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基因治疗病毒载体的研究进展 被引量:1
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作者 王振发 王烈 卫立辛 《福州总医院学报》 2009年第4期326-328,共3页
关键词 基因治疗病毒载体 免疫缺陷综合征 类白血病反应 疑难杂症 SCID 基因载体 主要表现 转染效率
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基于EBNA1和oriP的载体在基因治疗中的应用研究进展 被引量:3
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作者 何婕 张智清 《生物工程学报》 CAS CSCD 北大核心 2005年第3期507-510,共4页
非病毒载体用于基因治疗的主要问题是导入靶细胞的效率较低,目的基因表达水平低,疗效持续时间也较短。EBNA1和oriP元件使引入该元件的质粒在真核细胞内保持为游离体、转运入核、转录增强。质粒携带的目的基因能够获得较高的转染效率,高... 非病毒载体用于基因治疗的主要问题是导入靶细胞的效率较低,目的基因表达水平低,疗效持续时间也较短。EBNA1和oriP元件使引入该元件的质粒在真核细胞内保持为游离体、转运入核、转录增强。质粒携带的目的基因能够获得较高的转染效率,高水平、持续的表达。基于EBNA1 oriP的质粒在肿瘤、单基因缺陷先天性疾病、TNF相关的炎性疾病的基因治疗中显示了良好的应用前景。EBNA1 oriP元件用于构建人工染色体,携带目的基因的调控序列,可望实现可调控的基因治疗。 展开更多
关键词 基因治疗 EBNA1/oriP 病毒载体
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动态光散射法测定病毒载体基因治疗产品的平均粒径及粒径分布 被引量:2
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作者 李永红 毕华 +1 位作者 陈伟 饶春明 《现代生物医学进展》 CAS 2014年第34期6611-6613,共3页
目的:建立病毒载体基因产品的粒径及分布的测定方法。方法:采用马尔文Zetasizer Nano ZS型纳米粒度仪,以氯化钠注射液作为稀释剂对样品进行适当稀释后,用手动测量方式进行测定。结果:测定的60 nm标准纳米粒子平均粒径为61.24±0.68... 目的:建立病毒载体基因产品的粒径及分布的测定方法。方法:采用马尔文Zetasizer Nano ZS型纳米粒度仪,以氯化钠注射液作为稀释剂对样品进行适当稀释后,用手动测量方式进行测定。结果:测定的60 nm标准纳米粒子平均粒径为61.24±0.68nm,多分散指数(polydispersity index,PDI)为0.020±0.012 nm。重组人新型P53腺病毒注射液测定6次,平均粒径为122.42±1.80 nm,相对标准偏差(RSD)为1.4%;PDI值为0.056±0.014 nm,RSD为25.1%。重组人新型P53腺病毒注射液于37℃存放时,平均粒径和PDI值随时间延长有显著增加。至28天时,平均粒径增加到130.50,PDI值增加到0.265。结论:动态光散射法快速、简便、准确,可用于病毒载体基因治疗产品的粒径及分布的检测。 展开更多
关键词 动态光散射 粒径 粒径分布 病毒载体基因治疗产品 质量控制
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非病毒载体基因递送系统的研究进展 被引量:2
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作者 孙岚 张英鸽 《军事医学科学院院刊》 CSCD 北大核心 2003年第5期384-387,共4页
非病毒载体基因递送系统是相对于以病毒为载体的基因递送系统的另一种基因递送系统。具有高安全性、低免疫原性及易于生产的特性。本文就非病毒基因载体的种类、在疾病治疗中的应用前景及存在的主要问题作一综述。
关键词 病毒载体基因递送系统 研究进展 病毒载体基因治疗 基因疗法 病毒基因
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树状化合物在生物与医药中的应用 被引量:2
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作者 莫尊理 李俊 +2 位作者 高锦章 蒲陆梅 杨武 《化学试剂》 CAS CSCD 北大核心 2001年第2期84-85,共2页
简要综述了近几年来树状化合物在生物与医药中的应用进展
关键词 应用 树状化合物 医药 树状成像造影剂 硼中子捕获治疗 基因治疗病毒载体 免疫制剂 生物医学
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p53-expressing conditionally replicative adenovirus CNHK500-p53 against hepatocellular carcinoma in vitro 被引量:4
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作者 Hong-Chuan Zhao Qi Zhang Yang Yang Min-Qiang Lu Hua Li Chi Xu Gui-Hua Chen 《World Journal of Gastroenterology》 SCIE CAS CSCD 2007年第5期683-691,共9页
AIM: To develop a conditionally replicative gene-viral vector system called CNHK500-p53, which contains dual promoters within the E1 region, and combines the advantages of oncolytic virus and gene therapies for hepat... AIM: To develop a conditionally replicative gene-viral vector system called CNHK500-p53, which contains dual promoters within the E1 region, and combines the advantages of oncolytic virus and gene therapies for hepatocellular carcinoma (HCC). METHODS: CNHK500-p53 was constructed by using human telomerase reverse transcriptase (hTERT) promoter to drive adenovirus E1a gene and hypoxia response element (HRE) promoter to drive adenovirus E1b gene. p53 gene expressing cassette was inserted into the genome of replicative virus. Viral replication experiments, cytopathic effect (CPE) and methyl thiazolyl tetrazolium (MTT) assay were performed to test the selective replication and oncolytic efficacy of CNHK500-p53. RESULTS: Immunohistochemistry verified that infection with CNHK500-p53 was associated with selective replication of adenovirus and production of p53 protein in telomerase-positive and hypoxia-inducible factordependent HCC cells, p53 protein secreted from HepG2, infected with CNHK500-p53 was significantly higher than that infected with nonreplicative adenovirus Ad-p53 in vitro (388 ± 34.6 μg/L vs 76.3 ± 13.17 μg/L). Viral replication experiments showed that replication of CNHK500-p53 and CNHK500 or WtAd5, was much stronger than that of Ad-p53 in tested HCC cell lines. CPE and H1-F assay indicated that CNHK500-p53 selectively replicated in and killed HCC cells while leaving normal cells unaffected. CONCLUSION: A more efficient gene-viral system is developed by combining selective oncolysis with exogenous expression of p53 against HCC cells. 展开更多
