AIM: To investigate the killing efficiency of a recombinant plasmid containing a thymidine kinase (TK) domain insert driven by the vascular endothelial growth factor receptor 2 (VEGFR2) promoter (KDR) on vascular endo...AIM: To investigate the killing efficiency of a recombinant plasmid containing a thymidine kinase (TK) domain insert driven by the vascular endothelial growth factor receptor 2 (VEGFR2) promoter (KDR) on vascular endothelial cells.METHODS: The KDR-TK fragment was extracted from pBluescript Ⅱ KDR-TK plasmid by enzymatic digestion with Xho I and Sal I. The enhanced green fluorescence protein (EGFP) carrier was extracted from pEGFP by the same procedure. The KDR-TK was inserted into the pEGFP carrier to construct pEGFP-KDR-TK. Using ultrasound irradiation and microbubble, pEGFP-KDR-TK was transferred into human umbilical vein endothelial cells (HUVECs). The transient infection rate was estimated by green fluorescent protein (GFP) expression. Transfected HUVECs, non-transfected HUVECs, and HepG2 cells were cultured in the presence of different concentrations of ganciclovir (GCV), and the killing efficacy of HSV-TK/GCV was analyzed by 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyl tetrazolium bromide (MTT) assay. RESULTS: The recombinant pEGFP-KDR-TK was successfully constructed by inserting the KDR-TK fragment into the pEGFP carrier. Transfected HUVECs showed cytoplasmic green fluorescence, and the transient transfection rate was about 20.3%. Pools of G418-resistant cells exhibited a higher sensitivity to theprodrug/GCV compared to non-transfected HUVECs or non-transfected HepG2 cells, respectively. CONCLUSION: KDR promoter and the suicide gene/prodrug system mediated by diagnostic ultrasound combined with microbubble can significantly kill HUVECs. Such therapy may present a novel and attractive approach to target gene therapy on tumor vessels.展开更多
This study was carried out explore the mechanism underlying the inhibition of platelet activation by kelp fucoidans in deep venous thrombosis(DVT)mouse.In the control and sham mice,the walls of deep vein were regular ...This study was carried out explore the mechanism underlying the inhibition of platelet activation by kelp fucoidans in deep venous thrombosis(DVT)mouse.In the control and sham mice,the walls of deep vein were regular and smooth with intact intima,myometrium and adventitia.The blood vessel was wrapped with the tissue and there was no thrombosis in the lumen.In the DVT model,the wall was uneven with thicken intima,myometrium and adventitia.After treated with fucoidans LF1 and LF2,the thrombus was dissolved and the blood vessel was recanalized.Compared with the control group,the ROS content,ET-1 and VWF content and the expression of PKC-βand NF-κB in the model were significantly higher(P<0.05);these levels were significantly reduced following treatments with LF2 and LF1.Compared with H_(2)O_(2)treated-HUVECs,combined LF1 and LF2 treatment resulted in significant decrease in the expression of PKC-β,NF-κB,VWF and TM protein(P<0.05).It is clear that LF1 and LF2 reduces DVT-induced ET-1,VWF and TM expressions and production of ROS,thus inhibiting the activation of PKC-β/NF-κB signal pathway and the activation of coagulation system and ultimately reducing the formation of venous thrombus.展开更多
Leaf veins play an important role in plant growth and development,and the bundle sheath(BS)is believed to greatly improve the photosynthetic efficiency of C_(4) plants.The OBV mutation in tomato(Solanum lycopersicum)r...Leaf veins play an important role in plant growth and development,and the bundle sheath(BS)is believed to greatly improve the photosynthetic efficiency of C_(4) plants.The OBV mutation in tomato(Solanum lycopersicum)results in dark veins and has been used widely in processing tomato varieties.However,physiological performance has difficulty explaining fitness in production.In this study,we confirmed that this mutation was caused by both the increased chlorophyll content and the absence of bundle sheath extension(BSE)in the