情况。以MTT方法检测初步分析重组病毒对人Hela细胞的RNAi效果。结果限制性内切酶酶切和测序结果均表明成功构建了靶向人端粒酶逆转录酶的小干扰RNA的逆转录病毒表达载体pLXSN/EGFP-U6-siTERT。以磷酸钙共转染法制备的重组逆转录病毒滴定了病毒滴度后,重组病毒感染人Hela细胞24h时用流式细胞仪测得EGFP表达的阳性率为24.1%。重组逆转录病毒感染人Hela细胞48h时,MTT实验测得细胞死亡率为53.2%。结论成功构建了针对人端粒酶逆转录酶小干扰RNA的逆转录病毒表达载体pLXSN/EGFP-U6-siTERT,并制备了重组逆转录病毒,初步观察了效应,为进一步利用小干扰RNA技术研究肿瘤的基因治疗奠定了基础。
王全师王欣璐李华王巧愚王全颖杨广笑
关键词:RNA干扰逆转录病毒载体人端粒酶逆转录酶
PET/CT imaging of delayed radiation encephalopathy following radiotherapy for nasopharyngeal carcinoma被引量:9
2007年
Background With the significant improvement in the survival of patients with nasopharyngeal carcinoma (NPC) undergoing radiotherapy and the growing availability of the sophisticated imaging modalities, the number of radiation encephalopathy (RE) cases relating to NPC radiotherapy is increasing. In this study, we investigated the metabolic and density changes of the compromised brain tissues during delayed RE using a positron-emission tomography-computed tomography (PET/CT) to provide clinical evidences for the diagnosis of delayed RE following radiotherapy for NPC. Methods The PET/CT manifestations and the clinical data of 53 pathologically confirmed NPC patients with delayed RE following radical radiotherapy and 15 healthy volunteers were investigated. The standardized uptake values (SUV) of the bilateral temporal lobes, the occipital lobe and the brain stem were measured respectively; and then the metabolic reduction rate of 88 temporal lobes and 13 brain stems were calculated for a statistical comparison between the two groups. Results The earliest case of delayed RE in the investigated patients occurred 1.5 years after radiotherapy. Delayed RE frequently involved the inferior temporal lobe. For patients with delayed RE confirmed by clinical symptoms and imaging findings, PET maintained a 100% coincidence rate with CT; however, in the 25 temporal lobes of the 35 delayed RE patients, PET revealed obvious hypometabolic changes whereas CT displayed normal density. The incidence of brain stem metabolic reductions was 24.5% (13/53) in the investigated patients, including 4 patients with hypometabolic changes shown by PET and negative finding shown by CT. The incidence of granuloma adjacent to the hypometabolic region in the temporal lobe was 12.5% (11/88). Conclusion Delayed RE patients exhibit significant hypometabolic changes in the inferior temporal lobe, captured by PET much earlier than by CT. PET/CT offers a valuable means for the diagnosis of delayed RE in subacute stages and granulom
WANG Xin-luYIN Ji-linLI HuaLI Xiang-dongQUAN Jiang-tao
关键词:RADIOTHERAPY
PET/CT研究恶性淋巴瘤对肝脏和/或脾脏的侵犯
目的研究恶性淋巴瘤侵犯肝脏和/或脾脏的PETICT/CT影像学表现,观察肝脏和/或脾脏内病变累及的特点,为其正确诊断提供依据。方法分析累及肝脏和/或脾脏的恶性淋巴瘤患者41例(52次 PET/CT检查)的临床资料及PET...
王欣璐尹吉林李华李向东全江涛蔡超郑辉
关键词:恶性淋巴瘤肝脏脾脏
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RNAi靶向人端粒酶逆转录酶对人肝癌细胞的生长抑制作用被引量:11
2008年
目的利用RNAi技术降低人端粒酶逆转录酶(hTERT)基因的表达,抑制其活性,研究其对人肝癌细胞的增殖和凋亡影响。方法构建针对hTERT基因的siRNA真核表达载体pLXSN-EGFP-U6-siTERT,制备靶向hTERT的重组逆转录病毒;用重组逆转录病毒感染HepG2人肝癌细胞,以TRAP法检测端粒酶活性变化;流式细胞仪检测细胞凋亡率;MTT法检测肿瘤细胞生长抑制情况。结果成功制备出hTERTsiRNA逆转录病毒表达载体;重组病毒感染HepG2细胞24、48、72h后端粒酶活性分别下降了23.84%、58.03%和85.01%(P<0.05);感染后24h细胞凋亡率29.05%;MTT实验结果显示肿瘤生长受到抑制,并存在一定量效和时效关系。结论hTERTsiRNA能特异性使hTERT基因表达抑制,端粒酶活性下降,肝癌细胞增殖受到抑制。
李华王欣璐杨扬张剑汪根树姜楠陈规划
关键词:RNA干扰人端粒酶逆转录酶基因治疗
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