关键词 Conditionally replicative adenovirus Oncolytic virotherapy Gene therapy p53 gene Hepatocellular carcinoma
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Gene therapy for allergic rhinitis with recombinant adenovirus vector containing CTLA4Ig in mice 被引量:1
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作者 朱瑾 吴军 +3 位作者 陈希炜 易绍萱 罗高兴 贺伟峰 《Journal of Medical Colleges of PLA(China)》 CAS 2003年第1期15-18,共4页
Objective: To examine the role of recombinant adenovirus vector containing CTLA4Ig gene(Ad-CTLA4Ig) in the treatment of induced allergic rhinitis in mice.Methods: Allergic rhinitis was induced by sensitizing and chall... Objective: To examine the role of recombinant adenovirus vector containing CTLA4Ig gene(Ad-CTLA4Ig) in the treatment of induced allergic rhinitis in mice.Methods: Allergic rhinitis was induced by sensitizing and challenging with ovalbumin(OVA).Ad-CTLA4Ig was intraperitoneally injected 30 min before OVA challenge.Adenovirus vector without inserted CTLA4Ig cDNA served as the control.The symptoms and morphological changes of nasal mucosa of each group were observed, and the serum levels of IgE against OVA were detected with ELISA.Results: There were no obvious symptoms and pathological changes in Ad-CTLA4Ig treated group, in which the serum OVA-specific IgE levels were significantly lower than that in control groups(P< 0.05).Conclusion: Ad-CTLA4Ig prevents and treats allergic rhinitis of mice,implying the possibility of the usage of Ad-CTLA4Ig against allergic rhinitis in clinic in future. 展开更多
关键词 recombinant CTLA4Ig adenovirus vector allergic rhinitis IGE
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In vitro effect of p2l^(WAF-1/CIP1) gene on growth of human glioma cells mediated by EGFR targeted non-viral vector GE7 system
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作者 陈永新 许秀兰 +5 位作者 张光霁 王韦 金海英 卢亦成 朱诚 顾健人 《Journal of Medical Colleges of PLA(China)》 CAS 2003年第4期222-225,250,共5页
Objective: To construct the EGFR targeted non-viral vector GE7 system and explore the in vitro effect of p21WAF-1/CIPI gene on growth of human glioma cells mediated by the GE7 system. Methods: The EGFR targeted non-vi... Objective: To construct the EGFR targeted non-viral vector GE7 system and explore the in vitro effect of p21WAF-1/CIPI gene on growth of human glioma cells mediated by the GE7 system. Methods: The EGFR targeted non-viral vector GE7 gene delivery system was constructed. The malignant human glioma cell line U251MG was transfected in vitro with β-galactosidase gene ( reporter gene) and p21WAF-1/CIPI gee (therapeutic gene) using the GE7 system. By means of X-gal staining, MTS and FACS, the transfection efficiency of exogenous gene and apoptosis rate of tumor cells were examined. The expression of p21WAF-1/ CIPI gene in transfected U251MG cell was examined by immunohistochemis-try staining. Results: The highest transfer rate of exogenous gene was 70% . After transfection with p21WAF-1/CIPI gene, the expression of WAF-1 increased remarkably and steadily; the growth of U251MG cells were inhibited evidently. FACS examination showed G1 arrest. The average apoptosis rate was 25.2%. Conclusion: GE7 system has the ability to transfer exogenous gene to targeted cells efficiently, and expression of p21WAF-1/CIPI gene can induce apoptosis of glioma cell and inhibit its growth. 展开更多
关键词 GLIOMA EGFR targeted non-viral vector p21^(WAF-1CIPI) apoptosis IMMUNOHISTOCHEMISTRY
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Gene-viral vectors: a promising way to target tumor cells and express anticancer genes simultaneously
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作者 钱其军 岑信棠 +5 位作者 车小燕 徐建国 薛惠斌 崔贞福 朱斌 吴孟超 《Chinese Medical Journal》 SCIE CAS CSCD 2002年第8期1213-1217,154-155,共5页
