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载报告基因壳聚糖纳米粒制剂体外转染活性考察 被引量:5
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作者 周旭 黄伟 +3 位作者 贺俊峰 崔光华 全东琴 梅兴国 《解放军药学学报》 CAS 2003年第4期241-245,共5页
目的 考察壳聚糖纳米粒体外基因转染活性及壳聚糖纳米粒经表面修饰的体外转染活性变化。方法 用复凝聚法制备纳米粒 ,透射电镜观察粒子形态 ,体外基因转染实验评价纳米粒的体外转染活性 ,用倒置荧光显微镜观察和流式细胞仪测定转染结... 目的 考察壳聚糖纳米粒体外基因转染活性及壳聚糖纳米粒经表面修饰的体外转染活性变化。方法 用复凝聚法制备纳米粒 ,透射电镜观察粒子形态 ,体外基因转染实验评价纳米粒的体外转染活性 ,用倒置荧光显微镜观察和流式细胞仪测定转染结果。通过考察递送不同剂量的基因和转染后不同时间的转染效率 ,寻找本递送系统较好的转染条件。用纳米粒表面连接PEG以及半乳糖基白蛋白 ,对纳米粒进行修饰 ,通过体外转染实验 ,评价表面修饰对其转染活性的影响。结果 未经表面修饰的载基因纳米粒能够转染人胚胎肾细胞 (HEK2 93)和肝癌细胞 (HepG2 ) ,但转染效率仍不如脂质体转染试剂 ,且在转染实验后约 72h较高 ,递送基因较佳的剂量是 4 μg ,在以上两种细胞中 ,转染效率也不同。经PEG表面修饰的纳米粒仍保持纳米粒的体外转染活性。连接半乳糖基牛血清白蛋白的纳米粒转染活性却反而略有下降。结论 壳聚糖纳米粒能将基因递送到细胞内 ,并且报告基因能在细胞内表达。因此 ,可以用作基因递送的载体系统 ,值得进一步研究。经PEG表面修饰和冷冻干燥处理 ,保持生物活性 ,为基因药物制剂化的可能性提供了证据。对于靶向配基的选择 ,宜继续进行筛选。 展开更多
关键词 壳聚糖纳米粒制剂 基因转染活性 基因治疗 体外活性 基因药物
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Dual function of Arabidopsis A TAF1 in abiotic and biotic stress responses 被引量:60
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作者 Yaorong Wu Zhiyong Deng +8 位作者 Jianbin Lai Yiyue Zhang Cuiping Yang Bojiao Yin Qingzhen Zhao Ling Zhang Yin Li Chengwei Yang Qi Xie 《Cell Research》 SCIE CAS CSCD 2009年第11期1279-1290,共12页
NAC family genes encode plant-specific transcription factors involved in diverse biological processes. In this study, the Arabidopsis NAC gene ATAF1 was found to be induced by drought, high-salinity, abscisic acid (... NAC family genes encode plant-specific transcription factors involved in diverse biological processes. In this study, the Arabidopsis NAC gene ATAF1 was found to be induced by drought, high-salinity, abscisic acid (ABA), methyl jasmonate, mechanical wounding, and Botrytis cinerea infection. Significant induction of ATAF1 was found in an ABA-deficient mutant aba2 subjected to drought or high salinity, revealing an ABA-independent mechanism of expression. Arabidopsis ATAFl-overexpression lines displayed many altered phenotypes, including dwarfism and short primary roots. Furthermore, in vivo experiments indicate that ATAF1 is a bonafide regulator modulating plant responses to many abiotic stresses and necrotrophic-pathogen infection. Overexpression of ATAF1 in Arabidopsis increased plant sensitivity to ABA, salt, and oxidative stresses. Especially, ATAF1 overexpression plants, but not mutant lines, showed remarkably enhanced plant tolerance to drought. Additionally, ATAF1 overexpression enhanced plant susceptibility to the necrotrophic pathogen B. cinerea, but did not alter disease symptoms caused by avirulent or virulent strains of P. syringae pv tomato DC3000. Transgenic plants overexpressing ATAF1 were hypersensitive to oxidative stress, suggesting that reactive oxygen intermediates may be related to ATAFl-mediated signaling in response to both pathogen and abiotic stresses. 展开更多
关键词 abiotic stress ARABIDOPSIS ATAF1 biotic stress
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Osteogenic differentiation of mesenchymal stem cells promoted by overexpression of connective tissue growth factor 被引量:9
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作者 Jin-jing WANG Feng YE +6 位作者 Li-jia CHENG Yu-jun SHI Ji BAO Huai-qiang SUN Wei WANG Peng ZHANG Hong BU 《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》 SCIE CAS CSCD 2009年第5期355-367,共13页
Objective:Large segmental bone defect repair remains a clinical and scientific challenge with increasing interest focusing on combining gene transfection with tissue engineering techniques.The aim of this study is to ... Objective:Large segmental bone defect repair remains a clinical and scientific challenge with increasing interest focusing on combining gene transfection with tissue engineering techniques.The aim of this study is to investigate the effect of connective tissue growth factor(CTGF) on the proliferation and osteogenic differentiation of the bone marrow mesenchymal stem cells(MSCs).Methods:A CTGF-expressing plasmid(pCTGF) was constructed and transfected into MSCs.Then expressions of bone morphogenesis-related genes,proliferation rate,alkaline phosphatase activity,and mineralization were examined to evaluate the osteogenic potential of the CTGF gene-modified MSCs.Results:Overexpression of CTGF was confirmed in pCTGF-MSCs.pCTGF transfection significantly enhanced the proliferation rates of pCTGF-MSCs(P<0.05).CTGF induced a 7.5-fold increase in cell migration over control(P<0.05).pCTGF transfection enhanced the expression of bone matrix proteins,such as bone sialo-protein,osteocalcin,and collagen type I in MSCs.The levels of alkaline phosphatase(ALP) activities of pCTGF-MSCs at the 1st and 2nd weeks were 4.0-and 3.0-fold higher than those of MSCs cultured in OS-medium,significantly higher than those of mock-MSCs and normal control MSCs(P<0.05).Overexpression of CTGF in MSCs enhanced the capability to form mineralized nodules.Conclusion:Overexpression of CTGF could improve the osteogenic differentiation ability of MSCs,and the CTGF gene-modified MSCs are potential as novel cell resources of bone tissue engineering. 展开更多
关键词 Mesenchymal stem cells (MSCs) Connective tissue growth factor (CTGF) Osteogenic differentiation OSTEOBLASTS OVEREXPRESSION Gene modification
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