Objective To study the effect of phospholamban antisense RNA （asPLB） on sarcoplasmic reticulum Ca2＋-ATPase activity and cardiac function in rats with diabetes mellitus （DM） mediated by recombinant adeno-associate...Objective To study the effect of phospholamban antisense RNA （asPLB） on sarcoplasmic reticulum Ca2＋-ATPase activity and cardiac function in rats with diabetes mellitus （DM） mediated by recombinant adeno-associated virus （rAAV） vector. Methods Six weeks after the induction of DM by streptozotocin injected intraperitoneally, the rats were divided into three groups, namely： DM-rAAV-asPLB group, DM-saline group and DM group （control group）. The rats in the DM-rAAV-asPLB group were intramyocardially injected with rAAV-asPLB, the rats in the DM-saline group were injected with saline, and those in the control group did not receive any treatment. Six weeks after gene transfer, the expressions of PLB protein and PLB phosphorylation were detected by Western-blot, while the activity of sarcoplasmic reticulum （SR） Ca2＋-ATPase and left ventricular function were measured. Results The PLB protein expression level was significantly higher whereas the PLB phosphorylation, SR Ca2＋-ATPase activity and left ventricular function were significantly lower in the DM-saline group than in the control group. No significant difference was found in PLB protein expression level, PLB phosphorylation or SR Ca2＋-ATPase activity between the DM-rAAV-asPLB group and the control group. The left ventricular function in the DM-rAAV-asPLB group was poorer than in the control group and was better than in the DM-saline group. Conclusion rAAV-asPLB can down-regulate PLB protein expression and up-regulate phosphorylation and SR Ca2＋-ATPase activity, thus contributing to the improvement of in vivo ventricutar function. PLB left展开更多
In this study, we report that the insertion of a pseudoproline dipeptide for the solid-phase peptide synthesis of wild-type Phospholamban protein (WT-PLB) has two important advantages. First, it disrupts the formation...In this study, we report that the insertion of a pseudoproline dipeptide for the solid-phase peptide synthesis of wild-type Phospholamban protein (WT-PLB) has two important advantages. First, it disrupts the formation of different secondary structures, which is responsible for poor couplings during the preparation of highly aggregated sequences. Second, it enhances the purities and solubility of crude products leading to easier HPLC purification.展开更多
In this study, we report the microwave assisted synthesis of Phospholamban protein (WT-PLB) as a new gold standard in solid phase peptide synthesis. Microwave energy offers benefits for both the coupling and deprotect...In this study, we report the microwave assisted synthesis of Phospholamban protein (WT-PLB) as a new gold standard in solid phase peptide synthesis. Microwave energy offers benefits for both the coupling and deprotection reactions during peptide synthesis. The use of microwave energy for both the coupling and deprotection steps makes the microwave peptide synthesizers the most versatile and powerful systems available. It produces high yield and fast synthesis when compared to conventional peptide synthesizers.展开更多
Among orchids, Cymbidiums have got a very high demand in both cut flower and pot plant trade. In the present study the effect of some polysaccharides such as chitosan and NAG （n-acetyl-glucosamine） on organogenesis ...Among orchids, Cymbidiums have got a very high demand in both cut flower and pot plant trade. In the present study the effect of some polysaccharides such as chitosan and NAG （n-acetyl-glucosamine） on organogenesis in protocorm-like-bodies （PLBs） of C. insigne was studied. Synthetic phytohormones such as BA and TDZ （cytokinins） and NAA （auxin） were used for comparison. PLBs of C. insigne were explanted on modified Murashige and Skoog medium supplemented with the single addition of chitosan and NAG, and the combination of BA （benzyladenine） and NAA （1-naphthaleneacetic acid） also with the combination of NAA and TDZ （thidiazuron） among different concentrations. Combination treatments of auxin and cytokinins, the highest percentage of PLBs formation was 73% and shoot formation was 67% when cultured on the medium supplemented with 1.0 mg L1 BA without NAA. Combination treatment of NAA and TDZ, the PLBs formation was 90% and shoot formation was 60% obtained from medium supplemented with 1.0 mg Lt of NAA ＋ 0.1 mg L^-1 TDZ. Single addition of chitosan and NAG with modified MS medium was more effective for new PLBs and shoot formation. The highest percentage of PLBs formation was 87% and shoot formation was 80% obtained from the medium supplemented with 0.1 mg L^-1 chitosan. On the other hand, the PLBs formation rate reached 93% and shoot formation rate was 87% obtained from the medium supplemented with 0.01 mg L^-1 NAG. Application of polysaccharides to in vitro orchid PLBs allows developing new PLBs and shoot to form plantlet without synthetic phytohormones.展开更多
