检索结果-南通市图书馆

Construction of a bicistronic recombinant adenoviral vector for human interleukin-10 and enhanced green fluorescent protein expression in bone marrow mesenchymal stem cells

引用

Chinese Medical Journal 2012年第1期125卷 102-108页

作者： LIN Jian-qing LIN Cai-zhu LIN Xian-zhong ZENG Kai GAO You-guang Department of Anesthesiology First Affiliated Hospital of Fujian Medical UniversityInstitute of Technology and EngineeringFujian Medical UniversityFuzhouFujian 350005China

Background Human interleukin-10 （hlL-10） is a cytokine synthesis inhibitory factor,which is involved in various immune responses.The purpose of this study was to construct an adenoviral vector carrying the hlL-10 gene for expression of biologically active hlL-10 in rat bone marrow mesenchymal stem cells （rMSCs）.Methods A pSNAV2.0-hlL10 plasmid was used as a template to obtain a hlL-10 cDNA fragment that was subcloned by restriction enzyme digestion and ligation into a pDC316-IRES-EGFP-lacZ alpha plasmid carrying an enhanced green fluorescent protein （EGFP） marker gene.The pDC316-hlL-10-IRES-EGFP plasmid was linearized by Pmel digestion and used to transfect HEK293 packaging cells using the adenovirus packaging system AdMax.Virus particles were amplified by repeatedly infecting HEK293 cells with the seed virus and then purified by ion exchange.After the number of virus particles and titer was determined,rMSCs were infected with the adenoviral vector.The infection rate was determined by fluorescence microscopy and flow cytometry,and hlL-10 protein expression in rMSCs was measured by Western blotting.Results The virus particle concentration,OD260/280 value and virus titer of the amplified and purified recombinant adenovirus were 3.2×1011 VP/ml,approximately 2.0,and 1.1×1010 TCID50/ml,respectively.Bright green fluorescence was observed by fluorescence microscopy and flow cytometry in the recombinant adenovirus-infected rMSCs.GFP expression was considered the multiplicity of infection （MOI） and was time-dependent.The infection rate was 92.9% at 100 MOI.Conclusions A bicistrenic recombinant adenoviral vector for hlL-10 and EGFP gene expression were successfully constructed.The infection rate of rMSCs by the adenovirus was high （92.9% at 100 MOI） and the target gene hlL-10 was highly expressed in cells.The present study provides an experimental basis for further research of immunosuppressive therapy using hlL-10.The expression level of hlL-10 protein as detected by Western blotting was also MOI- and time-dependent.

关键词： human interleukin-10 adenoviral vector bone marrow mesenchymal stem cells

来源：

维普期刊数据库

同方期刊数据库评论

在线全文

学校读者我要写书评

暂无评论

TLR4 signaling imprints the quality of germinal center response to adenoviral vector-based vaccine

TLR4 signaling imprints the quality of germinal center respo...

引用

第十二届全国免疫学学术大会

作者： li ruihua Xu Junjie Chen Wei Chinese Academy of Military Science

Background:The hall mark of adenoviral(Ad)vector-based vaccine is the generation of rapid and sustained adaprive immune response after one shot of immunization.The germinal center(GC),as the primary site of affinity m... 详细信息

关键词： Toll-like receptor adenoviral vector germinal center

来源： cnki会议评论

在线全文

cnki会议

学校读者我要写书评

暂无评论

Adenovirus-mediated gene delivery:Potential applications for gene and cell-based therapies in the new era of personalized medicine

