Objective: To provide comprehensive data to understand mechanisms of vascular endothelial cell(VEC) response to hypoxia/re-oxygenation. Methods: Human umbilical vein endothelial cells(HUVECs) were employed to construc...Objective: To provide comprehensive data to understand mechanisms of vascular endothelial cell(VEC) response to hypoxia/re-oxygenation. Methods: Human umbilical vein endothelial cells(HUVECs) were employed to construct hypoxia/re-oxygenation-induced VEC transcriptome profiling. Cells incubated under 5% O2, 5% CO2, and 90% N2 for 3 h followed by 95% air and 5% CO2 for 1 h were used in the hypoxia/re-oxygenation group. Those incubated only under 95% air and 5% CO2 were used in the normoxia control group. Results: By using a well-established microarray chip consisting of 58 339 probes, the study identified 372 differentially expressed genes. While part of the genes are known to be VEC hypoxia/re-oxygenation-related, serving as a good control, a large number of genes related to VEC hypoxia/re-oxygenation were identified for the first time. Through bioinformatic analysis of these genes, we identified that multiple pathways were involved in the reaction. Subsequently, we applied real-time polymerase chain reaction(PCR) and western blot techniques to validate the microarray data. It was found that the expression of apoptosis-related proteins, like pleckstrin homology-like domain family A member 1(PHLDA1), was also consistently up-regulated in the hypoxia/re-oxygenation group. STRING analysis found that significantly differentially expressed genes SLC38A3, SLC5A5, Lnc-SLC36 A4-1, and Lnc-PLEKHJ1-1 may have physical or/and functional protein–protein interactions with PHLDA1. Conclusions: The data from this study have built a foundation to develop many hypotheses to further explore the hypoxia/re-oxygenation mechanisms, an area with great clinical significance for multiple diseases.展开更多
BACKGROUND： Cerebral hippocampal astrocytes are more sensitive.to ischemic injury than neurons. Hypoxic-ischemic brain injury induces profound astrocyte apoptosis, and propofol may protect against astrocyte apoptosis...BACKGROUND： Cerebral hippocampal astrocytes are more sensitive.to ischemic injury than neurons. Hypoxic-ischemic brain injury induces profound astrocyte apoptosis, and propofol may protect against astrocyte apoptosis. OBJECTIVE： To verify the protective effects of propofol against astrocyte apoptosis and to investigate anti-apoptotic Bcl-2 and pro-apoptotic Bax expression in primary cultures of rat hippocampal astrocytes exposed to hypoxia-reoxygenation for different periods of time following propofol treatment. DESIGN, TIME, AND SETTING： In vitro neural immunocytochemistry was performed at the Central Laboratory of Yunyang Medical College between September 2007 and March 2008.MATERIALS： A total of 30 Wistar rats, aged 1-3 days, wJth equal numbers of males and females, were included for isolation and culture of .hippocampal astrocytes. METHODS： Hippocampal astrocytes were purified and cultured for 3 weeks and treated with four culture conditions： 50 μL Hank＇s solution （normal control）; 0.2 mL/L Intralipid; 50 μL Hank＇s solution for 10 minutes followed by hypoxic incubation for 4 hours and normoxic incubation for 12, 24, 36, 48, 60 or 72 hours; propofol （250 μmol/L final） for 10 minutes followed by hypoxic incubation for 4 hours and normoxic incubation for 12, 24, 36, 48, 60 and 72 hours. MAIN OUTCOME MEASURES： （1） Morphologic changes in hippocampal astrocytes. （2） Levels of astrocyte apoptosis and Bcl-2 and Bax expression. RESULTS： Hypoxia and reoxygenation increased apoptosis over time, with Bcl-2 expression peaking at 24 hours and decreasing gradually （P 〈 0.01 ）; Bax expression peaked at 72 hours （P 〈 0.01）; the ratio of Bcl-2/Bax was 1.4, 0.8, and 0.6, respectively, at 24, 48 and 72 hours. Non-apoptotic astrocytes showed significant proliferation and swelling. Propofol treatment decreased apoptosis after hypoxia-reoxygenation （P 〈 0.01）, as well as Bct-2 and Bax expression （P 〈 0.05, P 〈 0.01）, with Bcl-2/Bax ratios of 1.6-1.8. Propofol treatmentalso blocked astrocyte proliferation and swelling. No apoptotic cells or Bcl-2/Bax expression was detected in astrocytes cultured in Hank＇s or Intralipid solution. CONCLUSION： Propofol protects astrocytes against injury caused by hypoxia and reoxygenation via a mechanism that involves maintaining high ratios of Bcl-2/Bax.展开更多
