To study the cartilage differentiation of mouse mesenchymal stem cells （MSCs） induced by cartilage-derived morphogenetic proteins-2 in vitro, the MSCs were isolated from mouse bone marrow and cultured in vitro. The ...To study the cartilage differentiation of mouse mesenchymal stem cells （MSCs） induced by cartilage-derived morphogenetic proteins-2 in vitro, the MSCs were isolated from mouse bone marrow and cultured in vitro. The cells in passage 3 were induced into chondrogenic differentiation with different concentrations of recombinant human cartilage-derived morphogenetic proteins-2 （0, 10, 20, 50 and 100 ng/mL）. After 14 days of induction, morphology of cells was observed under phase-contrast microscope. Collagen Ⅱ mRNA and protein were examined with RT-PCR, Western blotting and immunocytochemistry respectively and the sulfate glycosaminoglycan was measured by Alcian blue staining. RT-PCR showed that CDMP-2 could promote expression of collagen Ⅱ mRNA in an dose-dependant manner, especially at the concentration of 50 ng/mL and 100 ng/mL. Immunocytochemistry and Western blotting revealed a similar change. Alcian blue staining exhibited deposition of typical cartilage extracellular matrix. Our results suggest that mouse bone marrow mesencymal stem cells can differentiate into chondrogenic phonotype with the induction of CDMP-2 in vitro, which provides a basis for further research on the role of CDMP-2 in chondrogenesis.展开更多
Objective To analyze the excitotoxicity of monosodium glutamate (MSG) in the offspring cerebral cortex and hippocampal subregions after maternal oral administration of MSG. Methods Kunming mice were given per os MSG...Objective To analyze the excitotoxicity of monosodium glutamate (MSG) in the offspring cerebral cortex and hippocampal subregions after maternal oral administration of MSG. Methods Kunming mice were given per os MSG ( 4.0 g/kg ) at 17～21 days of pregnancy and their offspring behaviors were studied at 10, 20 , 30 days postnatally. By using immunohistochemical means, the involvement of Bcl-2 and Bax in the glutamate-induced cell death in cortical and hippocampal neurons were examined. Cell damage was assessed by direct cell counting. Results Administration of monosodium glutamate during the fetal period in mice resulted in a moderate increase in the expression of Bax in principal neurons in CA1, CA2, CA3, CA4 and in the cerebral cortex at postpartum 10, 20, 30 days in the offspring mice, whereas Bcl-2 protein expressions were reduced significantly in the same regions as compared with those of controls. Conclusion These findings suggest that glutamate toxicity results in cellular death via an apoptotic mechanism in which the Bcl-2/Bax-alpha molecular complex may be involved. The glutamate-induced apoptosis appears to be related to the modulation of Bcl-2 family gene products such as Bcl-2 and Bax.展开更多
Objective:To study the mechanisms of pancreatic cancer treatment with Kanglaite combined Gemcitabine by investigating the relationship between the apoptosis and the expression of bcl-2, Bax and VEGF in pancreatic canc...Objective:To study the mechanisms of pancreatic cancer treatment with Kanglaite combined Gemcitabine by investigating the relationship between the apoptosis and the expression of bcl-2, Bax and VEGF in pancreatic cancer cells.Methods:Nude mouse subcutaneous transplantation tumor model of Human PC-3 pancreatic cancer was established; the expressions of bcl-2, Bax and VEGF of transplantation tumor cell were determined; the earlier apoptosis rate of pancreatic cancer cell and the gross tumor volume were determined. Results:Kanglaite combined Gemcitabine remarkably decreased the protein expression of bcl-2,raised the expression of Bax,increased the apoptosis rate of the pancreatic cancer and contract the gross tumor volume. Kanglaite greatly decreased the protein expression of VEGF of the tumor cell. Conclusion:Therapeutic efficacy of Kanglaite combined Gemcitabine is far better than separate use of the two medicines in the pancreatic cancer transplantation tumor treatment.展开更多
AIMS: β-adrenergic augmentation of Ca2+ sparks and cardiac contractility has been functionally linked to phosphorylation-dependent dissociation of FK506 binding protein 12.
