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AIM:To study the protective effect of anti-aging Klotho protein on human umbilical vein endothe-lial cells (HUVECs) treated with high glucose (HG).METHODS:HUVECs were cultured in vitro, and divided into PBS control group, 5.5 mmol/L glucose group, 33.3 mmol/L glucose group, 0.1 μmol/L Klotho +33.3 mmol/L glucose group, 1 μmol/L Klotho+33.3 mmol/L glucose group , and 10μmol/L Klotho+33.3 mmol/L glucose group .The viabili-ty of the HUVECs was measured by MTT assay .The content of malondialdehyde ( MDA) , and the activities of lactate de-hydrogenase (LDH), superoxide dismutase (SOD) and glutathione (GSH) in cell culture supernatants were observed . The production of reactive oxygen species ( ROS) in HUVECs was analyzed by flow cytometry .The levels of nitric oxide ( NO) , endothelin ( ET-1 ) , intercellular adhesion molecule-1 ( ICAM-1 ) in HUVEC culture medium were detected by ELISA.The protein expression of nuclear factor-kappa B (NF-κB) in the HUVECs was determined by Western blot .RE-SULTS:Compared with PBS control group , 33.3 mmol/L glucose significantly decreased the HUVEC viability , increased ROS, LDH and MDA levels , reduced the activities of SOD and GSH , decreased the NO secretion , and induced the ET-1 and ICAM-1 secretion and the protein expression of NF-κB in HUVECs.When HUVECs were treated with Klotho protein at different concentrations combined with 33.3 mmol/L glucose, the cell viability was increased significantly , the ROS, LDH and MDA levels were decreased significantly , the antioxidant SOD and GSH activities were significantly increased , the se-cretion of NO was increased , but ET-1 and ICAM-1 releases and protein expression of NF-κB were significantly reduced . CONCLUSION:Anti-aging Klotho protein promotes the viability of HUVECs treated with HG , reduces the oxidative dam-age and ROS production , and restores the normal secretory function of HUVECs , thus playing a protective role in vascular endothelial cells through reducing the protein expression of NF-κB.
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[ ABSTRACT] AIM:To study the effects of anti-aging Klotho protein on neonatal rat myocardial cells with hypo-xia/reoxygenation ( H/R) injury.METHODS:The cardiomyocytes of neonatal SD rats were cultured to establish hypoxia/reoxygenation model.The myocardial cells were divided into normal control group, H/R model group, different concentra-tions of Klotho protein (0.1μmol/L, 1μmol/L and 10μmol/L) pretreatment groups.The myocardial cells pulse frequen-cy was observed before and after H/R.The cell viability was measured by MTT assay.The leakages of LDH, CK and AST, the content of MDA and the activity of SOD were detected.The apoptotic rate of the myocardial cells was analyzed by flow cytometry.The mRNA expression of endoplasmic reticulum stress markers and apoptosis-related molecules GRP78, CRT, CHOP and caspase-12 was measured by real-time PCR.The protein levels of CHOP, caspase-12 and phosphorylated Akt in the myocardial cells were determined by Western blot.RESULTS: Compared with normal control group, the pulse fre-quency, cell viability rate and SOD activity of myocardial cells were significantly decreased, the cell apoptotic rate as well as the contents of LDH, CK, AST and MDA were increased in H/R model group.The mRNA expressions of GRP78, CRT, CHOP and caspase-12 as well as the protein levels of CHOP and caspase-12 were increased, whereas p-Akt level was decreased obviously.Compared with H/R model group, the pulse frequency, cell viability rate and SOD activity were in-creased significantly, the cell apoptotic rate as well as the contents of LDH, CK, AST and MDA were decreased in Klotho pretreated group.The mRNA expression of GRP78, CRT, CHOP and caspase-12 as well as the protein levels of CHOP and caspase-12 were decreased, while p-Akt level increased significantly.CONCLUSION:Anti-aging Klotho protein improves the myocardial cell survival and inhibits the apoptosis by increasing the resistance of the cells to oxidative stress and exces-sive endoplasmic reticulum stress response, which is related with the activation of Akt phosphorylation in H/R-injured my-cardial cells.
