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Protein transduction domain of membrane penetrating peptide can efficiently deliver DNA and protein into mouse liver for gene therapy 被引量:4
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作者 Jun Xie, Bao-Feng Yu, Jun Xu, Yue-Hong Zhang, Niu-Liang Cheng, Bo Niu, Xiao-Nian Hu, Qian Xiang and Zheng-Guo Zhang Taiyuan, China Department of Biochemistry and Molecular Biology, Shanxi Medical University, Taiyuan 030001 , China Peking Union Medical College, Chi- nese Academy of Medical Sciences, Beijing 100005, China 《Hepatobiliary & Pancreatic Diseases International》 SCIE CAS 2005年第1期90-93,共4页
BACKGROUND: The development of a harmless and effi- cient nonviral gene delivery system that can facilitate the penetration of nucleic acids through the plasma membrane is a key to successful gene therapy. The aim of ... BACKGROUND: The development of a harmless and effi- cient nonviral gene delivery system that can facilitate the penetration of nucleic acids through the plasma membrane is a key to successful gene therapy. The aim of this study was to test a nonviral gene transferring vector's function of delivering DNA into liver cells to provide an important clue for gene transfer in liver gene therapy. METHODS: The complex of DNA and DNA delivering protein was injected into mice through their tail veins. Then the mice were killed and their liver tissue was sec- tioned. The gene transferring results were detected using a confocal laser scanning microscope. RESULTS: Fluorescence analysis indicated that both DNA- membrane penetrating peptide (MPP) complex and DNA- hepatocyte specific receptor binding domain ( HSRBD) - MPP complex could go into liver cells. The fluorescence value of liver cells in the DNA-HSRBD-MPP group was higher than that in the DNA-MPP group. CONCLUSIONS; MPP can successfully deliver DNA and protein into cells, and MPP with a HSRBD can specifically deliver DNA into liver cells. These have laid a foundation for further study on the nonviral liver cell gene delivering system. 展开更多
关键词 membrane penetrating peptide gene therapy gene delivering
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Protein trans-splicing based dual-vector delivery of the coagulation factor Ⅷ gene 被引量:27
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作者 ZHU FuXiang,LIU ZeLong,CHI XiaoYan & QU HuiGe Life Science College of Ludong University,Yantai 264025,China 《Science China(Life Sciences)》 SCIE CAS 2010年第6期683-689,共7页
A dual-vector system was explored for the delivery of the coagulation factor VIII gene,using intein-mediated protein trans-splicing as a means to produce intact functional factor VIII post-translationally.A pair of eu... A dual-vector system was explored for the delivery of the coagulation factor VIII gene,using intein-mediated protein trans-splicing as a means to produce intact functional factor VIII post-translationally.A pair of eukaryotic expression vectors,expressing Ssp DnaB intein-fused heavy and light chain genes of B-domain deleted factor VIII (BDD-FVIII),was constructed.With transient co-transfection of the two vectors into 293 and COS-7 cells,the culture supernatants contained (137±23) and (109±22) ng mL–1 spliced BDD-FVIII antigen with an activity of (1.05±0.16) and (0.79±0.23) IU mL–1 for 293 and COS-7 cells,respectively.The spliced BDD-FVIII was also detected in supernatants from a mixture of cells transfected with inteinfused heavy and light chain genes.The spliced BDD-FVIII protein bands from cell lysates were visualized by Western blotting.The data demonstrated that intein could be used to transfer the split factor VIII gene and provided valuable information on factor VIII gene delivery by dual-adeno-associated virus in hemophilia A gene therapy. 展开更多
