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Aim To study the effect of adiponectin ( APN) on myocardial ischemia reperfusion( IR) injury in diabetic rats and to explore the role of oxidation-an-tioxidation system. Methods Male Sprague Dawley rats were randomly divided into control group( NS) , IR group ( NIR ) , diabetes group ( DS ) , DS + IR group (DIR), DS +APN +IR group(APN). Experimental diabetes was induced in the animals by a single intrap-eritoneal injection of streptozotocin at a dose of 55 mg ·kg-1 . The IR and NIR group were subjected to myo-cardial I/R injury. APN group was administered APN through intravenous injection 10 min before the reper-fusion, the others were administered normal saline. The rats were considered diabetic and used for the study only if their glucose levels were higher than 15 mmol·L-1. Results All the diabetic rats exhibited increased levels of blood glucose and reduction of body weight ( P <0. 05 ) . Compared with those of NS and DS groups, the myocardial infarct size, cardiomyocyte apoptosis, MDA concentration and ROS in NIR and DIR groups were remarkably increased, activities of SOD and NO were decreased(P<0. 05 or P<0. 01). APN decreased oxidative stress product generation and myocardial apoptosis induced by diabetic myocardial I/R injury ( P <0. 05 ) . Conclusion APN exerts pro-tective effect on myocardial I/R injury in diabetic rats through anti-oxidative mechanism.
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Objective To investigate the effect of Ang-(1-7) on the apoptosis in human umbilical vein endothelial cells (HUVECs) induced by Ang Ⅱ.Methods HUVECs were isolated and cultured.Cultured HUVECs were incubated for 24 h with Ang-(1-7), Ang Ⅱ, Ang-(1-7) A-779, Ang-(1-7) + Ang Ⅱ, A-779 + Ang Ⅱ + Ang-( 1-7), respectively.Cultured HUVECs without incubating stimulator were chosen as controls.The apoptosis of endothelial cells were detected by flow cytometry.Results The apoptosis of endothelial cells in HUVECs were upregulated by AngⅡ ( 10-6 mol/L) (25.60% ±3.17% vs 2.32% ±0.24%, P ＜0.005).Compared with the AngⅡ group, Ang-(1-7) dose-dependently inhibited the apoptosis of endothelial cells in HUVECs ( (20.04% ± 2.21% ,16.04% ± 1.32 %, 10.04% ±2.05,7.79% ±1.50% vs AngⅡ group 25.60% ±3.17%, P ＜0.05 , P ＜0.05).The effects of Ang-(1-7) could be blocked by A-779 (23.37% ±0.75% vs 20.04% ± 2.21%, 16.04% ± 1.32,10.04% ± 2.05% ,7.79%± 1.50%, P ＜ 0.05 ).Conclusion Ang-(1-7) can attenuate the apoptosis of endothelial cells induced by Ang Ⅱ in HUVECs in a dose-dependent manner.The effects of Ang-(1-7) could be blocked by A-779( P＜0.05).
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Objective To induce adipose-derived mesenchymal stem cells (ADMSCs) differentiation into cardiomyocytes, so as to provide stem cells for myocardial regeneration.Method ADMSCs were isolated and cultured from adult adipose tissue in vitro.They were induced with 10μmol/L 5-azacytidine (5Aza) for 24h.At the 7th, 14th, 21st days after induction, the expression of α-sarcomeric actin, and myosin heavy chain were repeatedly detected by immunocytochemistry.The expression of ANP was detected by RT-PCR.Results At the 7th day after induction, there was no expression of α-sarcomeric actin and MHC.At the 14th day, there was a little expression of α-sarcomeric actin and MHC that could be seen in cells.At the 21st day, there was increased expression of α-sarcomeric and MHC, and the expression of ANP was positive results detected by RT-PCR.Conclusions 5-Aza can induced ADMSCs to differentiate into cardiomyocytes.ADMSCs might be a potential source of cell transplantation for myocardial.