veins.Using genome-wide association analysis and map-based cloning,we revealed that OBV encoded a C_(2)H_(2) L domain class transcription factor.It was localized in the nucleus and presented cell type-specific gene expression in the leaf veins.Furthermore,we verified the gene function by generating CRISPR/Cas9 knockout and overexpression mutants of the tomato gene.RNA sequencing analysis revealed that OBV was involved in regulating chloroplast development and photosynthesis,which greatly supported the change in chlorophyll content by mutation.Taken together,these findings demonstrated that OBV affected the growth and development of tomato by regulating chloroplast development in leaf veins.This study also provides a solid foundation to further decipher the mechanism of BSEs and to understand the evolution of photosynthesis in land plants.展开更多
The Wulasigou Cu-Pb-Zn deposit,located 15 km northwest of Altay city in Xinjiang,is one of many Cu-Pb-Zn polymetallic deposits in the Devonian Kelan volcanic-sedimentary basin in southern Altaids.Two mineralizing peri...The Wulasigou Cu-Pb-Zn deposit,located 15 km northwest of Altay city in Xinjiang,is one of many Cu-Pb-Zn polymetallic deposits in the Devonian Kelan volcanic-sedimentary basin in southern Altaids.Two mineralizing periods can be distinguished:the marine volcanic sedimentary PbZn mineralization period,and the metamorphic hydrothermal Cu mineralization period,which is further divided into an early bedded foliated quartz vein stage(Q1) and a late sulfide-quartz vein stage(Q2) crosscutting the foliation.Four types of fluid inclusions were recognized in the Q1 and Q2 quartz from the east orebodies of the Wulasigou deposit:H2O-CO2 inclusions,carbonic fluid inclusions,aqueous fluid inclusions,and daughter mineral-bearing fluid inclusions.Microthermometric studies show that solid CO2 melting temperatures(T(m,CO2)) of H2O-CO2 inclusions in Ql are from-62.3℃ to-58.5C,clathrate melting temperatures(T(m,clath)l) are from 0.5 C to 7.5 C,partial homogenization temperatures(T(h,CO2)) vary from 3.3℃ to 25.9℃(to liquid),and the total homogenization temperatures(T(h,tot)) vary from 285℃ to 378℃,with the salinities being 4.9%-15.1%NaCl eqv.and the CO2-phase densities being 0.50-0.86 g/cm-3.H2O-CO2 inclusions in Q2 have T(m,CO2) from-61.9℃ to-56.9℃,T(m,clath)from 1.3℃ to 9.5℃,T(h,CO2) from 3.4℃ to 28.7℃(to liquid),and T(h,tot) from 242℃ to 388℃,with the salinities being 1.0%-15.5%NaCl eqv.and the CO2-phase densities being 0.48-0.89 g/cm-3.The minimum trapping pressures of fluid inclusions in Q1 and Q2 are estimated to be 260-360 MPa and180-370 MPa,respectively.The δ-(34)S values of pyrite from the volcanic sedimentary period vary from2.3‰ to 2.8‰(CDT),and those from the sulfide-quartz veins fall in a narrow range of-1.9‰ to 2.6‰(CDT).The δD values of fluid inclusions in Q2 range from-121.0‰ to-100.8‰(SMOW),and theδ-(18)O(H2O) values calculated from δ-(18)O of quartz range from-0.2‰ to 8.3‰(SMOW).The δD-δ-(18)O(H2O)data are close to the magmatic and metamorphic fields.The fluid inclusion and stable isotope data documented in this study indicate that the vein-type copper mineralization in the Wulasigou Pb-Zn-Cu deposit took place in an orogenic-metamorphic enviroment.展开更多
Objective:This study aimed to analyze the effects of transcatheter arterial embolization(TAE)combined with portal venous embolization(PVE)on the expression of MMP-2 in residual VX2 liver tumor tissues,liver function a...Objective:This study aimed to analyze the effects of transcatheter arterial embolization(TAE)combined with portal venous embolization(PVE)on the expression of MMP-2 in residual VX2 liver tumor tissues,liver function and non-embolic lobe regeneration.Methods:A total of 72 rabbits were randomly divided into Sham,TAE,PVE and TAE+PVE groups(n=18/group).The tissue samples from each group were taken at 6 h,3 days and 7 days after interventional operation,respectively.MMP-2 expression was detected by immunohistochemistry,Real-time PCR,and Western-blotting.The main indicators(such as AST,ATL,and TBIL)of