By combining coral with recombinant human bone morphogenetic protein-2 (rhBMP-2), rhBMP-2/coral composite was obtained in this study. Following implantation of the composite into the muscle pouches of mice, cartilage ...By combining coral with recombinant human bone morphogenetic protein-2 (rhBMP-2), rhBMP-2/coral composite was obtained in this study. Following implantation of the composite into the muscle pouches of mice, cartilage growth was induced in the pores or on the surface of the implants at one week, woven bone at three week and lamellar bone with bone marrow at six week, and coral was absorbed partially. The induced formation of endochondral bone was time-related and rhBMP-2 dose-related. The results of this study indicate that the composite possesses a superior ability of osteogenesis, and coral acts as one of the most suitable rhBMP-2 slowrelease carriers currently available. The composite will be a new type of bone substitute to be used in orthopaedics and maxillofacial surgery.展开更多
Objective: To identify eukaryotic expression vector of human bone morphogenetic protein 2 pcDNA3/BMP2, verify its expression in transfected human mesenchymal stem cells (hMSCs) and the effect on hMSCs differentiation....Objective: To identify eukaryotic expression vector of human bone morphogenetic protein 2 pcDNA3/BMP2, verify its expression in transfected human mesenchymal stem cells (hMSCs) and the effect on hMSCs differentiation. Methods: The BMP2 gene was cloned into a eukaryotic expression vector pcDNA3. Transfected the recombinant into hMSCs by liposome. Immunnohistochemistry and in situ hybridization methods were used to identify the expression of BMP2 mRNA and protein; ALP and Von Kossa stains were performed to identify the BMP2 gene differentiated effect on the hMSCs. Results: The pcDNA3/BMP2 fragments were as large as theory. BMP2 mRNA and protein were expressed and synthesized both in 48 h and 4 weeks after transfection, the ALP and Ca deposit exhibition, which marked the osteogenic lineage of hMSCs, were enhanced and sped. Conclusion: Transfection of pcDNA3/BMP2 is able to provide transient and persistent expression in hMSCs, and promote the MSCs differentiation to osteogenic lineage.展开更多
To determine the ability of a new type of composite xenogeneic bone grafting to repair bone defect. Methods: The new type of composite xenogeneic bone was obtained by combining the chemically treated cance1lous bone w...To determine the ability of a new type of composite xenogeneic bone grafting to repair bone defect. Methods: The new type of composite xenogeneic bone was obtained by combining the chemically treated cance1lous bone with recombinant human bone morphogenetic protein-2 (rhBMP-2). It was implanted on the bone defect of rabbit. Results: There was a large amount of new bone formation within the combined material and the amount was increasing as the time elapsed. In contrast, there was a lot of fibrous tissue with a little new bone formed on the area of the bone defect when the treated cancellous bone was implanted alone. Conclusion: The results imply that the rhBMP-2 plays a very important role in new bone formation and the composite xenogeneic bone appear to be an ideal material for repair of bone defect.展开更多
To express die mature peptide of human bone morphogenetic protein-2 in Escherichia coil. Methods: TheDNA fragment encoding the mature peptide of human bone morphogenetic protein-2 (hBMP-2m) was inserted into expressio...To express die mature peptide of human bone morphogenetic protein-2 in Escherichia coil. Methods: TheDNA fragment encoding the mature peptide of human bone morphogenetic protein-2 (hBMP-2m) was inserted into expression vectorpDH in which foreign gene was controlled by PRPL promoters. E. coli DH5a transformed with recombinant plasmid pDHB2m wasinduced at 42℃to express the target protein. The expressed product was partially purified and refolded, and then implanted intorat thigh muscles to assay its bone inductive activity. Results: After induction, a protein band on SDS-PAGE gel with an apparentmol. wt. of 13kD was observed to anticipate in the strain carrying pDHB2m, but not in the control. The expressed hBMP-2m accounted for 45%-60% of the total bacterial protein. The expressed product existed in a form of inclusion body. After partially purified and refolded, rhBMP-2m could induce the formation of cartilage and bone tissue heterotopically. Conclusion: The maturepeptide of human bone morphogenetic protein-2 has ben successfully expressed in E. coli and the product has ectopic bone inductive activity.展开更多