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AIM: To study the characteristic of liver X receptor alpha (LXRα), its target gene expression and cholesterol efflux in human macrophages treated with atorvastatin. METHODS: Human monocyte-derived macrophages were collected and cultured. Macrophages were treated with or without atorvastatin. Apolipoprotein A-I mediated human monocyte-derived macrophage cholesterol efflux was detected by liquid scintillation counting method. Reverse transcription-polymerase chain reaction (RT-PCR) was used to evaluate the mRNA expression of LXRα and some of its target genes ABCA1, SREBP2, CETP, PLTP, apoE, MMP-9 and MIP-1α. The protein expression of LXRα, ABCA1, MMP-9 and MIP-1α was determined by Western blotting. RESULTS: Pre-incubation of human monocyte-derived macrophages with atorvastatin dose dependently (1-2 μmol/L) stimulated cholesterol efflux mediated by apolipoprotein A-I. Atorvastatin also increased the mRNA expression of LXRα, ABCA1, SREBP2, CETP, PLTP, and protein expression of LXRα, ABCA1, but decreased the expression of MMP-9 and MIP-1α at both mRNA and protein levels. CONCLUSION: Atorvastatin enhances the cholesterol efflux, upregulates LXR and some genes associated with cholesterol metabolism and inhibits inflammatory responses in macrophages, indicating that statins may affect the formation of foam cells by activating LXR signaling pathway.
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AIM: The present study was undertaken to investigate the effect of angiotensin II (AngⅡ) on expression of MMP-9 in THP-1 macrophages. METHODS: Macrophages converted from THP-1 monocytes by incubating with PMA (0.1 μmol/L) for 48 h were divided into PMA group; PMA+AngⅡ group (10-7mol/L, 1 h); PMA+AngⅡ+PDTC group (10 μmol/L, 30 min) and PDTC group. Western blotting was used to detect the MMP-9 and phosphorylation of NF-κB p65, and the expression of MMP-9 mRNA in THP-1 macrophages was measured by RT-PCR.RESULTS: Compared to control group, the expression of MMP-9 (1.06±0.11, P<0.05) and phosphorylation of NF-κB p65 (1.02±0.10, P<0.05) in THP-1 macrophages were expressed when treated with AngⅡ (10-7mol/L); and the expression of MMP-9 mRNA were upregulated (1.22±0.08, P<0.05). However, NF-κB inhibitor PDTC reduced the NF-κB p65 (0.99±0.12, P<0.01) and MMP-9 (1.04±0.14, P<0.01) expressions and decreased the expression of MMP-9 mRNA (0.90±0.06,P<0.01). CONCLUSION: NF-κB signaling pathway contributes to the expression of MMP-9 in THP-1 macrophage induced by AngⅡ.
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The technique of RNA interference has been used in the area of cardiovascular system. Chitosan nanoparticles(CS-NP) has been a hotspot of the research due to its good biological characteristics and has a great potential to be used as the carrier for gene delivery. This article gives a simple review the application of the techniques, the currently used preparation methods of CS-NP, the factors that affect the rate of complexing of plasmid with CS-NP, the transfection efficiency of plasmid CS-NP compounds, and its in vitro drug releasing behavior.
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The expression vector of shRNA targeted to the rat angiotensin Ⅱ receptor gene wasconstructed and the efficacy of siRNAs to modulate the expression of target gene in the in vitro cul-tured mammalian cells was investigated for antihypertensive therapy in spontaneous hypertensive rat(SHR) at post-transcriptional level. The sense and antisense RNA oligonucleotides strands targe-ting angiotensin Ⅱ receptor mRNA were synthesized individually according to the sequence of therat angiotensin Ⅱ receptor. For preparation of duplexes, sense- and antisense-stranded oligonucle-otides were mixed and annealed, and the annealed duplexes were cloned into the pGenesil-1 vector..The rat glioma cells were transfected with constructed pGenesil-1-shRNA plasmid and scrambled plasmid. The cultured cells were collected at different phases. RT-PCR and Western blot were performed. The AT1 mRNA and protein levels behaved ultimately same. Compared to control after 48 h, AT1 mRNA levels were decreased to 35.5 %±3.0 %, and the levels reached their lowest point after 72 h (20.7 %±4 % of control). At 24 and 48 h, AT1 protein was reduced to 46.9 %±4.2 %and 36.98 % ± 3.7 % respectively compared to control and a maximum reduction was observed after 72 h of incubation (28. 1% ±4 % compared to controls). Plasmid-based shRNA expression systems targeted against the rat angiotensin Ⅱ receptor gene were generated successfully. The shRNAs with a 22-nt stem and a short loop were cleaved into small interfering dsRNA (siRNA) by the Dicer. The in vitro transcribed siRNA enables the effective silencing of gene expression to the target mRNA and leads to effective inhibition of translation of proteins and will be lay the foundation of application of gene silencing technology to hypertensive rats.
