Background The present study aimed to investigate the detailed mode and specific sites for their binding as well as the functional relevance of this binding in the phenotypic proliferation of vascular smooth muscle ce...Background The present study aimed to investigate the detailed mode and specific sites for their binding as well as the functional relevance of this binding in the phenotypic proliferation of vascular smooth muscle cells(SMCs). Methods CREG knocked-down SMCs were employed to evaluate the biological activity of wtCREG and mCREG.Expressions of SMC differentiation markers SM myosin heavy chain(SM-MHC),SM-actin,heavy caldesmon and myocardin were determined by Western blotting using specific antibodies. Cellular growth of SMCs was assessed by bromide dewuridine (BrdU) incorporation and cell cycle analysis on fluorescence-activated cell sorting(FACS).A solid-phase binding assay was used to study the binding of CREG to extracellular domains of M6P/IGF2R.The cellular co-localization of the two recombinant CREGs with M6P/IGF2R was detected on SMC surface by immunoprecipitation and immunofluorescence analysis.Results The molecular weight of wtCREG was around 30 kD while that of the mCREG was～25 kD.Treatment of wtCREG with PNGase F reduced its molecular weight from～30 kD to～25 kD,whereas PNGase F treatment had no effect on the molecular weight of mCREG.Both wtCREG and mCREG proteins enhanced SMC differentiation,inhibited BrdU incorporation,and arrested cell cycle progression when added to the culture medium.In CREG knocked-down SMCs,the amount of CREG detected by immunoblotting in M6P/IGF2R immunoprecipitates was significantly reduced when compared to normal cells.Both recombinant CREGs co-immunoprecipitated with M6P/IGF2R, although slightly reduced amount of the mutant CREG was detected in M6P/IGF2R immunoprecipitates.Immunostaining revealed that His-tagged CREGs co-localized with IGF2R on the cell surface in a glycosylation-independent manner.In vitro binding assay showed that CREGs bound to M6P/ IGF2R extracellular domains 7-10 and 11-13 in a glycosylation -dependent and -independent manner,respectively.Further blocking experiments using soluble M6P/IGF2R fragments and M6P/IGF2R neutralizing antibody indicated that the biological activities of recombinant CREGs in SMC growth and the up-regulation of SMC differentiation markers were all abolished by treatment with the M6P/IGF2R neutralizing antibody. However,although the growth inhibitory effect of wtCREG was nearly abolished by D7-10 or D11-13,the effect of mCREG was only reversed by Dll-13,indicating that the binding to domains 11-13 is required for CREG to modulate the proliferation of SMCs.Conclusions These data suggest that solubleCREG proteins can exert their biological function via binding to the extracellular domains 7-10 and 11-13 of cell surface M6P/IGF2R in both a glycosylation-dependent and -independent manner.展开更多
Ⅰ.词汇A)根据句意及首字母提示,补全单词。1.The t—— month of the year is December.2.The w—— in China is quite different fromthat in Australia.3.It rained the whole day yesterday.The radiosays it’s w—— today than ...Ⅰ.词汇A)根据句意及首字母提示,补全单词。1.The t—— month of the year is December.2.The w—— in China is quite different fromthat in Australia.3.It rained the whole day yesterday.The radiosays it’s w—— today than yesterday.4.What kind of p——aye you going to givehim for his birthday?展开更多
利用苄硫醇、溴十一酸、4,7,10-三氧-1,13-十三烷二胺等原料,通过活化酯法和NH3/Na还原脱苄基,合成了一种含巯基的生物素衍生物——11-巯基十一酸-(13-生物素酰胺基-4,7,10-三氧十三烷基)酰胺,并用1 H NMR、IR对产物的分子结构进行了表...利用苄硫醇、溴十一酸、4,7,10-三氧-1,13-十三烷二胺等原料,通过活化酯法和NH3/Na还原脱苄基,合成了一种含巯基的生物素衍生物——11-巯基十一酸-(13-生物素酰胺基-4,7,10-三氧十三烷基)酰胺,并用1 H NMR、IR对产物的分子结构进行了表征.11-巯基十一酸-(13-生物素酰胺基-4,7,10-三氧十三烷基)酰胺可以在金表面自组装,而自组装膜在分子器件、生物与化学传感器、生物膜等很多方面有着广泛的应用.展开更多
During the past decades,early fetal ultrasound and diagnosis have increasingly gained attention in pregnancy care with the development of high-frequency transducers,which make it possible to obtain detailed images of ...During the past decades,early fetal ultrasound and diagnosis have increasingly gained attention in pregnancy care with the development of high-frequency transducers,which make it possible to obtain detailed images of the early fetus and its organs,and thus move part of the anatomy and anomaly scan from the second to the first-trimester.By transabdominal sonography or transvaginal sonography,or a combination of both approaches,it is now able to diagnose a wide variety of fetal abnormalities at 11-13^(+6)weeks.展开更多