liver function and the volume of non-embolized hepatic lobes were measured in each group after operation.One-way ANOVA and Kruskal-wallis method were used for statistical analysis.Results:The expression of MMP-2 mRNA and protein remained the highest in the Sham group,and the expression of MMP-2 mRNA and protein in TAE,PVE and TAE+PVE groups were successively increased,and the expression of MMP-2 in TAE+PVE group was always significantly higher than TAE group.The AST and ALT levels in each group on day 7 after operation showed a significant declination,and all groups have recovered to the preoperative baseline level and TBIL has a slight fluctuation in each group after operation with no statistical difference.On day 7 after operation,the increasing volume of non-embolized liver lobes in TAE+PVE group showed a more significant effect than those in PVE group,but there was no statistical significance(37.62±1.54 ml VS 36.18±1.15 ml,P=0.881),and its volume was significantly higher than those in the sham group(27.03±1.11 ml).Conclusion:TAE+PVE is considered to be an efficient and safe approach for treating rabbit VX2 liver transplantation tumor,but the expression of MMP-2 increased fastest after TAE+PVE,which might promote tumor cell invasion and metastasis.展开更多
Swarms of orthopyroxenite and websterite veins are found within Egiingol residual SSZ peridotite massif of Dzhida terrain(Central Asian Orogenic Belt,Northern Mongolia).The process of Egiingol pyroxenite veins formati...Swarms of orthopyroxenite and websterite veins are found within Egiingol residual SSZ peridotite massif of Dzhida terrain(Central Asian Orogenic Belt,Northern Mongolia).The process of Egiingol pyroxenite veins formation is investigated using new major and trace element analyses of pyroxenite minerals,calculations of closure temperatures and composition of equilibrium melt.The pyroxenites show abundant petrographic and geochemical evidence for replacement of the residual peridotite minerals by ortho-and clinopyroxene due to melt-rock interaction.Relics of peridotite olivines are found in pyroxenites,Cr#of spinel increases from peridotites to pyroxenites,and compositions of ortho-and clinopyroxene change from peridotite to pyroxenite.The authors show that calculated equilibrium melts for investigated pyroxenites are very similar to compositions of boninite lavas from the Dzhida terrain.Therefore,formation of pyroxenite veins most likely resulted from percolation of boninite melts through the Egiingol peridotites.Orthopyroxenite veins formed at first,followed by websterite veins.Thus,the authors assume that pyroxenite veins represent the channels for boninitic melts migration in supra-subduction environment.展开更多
The paper studies CO2 degassing and controlling factors under the condition of contact metamorphism in the Shuangshan area, southern Tan-Lu fault belt and the method of calculating the amount of CO2 degassing. The res...The paper studies CO2 degassing and controlling factors under the condition of contact metamorphism in the Shuangshan area, southern Tan-Lu fault belt and the method of calculating the amount of CO2 degassing. The results show that the amount of CO2 degassing is controlled by the characteristics of the country rocks, including the thermal conductivity, penetrability, porosity and connectivity. Compositions, size and depth of intrusive rock also have an important influence on CO2 degassing, i.e., they generated numerous cracks in the country rocks, and thus allowed the easy flow and accumulation of fluids. The amount of CO2 flux in contact metamorphism is calculated quantitatively based on the metamorphic reaction and time-integrated fluid flux. The value (0.729- 2.446×10^4 mol/cm^2) of CO2 flux suggests that CO2 was provided mainly by the contact metamorphic reaction. The generation and releasing of CO2 are positively correlated with the degree of metamorphism, and XCO2 in fluids gradually increases from dolomite zone to calcite zone, but in the zone of grossular, fluid flux is the largest and XCO2 sharply decreases due to involvement of magmatic water. This study presents evidence that a large amount of industrial-scale CO2 can be produced during contact metamorphism. On the basis of theoretical and practical studies, a cone model has been proposed to response CO2 degassing for the contact metamorphism, and it can be used to explore CO2 accumulations beyond the oil-gas basins. This model can also be applied to the study of inorganic genesis of CO2 accumulations.展开更多