Objective： We have previously found that mbr is a regulatory element of the bcl2 gene. The objective of this study is to isolate and identify the proteins binding to the 37 mbr in the 3 ＇ -end of the mbr. Methods： ...Objective： We have previously found that mbr is a regulatory element of the bcl2 gene. The objective of this study is to isolate and identify the proteins binding to the 37 mbr in the 3 ＇ -end of the mbr. Methods： Streptavidin magnetic particles were ligated to concatameric oligonucleotides of 37 mbr and incubated with the nuclear extracts of Jurkat cells. The DNA-binding proteins were eluted and then resolved by SDS-PAGE. After silver staining, the protein bands were excised and subjected to MALDI-TOF MS. Results： Several protein bands were detected after the isolation with magnetic particles, and Splicing factor, proline- and glutamine-rich（SFPQ）, Poly（ADP-ribose） polymerase I（PARP）, and promyelocytic leukemia protein（PML） were identified by MALDI-TOF MS. Conclusion： Several proteins were isolated and identified from the 37 mbr-protein complex. Results of this study establish a foundation for further study of the mechanisms by which mbr executes its regulatory function.展开更多
Familial acne inversa (AI) is an autoinflammatory disorder that affects hair follicles and is caused by loss-of-function mutations in y-secretase component genes.We and other researchers showed that nicastrin (NCSTN) ...Familial acne inversa (AI) is an autoinflammatory disorder that affects hair follicles and is caused by loss-of-function mutations in y-secretase component genes.We and other researchers showed that nicastrin (NCSTN) is the most frequently mutated gene in familial AI.In this study,we generated a keratin 5-Cre-driven epidermis-specific Ncstn conditional knockout mutant in mice.We determined that this mutant recapitulated the major phenotypes of AI,including hyperkeratosis of hair follicles and inflammation.In Ncstnflox/flox;K5-Cre mice,the IL-36a expression level markedly increased starting from postnatal day 0 (P0),and this increase occurred much earlier than those of TNF-α,IL-23A,IL-1 3,and TLR4.RNA-Seq analysis indicated that Sprr2d,a member of the small proline-rich protein 2 family,in the skin tissues of the Ncstnflox/flox,;K5-Cre mice was also upregulated on P0.Quantitative reverse-transcription polymerase chain reaction showed that other Sprr2 genes had a similar expression pattern.Our findings suggested that IL-36a might be a key inflammatory cytokine in the pathophysiology of AI and implicate malfunction of the skin barrier in the pathogenesis of AI.展开更多
目的:探讨石榴花水提物(pomegranate flower water extract,PFW)对2型糖尿病小鼠肝脏胰岛素信号传导的影响及机制。方法:将C57BL/6J随机分为正常组、模型组、二甲双胍组(Met)、石榴花水提物低剂量组(PFWL)和石榴花水提物高剂量组(PFWH)...目的:探讨石榴花水提物(pomegranate flower water extract,PFW)对2型糖尿病小鼠肝脏胰岛素信号传导的影响及机制。方法:将C57BL/6J随机分为正常组、模型组、二甲双胍组(Met)、石榴花水提物低剂量组(PFWL)和石榴花水提物高剂量组(PFWH)。连续给药11周后,称小鼠体质量,检测空腹血糖(FBG)、胰岛素(INS)、甘油三酯(TG)和总胆固醇(TC)的含量,计算胰岛素抵抗指数(HOMA-IR);苏木素-伊红(HE)染色观察肝组织病理变化;Western blot法检测肝组织中胰岛素受体底物1(IRS1)、p-IRS1(Ser307)、蛋白激酶B(AKT)、p-AKT(Ser473)、糖原合成酶激酶-3β(Gsk3β)、p-Gsk3β(S9)、芳香烃受体(AhR)、磷脂酰乙醇胺N-甲基转移酶(PEMT)、Bcl-2/腺病毒E1B-19kDa相互作用蛋白3(BNIP3)蛋白表达。结果:与模型组比较,PFWH组FBG、INS、HOMA-IR、TG和TC含量极显著降低(P<0.01);PFWH组小鼠肝细胞内脂肪滴明显减少;PFWH组极显著升高肝脏中IRS1、p-AKT(Ser473)/AKT、p-Gsk3β(S9)/Gsk3β、BNIP3蛋白表达(P<0.01),极显著降低p-IRS1(Ser307)/IRS1、AHR、PEMT蛋白表达(P<0.01)。结论:PFW可能通过调节AHR/BNIP3抑制肝脏脂质沉积,改善p-IRS1(Ser307)/p-AKT(Ser473)/p-GSK3β(S9)胰岛素信号通路转导。展开更多
基金This project was supported by a grant from the National Natural Sciences Foundation of China (No 30471753)
