Objective: To investigate the expression of CD147 on human ovarian neoplasm cell lines and its influence on production and activation of matrix metallproteinases(MMPs). Methods: The expression of CD147 on different hu...Objective: To investigate the expression of CD147 on human ovarian neoplasm cell lines and its influence on production and activation of matrix metallproteinases(MMPs). Methods: The expression of CD147 on different human ovarian neoplasm cell lines was studied by western blotting. Co-culture was carried out to investigate the stimulative effect of the positive expression CD147 cell HO-8910 on the production of MMPs of fibroblast cell in vitro. Zymography and immune blotting were used to study the production and activity of positive MMPs, at the time, to explore the relation between CD147 and MMPs. Results: CD147 was positively presented in 2 ovarian neoplasm cell lines(HO-8910,3-AO), but in SKOV3, TC-1,NIN3T3 cell was negative. MMP-2 and MMP-9 were detected by HO-8910 cell line, mouse fibroblast cell and co-culture cells; but the expression in co-culture cell is obviously higher than individual cultures of each type alone.CD147 stimulated MMPs in dose-dependent manner. Conclusion: CD147 causes increased production and activation of MMP-2, MMP-9.CD147 is probably a indirect marker of some ovarian cancer cells with invasion and metastasis.展开更多
Objective： To explore the influence of charred Gossamer urocteae （CGU） on the functions of primary cultured mouse oral fibroblasts and reveal its mechanism in wound healing. Methods： CGU was extracted with differe...Objective： To explore the influence of charred Gossamer urocteae （CGU） on the functions of primary cultured mouse oral fibroblasts and reveal its mechanism in wound healing. Methods： CGU was extracted with different solvents and ethanol extract （EE）, ethyl acetate fraction （EF）, n-butanol fraction （BF） and aqueous fraction （AF） were obtained. The effects of different fractions on the proliferation, matrix metaUoproteinase-2,9 （MMP-2,9） activities, synthesis of collagen and tissue inhibitor of metalloproteinase 1 （TIMP-1） in the mouse oral fibroblasts were determined by MTT, gelatin zymography, chloramine-T method, and enzyme-linked immunosorbent assay （ELISA） respectively. Results： EE, EF and BF at high concentrations could significantly inhibit proliferation of fibroblasts （P〈0.05 or P〈0.01）, and at low concentrations EF and BF could promote proliferation of fibroblasts, and BF and AF could significantly inhibit collagen synthesis （P〈0.05 or P〈0.01）. EE, EF and AF at high concentrations could significantly increase the MMP-9 activity, and BF and AF could significantly inhibit synthesis of TIMP-1. Conclusion： CGU at high concentrations can inhibit the proliferations of fibroblasts and synthesis of collagen, and in healing of wound, CGU at high concentrations possibly has the functions of anti-fibrosis and anti-scar, and the mechanism to promote degradation of collagen is possibly related to the increase in MMP-9 activity and the inhibition of TIMP-1 synthesis.展开更多
Mot-2 protein is shown to interact with p53 and inhibit its transcriptional activation function. Mot-2 overexpressing stable clones of NIH 3T3 cells were malignantly transformed, however, they had a high level of expr...Mot-2 protein is shown to interact with p53 and inhibit its transcriptional activation function. Mot-2 overexpressing stable clones of NIH 3T3 cells were malignantly transformed, however, they had a high level of expression of a p53 downstream gene, p21WAF1. The present study was undertaken to elucidate possible molecular mechanism(s) of such upregulation. An inCreased level of p21WAF1, expression was detected in sta- ble transfectants although an exogenous reporter gene driven by p21WAF1, promoter exhibited lower activity in these cells suggesting that some post-transcriptional mechanism contributes to upregulation. Western analyses of transient and stable clones revealed that upregulation of p21WAF1, in stable NIH 3T3/mot-2 cells may be mediated by cyclin D1 and cdk-2.展开更多