Microalgae has been consumed in human diet for thousands of years.It is an under-exploited crop for production of dietary foods.Microalgae cultivation does not compete with land and resources required for traditional ...Microalgae has been consumed in human diet for thousands of years.It is an under-exploited crop for production of dietary foods.Microalgae cultivation does not compete with land and resources required for traditional crops and has a superior yield compared to terrestrial crops.Its high protein content has exhibited a huge potential to meet the dietary requirements of growing population.Apart from being a source of protein,presence of various bio-active components in microalgae provide an added health benefit.This review describes various microalgal sources of proteins and other bio-active components.One of the heavily studied group of bio-active components are pigments due to their anticarcenogenic,antioxidative and antihypertensive properties.Compared to various plant and floral species,microalgae contain higher amounts of pigments.Microalgal derived proteins have complete Essential Amino Acids(EAA)profiles and their protein content is higher than conventional sources such as meat,poultry and dairy products.However,microalgal based functional foods have not flooded the market.The lack of awareness coupled with scarce incentives for producers result in under-exploitation of microalgal potential.Application of microalgal derived components as dietary and nutraceutical supplements is discussed comprehensively.展开更多
BACKGROUND： This study aimed to explore the effects of TNF-α on the expression of IP3R1 mRNA and protein in human mesangial cells （HMCs）, and to elucidate the mechanism of TNF-α relating to IP3R1 expression in th...BACKGROUND： This study aimed to explore the effects of TNF-α on the expression of IP3R1 mRNA and protein in human mesangial cells （HMCs）, and to elucidate the mechanism of TNF-α relating to IP3R1 expression in the occurrence of hepatorenal syndrome （HRS）.METHODS： HMCs were stimulated by tumor （TNF-α） with 100 ng/mL for different hours （2, 4, 8, and 24 hours）. The expression changes of IP3R1 mRNA and protein were detected by quantitative real-time polymerase chain reaction and immunoblotting. Several inhibitors including D609, U73122, PP1, safingol, rottlerin and non-radioactive protein kinase C （PKC） were used to examine the mechanism of signal transduction of TNF-α-regulated IP3R1 in HMCs.RESULTS： The levels of IP3R1 mRNA at 2 hours after TNF-α exposure were significantly enhanced and peaked at 8 hours in HMCs （P〈0.01）, then descended at 24 hours （P〈0.01）. The levels of IP3R1 protein at 4 hours after TNF-α exposure were obviously increased and peaked at 24 hours after TNF-α exposure （P〈0.01）. Compared to the control group, safingol （PKCa inhibitor） and D609 （phosphatidylcholine-specific phospholipase C inhibitor） significantly blocked the TNF-α- induced expression of IP3R1 mRNA （3.30±0.81 vs. 1.95±0.13, P〈0.05; 2.10±0.49, P〈0.01） and IP3R1 protein （3.09±0.13 vs. 1.86±0.39, P〈0.01; 1.98±0.02, P〈0.01）. TNF-α promoted PKCa activation with maximal PKCa phosphorylation that occurred 8 hours after stimulation measured by non-radioactive PKC assay, and the effect was markedly attenuated by pretreatment with D609 or safingol.CONCLUSION： TNF-α increased the expression of IP3R1 and this was mediated, at least in part, through the PC-PLC/PKCa signaling pathways in HMCs.展开更多
Gossypol-induced alterations of aminophospholipid composition in human sperm wereobserved and the effects of some factors on these alterations were investigated.The altera-tions of aminophospholipid composition of hum...Gossypol-induced alterations of aminophospholipid composition in human sperm wereobserved and the effects of some factors on these alterations were investigated.The altera-tions of aminophospholipid composition of human sperm induced by gossypoI included aprogressive decrease in the levels of phosphatidylethanolamine（PE）and phosphatidylserine（PS）when gossypol concentrations ranged from 5-500μmol/L,and a progressive increase inthe level of（LPE）at lower gossypol concentrations（5-50μmol/L）.The conversion of PEinto lysophosphatidylethanolamine（LPE）was strongly enhanced by Ca<sup>2+</sup>and inhibitedby 0.5mmol/L EDTA,while PMSF,NEM,iodoacetamide and Zn<sup>2+</sup>had no effect.Thisconversion was weaker in sperm with stripped surface proteins than in normal sperm.In addition,three surface proteins（MW 80,60 and 40 kD）were fixed on the plasmamembrane by gossypol treatment.The significance of gossypol action and the possibilitythat sperm PLA<sub>2</sub>, PE and.some surface proteins might play an important role in the physio-logical acrosome reaetion of human sperm,as well as in oocyte penetration,are discussed.展开更多
