OBJECTIVE To investigate the effect of polyethylene imine glycol （PEI-PEG）/siRNA nanocomposites in the in vitro transfection of human gastric cancer SGC7901 cell lines and the down-regulation of gene expression of t...OBJECTIVE To investigate the effect of polyethylene imine glycol （PEI-PEG）/siRNA nanocomposites in the in vitro transfection of human gastric cancer SGC7901 cell lines and the down-regulation of gene expression of the adherence factor CD44v6. METHODS PEI-PEG/siRNA nanoparticles, in different N/P ratios, were synthesized and transfected into gastric cancer cells. Lipo2000/siRNA was used in the control group. The transfection efficiencies were observed under fluorescence microscope. The cytotoxicity of the nanoparticles was measured using the MTT assay （mononuclear cell direct cytotoxicity assay）, and the down-regulation effect of siRNA on CD44v6 gene was evaluated by Western blot. Based on the different N/P ratios, PEI-PEG/siRNA composites were synthesized and transfected into gastric cancer cells. Lipo2000/siRNA was used in the controls. The transfection efficiency was observed under fluorescence microscope. The cytotoxicity of the nanoparticles was measured using the MTT assay and the down-regulation effect of siRNA on CD44v6 gene was evaluated by Western blot. RESULTS After transfection, the transfection efficiency of the PEI-PEG/siRNA nanocomposites increased incrementally in N/P ratio value. The transfection efficiency improved with an increase in N/P ratio. When the N/P value was 15, fluorescence became more intense in the PEI-PEG/siRNA group than in the Lipo2000/siRNA group. At the same time, cell viability was （80.4 ± 5.6）% in the MTT reduction assay, which was similar to that in the Lipo2000/siRNA group. The results of Western blot analysis showed that the expression level of CD44v6 protein decreased to （59.7 ± 3.0）% after siRNA-CD44v6 was inhibited. CONCLUSION PEI-PEG could effectively form the nanocomposite in combination with siRNA, be transfected into the SGC7901 gastric cancer cell lines and inhibit CD44v6 protein expression. Moreover, as a genetic carrier, PEI-PEG copolymer has greater advantages, including high transfection e. ciency, less cytotoxicity and an easily alterable vector structure.展开更多
The so-called comprehensive practice activities refer to that under teachers＇ guidance, students conduct independent comprehensive learning activities, and in activities, students can train practice ability, organica...The so-called comprehensive practice activities refer to that under teachers＇ guidance, students conduct independent comprehensive learning activities, and in activities, students can train practice ability, organically combining theory knowledge and practice knowledge together, in order to have a learning and practice effect. In current college English teaching, it exists module, single and dogmatic and a series of disadvantage trends, taking comprehensive practice activities as the carrier to improve college English teaching efficiency, which has a multiplier effect. This paper first analyzes the importance of comprehensive practical activities in college English teaching activities, and then it proposes a set of practicable activity strategies, hoping to play a valuable role.展开更多
Successful gene vectors should be with high transfection efficiency and minimal cytotoxicity. Natural polysaccharides, due to their good biocompatibility and biodegradability, have been widely studied and applied. Amy...Successful gene vectors should be with high transfection efficiency and minimal cytotoxicity. Natural polysaccharides, due to their good biocompatibility and biodegradability, have been widely studied and applied. Amylopectin is one of polysaccharides with dendritic structure and numerous hydroxyl groups that could be used for subsequent modification. In this work, a series of dendritic cationic gene vectors comprising amylopectin backbones and poly(2-(dimethylamino) ethyl methacrylate)(PDMAEMA) side chains with different lengths(termed as AMY-PDs) were readily prepared by atom transfer radical polymerization(ATRP). The gene condensation ability, cytotoxicity and gene transfection of AMY-PDs carriers were investigated. In comparison with "gold-standard" poly(ethyleneimine)(PEI, 25 k Da), the AMY-PDs exhibited higher transfection efficiency with lower cytotoxicity. AMY-PDs could be further modified with Au nanoparticles(termed as AMY-PD@Au). The potential of the AMY-PD@Au vectors to be utilized as a CT contrast agent for imaging of cancer cells was investigated. Such AMY-PD@Au vectors may realize gene therapy with the ability of real-time imaging.展开更多
基金This work was supported by the following funds： National Natural Science Foundation of China （No.30670951） Guangdong Provincial+5 种基金 Natural Science Foundation （No.06021322） Fund of Guangzhou Municipal Scientific Problem-Solving Program （No. 2003 Z 3-E0381） Fund of Guangdong Provincial Scientific Problem-Solving Program （No.2005 B31211002） Guangdong Provincial Government and Ministry of Education Project com- bining project initiation, study and research （No.2009B090300277）.