OBJECTIVE:To investigate the protective efficacy of Bunao Fuyuan decoction(BNFY)on cerebral Ischemia/reperfusion(I/R)injury.METHODS:The mouse PC12 cells were chosen,and the oxidative-glucose deprivation/re-oxygenation...OBJECTIVE:To investigate the protective efficacy of Bunao Fuyuan decoction(BNFY)on cerebral Ischemia/reperfusion(I/R)injury.METHODS:The mouse PC12 cells were chosen,and the oxidative-glucose deprivation/re-oxygenation(OGD/R)injury model were established to simulate cerebral I/R injury.Atorvastatin was selected as a positive drug,and a gradient dose of BNFY was given for 6,12 and 24 h.3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-di-phenytetrazoliumromide(MTT)assay were used to detect cell viability at each time point.Cell apoptosis was measured by terminal deoxynucleotidyl transferase-mediated d UTP-botin nick end labeling(TUNEL)staining.enzyme linked immunosorbent assay was used to detect the expression of tumor necrosis factor(TNF)-α,interleukin(IL)-6,IL-1βand platelet activating factor(PAF).Western blot assay were performed to detect the expression of key regulators[toll-like receptor 4(TLR4),nuclear factor kappa-B(NF-κB),p-p38 mito-gen-activated protein kinase(MAPK)and p-Akt(also known as protein kinase B,PKB)]of cell survival and inflammatory response.RESULTS:The results of MTT assay and TUNEL staining assay revealed that BNFY significantly increased cell viability and inhibited cell apoptosis of PC12 cells following OGD/R,respectively.Furthermore,the expression of TNF-α,1 L-6,1 L-1βand PAF were decreased after BNFY treatment.And the results of Western blot assay showed that BNFY downregulated TLR4,NF-κB,p-p38 MAPK expression and upregulated p-Akt expression.CONCLUSION:Our findings suggest that BNFY may play a role in protecting OGD/R injured PC12 cells through inhibiting the inflammatory response and cell apoptosis.展开更多
At hydraulic structures,some strong interactions may develop between fast flowing waters and the air adjacent to the water in motion that enhance the air-water transfer of atmospheric and volatile gases in the flow.In...At hydraulic structures,some strong interactions may develop between fast flowing waters and the air adjacent to the water in motion that enhance the air-water transfer of atmospheric and volatile gases in the flow.In turn,in-stream structures may contribute to the aeration and re-oxygenation during overflow.The present study aims to characterize the aeration performance of a steep stepped weir,based upon a detailed physical investigation of air-water interfacial properties across a relatively wide range of discharges.The data showed a strong fragmentation of the air-water flows,a very broad range of entrained bubbles and drops,and a large amount of particle clustering.The results implied a monotonic increase in re-aeration with increasing rate of energy dissipation,while the largest aeration efficiency was observed on the horizontal step weir chute,with the smallest on the 1V:2.33H inclined downward steps.Altogether,the study showed that a steep stepped chute can make a sizeable contribution to the re-oxygenation of the waters,although the downward inclined steps reduce the re-aeration performances.展开更多
基金Project supported by the National Natural Science Foundation of China(Nos.81801572 and 81272075)the Foundation of Key Discipline Construction of Zhejiang Province for Traditional Chinese Medicine(No.2017-XKA36)+5 种基金the Foundation of Key Research Project of Zhejiang Province for Traditional Chinese Medicine(No.2019ZZ014)the Medical and Health Science Foundation of Zhejiang Province(No.2019327552)the Key Research and Development Program of Zhejiang Province(No.2019C03076)the General Research Program of Zhejiang Provincial Department of Medical and Health(No.2013KYA066)the Opening Foundation of State Key Laboratory for the Diagnosis and Treatment of Infectious Diseases(Nos.2018KF02 and 2019KF06)the Program of Education Department of Zhejiang Province(No.Y201738150),China。
文摘Objective: To provide comprehensive data to understand mechanisms of vascular endothelial cell(VEC) response to hypoxia/re-oxygenation. Methods: Human umbilical vein endothelial cells(HUVECs) were employed to construct hypoxia/re-oxygenation-induced VEC transcriptome profiling. Cells incubated under 5% O2, 5% CO2, and 90% N2 for 3 h followed by 95% air and 5% CO2 for 1 h were used in the hypoxia/re-oxygenation group. Those incubated only under 95% air and 5% CO2 were used in the normoxia control group. Results: By using a well-established microarray chip consisting of 58 339 probes, the study identified 372 differentially expressed genes. While part of the genes are known to be VEC hypoxia/re-oxygenation-related, serving as a good control, a large number of genes related to VEC hypoxia/re-oxygenation were identified for the first time. Through bioinformatic analysis of these genes, we identified that multiple pathways were involved in the reaction. Subsequently, we applied real-time polymerase chain reaction(PCR) and western blot techniques to validate the microarray data. It was found that the expression of apoptosis-related proteins, like pleckstrin homology-like domain family A member 1(PHLDA1), was also consistently up-regulated in the hypoxia/re-oxygenation group. STRING analysis found that significantly differentially expressed genes SLC38A3, SLC5A5, Lnc-SLC36 A4-1, and Lnc-PLEKHJ1-1 may have physical or/and functional protein–protein interactions with PHLDA1. Conclusions: The data from this study have built a foundation to develop many hypotheses to further explore the hypoxia/re-oxygenation mechanisms, an area with great clinical significance for multiple diseases.