OBJECTIVE: To develop a new kind of vector system called gene-viral vector, which combines the advantages of gene and virus therapies. METHODS: Using recombinant technology, an anti-tumor gene was inserted into the ge... OBJECTIVE: To develop a new kind of vector system called gene-viral vector, which combines the advantages of gene and virus therapies. METHODS: Using recombinant technology, an anti-tumor gene was inserted into the genome of replicative virus specific for tumor cells. The cell killing effect, reporter gene expression of the green fluorescence protein, anti-tumor gene expression of mouse interleukin-12 (mIL-12) and replication of virus were observed by the methods of cell pathology, fluorescence microscopy, ELISA and electron microscopy, respectively. RESULTS: A new kind of gene-viral vector system of adenovirus, in which the E1b-55 kD gene was deleted but the E1a gene was preserved, was constructed. The vector system, like the replicative virus ONYX-015, replicated and proliferated in tumor cells but not in normal ones. Our vector had an advantage over ONYX-015 in that it carried different kinds of anti-tumor genes to enhance its therapeutic effect. The reporter gene expression of the green fluorescence protein in tumor cells was much better than the adenovirus vector employed in conventional gene the rapy, and the expression in our vector system was as low as or even less than that in the conventional adenovirus gene therapy system. Similar results were observed in experiments with this vector system carrying the anti-tumor gene mIL-12. Replication and proliferation of the virus carrying the mIL-12 gene in tumor cells were confirmed by electron microscopy. CONCLUSIONS: Gene-viral vectors are new vectors with an anti-tumor gene inserted into the genome of replicative virus specific for tumor cells. Because of the specific replication and proliferation of the virus in tumor cells, expression of the anti-tumor gene is increased hundreds to thousands of times. This approach takes full advantages of gene therapy and virus therapy to enhance the effect on the tumor. It overcomes the disadvantages of conventional gene therapy, such as low transfer rate, low gene expression, lack of target tropism, and low anti-tumor activity. We believe that this is a promising means for future tumor treatment. 展开更多
关键词 ADENOVIRIDAE Adenovirus E1A Proteins Adenovirus E1B Proteins Gene Therapy Genetic Vectors Humans INTERLEUKIN-12 Neoplasms Recombination Genetic Research Support Non-U.S. Gov't Tumor Cells Cultured Virus Replication
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Inhibition of allergic responsiveness in a murine asthma model via IFN-γ transgene expression
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作者 高占成 康禹 +3 位作者 徐钰 尚颖 改军 何权瀛 《Chinese Medical Journal》 SCIE CAS CSCD 2002年第10期30-34,144-145,共7页
To investigate adenoviral vector mediated exogenous gene expression in mouse lungs and the effect of mIFN γ transgene expression on allergen induced pulmonary eosinophil infiltration in a murine asthmatic model ... To investigate adenoviral vector mediated exogenous gene expression in mouse lungs and the effect of mIFN γ transgene expression on allergen induced pulmonary eosinophil infiltration in a murine asthmatic model Methods LacZ marker gene was transduced into CD 1 mouse airway epithelial cells by installation of a replication deficient adenovirus with LacZ gene (AdCMVLacZ) 5×10 9 plaque forming unit (pfu) in the intratrachea or nostril C57 mice were sensitized intraperitoneally and challenged by aerosol with ovalbumin (OVA) to produce an asthmatic model AdCMVmIFNγ 5×10 9 pfu was administered via nostril in asthmatic mice 48 h before OVA challenge Sera, bronchial alveolar lavage (BAL) and lungs were recovered 48 h after OVA challenge Results After administration with AdCMVLacZ by intratracheal installation or nose drop, the lungs revealed a high level of widespread LacZ transduction with X gal staining, mainly along airways IFN γ via adenoviral vector transduction could be overexpressed both in vitro and in vivo (1624 7±1321 5 pg/ml in BAL 96 h after AdCMVIFNγ infection) In AdCMVIFNγ treated asthmatic models, histological evaluation revealed marked suppression of eosinophil peribronchial and perivascular infiltration; the recoverable percentage of eosinophils in BAL was an average of 9 00%±4 58%, which was a statistically significant decrease versus that of the positive control group (75 13%±6 85%) ( P 【0 001) The total cell number in BAL ((145±55 6)×10 3 cells/ml) in AdCMVmIFNγ treated mice also was tremendously reduced compared to the positive control group ((216 6±71 1)×10 3 cells/ml) Conclusions Adenoviral vector was able to overexpress exogenous gene in murine lungs IFN γ overexpression via adenoviral vector in pulmonary epithelia in vivo can abrogate allergen induced eosinophilic infiltration in lungs in an asthmatic model, which may suggest a new preventively therapeutic method for cytokine immunogenetic transfer in allergic asthma 展开更多
关键词 gene therapy · adenoviral vector · inter feron type · asthma
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Adenovirus induced acute hepatitis in non-human primates after liver-directed gene therapy 被引量:1
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作者 鲁慧英 Deborah Sullivan +1 位作者 Michael A Gerbera Srikanta Dash 《Chinese Medical Journal》 SCIE CAS CSCD 2002年第5期726-731,153,共6页
OBJECTIVE: To define the mechanism of acute hepatitis in non-human primates after liver directed gene therapy. METHODS: Differences in immune response exhibited by 8 rhesus monkeys receiving adenovirus (Ad) or lipofec... OBJECTIVE: To define the mechanism of acute hepatitis in non-human primates after liver directed gene therapy. METHODS: Differences in immune response exhibited by 8 rhesus monkeys receiving adenovirus (Ad) or lipofectamine-mediated gene transfer by various routes, the time course, and the nature of the specific immune responses to both adenoviral vectors and transgene products were studied using HE staining (H&E) and immunohistochemical staining. RESULTS: The monkeys developed mild to moderate acute hepatitis 1 to 3 weeks after intravenous or intrabiliary injection of first generation replication-defective adenoviruses carrying the Escherichia coli lacZ gene. This was accompanied by adenovirus-mediated T-cell proliferation and neutralizing antibodies to the adenovirus. Increased numbers of CD3(+), CD4(+) and CD8(+) T-lymphocytes were detected in the diseased livers, while B-lymphocytes were absent. Hepatocytes demonstrated increased expression of beta 2-microglobulins (beta 2-MG) and HLA-DR antigens in the plasma membranes. The development of acute hepatitis and the accompanying immune abnormalities were delayed in immunosuppressed monkeys until after the discontinuation of immunosuppressive therapy. The monkeys infused with Ad. CMVluc showed more significant and longer durations of hepatitis than the monkeys infused with adenoviruses carrying the lacZ gene. Lipofectamine-mediated gene transfer was inefficient. There was neither lacZ expression nor significant immune response in the liver of monkeys infused with lipofectamine via the portal vein or the common bile duct. CONCLUSION: Immune response to the hepatocytes in liver directed gene therapy is MHC class I restricted and T-cell mediated. Both adenoviral vectors and foreign genes are related to the liver damage. Mild to moderate hepatic inflammation seen with the E-1 deleted vector is reversible. Immunosuppression regimens may prolong transgene expression and delay the development of acute adenoviral hepatitis. 展开更多
关键词 Acute Disease ADENOVIRIDAE Adenoviridae Infections Animals Antigens CD3 Antigens CD4 Antigens CD8 DNA Recombinant Gene Transfer Techniques HLA-DR Antigens Hepatitis Animal Immunohistochemistry LIVER Macaca mulatta beta 2-Microglobulin
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