We have cloned the E6 gene of human papillomavirus type 18 into anexpression plasmid pBD2.One of the recombinant plasmids (named pDV11) wasidentified by DNA analysis and protein product analysis.It could express a new...We have cloned the E6 gene of human papillomavirus type 18 into anexpression plasmid pBD2.One of the recombinant plasmids (named pDV11) wasidentified by DNA analysis and protein product analysis.It could express a newprotein whose molecular weight correspods well with the expected one.Afterpurification,the expressed protein showed a positive result in countercurrentimmuno-electrophoresis with anti-β-gal serum and was proved to be the expectedβ-gal/E6 fusion protein.The physical map of pDV11 was also prepared.展开更多
Objective: To investigate the effects of histamine on growth and apoptosis of human melanoma cells A375. Methods: The effect of histamine on growth of A375 cells in vitro was examined by MTT assay and Trypan blue excl...Objective: To investigate the effects of histamine on growth and apoptosis of human melanoma cells A375. Methods: The effect of histamine on growth of A375 cells in vitro was examined by MTT assay and Trypan blue exclusion assay. Cell cycle analysis, early apoptosis analysis by double staining with Annexin V-FITC and PI, and active caspase-3 analysis by staining FITC-conjugated monoclonal rabbit anti-active caspase-3 antibody were made by flow cytometer. StreptAvidin-Biotin Complex (SABC) immunocytochemical assays were adopted to detect Bax/Bcl-2 protein expressions.Results: Histamine inhibited proliferation of A375 cells in a dose- and time-dependent manner, and altered cell cycle distribution of A375 cells revealing an increase in G0/G1-phase population, a decrease in S-phase population and the inhibition of G1/S switching. Histamine induced apoptosis of A375 cells (P<0.05), elevated the cells population with detectable active caspase-3 (P<0.05), increased the number of cells forming Bax and decreased the number of cells forming Bcl-2 significantly (P<0.05). Conclusion: That histamine inhibits cell cycle progress of A375 cells is one of the possible mechanisms of proliferation arrest of A375 cells elicited by histamine. Histamine mediates apoptosis in A375 cells that may be caspase-dependent through mitochondria routine. Histamine with high concentration inhibits growth of A375 cells in vitro by interfering proliferation and inducing apoptosis of cells.展开更多
Bone morphogenetic protein (BMP) can promote the proliferation of dental pulp cells and induce reparative dentin formation. In this study, the inductive effect of BMP derived from bovine bone matrix on cultured human ...Bone morphogenetic protein (BMP) can promote the proliferation of dental pulp cells and induce reparative dentin formation. In this study, the inductive effect of BMP derived from bovine bone matrix on cultured human dental pulp tissue was observed under light microscope and transmission electron microscope. The results showed that. by the third day of culture, the proliferating star-shaped cells appeared with small cytoplasm and poorly-developed organelles; by the 7th day of the culture, the chondroblast-like cells with rich cytoplasm and well-developed organelles were seen embedded in hyaline matrix. This study suggests that BMP can induce dental pulp cells to differentiate from poorly differentiated state to well-differentiated state.展开更多