基金supported by Scientific Research Foundation of College of Medicine,Zhejiang UniversityFoundation of Zhejiang Science and Technology Bureau(No.2007B071)+1 种基金Foundation of Zhejiang Educational Committee(No.Y200908338)Foundation of Zhejiang Traditional Chinese Medicine Committee(No.2011B503103)
文摘Objective To study the effect of phospholamban antisense RNA （asPLB） on sarcoplasmic reticulum Ca2＋-ATPase activity and cardiac function in rats with diabetes mellitus （DM） mediated by recombinant adeno-associated virus （rAAV） vector. Methods Six weeks after the induction of DM by streptozotocin injected intraperitoneally, the rats were divided into three groups, namely： DM-rAAV-asPLB group, DM-saline group and DM group （control group）. The rats in the DM-rAAV-asPLB group were intramyocardially injected with rAAV-asPLB, the rats in the DM-saline group were injected with saline, and those in the control group did not receive any treatment. Six weeks after gene transfer, the expressions of PLB protein and PLB phosphorylation were detected by Western-blot, while the activity of sarcoplasmic reticulum （SR） Ca2＋-ATPase and left ventricular function were measured. Results The PLB protein expression level was significantly higher whereas the PLB phosphorylation, SR Ca2＋-ATPase activity and left ventricular function were significantly lower in the DM-saline group than in the control group. No significant difference was found in PLB protein expression level, PLB phosphorylation or SR Ca2＋-ATPase activity between the DM-rAAV-asPLB group and the control group. The left ventricular function in the DM-rAAV-asPLB group was poorer than in the control group and was better than in the DM-saline group. Conclusion rAAV-asPLB can down-regulate PLB protein expression and up-regulate phosphorylation and SR Ca2＋-ATPase activity, thus contributing to the improvement of in vivo ventricutar function. PLB left
文摘In this study, we report that the insertion of a pseudoproline dipeptide for the solid-phase peptide synthesis of wild-type Phospholamban protein (WT-PLB) has two important advantages. First, it disrupts the formation of different secondary structures, which is responsible for poor couplings during the preparation of highly aggregated sequences. Second, it enhances the purities and solubility of crude products leading to easier HPLC purification.
文摘In this study, we report the microwave assisted synthesis of Phospholamban protein (WT-PLB) as a new gold standard in solid phase peptide synthesis. Microwave energy offers benefits for both the coupling and deprotection reactions during peptide synthesis. The use of microwave energy for both the coupling and deprotection steps makes the microwave peptide synthesizers the most versatile and powerful systems available. It produces high yield and fast synthesis when compared to conventional peptide synthesizers.
文摘Among orchids, Cymbidiums have got a very high demand in both cut flower and pot plant trade. In the present study the effect of some polysaccharides such as chitosan and NAG （n-acetyl-glucosamine） on organogenesis in protocorm-like-bodies （PLBs） of C. insigne was studied. Synthetic phytohormones such as BA and TDZ （cytokinins） and NAA （auxin） were used for comparison. PLBs of C. insigne were explanted on modified Murashige and Skoog medium supplemented with the single addition of chitosan and NAG, and the combination of BA （benzyladenine） and NAA （1-naphthaleneacetic acid） also with the combination of NAA and TDZ （thidiazuron） among different concentrations. Combination treatments of auxin and cytokinins, the highest percentage of PLBs formation was 73% and shoot formation was 67% when cultured on the medium supplemented with 1.0 mg L1 BA without NAA. Combination treatment of NAA and TDZ, the PLBs formation was 90% and shoot formation was 60% obtained from medium supplemented with 1.0 mg Lt of NAA ＋ 0.1 mg L^-1 TDZ. Single addition of chitosan and NAG with modified MS medium was more effective for new PLBs and shoot formation. The highest percentage of PLBs formation was 87% and shoot formation was 80% obtained from the medium supplemented with 0.1 mg L^-1 chitosan. On the other hand, the PLBs formation rate reached 93% and shoot formation rate was 87% obtained from the medium supplemented with 0.01 mg L^-1 NAG. Application of polysaccharides to in vitro orchid PLBs allows developing new PLBs and shoot to form plantlet without synthetic phytohormones.