文摘To study the cartilage differentiation of mouse mesenchymal stem cells （MSCs） induced by cartilage-derived morphogenetic proteins-2 in vitro, the MSCs were isolated from mouse bone marrow and cultured in vitro. The cells in passage 3 were induced into chondrogenic differentiation with different concentrations of recombinant human cartilage-derived morphogenetic proteins-2 （0, 10, 20, 50 and 100 ng/mL）. After 14 days of induction, morphology of cells was observed under phase-contrast microscope. Collagen Ⅱ mRNA and protein were examined with RT-PCR, Western blotting and immunocytochemistry respectively and the sulfate glycosaminoglycan was measured by Alcian blue staining. RT-PCR showed that CDMP-2 could promote expression of collagen Ⅱ mRNA in an dose-dependant manner, especially at the concentration of 50 ng/mL and 100 ng/mL. Immunocytochemistry and Western blotting revealed a similar change. Alcian blue staining exhibited deposition of typical cartilage extracellular matrix. Our results suggest that mouse bone marrow mesencymal stem cells can differentiate into chondrogenic phonotype with the induction of CDMP-2 in vitro, which provides a basis for further research on the role of CDMP-2 in chondrogenesis.
基金ThisstudywassupportedbytheNationalNaturalScienceFoundationofChina (No .3 0 0 70 73 1 )
文摘Objective To analyze the excitotoxicity of monosodium glutamate (MSG) in the offspring cerebral cortex and hippocampal subregions after maternal oral administration of MSG. Methods Kunming mice were given per os MSG ( 4.0 g/kg ) at 17～21 days of pregnancy and their offspring behaviors were studied at 10, 20 , 30 days postnatally. By using immunohistochemical means, the involvement of Bcl-2 and Bax in the glutamate-induced cell death in cortical and hippocampal neurons were examined. Cell damage was assessed by direct cell counting. Results Administration of monosodium glutamate during the fetal period in mice resulted in a moderate increase in the expression of Bax in principal neurons in CA1, CA2, CA3, CA4 and in the cerebral cortex at postpartum 10, 20, 30 days in the offspring mice, whereas Bcl-2 protein expressions were reduced significantly in the same regions as compared with those of controls. Conclusion These findings suggest that glutamate toxicity results in cellular death via an apoptotic mechanism in which the Bcl-2/Bax-alpha molecular complex may be involved. The glutamate-induced apoptosis appears to be related to the modulation of Bcl-2 family gene products such as Bcl-2 and Bax.
文摘Objective:To study the mechanisms of pancreatic cancer treatment with Kanglaite combined Gemcitabine by investigating the relationship between the apoptosis and the expression of bcl-2, Bax and VEGF in pancreatic cancer cells.Methods:Nude mouse subcutaneous transplantation tumor model of Human PC-3 pancreatic cancer was established; the expressions of bcl-2, Bax and VEGF of transplantation tumor cell were determined; the earlier apoptosis rate of pancreatic cancer cell and the gross tumor volume were determined. Results:Kanglaite combined Gemcitabine remarkably decreased the protein expression of bcl-2,raised the expression of Bax,increased the apoptosis rate of the pancreatic cancer and contract the gross tumor volume. Kanglaite greatly decreased the protein expression of VEGF of the tumor cell. Conclusion:Therapeutic efficacy of Kanglaite combined Gemcitabine is far better than separate use of the two medicines in the pancreatic cancer transplantation tumor treatment.
文摘AIMS: β-adrenergic augmentation of Ca2+ sparks and cardiac contractility has been functionally linked to phosphorylation-dependent dissociation of FK506 binding protein 12.