基金New Century Distinguished Scholar Supporting Program of Ministry of Education (80000-3171404) The National Natural Science Foundation of China, No. 30300082, No. 30470467
文摘AIM: To investigate the killing efficiency of a recombinant plasmid containing a thymidine kinase (TK) domain insert driven by the vascular endothelial growth factor receptor 2 (VEGFR2) promoter (KDR) on vascular endothelial cells.METHODS: The KDR-TK fragment was extracted from pBluescript Ⅱ KDR-TK plasmid by enzymatic digestion with Xho I and Sal I. The enhanced green fluorescence protein (EGFP) carrier was extracted from pEGFP by the same procedure. The KDR-TK was inserted into the pEGFP carrier to construct pEGFP-KDR-TK. Using ultrasound irradiation and microbubble, pEGFP-KDR-TK was transferred into human umbilical vein endothelial cells (HUVECs). The transient infection rate was estimated by green fluorescent protein (GFP) expression. Transfected HUVECs, non-transfected HUVECs, and HepG2 cells were cultured in the presence of different concentrations of ganciclovir (GCV), and the killing efficacy of HSV-TK/GCV was analyzed by 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyl tetrazolium bromide (MTT) assay. RESULTS: The recombinant pEGFP-KDR-TK was successfully constructed by inserting the KDR-TK fragment into the pEGFP carrier. Transfected HUVECs showed cytoplasmic green fluorescence, and the transient transfection rate was about 20.3%. Pools of G418-resistant cells exhibited a higher sensitivity to theprodrug/GCV compared to non-transfected HUVECs or non-transfected HepG2 cells, respectively. CONCLUSION: KDR promoter and the suicide gene/prodrug system mediated by diagnostic ultrasound combined with microbubble can significantly kill HUVECs. Such therapy may present a novel and attractive approach to target gene therapy on tumor vessels.
基金supported by the Special Fund for Clinical Scientific Research of Shandong Medical Association(No.YXH2020ZX058).
文摘This study was carried out explore the mechanism underlying the inhibition of platelet activation by kelp fucoidans in deep venous thrombosis(DVT)mouse.In the control and sham mice,the walls of deep vein were regular and smooth with intact intima,myometrium and adventitia.The blood vessel was wrapped with the tissue and there was no thrombosis in the lumen.In the DVT model,the wall was uneven with thicken intima,myometrium and adventitia.After treated with fucoidans LF1 and LF2,the thrombus was dissolved and the blood vessel was recanalized.Compared with the control group,the ROS content,ET-1 and VWF content and the expression of PKC-βand NF-κB in the model were significantly higher(P<0.05);these levels were significantly reduced following treatments with LF2 and LF1.Compared with H_(2)O_(2)treated-HUVECs,combined LF1 and LF2 treatment resulted in significant decrease in the expression of PKC-β,NF-κB,VWF and TM protein(P<0.05).It is clear that LF1 and LF2 reduces DVT-induced ET-1,VWF and TM expressions and production of ROS,thus inhibiting the activation of PKC-β/NF-κB signal pathway and the activation of coagulation system and ultimately reducing the formation of venous thrombus.
基金This research was supported by The National Key Research and Development Program of China(Grant No.2019YFD1000301)the Fundamental Research Funds for Central Nonprofit Scientific Institution(Grant No.IVF-BRF2018006)+1 种基金the Key Laboratory of Biology and Genetic Improvement of Horticultural Crops,Ministry of Agriculture,Chinathe Science and Technology Innovation Program of the Chinese Academy of Agricultural Sciences(Grant No.CAAS-ASTIP-IVFCAAS).