The bioactive secondary metabolites from the endophytic fungus Fusarium napiforme was evaluated for the cytotoxic effect and antioxidant activity.The total antioxidant capacity(TAC)of the extract was determined by 2,2...The bioactive secondary metabolites from the endophytic fungus Fusarium napiforme was evaluated for the cytotoxic effect and antioxidant activity.The total antioxidant capacity(TAC)of the extract was determined by 2,2-diphenyl-1-picrylhydrazyl(DPPH),phosphomolybdate,and reducing power assay methods.The cytotoxicity of the extract was evaluated against lung adenocarcinoma(A549)cells and mouse embryo fibroblast(NIH3T3)cells by methyl thiazolyl tetrazolium(MTT)method.The major composition of the crude extract was identified by the Gas Chromatography-Mass Spectrometry(GC-MS)analysis.Estimation of the endophyte crude extract revealed a high amount of the total flavonoid content(TFC)and total phenolic content(TPC).The extract showed high cytotoxic activity against the A549 cell line with the mean cytotoxicity of 69.74±0.49%.The extract did not show any cytotoxic effect against the NIH3T3 cell line.The extract exhibited high antioxidant activity as a function of the concentrations.At a test concentration of 1 mg ml-1,the extract showed the highest inhibition against DPPH radical at 75.4607%±0.47688,ferric ion reducing power at 0.882±0.0120,and 255.434±21.404 AAE/g of the extract by phosphomolybdenum assay(PMA).There is a significant correlation between TPC and antioxidant activity at p<0.05.The correlation between reducing power and DPPH is significant at p<0.01.The major types of bioactive compounds identified by the GC-MS have shown the presence of nine major compounds.This result strongly exhibits that the endophyte F.napiforme can be a potential source for the formulation of natural anticancer drugs and protecting the body from oxidative damages.展开更多
Bioengineered scaffolds are crucial components in artificial tissue construction.In general,these scaffolds provide inert three-dimensional(3D)surfaces supporting cell growth.However,some scaffolds can affect the phen...Bioengineered scaffolds are crucial components in artificial tissue construction.In general,these scaffolds provide inert three-dimensional(3D)surfaces supporting cell growth.However,some scaffolds can affect the phenotype of cultured cells,especially,adherent stromal cells,such as fibroblasts.Here we report on unique properties of 3D fibroin/gelatin materials,which may rapidly induce expression of adhesion molecules,such as ICAM-1 and VCAM-1,in cultured primary murine embryonic fibroblasts(MEFs).In contrast,two-dimensional(2D)fibroin/gelatin films did not show significant effects on gene expression profiles in fibroblasts as compared to 3D culture conditions.Interestingly,TNF expression was induced in MEFs cultured in 3D fibroin/gelatin scaffolds,while genetic or pharmacological TNF ablation resulted in diminished ICAM-1 and VCAM-1 expression by these cells.Using selective MAPK inhibitors,we uncovered critical contribution of JNK to 3D-induced upregulation of these adhesion molecules.Moreover,we observed ICAM-1/VCAM-1-dependent adhesion of lymphocytes to fibroblasts cultured in 3D fibroin/gelatin scaffolds,but not on 2D fibroin/gelatin films,suggesting functional reprogramming in stromal cells,when exposed to 3D environment.Finally,we observed significant infiltration of lymphocytes into 3D fibroin/gelatin,but not into collagen scaffolds in vivo upon subcapsular kidney implantation in mice.Together our data highlight the important features of fibroin/gelatin scaffolds,when they are produced as 3D sponges rather than 2D films,which should be considered when using these materials for tissue engineering.展开更多
The lifetime of orthopaedic implants can be extended by coating the softer Ti6Al4V alloy with harder biocompatible thin films.In this work,thin films of Ti_((1-x))Au_((x))are grown on Ti_(6)Al_(4)V and glass substrate...The lifetime of orthopaedic implants can be extended by coating the softer Ti6Al4V alloy with harder biocompatible thin films.In this work,thin films of Ti_((1-x))Au_((x))are grown on Ti_(6)Al_(4)V and glass substrates by magnetron sputtering in the entire x=0-1 range,before their key biomechanical properties are performance tuned by thermal activation.For the first time,we explore the effect of in-situ substrate heating versus ex-situ post-deposition heat-treatment,on development of mechanical and biocompatibility performance in Ti-Au films.A~250% increase in hardness is achieved for Ti-Au films compared to bulk Ti6Al4V and a~40%improvement from 8.8 GPa as-grown to 11.9 and 12.3 GPa with in-situ and ex-situ heat-treatment respectively,is corelated to changes