基金supported by grants from the National Natural Science Foundation of China(30500585)the Natural Science Foundation of Jiangsu Province(BK2008450)
文摘Objective： We have previously found that mbr is a regulatory element of the bcl2 gene. The objective of this study is to isolate and identify the proteins binding to the 37 mbr in the 3 ＇ -end of the mbr. Methods： Streptavidin magnetic particles were ligated to concatameric oligonucleotides of 37 mbr and incubated with the nuclear extracts of Jurkat cells. The DNA-binding proteins were eluted and then resolved by SDS-PAGE. After silver staining, the protein bands were excised and subjected to MALDI-TOF MS. Results： Several protein bands were detected after the isolation with magnetic particles, and Splicing factor, proline- and glutamine-rich（SFPQ）, Poly（ADP-ribose） polymerase I（PARP）, and promyelocytic leukemia protein（PML） were identified by MALDI-TOF MS. Conclusion： Several proteins were isolated and identified from the 37 mbr-protein complex. Results of this study establish a foundation for further study of the mechanisms by which mbr executes its regulatory function.
基金This work was financially supported by the National Key Research and Development Program of China(No.2016Y FC0905100)the CAMS Innovation Fund for Medical Sciences(No.2016-I2M-1-002)+3 种基金the National Natural Science Foundation of China(NSFCNos.81788101 and 81230015)the Beijing Municipal Science and Technology Commission(No.Z151100003915078)for Xue Zhangby the National NSFC(No.31271345)for Yaping Liu.
文摘Familial acne inversa (AI) is an autoinflammatory disorder that affects hair follicles and is caused by loss-of-function mutations in y-secretase component genes.We and other researchers showed that nicastrin (NCSTN) is the most frequently mutated gene in familial AI.In this study,we generated a keratin 5-Cre-driven epidermis-specific Ncstn conditional knockout mutant in mice.We determined that this mutant recapitulated the major phenotypes of AI,including hyperkeratosis of hair follicles and inflammation.In Ncstnflox/flox;K5-Cre mice,the IL-36a expression level markedly increased starting from postnatal day 0 (P0),and this increase occurred much earlier than those of TNF-α,IL-23A,IL-1 3,and TLR4.RNA-Seq analysis indicated that Sprr2d,a member of the small proline-rich protein 2 family,in the skin tissues of the Ncstnflox/flox,;K5-Cre mice was also upregulated on P0.Quantitative reverse-transcription polymerase chain reaction showed that other Sprr2 genes had a similar expression pattern.Our findings suggested that IL-36a might be a key inflammatory cytokine in the pathophysiology of AI and implicate malfunction of the skin barrier in the pathogenesis of AI.
文摘目的:探讨石榴花水提物(pomegranate flower water extract,PFW)对2型糖尿病小鼠肝脏胰岛素信号传导的影响及机制。方法:将C57BL/6J随机分为正常组、模型组、二甲双胍组(Met)、石榴花水提物低剂量组(PFWL)和石榴花水提物高剂量组(PFWH)。连续给药11周后,称小鼠体质量,检测空腹血糖(FBG)、胰岛素(INS)、甘油三酯(TG)和总胆固醇(TC)的含量,计算胰岛素抵抗指数(HOMA-IR);苏木素-伊红(HE)染色观察肝组织病理变化;Western blot法检测肝组织中胰岛素受体底物1(IRS1)、p-IRS1(Ser307)、蛋白激酶B(AKT)、p-AKT(Ser473)、糖原合成酶激酶-3β(Gsk3β)、p-Gsk3β(S9)、芳香烃受体(AhR)、磷脂酰乙醇胺N-甲基转移酶(PEMT)、Bcl-2/腺病毒E1B-19kDa相互作用蛋白3(BNIP3)蛋白表达。结果:与模型组比较,PFWH组FBG、INS、HOMA-IR、TG和TC含量极显著降低(P<0.01);PFWH组小鼠肝细胞内脂肪滴明显减少;PFWH组极显著升高肝脏中IRS1、p-AKT(Ser473)/AKT、p-Gsk3β(S9)/Gsk3β、BNIP3蛋白表达(P<0.01),极显著降低p-IRS1(Ser307)/IRS1、AHR、PEMT蛋白表达(P<0.01)。结论:PFW可能通过调节AHR/BNIP3抑制肝脏脂质沉积,改善p-IRS1(Ser307)/p-AKT(Ser473)/p-GSK3β(S9)胰岛素信号通路转导。