A ftrst facile total synthesis of 12-hydroxy- 13-methylpodocarpa-9, 11, 13-trien-3-one 9 was accomplished via a strategy of AC→ABC. In the synthetic process, compound 5 was synthesized through Reimer-Teimann reaction...A ftrst facile total synthesis of 12-hydroxy- 13-methylpodocarpa-9, 11, 13-trien-3-one 9 was accomplished via a strategy of AC→ABC. In the synthetic process, compound 5 was synthesized through Reimer-Teimann reaction, followed by methylation, reduction, dehydration, addition to ethylene epoxide and chlorination. The target compound 9 was finally obtained through the condensation of Grignard reagent of compound 5 with 6, followed by intramolecular cyclization, reduction and demethylation.展开更多
文摘Background The present study aimed to investigate the detailed mode and specific sites for their binding as well as the functional relevance of this binding in the phenotypic proliferation of vascular smooth muscle cells(SMCs). Methods CREG knocked-down SMCs were employed to evaluate the biological activity of wtCREG and mCREG.Expressions of SMC differentiation markers SM myosin heavy chain(SM-MHC),SM-actin,heavy caldesmon and myocardin were determined by Western blotting using specific antibodies. Cellular growth of SMCs was assessed by bromide dewuridine (BrdU) incorporation and cell cycle analysis on fluorescence-activated cell sorting(FACS).A solid-phase binding assay was used to study the binding of CREG to extracellular domains of M6P/IGF2R.The cellular co-localization of the two recombinant CREGs with M6P/IGF2R was detected on SMC surface by immunoprecipitation and immunofluorescence analysis.Results The molecular weight of wtCREG was around 30 kD while that of the mCREG was～25 kD.Treatment of wtCREG with PNGase F reduced its molecular weight from～30 kD to～25 kD,whereas PNGase F treatment had no effect on the molecular weight of mCREG.Both wtCREG and mCREG proteins enhanced SMC differentiation,inhibited BrdU incorporation,and arrested cell cycle progression when added to the culture medium.In CREG knocked-down SMCs,the amount of CREG detected by immunoblotting in M6P/IGF2R immunoprecipitates was significantly reduced when compared to normal cells.Both recombinant CREGs co-immunoprecipitated with M6P/IGF2R, although slightly reduced amount of the mutant CREG was detected in M6P/IGF2R immunoprecipitates.Immunostaining revealed that His-tagged CREGs co-localized with IGF2R on the cell surface in a glycosylation-independent manner.In vitro binding assay showed that CREGs bound to M6P/ IGF2R extracellular domains 7-10 and 11-13 in a glycosylation -dependent and -independent manner,respectively.Further blocking experiments using soluble M6P/IGF2R fragments and M6P/IGF2R neutralizing antibody indicated that the biological activities of recombinant CREGs in SMC growth and the up-regulation of SMC differentiation markers were all abolished by treatment with the M6P/IGF2R neutralizing antibody. However,although the growth inhibitory effect of wtCREG was nearly abolished by D7-10 or D11-13,the effect of mCREG was only reversed by Dll-13,indicating that the binding to domains 11-13 is required for CREG to modulate the proliferation of SMCs.Conclusions These data suggest that solubleCREG proteins can exert their biological function via binding to the extracellular domains 7-10 and 11-13 of cell surface M6P/IGF2R in both a glycosylation-dependent and -independent manner.
文摘Ⅰ.词汇A)根据句意及首字母提示,补全单词。1.The t—— month of the year is December.2.The w—— in China is quite different fromthat in Australia.3.It rained the whole day yesterday.The radiosays it’s w—— today than yesterday.4.What kind of p——aye you going to givehim for his birthday?
文摘利用苄硫醇、溴十一酸、4,7,10-三氧-1,13-十三烷二胺等原料,通过活化酯法和NH3/Na还原脱苄基,合成了一种含巯基的生物素衍生物——11-巯基十一酸-(13-生物素酰胺基-4,7,10-三氧十三烷基)酰胺,并用1 H NMR、IR对产物的分子结构进行了表征.11-巯基十一酸-(13-生物素酰胺基-4,7,10-三氧十三烷基)酰胺可以在金表面自组装,而自组装膜在分子器件、生物与化学传感器、生物膜等很多方面有着广泛的应用.
文摘During the past decades,early fetal ultrasound and diagnosis have increasingly gained attention in pregnancy care with the development of high-frequency transducers,which make it possible to obtain detailed images of the early fetus and its organs,and thus move part of the anatomy and anomaly scan from the second to the first-trimester.By transabdominal sonography or transvaginal sonography,or a combination of both approaches,it is now able to diagnose a wide variety of fetal abnormalities at 11-13^(+6)weeks.
文摘A ftrst facile total synthesis of 12-hydroxy- 13-methylpodocarpa-9, 11, 13-trien-3-one 9 was accomplished via a strategy of AC→ABC. In the synthetic process, compound 5 was synthesized through Reimer-Teimann reaction, followed by methylation, reduction, dehydration, addition to ethylene epoxide and chlorination. The target compound 9 was finally obtained through the condensation of Grignard reagent of compound 5 with 6, followed by intramolecular cyclization, reduction and demethylation.