关键词 INTEIN protein TRANS-SPLICING coagulation factor VIII dual-vector gene delivery
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超声微泡介导靶向治疗研究进展 被引量:2
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作者 王顺义 李怡 《医学与哲学(B)》 2017年第1期57-60,共4页
载药超声微泡的靶向治疗是当前医药领域中的研究热点。超声微泡破坏技术介导的靶向药物和基因治疗是一种最新的靶向治疗方法。超声微泡不仅可以作为一种超声造影剂增强成像对比度,更可以通过各种物理、化学修饰携带各种药物或基因在超... 载药超声微泡的靶向治疗是当前医药领域中的研究热点。超声微泡破坏技术介导的靶向药物和基因治疗是一种最新的靶向治疗方法。超声微泡不仅可以作为一种超声造影剂增强成像对比度,更可以通过各种物理、化学修饰携带各种药物或基因在超声场强破坏作用下进行靶向治疗。超声微泡造影剂靶向治疗包括超声介导载药微泡的治疗及超声介导靶向载药微泡的治疗。本文就超声微泡特性、超声靶向微泡破坏(UTMD)技术机制、超声微泡介导靶向治疗三方面进行综述。 展开更多
关键词 超声 微泡 携基因或药物 靶向治疗
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初进高原者耐缺氧机制与高原条件下的输血策略 被引量:5
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作者 李翠莹 李小薇 《中国输血杂志》 北大核心 2017年第8期859-862,共4页
高原环境中的低压低氧、寒冷干燥及紫外线强等均是对机体不利的因素,平原人群进入高原难免发生一系列生理改变。平原人群在高原地区负(战、创)伤时,不仅表现为失血耐受能力降低、易发生休克,而且对液体耐受能力更小,使得脑水肿、肺水肿... 高原环境中的低压低氧、寒冷干燥及紫外线强等均是对机体不利的因素,平原人群进入高原难免发生一系列生理改变。平原人群在高原地区负(战、创)伤时,不仅表现为失血耐受能力降低、易发生休克,而且对液体耐受能力更小,使得脑水肿、肺水肿、右心功能不全以及多器官衰竭早等较易发生。失血是导致伤者的这些病状,乃至死亡的关键原因,而输血治疗也成为高原战创伤救治中更重要的手段。本文首先对本期"高原输血专题"中涉及的平原人群进入高原后血常规变化、高原适应基因表达改变、红细胞形态及携氧功能差异等方面研究作了评述,并分析总结了其中的变化规律及意义,以提高平原人群高原习服适应能力;其次针对高原低氧环境下藏、汉族人群发生高原战创伤时异于平原地区人群的特征,概述了有关高原失血性休克患者的输血策略、特点及其救治的研究进展,分析了高原适应机制与之的相关性;最后针对高原输血目前存在的困窘与亟待解决的问题,提出并探讨了自己的高原的输血策略。 展开更多
关键词 高原输血策略 高原低氧适应 失血性休克 失血耐受 高原适应基因 红细胞形态 红细胞携氧
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辐射制备的M-PEIs基因转导载体细胞生物学研究
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作者 董黎 徐冬梅 +2 位作者 洪军 宫培军 姚思德 《核技术》 EI CAS CSCD 北大核心 2008年第6期465-468,共4页
将辐射制备的M-PEIs纳米凝胶转导质粒短发夹状RNA(shRNA),研究了载体复合物的细胞毒性、shRNA转染效率及对靶基因的表达抑制。结果表明M-PEIs纳米凝胶的细胞毒性较低,可以较高基因转染效率将shRNA转移到细胞内,同时携带的质粒shRNA在靶... 将辐射制备的M-PEIs纳米凝胶转导质粒短发夹状RNA(shRNA),研究了载体复合物的细胞毒性、shRNA转染效率及对靶基因的表达抑制。结果表明M-PEIs纳米凝胶的细胞毒性较低,可以较高基因转染效率将shRNA转移到细胞内,同时携带的质粒shRNA在靶细胞内具有较长的释放期,转染的shRNA能使肝癌细胞的靶基因失活,有效抑制了靶基因的表达,初步证明辐射制备的M-PEIs纳米凝胶是一种有效的基因治疗载体。 展开更多
关键词 辐射制备 聚乙烯亚胺 基因载体 短发夹状RNA
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Polycation-functionalized gold nanodots with tunable near-infrared fluorescence for simultaneous gene delivery and cell imaging 被引量:1
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作者 Yuanqing Sun Dandan Wang +6 位作者 Yueqi Zhao Tianxin Zhao Hongchen Sun Xiangwei Li Chuanxi Wang Bai Yang Quan Lin 《Nano Research》 SCIE EI CAS CSCD 2018年第5期2392-2404,共13页
Near-infrared (NIR) fluorescent metal nanodots may have significant advantages in biological detection and bioimaging. Herein, we introduce tunable near-infrared fluorescent gold nanodots (AuNDs) protected by bran... Near-infrared (NIR) fluorescent metal nanodots may have significant advantages in biological detection and bioimaging. Herein, we introduce tunable near-infrared fluorescent gold nanodots (AuNDs) protected by branched polyethylenimine (PEI) modified by surface segmental attachment of sulfhydryl groups (PEI-SH), abbreviated as PEI-SH-AuNDs, for simultaneous gene delivery and cell imaging. The modified PEI endows the resultant PEI-SH-AuNDs with the following excellent advantages. Sulfhydryl groups of PEI-SH anchor to the surface of AuNDs, and such polycations with amine groups give PEI-SH-AuNDs remarkable stability. The cationic polymer PEI-SH with positive charges enables PEI-SH-AuNDs