in structural,morphological and chemical properties,providing insights into the origins of super-hardness in the Ti rich regions of these materials.X-ray diffraction reveals that as-grown films are in nanocrystalline states of Ti-Au intermetallic phases and thermal activation leads to emergence of mechanically hard Ti-Au intermetallics,with films prepared by in-situ substrate heating having enhanced crystalline quality.Surface morphology images show clear changes in grain size,shape and surface roughness following thermal activation,while elemental analysis reveals that in-situ substrate heating is better for development of oxide free Ti3Auβ-phases.All tested Ti-Au films are non-cytotoxic against L929 mouse fibroblast cells,while extremely low leached ion concentrations confirm their biocompatibility.With peak hardness performance tuned to>12 GPa and excellent biocompatibility,Ti-Au films have potential as a future coating technology for load bearing medical implants.展开更多
文摘Objective: To investigate the expression of CD147 on human ovarian neoplasm cell lines and its influence on production and activation of matrix metallproteinases(MMPs). Methods: The expression of CD147 on different human ovarian neoplasm cell lines was studied by western blotting. Co-culture was carried out to investigate the stimulative effect of the positive expression CD147 cell HO-8910 on the production of MMPs of fibroblast cell in vitro. Zymography and immune blotting were used to study the production and activity of positive MMPs, at the time, to explore the relation between CD147 and MMPs. Results: CD147 was positively presented in 2 ovarian neoplasm cell lines(HO-8910,3-AO), but in SKOV3, TC-1,NIN3T3 cell was negative. MMP-2 and MMP-9 were detected by HO-8910 cell line, mouse fibroblast cell and co-culture cells; but the expression in co-culture cell is obviously higher than individual cultures of each type alone.CD147 stimulated MMPs in dose-dependent manner. Conclusion: CD147 causes increased production and activation of MMP-2, MMP-9.CD147 is probably a indirect marker of some ovarian cancer cells with invasion and metastasis.
文摘Objective： To explore the influence of charred Gossamer urocteae （CGU） on the functions of primary cultured mouse oral fibroblasts and reveal its mechanism in wound healing. Methods： CGU was extracted with different solvents and ethanol extract （EE）, ethyl acetate fraction （EF）, n-butanol fraction （BF） and aqueous fraction （AF） were obtained. The effects of different fractions on the proliferation, matrix metaUoproteinase-2,9 （MMP-2,9） activities, synthesis of collagen and tissue inhibitor of metalloproteinase 1 （TIMP-1） in the mouse oral fibroblasts were determined by MTT, gelatin zymography, chloramine-T method, and enzyme-linked immunosorbent assay （ELISA） respectively. Results： EE, EF and BF at high concentrations could significantly inhibit proliferation of fibroblasts （P〈0.05 or P〈0.01）, and at low concentrations EF and BF could promote proliferation of fibroblasts, and BF and AF could significantly inhibit collagen synthesis （P〈0.05 or P〈0.01）. EE, EF and AF at high concentrations could significantly increase the MMP-9 activity, and BF and AF could significantly inhibit synthesis of TIMP-1. Conclusion： CGU at high concentrations can inhibit the proliferations of fibroblasts and synthesis of collagen, and in healing of wound, CGU at high concentrations possibly has the functions of anti-fibrosis and anti-scar, and the mechanism to promote degradation of collagen is possibly related to the increase in MMP-9 activity and the inhibition of TIMP-1 synthesis.
文摘Mot-2 protein is shown to interact with p53 and inhibit its transcriptional activation function. Mot-2 overexpressing stable clones of NIH 3T3 cells were malignantly transformed, however, they had a high level of expression of a p53 downstream gene, p21WAF1. The present study was undertaken to elucidate possible molecular mechanism(s) of such upregulation. An inCreased level of p21WAF1, expression was detected in sta- ble transfectants although an exogenous reporter gene driven by p21WAF1, promoter exhibited lower activity in these cells suggesting that some post-transcriptional mechanism contributes to upregulation. Western analyses of transient and stable clones revealed that upregulation of p21WAF1, in stable NIH 3T3/mot-2 cells may be mediated by cyclin D1 and cdk-2.