Human bone morphogenetic protein 2A (hBMP2A) cDNA terminal 567 nucleotides were cloned and expressed in a phage display vector pCSM21. Human BMP2A C-terminal peptide displayed on the surface of the phage can bind spec...Human bone morphogenetic protein 2A (hBMP2A) cDNA terminal 567 nucleotides were cloned and expressed in a phage display vector pCSM21. Human BMP2A C-terminal peptide displayed on the surface of the phage can bind specifically to the surface of mouse osteoblastic cell (MC3T3) membrane. ELISA assay showed a positive signal of the binding by using antibody against M13 phage gene 8 protein. After labeling with 3HTdR,the counts of the binding groups were 3 to 10 times higher than the control groups. It suggests that the surface of MC3T3 cells exist the receptor for hBMP2A.展开更多
Background:In sepsis,vitamin D binding protein(VDBP)has been shown to be low-expressed.The current study examined the relationship between serum VDBP level and liver injury in sepsis patients,as well as in a mouse mod...Background:In sepsis,vitamin D binding protein(VDBP)has been shown to be low-expressed.The current study examined the relationship between serum VDBP level and liver injury in sepsis patients,as well as in a mouse model for sepsis and in cultured liver epithelial cell line exposed to lipopolysaccharide(LPS).Methods:The human study included 78 sepsis patients and 50 healthy volunteers.Sepsis patients were categorized into sepsis survivor group(n=43)and sepsis non-survivor group(n=35)based on 28-day mortality for data analysis.Adult male C57BL/6 mice were subjected to cecal ligation and puncture(CLP).Serum samples were collected on day 1,3,5 and 7 to determine the levels of VDBP,25-hydroxyvitamin D[25(OH)D_(3)],1,25-dihydroxyvitamin D[1,25(OH)_(2)D_(3)],interleukin-6(IL-6)and tumor necrosis factor alpha(TNF-α).Potential protective effects of VDBP overexpression against LPS-induced liver damage were examined in cultured THLE2 cells.Results:Serum levels of VDBP,25(OH)D_(3),and 1,25(OH)_(2)D_(3)were significantly lower in sepsis patients vs.the healthy control(P<0.001),as well as in the sepsis non-survivor group vs.the sepsis survivor group(P<0.001,P=0.0338,or P=0.0013,respectively).Lower serum VDBP level was associated with higher Acute Physiology and Chronic Health Evaluation(APACHE)II score(r=−0.2565,P=0.0234)and Sequential Organ Failure Assessment score(r=−0.3522,P=0.0016),but lower serum albumin(ALB,r=0.4628,P<0.001)and total protein(TP,r=0.263,P=0.02).In CLP mice,there was a 5-day period of serum VDBP reduction,followed by return towards the baseline on day 7.VDBP was also decreased in LPS-treated THLE2 cells(P<0.001).VDBP overexpression reduced LPS-induced THLE2 damage.Reduced damage was associated with decreased oxidative stress and inactivation of the c-Jun N-terminal kinase signaling pathway.Conclusion:VDBP may be protective against sepsis-induced liver injury.展开更多
文摘By combining coral with recombinant human bone morphogenetic protein-2 (rhBMP-2), rhBMP-2/coral composite was obtained in this study. Following implantation of the composite into the muscle pouches of mice, cartilage growth was induced in the pores or on the surface of the implants at one week, woven bone at three week and lamellar bone with bone marrow at six week, and coral was absorbed partially. The induced formation of endochondral bone was time-related and rhBMP-2 dose-related. The results of this study indicate that the composite possesses a superior ability of osteogenesis, and coral acts as one of the most suitable rhBMP-2 slowrelease carriers currently available. The composite will be a new type of bone substitute to be used in orthopaedics and maxillofacial surgery.
文摘Objective: To identify eukaryotic expression vector of human bone morphogenetic protein 2 pcDNA3/BMP2, verify its expression in transfected human mesenchymal stem cells (hMSCs) and the effect on hMSCs differentiation. Methods: The BMP2 gene was cloned into a eukaryotic expression vector pcDNA3. Transfected the recombinant into hMSCs by liposome. Immunnohistochemistry and in situ hybridization methods were used to identify the expression of BMP2 mRNA and protein; ALP and Von Kossa stains were performed to identify the BMP2 gene differentiated effect on the hMSCs. Results: The pcDNA3/BMP2 fragments were as large as theory. BMP2 mRNA and protein were expressed and synthesized both in 48 h and 4 weeks after transfection, the ALP and Ca deposit exhibition, which marked the osteogenic lineage of hMSCs, were enhanced and sped. Conclusion: Transfection of pcDNA3/BMP2 is able to provide transient and persistent expression in hMSCs, and promote the MSCs differentiation to osteogenic lineage.