to perform gene delivery, and the gene transfection efficiency can reach 22.8%. Moreover, the fluorescence of PEI-SH-AuNDs is tunable from visible red light (wavelength 609 nm) to NIR light (wavelength 811 run) via an increase in the size of AuNDs. PEI-SH-AuNDs yielded gene transfection efficiency similar to that of commercial PEI, but showed much lower cytotoxicity and much greater red-shift fluorescence. With excellent photoluminescent properties, such multifunctional fluorescent PEI-SH-AuNDs hold promise in applications to bioimaging and as ideal fluorescent probes for tracking gene transfection behavior. 展开更多
关键词 near-infrared fluorescent material Au nanodot tunable fluorescence gene deliver BIOIMAGING
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Inter-chain disulfide bond improved protein trans-splicing increases plasma coagulation activity in C57BL/6 mice following portal vein FVIII gene delivery by dual vectors 被引量:1
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作者 ZHU FuXiang LIU ZeLong +3 位作者 WANG XiaoLei MIAO Jing QU HuiGe CHI XiaoYan 《Science China(Life Sciences)》 SCIE CAS 2013年第3期262-267,共6页
Protein trans-splicing based dual-vector factor VIII (FVIII) gene delivery is adversely affected by less efficiency of protein splicing. We sought to increase the amount of spliced FVIII protein and plasma coagulati... Protein trans-splicing based dual-vector factor VIII (FVIII) gene delivery is adversely affected by less efficiency of protein splicing. We sought to increase the amount of spliced FVIII protein and plasma coagulation activity in dual-vector FVIII transgene in mice by means of strengthening the interaction of inteins, protein splicing elements, thereby facilitating protein trans-splicing. Dual-vector delivery of the FVIII gene in cultured cells showed that replacement of Met226 in the heavy chain and Asp1828 in the light chain with Cys residues could facilitate inter-chain disulfide linking and improve protein trans-splicing, increasing the levels of spliced FVIII protein. In this study, C57BL/6 mice were coadministered dual vectors of intein-fused human FVIII heavy chain and light chain with Cys mutations via portal vein injection into the liver. Forty-eight hours post-injection, plasma was collected and analyzed for FVIII antigen concentration and coagulation activity. These mice showed increased circulating FVIII heavy chain polypeptide (442± 151 ng mL i vs. 305±103 ng mL-1) and coagulation activi- ty (1.46±0.37 IU mL i vs. 0.85±0.23 IU mL-1) compared with control mice co-administered dual vectors expressing the heavy and light chains of wild-type FVIII. Moreover, coagulation activity was similar to that of mice receiving a single vector ex- pressing FVIII (1.79_+0.59 IU mL-l). These findings indicate that improving protein trans-splicing by inter-chain disulfide bonding is a promising approach for increasing the efficacy of dual-vector based FVIII gene transfer. 展开更多
关键词 coagulation factor VIII protein trans-splicing dual-vector gene delivery plasma coagulation activity
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Nobel Prize Winner Craig Mello Delivers Speech at Tsinghua University
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《Tsinghua Science and Technology》 SCIE EI CAS 2007年第3期301-301,共1页
Dr. Craig Mello, the Nobel Prize Winner of 2006 in Physiology or Medicine, delivered a speech entitled Return to The RNA World: Rethinking Gene Expression, Evolution, and Medicine at Tsinghua on March 26, 2007.