文摘Leaf veins play an important role in plant growth and development,and the bundle sheath(BS)is believed to greatly improve the photosynthetic efficiency of C_(4) plants.The OBV mutation in tomato(Solanum lycopersicum)results in dark veins and has been used widely in processing tomato varieties.However,physiological performance has difficulty explaining fitness in production.In this study,we confirmed that this mutation was caused by both the increased chlorophyll content and the absence of bundle sheath extension(BSE)in the veins.Using genome-wide association analysis and map-based cloning,we revealed that OBV encoded a C_(2)H_(2) L domain class transcription factor.It was localized in the nucleus and presented cell type-specific gene expression in the leaf veins.Furthermore,we verified the gene function by generating CRISPR/Cas9 knockout and overexpression mutants of the tomato gene.RNA sequencing analysis revealed that OBV was involved in regulating chloroplast development and photosynthesis,which greatly supported the change in chlorophyll content by mutation.Taken together,these findings demonstrated that OBV affected the growth and development of tomato by regulating chloroplast development in leaf veins.This study also provides a solid foundation to further decipher the mechanism of BSEs and to understand the evolution of photosynthesis in land plants.
基金funded by National Nature Science Foundation of China(41372096)
文摘The Wulasigou Cu-Pb-Zn deposit,located 15 km northwest of Altay city in Xinjiang,is one of many Cu-Pb-Zn polymetallic deposits in the Devonian Kelan volcanic-sedimentary basin in southern Altaids.Two mineralizing periods can be distinguished:the marine volcanic sedimentary PbZn mineralization period,and the metamorphic hydrothermal Cu mineralization period,which is further divided into an early bedded foliated quartz vein stage(Q1) and a late sulfide-quartz vein stage(Q2) crosscutting the foliation.Four types of fluid inclusions were recognized in the Q1 and Q2 quartz from the east orebodies of the Wulasigou deposit:H2O-CO2 inclusions,carbonic fluid inclusions,aqueous fluid inclusions,and daughter mineral-bearing fluid inclusions.Microthermometric studies show that solid CO2 melting temperatures(T(m,CO2)) of H2O-CO2 inclusions in Ql are from-62.3℃ to-58.5C,clathrate melting temperatures(T(m,clath)l) are from 0.5 C to 7.5 C,partial homogenization temperatures(T(h,CO2)) vary from 3.3℃ to 25.9℃(to liquid),and the total homogenization temperatures(T(h,tot)) vary from 285℃ to 378℃,with the salinities being 4.9%-15.1%NaCl eqv.and the CO2-phase densities being 0.50-0.86 g/cm-3.H2O-CO2 inclusions in Q2 have T(m,CO2) from-61.9℃ to-56.9℃,T(m,clath)from 1.3℃ to 9.5℃,T(h,CO2) from 3.4℃ to 28.7℃(to liquid),and T(h,tot) from 242℃ to 388℃,with the salinities being 1.0%-15.5%NaCl eqv.and the CO2-phase densities being 0.48-0.89 g/cm-3.The minimum trapping pressures of fluid inclusions in Q1 and Q2 are estimated to be 260-360 MPa and180-370 MPa,respectively.The δ-(34)S values of pyrite from the volcanic sedimentary period vary from2.3‰ to 2.8‰(CDT),and those from the sulfide-quartz veins fall in a narrow range of-1.9‰ to 2.6‰(CDT).The δD values of fluid inclusions in Q2 range from-121.0‰ to-100.8‰(SMOW),and theδ-(18)O(H2O) values calculated from δ-(18)O of quartz range from-0.2‰ to 8.3‰(SMOW).The δD-δ-(18)O(H2O)data are close to the magmatic and metamorphic fields.The fluid inclusion and stable isotope data documented in this study indicate that the vein-type copper mineralization in the Wulasigou Pb-Zn-Cu deposit took place in an orogenic-metamorphic enviroment.