引用

Genes & Diseases 2017年第2期4卷 43-63页

作者： Cody S.Lee Elliot S.Bishop Ruyi Zhang Xinyi Yu Evan M.Farina Shujuan Yan Chen Zhao Zongyue Zeng Yi Shu Xingye Wu Jiayan Lei Yasha Li Wenwen Zhang Chao Yang Ke Wu Ying Wu Sherwin Ho Aravind Athiviraham Michael J.Lee Jennifer Moriatis Wolf Russell R.Reid Tong-Chuan He The University of Chicago Pritzker School of Medicine ChicagoIL 60637USA Laboratory of Craniofacial Biology and Development Section of Plastic and Reconstructive SurgeryDepartment of SurgeryThe University of Chicago Medical CenterChicagoIL 60637USA Molecular Oncology Laboratory Department of Orthopaedic Surgery and Rehabilitation MedicineThe University of Chicago Medical CenterChicagoIL 60637USA Ministry of Education Key Laboratory of Diagnostic Medicine and the Affiliated Hospitals of Chongqing Medical UniversityChongqing 400016China Department of Laboratory Medicine and Clinical Diagnostics The Affiliated Yantai HospitalBinzhou Medical UniversityYantai 264100China Department of Immunology and Microbiology Beijing University of Chinese MedicineBeijing 100029China

With rapid advances in understanding molecular pathogenesis of human diseases in the era of genome sciences and systems biology,it is anticipated that increasing numbers of therapeutic genes or targets will become available for targeted therapies.Despite numerous setbacks,efficacious gene and/or cell-based therapies still hold the great promise to revolutionize the clinical management of human diseases.It is wildly recognized that poor gene delivery is the limiting factor for most in vivo gene therapies.There has been a long-lasting interest in using viral vectors,especially adenoviral vectors,to deliver therapeutic genes for the past two decades.Among all currently available viral vectors,adenovirus is the most efficient gene delivery system in a broad range of cell and tissue types.The applications of adenoviral vectors in gene delivery have greatly increased in number and efficiency since their initial development.In fact,among over 2000 gene therapy clinical trials approved worldwide since 1989,a significant portion of the trials have utilized adenoviral vectors.This review aims to provide a comprehensive overview on the characteristics of adenoviral vectors,including adenoviral biology,approaches to engineering adenoviral vectors,and their applications in clinical and preclinical studies with an emphasis in the areas of cancer treatment,vaccination and regenerative medicine.Current challenges and future directions regarding the use of adenoviral vectors are also discussed.It is expected that the continued improvements in adenoviral vectors should provide great opportunities for cell and gene therapies to live up to its enormous potential in personalized medicine.

关键词： Adenovirus adenoviral vector Cell therapy Gene transfer Gene therapy Oncolytic virus Regenerative medicine Vaccine development

来源：

维普期刊数据库评论

在线全文

维普期刊数据库

学校读者我要写书评

暂无评论

Inhibition of KIT RNAi mediated with adenovirus in gastrointestinal stromal tumor xenograft

引用

World Journal of Gastroenterology 2010年第40期16卷 5122-5129页

作者： Tian-Bao Wang Wen-Sheng Huang Wei-Hao Lin HanPing Shi Wen-Guang Dong Department of Surgery the First Affiliated Hospital Sun Yat-sen University Guangzhou 510080 Guangdong Province China

AIM: To investigate a therapeutic method for gastrointestinal stromal tumor (GIST) based on KIT RNA interference (RNAi) with AdMax adenovirus. METHODS: KIT short hairpin RNA (shRNA), whose lateral sides were decorated with restriction endonuclease sequences, was designed. T 4 DNA ligase catalyzed the joint of the KIT shRNA and the green fluorescent protein-containing PDC316-EGFP-U6 to form PDC316EGFP-U6-KIT. Homologous recombination of AdEGFPU6-KIT was performed with the AdMax system. Heterotopically transplanted GISTs were established in nude mice. AdEGFP-U6-KIT was intratumorally injected. The volume, inhibition ratio of tumor and CD117 expression of GIST graft tumor in nude mice were compared between test and control groups. RESULTS: The length of KIT shRNA was determined to be about 50bp by agarose electrophoresis. Gene se-quencing detected the designed KIT RNAi sequence in PDC316-EGFP-U6-KIT. After transfection with AdEGFPU6-KIT, 293 cells displayed green fluorescence. The physical and infective titers of AdEGFP-U6-KIT were 5 × 10 11 viral particles/mL and 5.67 × 10 7 plaque forming units/mL, respectively. The mean volume of the grafted tumor was significantly smaller in test mice than in control mice (75.3 ± 22.9 mm 3 vs 988.6 ± 30.5 mm 3 , t = -18.132, P vector system can effectively inhibit the expression of the KIT gene and the growth of GIST in nude mice.