文摘OBJECTIVE To investigate the effect of polyethylene imine glycol （PEI-PEG）/siRNA nanocomposites in the in vitro transfection of human gastric cancer SGC7901 cell lines and the down-regulation of gene expression of the adherence factor CD44v6. METHODS PEI-PEG/siRNA nanoparticles, in different N/P ratios, were synthesized and transfected into gastric cancer cells. Lipo2000/siRNA was used in the control group. The transfection efficiencies were observed under fluorescence microscope. The cytotoxicity of the nanoparticles was measured using the MTT assay （mononuclear cell direct cytotoxicity assay）, and the down-regulation effect of siRNA on CD44v6 gene was evaluated by Western blot. Based on the different N/P ratios, PEI-PEG/siRNA composites were synthesized and transfected into gastric cancer cells. Lipo2000/siRNA was used in the controls. The transfection efficiency was observed under fluorescence microscope. The cytotoxicity of the nanoparticles was measured using the MTT assay and the down-regulation effect of siRNA on CD44v6 gene was evaluated by Western blot. RESULTS After transfection, the transfection efficiency of the PEI-PEG/siRNA nanocomposites increased incrementally in N/P ratio value. The transfection efficiency improved with an increase in N/P ratio. When the N/P value was 15, fluorescence became more intense in the PEI-PEG/siRNA group than in the Lipo2000/siRNA group. At the same time, cell viability was （80.4 ± 5.6）% in the MTT reduction assay, which was similar to that in the Lipo2000/siRNA group. The results of Western blot analysis showed that the expression level of CD44v6 protein decreased to （59.7 ± 3.0）% after siRNA-CD44v6 was inhibited. CONCLUSION PEI-PEG could effectively form the nanocomposite in combination with siRNA, be transfected into the SGC7901 gastric cancer cell lines and inhibit CD44v6 protein expression. Moreover, as a genetic carrier, PEI-PEG copolymer has greater advantages, including high transfection e. ciency, less cytotoxicity and an easily alterable vector structure.
文摘The so-called comprehensive practice activities refer to that under teachers＇ guidance, students conduct independent comprehensive learning activities, and in activities, students can train practice ability, organically combining theory knowledge and practice knowledge together, in order to have a learning and practice effect. In current college English teaching, it exists module, single and dogmatic and a series of disadvantage trends, taking comprehensive practice activities as the carrier to improve college English teaching efficiency, which has a multiplier effect. This paper first analyzes the importance of comprehensive practical activities in college English teaching activities, and then it proposes a set of practicable activity strategies, hoping to play a valuable role.
基金supported by the National Natural Science Foundation of China(51173014,51221002,51325304,51373017,51302009,51473014)the Research Fund for Doctoral Program of Higher Education of China(20120010120007)Collaborative Innovation Center for Cardiovascular Disorders,Beijing Anzhen Hospital Affiliated to the Capital Medical University
文摘Successful gene vectors should be with high transfection efficiency and minimal cytotoxicity. Natural polysaccharides, due to their good biocompatibility and biodegradability, have been widely studied and applied. Amylopectin is one of polysaccharides with dendritic structure and numerous hydroxyl groups that could be used for subsequent modification. In this work, a series of dendritic cationic gene vectors comprising amylopectin backbones and poly(2-(dimethylamino) ethyl methacrylate)(PDMAEMA) side chains with different lengths(termed as AMY-PDs) were readily prepared by atom transfer radical polymerization(ATRP). The gene condensation ability, cytotoxicity and gene transfection of AMY-PDs carriers were investigated. In comparison with "gold-standard" poly(ethyleneimine)(PEI, 25 k Da), the AMY-PDs exhibited higher transfection efficiency with lower cytotoxicity. AMY-PDs could be further modified with Au nanoparticles(termed as AMY-PD@Au). The potential of the AMY-PD@Au vectors to be utilized as a CT contrast agent for imaging of cancer cells was investigated. Such AMY-PD@Au vectors may realize gene therapy with the ability of real-time imaging.
