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Design,delivery and efficacy testing of therapeutic nucleic acids used to inhibit hepatitis C virus gene expression in vitro and in vivo 被引量:9
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作者 Wolfgang H.Caselmann Matthias Serwe +3 位作者 Thomas Lehmann János Ludwig Brian S.Sproat Joachim W.Engels 《World Journal of Gastroenterology》 SCIE CAS CSCD 2000年第5期626-629,共4页
Despite major achievements in the treatment ofchronic hepatitis C with the combination ofinterferons and the nucleoside analog ribavirin themajority of patients with chronic hepatitis C virus(HCV) infection cannot be ... Despite major achievements in the treatment ofchronic hepatitis C with the combination ofinterferons and the nucleoside analog ribavirin themajority of patients with chronic hepatitis C virus(HCV) infection cannot be treated effectively.Toimprove this response rate we used antisensetechnologies to inhibit HCV translation as possibleadditional option for experimental treatment.Antisense oligodeoxynucleotides(ODN) are 展开更多
关键词 hepatitis C-like viruses/therapy gene expression in VITRO in vivo nucleic acids/therapeutic use CYTOMEGALOVIRUS
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自杀基因系统HSV-TK/GCV联合声动力疗法对人肺癌细胞NCI-446的体外杀伤实验 被引量:2
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作者 李洋 马程远 +1 位作者 里程楠 钱东华 《肿瘤防治研究》 CAS CSCD 北大核心 2013年第3期226-231,共6页
目的探讨自杀基因系统HSV-TK/GCV联合声动力疗法对人小细胞肺癌NCI-446细胞的体外杀伤作用。方法利用荧光和RT-PCR检测方法,筛选出含HSV-TK基因的阳性克隆株NCI-446/TK。用MTT法检测不同浓度的GCV对NCI-446/TK细胞的杀伤效应,以及不同... 目的探讨自杀基因系统HSV-TK/GCV联合声动力疗法对人小细胞肺癌NCI-446细胞的体外杀伤作用。方法利用荧光和RT-PCR检测方法,筛选出含HSV-TK基因的阳性克隆株NCI-446/TK。用MTT法检测不同浓度的GCV对NCI-446/TK细胞的杀伤效应,以及不同浓度混合NCI-446和NCI-446/TK细胞后的旁观者效应。同时,将HSV-TK/GCV自杀基因系统与声动力疗法进行联合,通过MTT法测定细胞存活率,并以流式细胞术检测细胞周期及凋亡率。结果当GCV浓度为0.2 mg/L时,筛选出的阳性克隆株NCI-446/TK细胞的生长受到明显的抑制(P<0.01),并且随着GCV浓度的递增,抑制效应越明显。当NCI-446/TK细胞占总细胞的10%时,有46%的混合培养细胞生长受到抑制(P<0.01),其抑制效应随NCI-446/TK细胞所占比例的增加而增加。将HSV-TK/GCV自杀基因系统与声动力疗法进行联合后,其细胞存活率明显低于单一治疗组(P<0.01),并且G0/G1期细胞所占比例及凋亡率均高于单一治疗组。结论 HSV-TK/GCV自杀基因系统的杀伤效应随着GCV浓度的递增而增强,并且有明显的旁观者效应。与声动力疗法联合后,其作用效果优于单一治疗组。 展开更多
关键词 自杀基因系统 声动力疗法 肺癌细胞 联合治疗
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HSV1-tk自杀基因系统对小鼠黑色素瘤的杀伤和抑制效应 被引量:1
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作者 刘晶晶 杜标炎 +3 位作者 谭宇蕙 吴映雅 周瑶 刘喜娟 《广州中医药大学学报》 CAS 2010年第4期389-392,438,439,共6页
【目的】检测自杀基因系统单纯疱疹病毒Ⅰ型胸苷激酶/丙氧鸟苷(HSV1-tk/GCV)对小鼠黑色素瘤的杀伤和抑制效应。【方法】将B16/tk(tk+)细胞和未经修饰的B16(tk-)细胞按tk+细胞占总细胞数的0%、10%、20%、40%、80%比例分别进行混合,各组... 【目的】检测自杀基因系统单纯疱疹病毒Ⅰ型胸苷激酶/丙氧鸟苷(HSV1-tk/GCV)对小鼠黑色素瘤的杀伤和抑制效应。【方法】将B16/tk(tk+)细胞和未经修饰的B16(tk-)细胞按tk+细胞占总细胞数的0%、10%、20%、40%、80%比例分别进行混合,各组细胞于96孔板中培养,加入丙氧鸟苷(GCV)至15.7μmol/L,采用四甲基偶氮唑盐(MTT)法检测不同tk+细胞比例的HSV1-tk/GCV系统在体外对肿瘤细胞的杀伤率。各组混合细胞分别接种于小鼠腋下,以GCV注射液按100 mg.kg-1.d-1进行治疗,检测瘤体体积、湿质量,测定抑瘤效应。【结果】tk+比例10%以上治疗组,HSV1-tk/GCV系统对黑色素瘤B16细胞均有明显杀伤效应,但旁杀伤作用不明显;从移植瘤体积看,20%以上tk/GCV治疗组肿瘤呈现明显生长抑制状态(P<0.05),以80%tk/GCV治疗组作用最显著(P<0.01);从移植瘤质量看,80%tk/GCV治疗组肿瘤呈现明显生长抑制状态(P<0.01),其他组虽随着tk+细胞比例的增高瘤体平均质量下降,但差异无显著性意义。【结论】HSV1-tk/GCV系统对小鼠黑色素瘤的杀伤和抑制效应均随着B16/tk细胞比例的增高而增强,已建立显示出体外杀伤效应和体内抑瘤活性的小鼠黑色素瘤自杀基因系统。 展开更多
关键词 黑色素瘤 自杀基因系统/治疗应用 基因疗法 细胞培养
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Mechanism of exogenous nucleic acids and their precursors improving the repair of intestinal epithelium after 7-irradiation in mice 被引量:8
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作者 Da Xiang Cui~1 Guei Ying Zeng~2 Feng Wang~1 Jun Rong Xu~1 Dong Qing Ren~2 Yan Hai Guo~1 Fu Rong Tian~2 Xiao Jun Yan~1 Yu Hou~1 Cheng Zhi Su~1 1 Institute of Genetic Diagnosis of the Fourth Military Medical University,Xi’an 710032,China 2 Department of Irradiation Medicine of the Fourth Military Medical University,Xi’an 710032,China 《World Journal of Gastroenterology》 SCIE CAS CSCD 2000年第5期709-717,共9页
AIM To clone expressed genes associated withrepair of irradiation-damaged mice intestinalgland cells treated by small intestinal RNA,andto explore the molecular mechanism ofexogenous nucleic acids improving repair ofi... AIM To clone expressed genes associated withrepair of irradiation-damaged mice intestinalgland cells treated by small intestinal RNA,andto explore the molecular mechanism ofexogenous nucleic acids improving repair ofintestinal crypt.METHODS The animal mode of test group andcontrol group was established,forty-five micebeing irradiated by γ ray were treated with smallintestinal RNA as test group,forty mice beingirradiated by γ ray were treated withphysiological saline as control group,five micewithout irradiation were used as normal control,their jejunal specimens were collectedrespectively at 6h,12h,24h,4d and 8d afterirradiation.Then by using LD-PCR based onsubtractive hybridization,these gene fragmentsdifferentially expressed between test group andcontrol group were obtained,and then werecloned into T vectors as well as beingsequenced.Obtained sequences were screenedagainst.GeneBank,if being new sequences,they were submitted to GeneBank.RESULTS Ninety clones were associated withrepair of irradiation-damaged intestinal glandcells treated by intestinal RNA.These clonesfrom test group of 6h,12h,24h,4d and 8dwere respectively 18,22,25,13,12.By screening against GeneBank,18 of which werenew sequences,the others were dramaticallysimilar to the known sequences,mainly similarto hsp,Nmi,Dutt1,alkaline phosphatase,homeobox,anti-CEA ScFv antibody,arginine/serine kinase and BMP-4,repA.Eighteen genefragments were new sequences,their acceptnumbers in GeneBank were respectivelyAF240164-AF240181.CONCLUSION Ninety clones were obtained tobe associated with repair of irradiation-damagedmice intestinal gland cells treated by smallintestinal RNA,which may be related toabnormal expression of genes and matchedproteins of hsp,Nmi,Duttl,Na,K-ATPase,alkalineph-osphatase,glkA,single strandedreplicative centromeric gene as well as 18 newsequences. 展开更多
关键词 radiation ionizing INTESTINE small/injuries RNA gene expression nucleic acids/therapeutic use POLYMERASE chain reaction REPAIR intestinal EPITHELIUM MICE
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胶质瘤自杀基因治疗模式优化的研究
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作者 杨学军 浦佩玉 +4 位作者 曹亚 姚开泰 马先勇 张云亭 刘松龄 《中国临床神经科学》 1998年第4期191-194,共4页
应用携带HSV-tk基因的逆转录病毒载体pMV_7(tk)及其包装细胞PA317tk,研究在SD大鼠颅内C6胶质瘤中的HSV-tk基因转移效率;在皮下肿瘤模型中进行治疗模式研究.结果表明,大鼠颅内胶质瘤模型呈典型的肿瘤病理表现,包装细胞种植后1周HSV-tk m... 应用携带HSV-tk基因的逆转录病毒载体pMV_7(tk)及其包装细胞PA317tk,研究在SD大鼠颅内C6胶质瘤中的HSV-tk基因转移效率;在皮下肿瘤模型中进行治疗模式研究.结果表明,大鼠颅内胶质瘤模型呈典型的肿瘤病理表现,包装细胞种植后1周HSV-tk mRNA的阳性率为30%~60%;肿瘤内多点注射包装细胞,肿瘤消退速度明显快于单点注射(P<0.05).提示:优化胶质瘤基因治疗模式可进一步提高基因治疗效果. 展开更多
关键词 胶质瘤 自杀基因 治疗 模式 基因疗法
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