关键词： Gastrointestinal stromal tumor RNA interference KIT adenoviral vector Nude mice

来源：

维普期刊数据库评论

在线全文

维普期刊数据库

学校读者我要写书评

暂无评论

Adenovirus-mediated expression of pig α(1,3) galactosyltransferase reconstructs Gal α(1,3) Gal epitope on the surface of human tumor cells

引用

Cell Research 2001年第2期11卷 116-124页

作者： XING LI, GUO HONG XIA, JIAN FEI, FANG HUANG, LI HE GUO Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai China

Gal α(1, 3) Gal (gal epitope) is a carbohydrate epitope and synthesized in large amount by α(1, 3) galactosyltransferase [α(1, 3) GT] enzyme on the cells of lower mammalian animals such as pigs and mice. Human has no gal epitope due to the inactivation of α(1, 3) GT gene but produces a large amount of antibodies (anti-Gal) which recognize Gal α(1, 3) Gal structures specifically. In this study, a replication deficient recombinant adenoviral vector Ad5sGT containing pig α(1, 3) GT cDNA was constructed and characterized. adenoviral vector-mediated transfer of pig α(1, 3) GT gene into human tumor cells such as malignant melanoma A375, stomach cancer SGC-7901, and lung cancer SPC-A-1 was reported for the first time. Results showed that Gal epitope did not increase the sensitivity of human tumor cells to human complement-mediated lysis, although human complement activation and the binding of human IgG and IgM natural antibodies to human tumor cells were enhanced significantly after Ad5sGT transduction. Appearance of gal epitope on the human tumor cells changed the expression of cell surface carbohydrates reacting with Ulex europaeus I (UEA I) lectins, Vicia villosa agglutinin (VVA), Arachis hypogaea agglutinin (PNA), and Glycine max agglutinin (SBA) to different degrees. In addition, no effect of gal epitope on the growth in vitro of human tumor cells was observed in MTT assay.

关键词： adenoviral vector galactosyltransferase Gal α(1 3) Gal gene expression human tumor cell.

来源：

同方期刊数据库评论

在线全文

同方期刊数据库

学校读者我要写书评

暂无评论

Combination of epidural electrical stimulation with ex vivo triple gene therapy for spinal cord injury:a proof of principle study

引用

Neural Regeneration Research 2021年第3期16卷 550-560页

作者： Filip Olegovich Fadeev Farid Vagizovich Bashirov Vahe Arshaluysovich Markosyan Andrey Alexandrovich Izmailov Tatyana Vyacheslavovna Povysheva Mikhail Evgenyevich Sokolov Maxim Sergeevich Kuznetsov Anton Alexandrovich Eremeev Ilnur Ildusovich Salafutdinov Albert Anatolyevich Rizvanov Hyun Joon Lee Rustem Robertovich Islamov Department of Medical Biology and Genetics Kazan State Medical UniversityKazanRussia Federal University KazanRussia Department of Neurology University of Mississippi Medical CenterJacksonMSUSA Research Service G.V.(Sonny)Montgomery VA Medical CenterJacksonMSUSA