文摘BACKGROUND： Cerebral hippocampal astrocytes are more sensitive.to ischemic injury than neurons. Hypoxic-ischemic brain injury induces profound astrocyte apoptosis, and propofol may protect against astrocyte apoptosis. OBJECTIVE： To verify the protective effects of propofol against astrocyte apoptosis and to investigate anti-apoptotic Bcl-2 and pro-apoptotic Bax expression in primary cultures of rat hippocampal astrocytes exposed to hypoxia-reoxygenation for different periods of time following propofol treatment. DESIGN, TIME, AND SETTING： In vitro neural immunocytochemistry was performed at the Central Laboratory of Yunyang Medical College between September 2007 and March 2008.MATERIALS： A total of 30 Wistar rats, aged 1-3 days, wJth equal numbers of males and females, were included for isolation and culture of .hippocampal astrocytes. METHODS： Hippocampal astrocytes were purified and cultured for 3 weeks and treated with four culture conditions： 50 μL Hank＇s solution （normal control）; 0.2 mL/L Intralipid; 50 μL Hank＇s solution for 10 minutes followed by hypoxic incubation for 4 hours and normoxic incubation for 12, 24, 36, 48, 60 or 72 hours; propofol （250 μmol/L final） for 10 minutes followed by hypoxic incubation for 4 hours and normoxic incubation for 12, 24, 36, 48, 60 and 72 hours. MAIN OUTCOME MEASURES： （1） Morphologic changes in hippocampal astrocytes. （2） Levels of astrocyte apoptosis and Bcl-2 and Bax expression. RESULTS： Hypoxia and reoxygenation increased apoptosis over time, with Bcl-2 expression peaking at 24 hours and decreasing gradually （P 〈 0.01 ）; Bax expression peaked at 72 hours （P 〈 0.01）; the ratio of Bcl-2/Bax was 1.4, 0.8, and 0.6, respectively, at 24, 48 and 72 hours. Non-apoptotic astrocytes showed significant proliferation and swelling. Propofol treatment decreased apoptosis after hypoxia-reoxygenation （P 〈 0.01）, as well as Bct-2 and Bax expression （P 〈 0.05, P 〈 0.01）, with Bcl-2/Bax ratios of 1.6-1.8. Propofol treatmentalso blocked astrocyte proliferation and swelling. No apoptotic cells or Bcl-2/Bax expression was detected in astrocytes cultured in Hank＇s or Intralipid solution. CONCLUSION： Propofol protects astrocytes against injury caused by hypoxia and reoxygenation via a mechanism that involves maintaining high ratios of Bcl-2/Bax.
文摘OBJECTIVE:To investigate the protective efficacy of Bunao Fuyuan decoction(BNFY)on cerebral Ischemia/reperfusion(I/R)injury.METHODS:The mouse PC12 cells were chosen,and the oxidative-glucose deprivation/re-oxygenation(OGD/R)injury model were established to simulate cerebral I/R injury.Atorvastatin was selected as a positive drug,and a gradient dose of BNFY was given for 6,12 and 24 h.3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-di-phenytetrazoliumromide(MTT)assay were used to detect cell viability at each time point.Cell apoptosis was measured by terminal deoxynucleotidyl transferase-mediated d UTP-botin nick end labeling(TUNEL)staining.enzyme linked immunosorbent assay was used to detect the expression of tumor necrosis factor(TNF)-α,interleukin(IL)-6,IL-1βand platelet activating factor(PAF).Western blot assay were performed to detect the expression of key regulators[toll-like receptor 4(TLR4),nuclear factor kappa-B(NF-κB),p-p38 mito-gen-activated protein kinase(MAPK)and p-Akt(also known as protein kinase B,PKB)]of cell survival and inflammatory response.RESULTS:The results of MTT assay and TUNEL staining assay revealed that BNFY significantly increased cell viability and inhibited cell apoptosis of PC12 cells following OGD/R,respectively.Furthermore,the expression of TNF-α,1 L-6,1 L-1βand PAF were decreased after BNFY treatment.And the results of Western blot assay showed that BNFY downregulated TLR4,NF-κB,p-p38 MAPK expression and upregulated p-Akt expression.CONCLUSION:Our findings suggest that BNFY may play a role in protecting OGD/R injured PC12 cells through inhibiting the inflammatory response and cell apoptosis.
基金This work was supported by the Swiss National Science Foundation(Grant No.P2ELP2_181794),the School of Civil Engineering,University of Queensland.
文摘At hydraulic structures,some strong interactions may develop between fast flowing waters and the air adjacent to the water in motion that enhance the air-water transfer of atmospheric and volatile gases in the flow.In turn,in-stream structures may contribute to the aeration and re-oxygenation during overflow.The present study aims to characterize the aeration performance of a steep stepped weir,based upon a detailed physical investigation of air-water interfacial properties across a relatively wide range of discharges.The data showed a strong fragmentation of the air-water flows,a very broad range of entrained bubbles and drops,and a large amount of particle clustering.The results implied a monotonic increase in re-aeration with increasing rate of energy dissipation,while the largest aeration efficiency was observed on the horizontal step weir chute,with the smallest on the 1V:2.33H inclined downward steps.Altogether,the study showed that a steep stepped chute can make a sizeable contribution to the re-oxygenation of the waters,although the downward inclined steps reduce the re-aeration performances.