关键词 RNA World Nobel Prize Winner Craig Mello delivers Speech at Tsinghua University gene
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TMD2前断裂CFTR翻译后的连接及氯离子通道功能 被引量:1
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作者 朱甫祥 宫贤弟 +3 位作者 刘泽隆 杨树德 屈慧鸽 迟晓艳 《生物工程学报》 CAS CSCD 北大核心 2010年第12期1710-1716,共7页
CFTR基因突变导致一种常染色体隐性遗传疾病——囊性纤维化(CF)。利用split Ssp DnaB intein的蛋白质反式剪接技术的真核细胞双载体转CFTR基因,旨在研究翻译后水平CFTR的连接,以及由其建立的氯离子通道功能。于CFTR膜内第2个跨膜结构域(... CFTR基因突变导致一种常染色体隐性遗传疾病——囊性纤维化(CF)。利用split Ssp DnaB intein的蛋白质反式剪接技术的真核细胞双载体转CFTR基因,旨在研究翻译后水平CFTR的连接,以及由其建立的氯离子通道功能。于CFTR膜内第2个跨膜结构域(TMD2)前的Glu838密码子后将其cDNA断裂为N端和C端两部分,与具有蛋白质反式剪接作用的split Ssp DnaB intein编码序列融合,分别插入到载体pEGFP-N1和pEYFP-N1,构建一对真核表达载体pEGFP-NInt和pEYFP-IntC。用脂质体将这对载体共转染至幼年仓鼠肾细胞(BHK),瞬时表达实验用Western blotting观察CFTR蛋白质的连接,并用膜片钳技术记录Cl-通道电流。结果显示,基因共转染细胞呈现完整的CFTR蛋白条带,膜片钳记录到全细胞Cl-电流和单个Cl-通道开放活性。结果表明split Ssp DnaB intein的蛋白质反式剪接技术可用于双载体共转移CFTR基因,为CF基因治疗应用双腺相关病毒载体(AAV)转运CFTR基因,克服AAV的容量限制提供了依据。 展开更多
关键词 CFTR Cl-通道 SPLIT SSP DNAB INTEIN 蛋白质反式剪接 双载体基因转移
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新型基因载体纳米儿茶素-Fe^(2+)对人血管平滑肌细胞的转染效率及毒性评价
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作者 张德元 张展强 +2 位作者 胡黎平 梁天文 罗小跑 《热带医学杂志》 CAS 2017年第11期1429-1432,共4页
目的探讨纳米儿茶素-Fe^(2+)对人心血管平滑肌细胞的基因转染效率及细胞毒性,为后续对其转染机制及转染条件优化的研究提供依据。方法制备纳米儿茶素-Fe^(2+)的基因导入材料,将质量比为50∶1的儿茶素-Fe^(2+)/pEGFP-C1复合物转染人心血... 目的探讨纳米儿茶素-Fe^(2+)对人心血管平滑肌细胞的基因转染效率及细胞毒性,为后续对其转染机制及转染条件优化的研究提供依据。方法制备纳米儿茶素-Fe^(2+)的基因导入材料,将质量比为50∶1的儿茶素-Fe^(2+)/pEGFP-C1复合物转染人心血管平滑肌细胞,并设Lipo2000为对照,在荧光显微镜下观察细胞转染情况,采用MTT法检测儿茶素-Fe^(2+)/pEGFP-C1复合物转染对人心血管平滑肌细胞生长的影响,评价细胞毒性。结果纳米儿茶素-Fe^(2+)与pEGFP-C1最佳结合效率为50∶1;纳米儿茶素-Fe^(2+)组与Lipo2000组转染率差异无统计学意义(P>0.05);儿茶素-Fe^(2+)纳米粒子对转染细胞无明显细胞毒性;儿茶素-Fe^(2+)/pEGFP-C1纳米粒子转染后细胞荧光及EGFP荧光强度与Lipo2000组比较,转染效率相当(P>0.05)。结论儿茶素-Fe^(2+)是一种低细胞毒性,高转染率的纳米材料,具备良好的应用前景。 展开更多
关键词 纳米 儿茶素 基因导入材料
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昆虫翅发育研究的现状与前景 被引量:3
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作者 郭韶堃 罗丹 +1 位作者 李志红 沈杰 《植物保护学报》 CAS CSCD 北大核心 2017年第2期185-195,共11页
翅的形成有利于昆虫的迁移,一些害虫依靠强大的飞行能力扩散为害,致使农林业产业经济损失严重。目前,昆虫翅发育研究在遗传调控机理方面取得了大量突破性进展,作为生物器官的研究模型,昆虫翅为生物医学提供了良好的范本。为深入了解昆... 翅的形成有利于昆虫的迁移,一些害虫依靠强大的飞行能力扩散为害,致使农林业产业经济损失严重。目前,昆虫翅发育研究在遗传调控机理方面取得了大量突破性进展,作为生物器官的研究模型,昆虫翅为生物医学提供了良好的范本。为深入了解昆虫翅发育的研究进展和在植物保护及生物医学领域中的应用,本文介绍了昆虫翅发育的普遍调控机理,对比了不同变态发育类型代表性昆虫的翅发育关键调控基因及其作用机理,并结合最新涌现的RNA农药、纳米材料转染生物分子、基因编辑等技术,分析并展望了农业害虫防治中的翅模型研究的应用前景。 展开更多