Despite emerging contemporary biotechnological methods such as gene-and stem cell-based therapy,there are no clinically established therapeutic strategies for neural regeneration after spinal cord injury.Our previous studies have demonstrated that transplantation of genetically engineered human umbilical cord blood mononuclear cells producing three recombinant therapeutic molecules,including vascular endothelial growth factor(VEGF),glial cell-line derived neurotrophic factor(GDNF),and neural cell adhesion molecule(NCAM)can improve morpho-functional recovery of injured spinal cord in rats and mini-pigs.To investigate the efficacy of human umbilical cord blood mononuclear cells-mediated triple-gene therapy combined with epidural electrical stimulation in the treatment of spinal cord injury,in this study,rats with moderate spinal cord contusion injury were intrathecally infused with human umbilical cord blood mononuclear cells expressing recombinant genes VEGF165,GDNF,NCAM1 at 4 hours after spinal cord injury.Three days after injury,epidural stimulations were given simultaneously above the lesion site at C5(to stimulate the cervical network related to forelimb functions)and below the lesion site at L2(to activate the central pattern generators)every other day for 4 weeks.Rats subjected to the combined treatment showed a limited functional improvement of the knee joint,high preservation of muscle fiber area in tibialis anterior muscle and increased H/M ratio in gastrocnemius muscle 30 days after spinal cord injury.However,beneficial cellular outcomes such as reduced apoptosis and increased sparing of the gray and white matters,and enhanced expression of heat shock and synaptic proteins were found in rats with spinal cord injury subjected to the combined epidural electrical stimulation with gene therapy.This study presents the first proof of principle study of combination of the multisite epidural electrical stimulation with ex vivo triple gene therapy(VEGF,GDNF and NCAM)for treatment of spinal cord injury in rat mode

关键词： adenoviral vector epidural electrical stimulation gene therapy glial cell-line derived neurotrophic factor human umbilical cord blood mononuclear cell neural cell adhesion molecule spinal cord injury vascular endothelial growth factor

来源：

维普期刊数据库

同方期刊数据库评论

在线全文

学校读者我要写书评

暂无评论

Construction of recombinant human nerve growth factor beta adenovirus and evaluation of its function An in vitro and in vivo study

引用

Neural Regeneration Research 2010年第16期5卷 1261-1269页

作者： En-Feng Gao Si-Ho Choi Mi-Ae Sung Bo-Han Li Samir Jabaiti Sang Bae Yoo Sung-June Kim Soung-Min Kim Jeong Won Jahng Jong-Ho Lee Department of Stomatology China-Japan Friendship Hospital Beijing 100029 China Center for Stem Cell and Regenerative Medicine Keck School of Medicine University of Sourthern Californina USA Department of Oral and Maxillofacial Surgery & Dental Research Institute Seoul National University Seoul Korea Department of Plastic and Reconstructive Surgery Jordan University Hospital Amman Jordan Department of Electrical Engineering School of Electrical and Computer Science Seoul National University Seoul Korea

Exogenous delivery of nerve growth factor （NGF） promotes neural regeneration. However, the short half-life limits delivery efficacy. Therefore, a long-term, efficient, local delivery tool or scheme is needed. The purpose of this study was to construct a functioning, recombinant, adenoviral vector carrying human NGF-β （hNGF-β） DNA, and to measure expression of the constructed vector in vitro and in vivo. rhNGF-β adenoviral vector containing full-length hNGF-β cDNA was generated by homologous recombination in Escherichia CoIL The rhNGF-β adenovirus was packaged and amplified in human embryonic kidney HEK293 cells. Transformation efficiency, expression and function of rhNGF-β adenovirus for primary Schwann cells, Schwann cell lines, human embryonic kidney HEK 293 cells, CRH myoblasts, and NIH3T3 fibroblasts were evaluated. Subsequently, expression of rhNGF-β adenovirus at the peripheral nerve of rat was also assessed. Recombinant adenoviral vector carrying hNGF-β was successfully constructed and confirmed by restriction endonuclease analysis and DNA sequence analysis. Green fluorescent protein expression was observed in 90% of rhNGF-β adenovirus-infected cells （primary Schwann cells, Schwann cell line, human embryonic kidney HEK 293 cells, CRH myoblasts, and NIH3T3 fibroblasts） compared with non-infected cells. Total mRNA isolated from rhNGF-β adenovirus-infected cells exhibited strong expression. Maximum NGF release was induced by primary cultured Schwann cells at 4 days after infection, which steadily continued for 14 days. PC-12 cells exposed to media conditioned with rhNGF-β adenovirus-infected Schwann cells exhibited increased neurite extension. In vivo experiment revealed that the injected rhNGF-β adenovirus was transfected into the cells at the injected site and promoted expression of NGF, p75NTR and brain derived neurotrophic factor at the sciatic nerve and dorsal root ganglia.