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AIM:To explore the effects of transforming growth factor-α( TGF-α) in the monoclonal formation , proliferation, migration and adhesiveness of human endothelial progenitor cells ( EPCs).METHODS: The isolated and cultured EPCs were treated with various concentrations of TGF-α(final concentrations of 1, 5, 10μg/L, respectively).At the same time, the PBS control and epidermal growth factor receptor-tyrosine kinase inhibitor (EGFR-TKI) group (10μg/L TGF-αplus 1∶1 000 EGFR-TKI) were set.The effects of TGF-αon monoclonal formation , proliferation, migration and adhesiveness of EPCs were determined by clone formation experiment , thiazolyl blue tetrazolium bromide (MTT), EdU, Transwell and adhesion assays , respectively.The expression of epithelial growth receptor (EGFR) and vascular endothelial growth factor ( VEGF) were measured by Western blotting .RESULTS:Different concentrations of TGF-αall significantly induced the monoclonal formation , proliferation, migration and adhesiveness of EPCs (P<0.01), which were inhibited by EGFR-TKI.The results of Western blotting showed that TGF-αalso induced the expression of EGFR and VEGF with a cer-tain concentration effect ( P<0.01) .CONCLUSION:By combining with EGFR induced the expression of VEGF , TGF-αsignificantly promotes the related cell function of monoclonal formation , proliferation, migration, adhesiveness in EPCs.
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Objective To study the quality of life(QOL) of patients with mild to moderate primary hypertension after treatment adjusted medication time by ambulatory blood pressure .Methods All 128 cases were divided into two groups randomly including chronos medication group and normal medication group .We tested ambulatory blood pressure ,QOL and influencing factors before medication and 6 months later .Results It′s observed the parameters of the ambulatory blood pressure improved significantly in all of the groups .There existed significant difference in nSBP ,nDBP ,nMAP ,SBP in dangerous night time ,and nocturnal blood pres‐sure decline rate between two groups (P<0 .05) .There existed very significant difference in DBP and MAP in dangerous night time (P<0 .01) .There were significant difference in the fields of MH (mental health) and RE (role limitation because of emotional health problem) (P<0 .05) ,and very significant difference in the fields of PF (physical functioning) ,RP (role limitation because of physical health) ,VT(vitality) (P<0 .01) .Multiple regression analysis indicate the influencing factors of the quality of life about patients with hypertension are age ,BMI index number ,medication time ,average family income ,medical insurance type ,culture de‐gree and job title .Age ,BMI index number and medication time depress QOL of patient with hypertension while average family in‐come ,better medical insurance type ,culture degree and job title improve it .Conclusion It′s possible to improved the parameters of the ambulatory blood pressure for patients with mild to moderate primary hypertension by chronos medication compared with nor‐mal medication more significantly .It′s useful to improve the QOL of patient with hypertension by improving average family income , medical insurance type and culture degree .
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Objective To assess the clinical diagnostic value of left atrial function index (LAFI) for heart failure with normal left ventricular ejection fraction (HFNEF).Methods One hundred and ten patients with HFNEF were divided into 3 groups by New York Heart Association (NYHA) cardiac functional grading:NYHA Ⅱ grade group (43 cases),NYHA Ⅲ grade group (40 cases) and NYHA Ⅳ grade group (27 cases),and another 33 healthy subjects were selected as control group.The 4 groups were examined by ultrasoundcardiogram,LAFI and E/E' were calculated,and the plasma brain natriuretic peptide (BNP)levels were determined.The relation between LAFI and NYHA cardiac functional grading and the correlation between LAFI and plasma BNP,E/E' were observed.Results The worse heart function,the higher plasma BNP and E/E',and the lower LAFI,and there were statistical differences [control group:(40.9 ± 26.1) ng/L,6.4 ± 2.0,0.73 ± 0.10,NYHA Ⅱ grade group:(230.1 ± 85.9) ng/L,10.0 ± 3.7,0.46 ± 0.30,NYHA Ⅲ grade group:(398.6 ±98.7) ng/L,16.9 ±4.0,0.26 ± 0.30,NYHA Ⅳ grade group:(680.3 ± 146.6) ng/L,23.8 ± 3.9,0.17 ± 0.20,P ＜ 0.05].The correlation analysis results showed that LAFI and plasma BNP,E/E' were negatively correlated in patients with HFNEF (r =-0.868,-0.873,P ＜ 0.01).Conclusions In HFNEF patients,there is significant correlation between LAFI and left ventricular diastolic dysfunction.LAFI can well diagnose diastolic heart failure early and evaluate the diastolic function of the left ventricle.