关键词 昆虫 翅发育 RNA农药 纳米材料转染生物分子 基因编辑
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Enhanced plasma factor Ⅷ activity in mice via cysteine mutation using dual vectors 被引量:2
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作者 ZHU FuXiang LIU ZeLong +2 位作者 MIAO Jing QU HuiGe CHI XiaoYan 《Science China(Life Sciences)》 SCIE CAS 2012年第6期521-526,共6页
Hemophilia A is caused by a genetic mutation in coagulation factor VIII (FVIII) gene and gene therapy is considered to be a promising strategy for its treatment. We recently demonstrated that co-delivery of two vect... Hemophilia A is caused by a genetic mutation in coagulation factor VIII (FVIII) gene and gene therapy is considered to be a promising strategy for its treatment. We recently demonstrated that co-delivery of two vectors expressing M662C mutated heavy and D1828C mutated light chain genes of B-domain-deleted coagulation factor VIII (BDD-FVIII) leads to inter-chain disulfide cross-linking and improved heavy chain secretion in vitro. In this study, co-injection of both M662C and D1828C mutated BDD-FVIII gene expression vectors into mice resulted in increased heavy chain secretion and coagulation activity in plasma in vivo. Approximately (239+_56) ng mL-1 above endogenous levels of transgenic FVIII heavy chain was found in mouse plasma using a chain-specific ELISA. For FVIII coagulation activity, approximately (1.09+_0.25) IU mL-1 above en- dogenous levels were detected in co-injected transgenic mouse plasma using a chromogenic assay. These data demonstrate that inter-chain disulfide bonds likely increase heavy chain secretion and coagulation activity in the plasma of transgenic mice with an improved efficacy of a dual-vector delivery of BDD-FVIII gene. These findings support our ongoing efforts to develop a gene therapy for hemophilia A treatment using dual-AAV vectors. 展开更多
关键词 coagulation factor VIII dual-vector gene delivery inter-chain disulfide bonding heavy chain secretion coagulation ac-tivity
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