关键词： nerve growth factor Schwann cell peripheral nerve regeneration adenoviral vector HEK293 cells myoblasts fibroblasts

来源：

维普期刊数据库

同方期刊数据库评论

在线全文

学校读者我要写书评

暂无评论

Overexpression of the mTERT gene by adenoviral vectors promotes the proliferation of neuronal stem cells in vitro and stimulates neurogenesis in the hippocampus of mice

引用

The Journal of Biomedical Research 2012年第5期26卷 381-388页

作者： Mengying Liu Yao Hu Lijuan Zhu Chen Chen Yu Zhang Weixiang Sun Qigang Zhou Department of Pharmacology Pharmacy CollegeNanjing Medical UniversityNanjingJiangsu 210029China

We sought to construct the adenoviral vector carrying the gene encoding mouse telomerase reverse transcriptase（mTERT）,as well as detect its expression and effect on the proliferation of neuronal stem cells.mTERT was am-plified by RT-PCR and then the eukaryotic expression vector of pDC-EGFP-TERT was constructed.After DNA sequence analysis,we detected that there were 293 cells transfected with pDC-EGFP-TERT and helper adenovirus plasmid pBHG lox ΔE1,and three Cre using Lipofectamine 2000 mediation,named Ad-mTERT-GFP,to pack-age adenoviral particles.The Ad-mTERT-GFP was used to infect neuronal stem cells and then the expression and activity of mTERT were detected.In addition,Bromodeoxyuridine labeling test identified the impact of mTERT overexpression on proliferation of neuronal stem cells.The recombinant adenoviral vector confirmed that mTERT was successfully constructed.Overexpression of mTERT stimulated the proliferation of neuronal stem cells both in vitro and in vivo.mTERT overexpression via adenoviral vector carrying mTERT cDNA upregulated the ability of proliferation in neuronal stem cells.

关键词： telomerase construct eukaryotic expression vector adenoviral vector proliferation neuronal stemcells

来源：

维普期刊数据库

同方期刊数据库评论

在线全文

学校读者我要写书评

暂无评论

EFFECT ON BIOLOGICAL BEHAVIOR OF CHEMOTHERAPY-RESISTANT TUMOR CELLS BY HUMAN WILD-TYPE p53,GM-CSF AND B7-1 GENES VIA RECOMBINANTADENOVIRUS

引用

Chinese Journal of Cancer Research 2000年第2期12卷 105-114页

作者：范国昌吴祖泽王艳飞吴世凯邱兆华 Department of Experimental Hematology Beijing Institute of Radiation Medicine Beijing China

Objective: To explore the effect on biological behavior of chemotherapy-resistant tumor cells by human wild-type p53, GM-CSF and B7-1 genes mediated via recombinant adenovirus. Methods: p53-abnormal KB-v200 (VCR resistant) and KB-s (VCR sensitive) cell lines were used as model tumor cells, which are resistant and sensitive to chemotherapeutic drugs respectively. After infected with recombinant adenovirus carrying human wild-type p53, GM-CSF and B7-1 genes, changes in biological behavior (including drug sensitivity) of these two kinds of gene-transduced cancer cells were observed. Results: Both of the cell lines were susceptible to adenovirus, all of three exogenous genes (p53, GM-CSF and B7-1) could be effectively expressed in these cell lines, their growth was suppressed, and apoptosis was induced. The drug-pumping-out function of Pgp glycoprotein on the cytomembrane of drug-resistant KB-v200 cells was markedly affected 48h after transfection of the recombinant adenovirus, revealed by increase of the amount of rhodamine 123 accumulation in the cells. The MTT assay also indicated the reversal of their sensitivity to VCR drugs.In vivo experiment in nude mice it was demonstrated reduction of tumorigenicity of the KB-v200 cells or KB-s cells infected with the recombinant adenovirus, and increase of their sensitivity to VCR. Conclusion: The clinical application of this recombinant adenovirus carrying agents might be more effective in treatment of tumors with multidrug resistance(MDR).