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Objective To compare the quality of life before ischemia postconditioning and 6 months after ischemia postconditioning in patients with acute ST segment elevation myocardial infarction,and investigate the related risk factors.Methods One hundred and twenty-nine patients with acute ST segment elevation myocardial infarction were given the ischemie postconditioning.The quality of life was evaluated before ischemia postconditioning and 6 months after ischemia postconditioning by 36-item short-form health survey (SF-36) and seattle angina questionnaire (SAQ).Results In acute ST segment elevation myocardial infarction patients treated with ischemia postconditioning,the scores of SF-36 and SAQ were significantly lower at 6 months after ischemia postconditioning than those before ischemia postconditioning [SF-36:total score of body health (76.4 ± 17.3) scores vs.(56.3 ± 16.8) scores,physiologic function (75.3 ± 18.1) scores vs.(52.4 ± 19.2) scores,physiologic functional authority (75.6 ± 16.5) scores vs.(48.5 ± 20.3) scores,body pain (77.2 ± 15.4) scores vs.(58.7 ± 16.6) scores,total health state (73.6 ± 17.8) scores vs.(50.6 ± 14.7)scores,total score of mental status (77.5 ± 15.3) scores vs.(55.3 ± 17.3) scores,vitality (69.3 ± 18.1)scores vs.(43.2 ± 17.4) scores,society function (70.3 ± 17.5) scores vs.(41.3 ± 14.2)scores,affection functional authority (80.2 ± 17.4)scores vs.(63.5 ± 14.3)scores,mental health (77.6 ± 18.4) scores vs.(55.6 ± 20.1) scores,health change (76.3 ± 17.4) scores vs.(53.4 ± 16.6) scores;SAQ:body limitation of activity (78.46 ± 12.21) scores vs.(35.34 ± 15.33)scores,angina stable state (74.23 ± 8.53) scores vs.(30.12 ± 5.38) scores,angina episode state (72.34 ± 10.33) scores vs.(27.33 ± 9.12) scores,satisfaction degree of treatment (76.42 ± 12.13)scores vs.(30.56 ± 15.57) scores,knowledge of diseases (74.22 ± 9.35)scores vs.(37.25 ± 20.32) scores] (P ＜ 0.05).The results of multiple linear regression equation showed that the older,more severe vascular lesions,and lower total score of body health score before ischemia postconditioning indicated lower total score of body health,and worse quality of life after ischemia postconditioning.Conclusions The quality of life of patients with acute ST segment elevation myocardial infarction after ischemia postconditioning is significantly improved,compared with that before ischemia postconditioning.Advanced age,vascular disease and lower total scores of body health before ischemia postconditioning are independent risk factors of total scores of body health after ischemia postconditioning.
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Objective To study the effect of 60 Co radiotherapy on the gene expession of fibronectin (FN) and matrix metalloproteinase-2 (MMP-2) in the cultured rat vascular smooth muscle cells (VSMCs). Methods Rat VSMCs were cultured in DMEM containing 10% FBS, and were radiated by 60 Co at the doses of 0,7,14 and 28Gy, respectively. mRNA level of FN and MMP-2 was measured by RT-PCR. Results The expression level of MMP-2 mRNA decreased from 82.52?6.50 to 64.25?3.81 and 42.17?3.16 in VSMCs with 14,25Gy radiation, respectively(P