文摘To determine the ability of a new type of composite xenogeneic bone grafting to repair bone defect. Methods: The new type of composite xenogeneic bone was obtained by combining the chemically treated cance1lous bone with recombinant human bone morphogenetic protein-2 (rhBMP-2). It was implanted on the bone defect of rabbit. Results: There was a large amount of new bone formation within the combined material and the amount was increasing as the time elapsed. In contrast, there was a lot of fibrous tissue with a little new bone formed on the area of the bone defect when the treated cancellous bone was implanted alone. Conclusion: The results imply that the rhBMP-2 plays a very important role in new bone formation and the composite xenogeneic bone appear to be an ideal material for repair of bone defect.
文摘To express die mature peptide of human bone morphogenetic protein-2 in Escherichia coil. Methods: TheDNA fragment encoding the mature peptide of human bone morphogenetic protein-2 (hBMP-2m) was inserted into expression vectorpDH in which foreign gene was controlled by PRPL promoters. E. coli DH5a transformed with recombinant plasmid pDHB2m wasinduced at 42℃to express the target protein. The expressed product was partially purified and refolded, and then implanted intorat thigh muscles to assay its bone inductive activity. Results: After induction, a protein band on SDS-PAGE gel with an apparentmol. wt. of 13kD was observed to anticipate in the strain carrying pDHB2m, but not in the control. The expressed hBMP-2m accounted for 45%-60% of the total bacterial protein. The expressed product existed in a form of inclusion body. After partially purified and refolded, rhBMP-2m could induce the formation of cartilage and bone tissue heterotopically. Conclusion: The maturepeptide of human bone morphogenetic protein-2 has ben successfully expressed in E. coli and the product has ectopic bone inductive activity.
基金the Fundamental Research Grant Scheme,Malaysia[FRGS/1/2015/SG05/UNIM/03/1]the Ministry of Science and Technology,Malaysia[MOSTI02-02-12-SF0256]+1 种基金the Prototype Research Grant Scheme,Malaysia[PRGS/2/2015/SG05/UNIM/03/1]International Cooperation Seeds Funding of Nanjing Agricultural University(Grant number:2018-AH-04).
文摘Microalgae has been consumed in human diet for thousands of years.It is an under-exploited crop for production of dietary foods.Microalgae cultivation does not compete with land and resources required for traditional crops and has a superior yield compared to terrestrial crops.Its high protein content has exhibited a huge potential to meet the dietary requirements of growing population.Apart from being a source of protein,presence of various bio-active components in microalgae provide an added health benefit.This review describes various microalgal sources of proteins and other bio-active components.One of the heavily studied group of bio-active components are pigments due to their anticarcenogenic,antioxidative and antihypertensive properties.Compared to various plant and floral species,microalgae contain higher amounts of pigments.Microalgal derived proteins have complete Essential Amino Acids(EAA)profiles and their protein content is higher than conventional sources such as meat,poultry and dairy products.However,microalgal based functional foods have not flooded the market.The lack of awareness coupled with scarce incentives for producers result in under-exploitation of microalgal potential.Application of microalgal derived components as dietary and nutraceutical supplements is discussed comprehensively.