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Objective To investigate Tpeak - Tend interval and Tpeak - Tend/Q - T interval ratio in healthy people. Methods Q - T interval and Q - Tpeak interval (time interval from QRS - wave starting to T - wave peak) were measured on electrocardiogram in 100 healthy people, rectrospectively. Then , Tpeak - Tend was calculated by Q - T interval substrating Q - T peak, and T peak - Tend /Q -T interval ratio was calculated. Results T peak - T end /Q - T ratio and T peak - T end interval were greatest in lead V2 (0.24?0.04, 93.20?16.07)(ms) and smallest in lead avL (0.16?0.03 ,58.06?13.70) (ms). Tpeak - Tend interval in Ⅱ,Ⅲ,avR, V2～V6 were significantly greater in male than those in female. Tpeak - Tend /Q - T ratio in Ⅱ, Ⅲ, avR, avF, V2～V6 were significantly greater in male than those in female. Total 95% confidence interval of Tpeak - Tend interval and Tpeak - Tend /Q - T ratio in lead V2 were (90.01 , 96.39) (ms) and (0.23 , 0.25) /respectively. 95% confidence interval of Tpeak - Tend interval of Male and female subjects in lead V2 were (93.67,103.07) (ms) and (85.13,93.46) (ms) , respectively. 95% confidence interval of Tpeak -Tend interval of Male and female subjects in lead V2 were(0.24 ,0.26) and (0.21 ,0.24) , respectively. Conclusion Our study provides values of T peak -T end interval and T peak - T end/Q - T ratio in healthy people.
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Objective To compare the relativity between Tp-Te interval,Tp-Te /QT ratio with heart rate(HR)and explore their clinical significance retrospectively.Methods 200 normal individuals' ECG were randomly selected and the Tp-Te interval and Tp-Te/QT ratio were measured in precordial lead V6 retrospectively.The relativity between Tp-Te interval and Heart Rate,Tp-Te/QT ratio and Heart Rate were calculated retrospectively.Results The Tp-Te interval was about(83.17?12.64)ms in precordial lead V6 of normal individuals.The Tp-Te interval decreased linearly with the increase in HR with the range of Tp-Te interval being 56 to 114 milliseconds(Pearson's correlation coefficient r was-0.239,P=0.007 0.01).Conclusion There is an inverse correlation between Tp-Te interva and HR while no correlation between Tp-Te/QT ratio and HR.
