This study investigated the abnormal expression of ATP synthase β-subunit（ATPsyn-β） in pancreas islets of rat model of polycystic ovary syndrome（PCOS） with type 2 diabetes mellitus（T2DM）,and the secretion func...This study investigated the abnormal expression of ATP synthase β-subunit（ATPsyn-β） in pancreas islets of rat model of polycystic ovary syndrome（PCOS） with type 2 diabetes mellitus（T2DM）,and the secretion function changes after up-regulation of ATP5 b.Sixty female SD rats were divided into three groups randomly and equally.The rat model of PCOS with T2 DM was established by free access to the high-carbohydrate/high-fat diet,subcutaneous injections of DHEA,and a single injection of streptozotocin.The pancreas was removed for the detection of the ATPsyn-β expression by immunohistochemical staining,Western blotting and reverse transcription-PCR（RT-PCR）.The pancreas islets of the rats were cultured,isolated with collagenase Ⅴ and purified by gradient centrifugation,and the insulin secretion after treatment with different glucose concentrations was tested.Lentivirus ATP5 b was successfully constructed with the vector of GV208 and transfected into the pancreas islets for the over-expression of ATPsyn-β.The insulin secretion and intracellular ATP content were determined after transfection of the PCOS-T2 DM pancreas islets with Lenti-ATP5 b.The results showed that the expression of ATPsyn-β protein and m RNA was significantly decreased in the pancreas of PCOS-T2 DM rats.The ATP content in the pancreas islets was greatly increased and the insulin secretion was improved after the up-regulation of ATPsyn-β in the pancreas islets transfected with lenti-ATP5 b.These results indicated that for PCOS,the ATPsyn-β might be one of the key factors for the attack of T2 DM.展开更多
In this study, the immuno-response of recombinant hCG-β and natural hCGβ was comparatively investigated by using Freund's adjuvant. The results showed that, the properties and merits of the antibodies elicited b...In this study, the immuno-response of recombinant hCG-β and natural hCGβ was comparatively investigated by using Freund's adjuvant. The results showed that, the properties and merits of the antibodies elicited by both kinds of hCG-β were similar. The antisera had high affinity for binding with hCG (Kαγβ≈5.86×108/mol/L, Kαβ≈8.18×108/mol/L), and were found to be effective in inhibiting the binding of 125I-hCG to receptors in rat testes. Results also indicated that, similar to the antisera induced by natural hCG-β, the recombinant hCG-β induced antisera had capacity of neutralizing the biological activities of hCG. Recombinant hCG-β could be used as an immunogen for contraceptive vaccine.展开更多
EFR is a plasma-membrane resident receptor responsible for recognition of microbial elongation factorTu （EF-Tu） and thus triggering plant innate immunity to fend off phytopathogens. Functional EFR must be subject to...EFR is a plasma-membrane resident receptor responsible for recognition of microbial elongation factorTu （EF-Tu） and thus triggering plant innate immunity to fend off phytopathogens. Functional EFR must be subject to the endoplasmic reticulum quality control （ERQC） machinery for the correct folding and proper assembly in order to reach its final destination. Genetic studies have demonstrated that ERD2b, a counterpart of the yeast or mammalian HDEL receptor ERD2 for retaining proteins in the endoplasmic reticulum （ER） lumen, is required for EFR function in plants （Li et al., 2009）. In this study, we characterized the Arabidopsis glucosidase Ⅱ β--subunit via the H DEL motif against the non-redundant protein database. Data mining also revealed that the glucosidase Ⅱ β--subunit gene has a highly similar expression pattern to ERD2b and the other known ERQC components involved in EFR biogenesis. Importantly, the T-DNA insertion lines of the glucosidase Ⅱ β-subunit gene showed that EFR-controlled responses were substantially reduced or completely blocked in these mutants. The responses include seedling growth inhibition, induction of marker genes, MAP kinase activation, and callose deposition, trigged by peptide elf18, a full mimic of E F-Tu. Taken together, ourdata indicate a requirement of the glucosidase Ⅱ β-subunitfor EFR function.展开更多
文摘This study investigated the abnormal expression of ATP synthase β-subunit（ATPsyn-β） in pancreas islets of rat model of polycystic ovary syndrome（PCOS） with type 2 diabetes mellitus（T2DM）,and the secretion function changes after up-regulation of ATP5 b.Sixty female SD rats were divided into three groups randomly and equally.The rat model of PCOS with T2 DM was established by free access to the high-carbohydrate/high-fat diet,subcutaneous injections of DHEA,and a single injection of streptozotocin.The pancreas was removed for the detection of the ATPsyn-β expression by immunohistochemical staining,Western blotting and reverse transcription-PCR（RT-PCR）.The pancreas islets of the rats were cultured,isolated with collagenase Ⅴ and purified by gradient centrifugation,and the insulin secretion after treatment with different glucose concentrations was tested.Lentivirus ATP5 b was successfully constructed with the vector of GV208 and transfected into the pancreas islets for the over-expression of ATPsyn-β.The insulin secretion and intracellular ATP content were determined after transfection of the PCOS-T2 DM pancreas islets with Lenti-ATP5 b.The results showed that the expression of ATPsyn-β protein and m RNA was significantly decreased in the pancreas of PCOS-T2 DM rats.The ATP content in the pancreas islets was greatly increased and the insulin secretion was improved after the up-regulation of ATPsyn-β in the pancreas islets transfected with lenti-ATP5 b.These results indicated that for PCOS,the ATPsyn-β might be one of the key factors for the attack of T2 DM.
文摘In this study, the immuno-response of recombinant hCG-β and natural hCGβ was comparatively investigated by using Freund's adjuvant. The results showed that, the properties and merits of the antibodies elicited by both kinds of hCG-β were similar. The antisera had high affinity for binding with hCG (Kαγβ≈5.86×108/mol/L, Kαβ≈8.18×108/mol/L), and were found to be effective in inhibiting the binding of 125I-hCG to receptors in rat testes. Results also indicated that, similar to the antisera induced by natural hCG-β, the recombinant hCG-β induced antisera had capacity of neutralizing the biological activities of hCG. Recombinant hCG-β could be used as an immunogen for contraceptive vaccine.
文摘EFR is a plasma-membrane resident receptor responsible for recognition of microbial elongation factorTu （EF-Tu） and thus triggering plant innate immunity to fend off phytopathogens. Functional EFR must be subject to the endoplasmic reticulum quality control （ERQC） machinery for the correct folding and proper assembly in order to reach its final destination. Genetic studies have demonstrated that ERD2b, a counterpart of the yeast or mammalian HDEL receptor ERD2 for retaining proteins in the endoplasmic reticulum （ER） lumen, is required for EFR function in plants （Li et al., 2009）. In this study, we characterized the Arabidopsis glucosidase Ⅱ β--subunit via the H DEL motif against the non-redundant protein database. Data mining also revealed that the glucosidase Ⅱ β--subunit gene has a highly similar expression pattern to ERD2b and the other known ERQC components involved in EFR biogenesis. Importantly, the T-DNA insertion lines of the glucosidase Ⅱ β-subunit gene showed that EFR-controlled responses were substantially reduced or completely blocked in these mutants. The responses include seedling growth inhibition, induction of marker genes, MAP kinase activation, and callose deposition, trigged by peptide elf18, a full mimic of E F-Tu. Taken together, ourdata indicate a requirement of the glucosidase Ⅱ β-subunitfor EFR function.