基金supported by a grant from Health Bureauof Jiangxi Province
文摘BACKGROUND： This study aimed to explore the effects of TNF-α on the expression of IP3R1 mRNA and protein in human mesangial cells （HMCs）, and to elucidate the mechanism of TNF-α relating to IP3R1 expression in the occurrence of hepatorenal syndrome （HRS）.METHODS： HMCs were stimulated by tumor （TNF-α） with 100 ng/mL for different hours （2, 4, 8, and 24 hours）. The expression changes of IP3R1 mRNA and protein were detected by quantitative real-time polymerase chain reaction and immunoblotting. Several inhibitors including D609, U73122, PP1, safingol, rottlerin and non-radioactive protein kinase C （PKC） were used to examine the mechanism of signal transduction of TNF-α-regulated IP3R1 in HMCs.RESULTS： The levels of IP3R1 mRNA at 2 hours after TNF-α exposure were significantly enhanced and peaked at 8 hours in HMCs （P〈0.01）, then descended at 24 hours （P〈0.01）. The levels of IP3R1 protein at 4 hours after TNF-α exposure were obviously increased and peaked at 24 hours after TNF-α exposure （P〈0.01）. Compared to the control group, safingol （PKCa inhibitor） and D609 （phosphatidylcholine-specific phospholipase C inhibitor） significantly blocked the TNF-α- induced expression of IP3R1 mRNA （3.30±0.81 vs. 1.95±0.13, P〈0.05; 2.10±0.49, P〈0.01） and IP3R1 protein （3.09±0.13 vs. 1.86±0.39, P〈0.01; 1.98±0.02, P〈0.01）. TNF-α promoted PKCa activation with maximal PKCa phosphorylation that occurred 8 hours after stimulation measured by non-radioactive PKC assay, and the effect was markedly attenuated by pretreatment with D609 or safingol.CONCLUSION： TNF-α increased the expression of IP3R1 and this was mediated, at least in part, through the PC-PLC/PKCa signaling pathways in HMCs.
文摘Gossypol-induced alterations of aminophospholipid composition in human sperm wereobserved and the effects of some factors on these alterations were investigated.The altera-tions of aminophospholipid composition of human sperm induced by gossypoI included aprogressive decrease in the levels of phosphatidylethanolamine（PE）and phosphatidylserine（PS）when gossypol concentrations ranged from 5-500μmol/L,and a progressive increase inthe level of（LPE）at lower gossypol concentrations（5-50μmol/L）.The conversion of PEinto lysophosphatidylethanolamine（LPE）was strongly enhanced by Ca<sup>2+</sup>and inhibitedby 0.5mmol/L EDTA,while PMSF,NEM,iodoacetamide and Zn<sup>2+</sup>had no effect.Thisconversion was weaker in sperm with stripped surface proteins than in normal sperm.In addition,three surface proteins（MW 80,60 and 40 kD）were fixed on the plasmamembrane by gossypol treatment.The significance of gossypol action and the possibilitythat sperm PLA<sub>2</sub>, PE and.some surface proteins might play an important role in the physio-logical acrosome reaetion of human sperm,as well as in oocyte penetration,are discussed.
基金The project was supported by National Natural Science Foundation of China
文摘We have cloned the E6 gene of human papillomavirus type 18 into anexpression plasmid pBD2.One of the recombinant plasmids (named pDV11) wasidentified by DNA analysis and protein product analysis.It could express a newprotein whose molecular weight correspods well with the expected one.Afterpurification,the expressed protein showed a positive result in countercurrentimmuno-electrophoresis with anti-β-gal serum and was proved to be the expectedβ-gal/E6 fusion protein.The physical map of pDV11 was also prepared.
文摘Objective: To investigate the effects of histamine on growth and apoptosis of human melanoma cells A375. Methods: The effect of histamine on growth of A375 cells in vitro was examined by MTT assay and Trypan blue exclusion assay. Cell cycle analysis, early apoptosis analysis by double staining with Annexin V-FITC and PI, and active caspase-3 analysis by staining FITC-conjugated monoclonal rabbit anti-active caspase-3 antibody were made by flow cytometer. StreptAvidin-Biotin Complex (SABC) immunocytochemical assays were adopted to detect Bax/Bcl-2 protein expressions.Results: Histamine inhibited proliferation of A375 cells in a dose- and time-dependent manner, and altered cell cycle distribution of A375 cells revealing an increase in G0/G1-phase population, a decrease in S-phase population and the inhibition of G1/S switching. Histamine induced apoptosis of A375 cells (P<0.05), elevated the cells population with detectable active caspase-3 (P<0.05), increased the number of cells forming Bax and decreased the number of cells forming Bcl-2 significantly (P<0.05). Conclusion: That histamine inhibits cell cycle progress of A375 cells is one of the possible mechanisms of proliferation arrest of A375 cells elicited by histamine. Histamine mediates apoptosis in A375 cells that may be caspase-dependent through mitochondria routine. Histamine with high concentration inhibits growth of A375 cells in vitro by interfering proliferation and inducing apoptosis of cells.