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Objective To study the genome DNA methylation in SLE and the related factors of DNA methylation. Methods Twenty-six cases with SLE and 20 controls were recruited to participate the study. Plasma Hcy, SAM, SAH and the MTHFR gene C677T polymorphism were measured in all patients and controls. Results {1} The SAM levels were lower significantly in SLE groups than in controls. The SAH levels were higher significantly in SLE groups than in controls. {2} There was significant inverse correlation between plasma Hcy level and SAM level (r=-0.897, P
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AIM:The present study was undertaken to investigate the effect of angiotensin II (AngⅡ) on expression of MMP-9 in THP-1 macrophages. METHODS:Macrophages converted from THP-1 monocytes by incubating with PMA (0.1 ?mol/L) for 48 h were divided into PMA group; PMA+AngⅡ group (10-7mol/L,1 h); PMA+AngⅡ+PDTC group (10 ?mol/L,30 min) and PDTC group. Western blotting was used to detect the MMP-9 and phosphorylation of NF-?B p65,and the expression of MMP-9 mRNA in THP-1 macrophages was measured by RT-PCR.RESULTS:Compared to control group,the expression of MMP-9 (1.06?0.11,P
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AIM:To observe the effects of adiponectin on injury of the 3-4 d SD rat cardiomyocytes induced by the intervention of H2O2.METHODS:Primary cardiomyocytes were obtained from neonatal rat and were cultured by enzymatic digestion methods.The molecular marker was observed by ?-actin immunocytochemistry.Primary cultured 3-4 d cells were used in experiment,and the injury model was established by H2O2,and adiponectin and Ara-A were used for pre-treatment before cell culture.The morphological change of cardiomyocytes was observed under electron microscope.The contents of LDH,MDA and the activity of SOD were measured.The apoptosis of cardiomyocytes was detected by agarose gel electrophoresis and Annexin V/PI staining with flow cytometry.RESULTS:Adiponectin pretreatment significantly decreased the release of LDH(P
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0.05).CONCLUSION:Ang-(1-7) inhibits the secretion of MCP-1 and ICAM-1 in cultured HUVEC in a concentration dependent matter induced by AngⅡ. The inhibitory effect of Ang-(1-7) may be through its specific receptor.
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Objective By Comparing the early and late coronary angiograms results between cutting balloon angioplasty (CBA) and plain old balloon angioplasty (POBA), to evaluate the efficacy of CBA on coronary in stent restenosis Methods 166 patients with in stent restenosis after PTCA were randomized into two groups: CBA group (98 cases) and POBA group (68 cases) according to the balloon used Its instant and late mimimal lumen diameters (MLD) diameter stenoses (DS) and restenosis rates after PTCA in above two groups were compared Results Although no significant difference in mimimal lumen diameters (MLD) ([2 6?0 6]mm vs [2 7?0 4]mm), diameter stenosis (DS) (% vs %) was observed instantly after the procedure between the two groups, the required balloon inflation pressure was significantly lower with CBA ([8 3?0 9]atm vs [14 7?4 6]atm, P
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AIM: Using simvastatin and vitamine E (Vit-E) treatment to coronary artery disease (CAD) patients with low HDL, to investigate the relationship between Ox-LDL, platelet activation and HDL. METHODS: 40 CAD patients with low HDL were divided into two groups (A and B): A group oral simvastatin, B group oral simvastatin and Vit-E. The level of serum Ox-LDL, TXB 2 and GMP-140 were measured before and after treatment. The relationship between Ox-LDL, TXB 2, GMP-140 and HDL were analysed. RESULTS:The level of serum HDL was significantly increased in A and B group after treatment and attained normal level. The level of serum Ox-LDL, TXB 2 and GMP-140 were decreased significantly after simvastatin and Vit-E treatment and neared normal. CONCLUSIONS:This study confirmed that HDL can effectively refrain LDL oxidation. It also revealed that Vit-E and simvastatin treatment were more effectively refrained platelet activation by increasement of HDL and decreasement of Ox-LDL.
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AIM: To investigate the effect of hyperhomo cysteine on endothelial cell function. METHODS: By establishing hyperhomocysteinemia model, 18 male New Zealand rabbits were divided into control group (control group) and high-methi o nine-diet group (M group). At the end of 3 weeks, M group was divided again into M+0 group (continuing high methionine-diet) and M+F group (high-methionine-diet plus folic acid, vitamin B 12). At the end of 6 weeks, isolated aortic rin g s were made and the maximum vasodilation of the aortic rings to Ach was investig ated. Meanwhile, the plasma concentrations of Hcy, NO, ET-1, Ang II at 0 week, 3 weeks and 6 weeks and the contents of NO 2-/NO 3-, Ang Ⅱ, ET-1, NOS i n regional vascular tissue at 8 weeks were also measured. RESULTS: (1) In contrast to M+F group and control group, the max imum vasodilation to Ach were decreased (E max=26.73?4.51 vs 47.84 ?5.62, 56.42?7.82, P