文摘The bioactive secondary metabolites from the endophytic fungus Fusarium napiforme was evaluated for the cytotoxic effect and antioxidant activity.The total antioxidant capacity(TAC)of the extract was determined by 2,2-diphenyl-1-picrylhydrazyl(DPPH),phosphomolybdate,and reducing power assay methods.The cytotoxicity of the extract was evaluated against lung adenocarcinoma(A549)cells and mouse embryo fibroblast(NIH3T3)cells by methyl thiazolyl tetrazolium(MTT)method.The major composition of the crude extract was identified by the Gas Chromatography-Mass Spectrometry(GC-MS)analysis.Estimation of the endophyte crude extract revealed a high amount of the total flavonoid content(TFC)and total phenolic content(TPC).The extract showed high cytotoxic activity against the A549 cell line with the mean cytotoxicity of 69.74±0.49%.The extract did not show any cytotoxic effect against the NIH3T3 cell line.The extract exhibited high antioxidant activity as a function of the concentrations.At a test concentration of 1 mg ml-1,the extract showed the highest inhibition against DPPH radical at 75.4607%±0.47688,ferric ion reducing power at 0.882±0.0120,and 255.434±21.404 AAE/g of the extract by phosphomolybdenum assay(PMA).There is a significant correlation between TPC and antioxidant activity at p<0.05.The correlation between reducing power and DPPH is significant at p<0.01.The major types of bioactive compounds identified by the GC-MS have shown the presence of nine major compounds.This result strongly exhibits that the endophyte F.napiforme can be a potential source for the formulation of natural anticancer drugs and protecting the body from oxidative damages.
基金This work was supported by the Russian Science Foundation grant#19-75-30032Genotyping of mice and primary cell cultures was supported by grant 075-15-2019-1660 from the Ministry of Science and Higher Education of the Russian Federation.Generation of TNF KO and IL-6 KO MEFs was supported by the Russian Foundation for Basic Research grant#19-04-01094.
文摘Bioengineered scaffolds are crucial components in artificial tissue construction.In general,these scaffolds provide inert three-dimensional(3D)surfaces supporting cell growth.However,some scaffolds can affect the phenotype of cultured cells,especially,adherent stromal cells,such as fibroblasts.Here we report on unique properties of 3D fibroin/gelatin materials,which may rapidly induce expression of adhesion molecules,such as ICAM-1 and VCAM-1,in cultured primary murine embryonic fibroblasts(MEFs).In contrast,two-dimensional(2D)fibroin/gelatin films did not show significant effects on gene expression profiles in fibroblasts as compared to 3D culture conditions.Interestingly,TNF expression was induced in MEFs cultured in 3D fibroin/gelatin scaffolds,while genetic or pharmacological TNF ablation resulted in diminished ICAM-1 and VCAM-1 expression by these cells.Using selective MAPK inhibitors,we uncovered critical contribution of JNK to 3D-induced upregulation of these adhesion molecules.Moreover,we observed ICAM-1/VCAM-1-dependent adhesion of lymphocytes to fibroblasts cultured in 3D fibroin/gelatin scaffolds,but not on 2D fibroin/gelatin films,suggesting functional reprogramming in stromal cells,when exposed to 3D environment.Finally,we observed significant infiltration of lymphocytes into 3D fibroin/gelatin,but not into collagen scaffolds in vivo upon subcapsular kidney implantation in mice.Together our data highlight the important features of fibroin/gelatin scaffolds,when they are produced as 3D sponges rather than 2D films,which should be considered when using these materials for tissue engineering.
基金funded and supported by the Leverhulme Trust Research Project Grant(RPG-2018-344)。
文摘The lifetime of orthopaedic implants can be extended by coating the softer Ti6Al4V alloy with harder biocompatible thin films.In this work,thin films of Ti_((1-x))Au_((x))are grown on Ti_(6)Al_(4)V and glass substrates by magnetron sputtering in the entire x=0-1 range,before their key biomechanical properties are performance tuned by thermal activation.For the first time,we explore the effect of in-situ substrate heating versus ex-situ post-deposition heat-treatment,on development of mechanical and biocompatibility performance in Ti-Au films.A~250% increase in hardness is achieved for Ti-Au films compared to bulk Ti6Al4V and a~40%improvement from 8.8 GPa as-grown to 11.9 and 12.3 GPa with in-situ and ex-situ heat-treatment respectively,is corelated to changes in structural,morphological and chemical properties,providing insights into the origins of super-hardness in the Ti rich regions of these materials.X-ray diffraction reveals that as-grown films are in nanocrystalline states of Ti-Au intermetallic phases and thermal activation leads to emergence of mechanically hard Ti-Au intermetallics,with films prepared by in-situ substrate heating having enhanced crystalline quality.Surface morphology images show clear changes in grain size,shape and surface roughness following thermal activation,while elemental analysis reveals that in-situ substrate heating is better for development of oxide free Ti3Auβ-phases.All tested Ti-Au films are non-cytotoxic against L929 mouse fibroblast cells,while extremely low leached ion concentrations confirm their biocompatibility.With peak hardness performance tuned to>12 GPa and excellent biocompatibility,Ti-Au films have potential as a future coating technology for load bearing medical implants.