文摘Bone morphogenetic protein (BMP) can promote the proliferation of dental pulp cells and induce reparative dentin formation. In this study, the inductive effect of BMP derived from bovine bone matrix on cultured human dental pulp tissue was observed under light microscope and transmission electron microscope. The results showed that. by the third day of culture, the proliferating star-shaped cells appeared with small cytoplasm and poorly-developed organelles; by the 7th day of the culture, the chondroblast-like cells with rich cytoplasm and well-developed organelles were seen embedded in hyaline matrix. This study suggests that BMP can induce dental pulp cells to differentiate from poorly differentiated state to well-differentiated state.
文摘Human bone morphogenetic protein 2A (hBMP2A) cDNA terminal 567 nucleotides were cloned and expressed in a phage display vector pCSM21. Human BMP2A C-terminal peptide displayed on the surface of the phage can bind specifically to the surface of mouse osteoblastic cell (MC3T3) membrane. ELISA assay showed a positive signal of the binding by using antibody against M13 phage gene 8 protein. After labeling with 3HTdR,the counts of the binding groups were 3 to 10 times higher than the control groups. It suggests that the surface of MC3T3 cells exist the receptor for hBMP2A.
基金the Clinical Research Support Fund of Chinese PLA General Hospital(2018FC-WJFWZX-1-03)Youth Talents Promotion Project of China(17-JCJQ-QT-036)Natural Science Foundation of Beijing(7214254).
文摘Background:In sepsis,vitamin D binding protein(VDBP)has been shown to be low-expressed.The current study examined the relationship between serum VDBP level and liver injury in sepsis patients,as well as in a mouse model for sepsis and in cultured liver epithelial cell line exposed to lipopolysaccharide(LPS).Methods:The human study included 78 sepsis patients and 50 healthy volunteers.Sepsis patients were categorized into sepsis survivor group(n=43)and sepsis non-survivor group(n=35)based on 28-day mortality for data analysis.Adult male C57BL/6 mice were subjected to cecal ligation and puncture(CLP).Serum samples were collected on day 1,3,5 and 7 to determine the levels of VDBP,25-hydroxyvitamin D[25(OH)D_(3)],1,25-dihydroxyvitamin D[1,25(OH)_(2)D_(3)],interleukin-6(IL-6)and tumor necrosis factor alpha(TNF-α).Potential protective effects of VDBP overexpression against LPS-induced liver damage were examined in cultured THLE2 cells.Results:Serum levels of VDBP,25(OH)D_(3),and 1,25(OH)_(2)D_(3)were significantly lower in sepsis patients vs.the healthy control(P<0.001),as well as in the sepsis non-survivor group vs.the sepsis survivor group(P<0.001,P=0.0338,or P=0.0013,respectively).Lower serum VDBP level was associated with higher Acute Physiology and Chronic Health Evaluation(APACHE)II score(r=−0.2565,P=0.0234)and Sequential Organ Failure Assessment score(r=−0.3522,P=0.0016),but lower serum albumin(ALB,r=0.4628,P<0.001)and total protein(TP,r=0.263,P=0.02).In CLP mice,there was a 5-day period of serum VDBP reduction,followed by return towards the baseline on day 7.VDBP was also decreased in LPS-treated THLE2 cells(P<0.001).VDBP overexpression reduced LPS-induced THLE2 damage.Reduced damage was associated with decreased oxidative stress and inactivation of the c-Jun N-terminal kinase signaling pathway.Conclusion:VDBP may be protective against sepsis-induced liver injury.