关键词： Tumor gene therapy multidrug resistance adenoviral vector

来源：

维普期刊数据库

同方期刊数据库评论

在线全文

学校读者我要写书评

暂无评论

Long term survival and limited migration of genetically modified monocytes/macrophages grafted into the mouse brain

引用

Journal of Biomedical Science and Engineering 2013年第5期6卷 561-571页

作者： Chamsy Sarkis Gabriel Gras Francoise Sanchez Jacques Mallet Che Serguera Centre de Recherche de l’Institut du Cerveau et de la Moelle Epiniere Equipe de Biotechnologie et BiothérapieHopital Pitié-SalpetriereParisFrance CEA Institute of Emerging Diseases and Innovative TherapiesDivision of Immuno-VirologyUMR E1 CEA and University Paris SouthFontenay-aux RosesFrance Centre d’Etudes d’Agents Pathogenes et Biotechnologies pour la Sante(CPBS) UMR 5236MontpellierFrance Laboratoire de Modélisation des Biotherapies Groupe INSERM U986MIRCenCEAFontenay-aux-RosesFrance

In mammals, myeloid progenitors infiltrate the developing central nervous system (CNS), through the immature blood-brain barrier (BBB), the ventricular layer or the pial surface migrate and give rise to resident microglia. In the mature brain, however, the BBB hampers such recruitment from the blood-stream and long-term establishment of blood borne myeloid cells in the CNS thus appears at best limited. Hematopoietic stem cell-derived microglia, nevertheless, represents a promising tool for the correction of genetic deficits in the brain. We thus investigated the fate of primary human monocytes, and monocyte-derived macrophages, following transplantation into the adult mouse brain overpassing the BBB. Furthermore, we documented the ability of such cells to deliver a lysosomal enzyme into the brain following genetic modification with a recombinant adenoviral vector carrying the human β-glucuronidase cDNA. When implanted into the mouse striatum, the engineered primary cells survived and expressed the transgene for as much as 8 months. Moreover, the donor cells could migrate out of the grafting site and settle along blood vessels or myelin tracts although at limited distance. Migrating donor cells down-regulated the expression of CD14 andHLA DR, suggesting the adoption of a deactivated microglia-like phenotype. Our observations establish the ability of circulating mononuclear phagocytes to integrate into the brain after transplantation and express a transgene on the long term. These cells might thus be employed for autologous transplantation for the delivery of secreted therapeutic proteins in the context of a wide range of brain affections.

关键词： adenoviral vector Human Myeloid Cells β-Glucuronidase Ex Vivo Gene Therapy Neurotransplantation

来源：

维普期刊数据库评论

在线全文

维普期刊数据库

学校读者我要写书评

暂无评论

欢迎您,

建议与咨询留下您的常用邮箱和电话号码，以便我们向您反馈解决方案和替代方法

时间限定

文献类型

馆藏选择

核心期刊

语言

文献类型

帮助

文字说明：

检索规则说明：

检索范例：

分类表

所选分类

限定检索结果

文献类型

馆藏范围

日期分布

学科分类号

主题

机构

作者

语言

在线全文

在线全文

在线全文

在线全文

在线全文

在线全文

在线全文

在线全文

在线全文

在线全文

请选择保存的检索档案：

请选择收藏分类：

通借通还

欢迎您,

建议与咨询 留下您的常用邮箱和电话号码，以便我们向您反馈解决方案和替代方法

时间限定

文献类型

馆藏选择

核心期刊

语言

文献类型

帮助

文字说明：

检索规则说明：

检索范例：

分类表

所选分类

限定检索结果

文献类型

馆藏范围

日期分布

学科分类号

主题

机构

作者

语言

在线全文

在线全文

在线全文

在线全文

在线全文

在线全文

在线全文

在线全文

在线全文

在线全文

请选择保存的检索档案： 新增检索档案 确定 取消

请选择收藏分类： 新增自定义分类 确定 取消

通借通还

建议与咨询留下您的常用邮箱和电话号码，以便我们向您反馈解决方案和替代方法

请选择保存的检索档案：

请选择收藏分类：