基金supported by Natural Science Foundation of Anhui Province(NO.1408085MH162)
文摘Objective:This study aimed to analyze the effects of transcatheter arterial embolization(TAE)combined with portal venous embolization(PVE)on the expression of MMP-2 in residual VX2 liver tumor tissues,liver function and non-embolic lobe regeneration.Methods:A total of 72 rabbits were randomly divided into Sham,TAE,PVE and TAE+PVE groups(n=18/group).The tissue samples from each group were taken at 6 h,3 days and 7 days after interventional operation,respectively.MMP-2 expression was detected by immunohistochemistry,Real-time PCR,and Western-blotting.The main indicators(such as AST,ATL,and TBIL)of liver function and the volume of non-embolized hepatic lobes were measured in each group after operation.One-way ANOVA and Kruskal-wallis method were used for statistical analysis.Results:The expression of MMP-2 mRNA and protein remained the highest in the Sham group,and the expression of MMP-2 mRNA and protein in TAE,PVE and TAE+PVE groups were successively increased,and the expression of MMP-2 in TAE+PVE group was always significantly higher than TAE group.The AST and ALT levels in each group on day 7 after operation showed a significant declination,and all groups have recovered to the preoperative baseline level and TBIL has a slight fluctuation in each group after operation with no statistical difference.On day 7 after operation,the increasing volume of non-embolized liver lobes in TAE+PVE group showed a more significant effect than those in PVE group,but there was no statistical significance(37.62±1.54 ml VS 36.18±1.15 ml,P=0.881),and its volume was significantly higher than those in the sham group(27.03±1.11 ml).Conclusion:TAE+PVE is considered to be an efficient and safe approach for treating rabbit VX2 liver transplantation tumor,but the expression of MMP-2 increased fastest after TAE+PVE,which might promote tumor cell invasion and metastasis.
基金The reported study was funded by RFBR according to the research project(18-35-00535)IGC State Assignment Project(0350-2019-0008).
文摘Swarms of orthopyroxenite and websterite veins are found within Egiingol residual SSZ peridotite massif of Dzhida terrain(Central Asian Orogenic Belt,Northern Mongolia).The process of Egiingol pyroxenite veins formation is investigated using new major and trace element analyses of pyroxenite minerals,calculations of closure temperatures and composition of equilibrium melt.The pyroxenites show abundant petrographic and geochemical evidence for replacement of the residual peridotite minerals by ortho-and clinopyroxene due to melt-rock interaction.Relics of peridotite olivines are found in pyroxenites,Cr#of spinel increases from peridotites to pyroxenites,and compositions of ortho-and clinopyroxene change from peridotite to pyroxenite.The authors show that calculated equilibrium melts for investigated pyroxenites are very similar to compositions of boninite lavas from the Dzhida terrain.Therefore,formation of pyroxenite veins most likely resulted from percolation of boninite melts through the Egiingol peridotites.Orthopyroxenite veins formed at first,followed by websterite veins.Thus,the authors assume that pyroxenite veins represent the channels for boninitic melts migration in supra-subduction environment.
基金supported by grants from the Natural Science Foundation of China(grant Nos:40473021 and 40472074)the National Basic Research Program of China(grant No:2003CB214600).
文摘The paper studies CO2 degassing and controlling factors under the condition of contact metamorphism in the Shuangshan area, southern Tan-Lu fault belt and the method of calculating the amount of CO2 degassing. The results show that the amount of CO2 degassing is controlled by the characteristics of the country rocks, including the thermal conductivity, penetrability, porosity and connectivity. Compositions, size and depth of intrusive rock also have an important influence on CO2 degassing, i.e., they generated numerous cracks in the country rocks, and thus allowed the easy flow and accumulation of fluids. The amount of CO2 flux in contact metamorphism is calculated quantitatively based on the metamorphic reaction and time-integrated fluid flux. The value (0.729- 2.446×10^4 mol/cm^2) of CO2 flux suggests that CO2 was provided mainly by the contact metamorphic reaction. The generation and releasing of CO2 are positively correlated with the degree of metamorphism, and XCO2 in fluids gradually increases from dolomite zone to calcite zone, but in the zone of grossular, fluid flux is the largest and XCO2 sharply decreases due to involvement of magmatic water. This study presents evidence that a large amount of industrial-scale CO2 can be produced during contact metamorphism. On the basis of theoretical and practical studies, a cone model has been proposed to response CO2 degassing for the contact metamorphism, and it can be used to explore CO2 accumulations beyond the oil-gas basins. This model can also be applied to the study of inorganic genesis of CO2 accumulations.