Objective: To investigate the role of miR-19a/b in myocardial ischemia reperfusion injury (R/I) with its clinical significance. Methods: Our research included in 2 parts. Part 1: in H9c2 cells. miR-19a/b expression in H9c2 cells with 10 h hypoxia and 2 h re-oxygenation was detected by real-time PCR, miR-19a/b potential target gene was assessed by luciferase reporter activity assay. Part 2: in natural person. Control group,n=40 healthy subjects and AMI (acute myocardial infarction) group,n=40 relevant patients. Peripheral blood levels of miR-19a/b were detected as Part 1, cTnI were measured by chemiluminescence immune analysis and CK-MB were assessed by immune inhibition method. Results: Part 1: Compared with normal H9c2 cells, miR-19a/b expressions were increased 5.876 times and 2.761 times in H9c2 cells with 10 h hypoxia and 2 h re-oxygenation, bothP<0.01. With respectively transfected miR-19a/b mimic and inhibitor, miR-19a/b expression was up-regulated and down-regulated respectively. miR-19a/b and chromosome-10 deleted phosphatase, tensing homolog gene (PTEN) had the targeting effect. With up-regulated miR-19a/b expression, PTEN level was decreased and with down-regulated miR-19a/b expression, PTEN level was increased. Part 2: Compared with Control group, AMI group had elevated blood level of miR-19a/b,P<0.01. In AMI patients, miR-19a/b was increasing at 0-3 h of chest pain and reaching the peak at 6-12 h; CK-MB enzyme activity and cTnI content were elevating at 2-6 h of onset and reaching the peak at 24 h. Conclusion: miR-19a/b expression was up-regulated by myocardial ischemia reperfusion in H9c2 cells, PTEN was the potential target gene of miR-19a/b. Circulating miR-19a/b might be used as a new non-invasive biological marker for myocardial ischemia R/I diagnosis.

Aim To explore the effects of Puerarin on the Apelin and its receptor APJ of lung tissue in rats with hypoxia pulmonary hypertension.Methods Thirty SD rats were randomly divided into normal control group(N),and hypoxia hypercapnia group(F),and hypoxia hypercapnia+Puerarin group(P).The levels of Apelin-36 in serum and in lung tissue were measured by radioimmunity,the expressions of Apelin and APJ in pulmonary tissuse were observed by RT-PCR.Results ①Mean pulmonary arterial pressure(mPAP),weight ratio of RV to LV+S,the levels of Apelin in serum and in lung tissue of group F were significantly higher than those of group N(P

Objective To investigate the changes of PTX3,NT -ProBNP level and the correlation between them and the prognosis of the patients with AMI combined T2DM.Methods 143 patients with primary AMI were enrolled,of which 63 patients with type 2 diabetes (observation group)and 80 patients with no diabetes (control group).Plasma PTX3 values were measured with ELISA method and NT -ProBNP was detected by the electrochemi luminescence method.All patients were observed for the occurrence of MACE during hospitalization and 12 months after discharge.The correlation between PTX3,NT -ProBNP concentration and occurrence of MACE in observation group were analyzed.Results The PTX3,NT -ProBNP concentration[(8.95 ±5.06)ng/mL,(1 609 ±1 049)pg/mL]in the observation group were significantly higher than those in the control group[(7.03 ±3.70)ng/mL,(1 198 ± 809)pg/mL](P =0.010,P =0.009);The number(n =26)of patients with occurrence of MACE in the observation group was significantly higher than 15 cases in the control group(P =0.003).In the observation group,the PTX3, NT -ProBNP concentration[(10.98 ±5.45)ng/mL,(2 007 ±1 097)ng/mL]in patients with MACE were signifi-cantly higher than those in patients without MACE[(7.53 ±4.28)ng/mL,(1330 ±930)ng/mL](P =0.007,P =0.010),and in MACE group,the PTX3,NT -ProBNP concentration[(13.88 ±6.84)ng/mL,(2 596 ±1 333)ng/mL] in patients with death weresignificantly higher than those in patients without death[(9.18 ±3.52)ng/mL,(1 639 ± 751)ng/mL](P =0.029,P =0.023).Conclusion More strong chronic inflammatory reaction and serious myocar-dial injury might occur in the patients with AMI combined and T2DM,and those patients would have poorer prognosis. The combined detection of PTX3,NT -ProBNP has an important significance in evaluating of the degree of myocardial damage and the prognosis of the patients with AMI combined T2DM.

AIM: To investigate the effect of chronic hypoxia-hypercapnia and L-arginine (L-Arg) liposome on L-Arg transport in rats pulmonary artery. METHODS: Forty Sprague-Dawley rats were randomly divided into four groups, normal control group (NC), chronic hypoxia-hypercapnia group (HH), chronic hypoxia- hypercapnia group+L-Arg (HL) and chronic hypoxia-hypercapnia group+L-Arg liposome (HP). Changes in pulmonary artery L-Arg transport and pulmonary arterial microscopy were observed. RESULTS: (1) The mean pulmonary artery pressure (mPAP) and weight ratio of right ventricle to left ventricle and septum (RV/LV+S) in HH group were higher than those in NC group, and in HP group was lower than that in HH group and HL group, but there was no significant difference between HL group and HH group; (2) At 0.005 mmol/L, 0.01mmol/L, 0.02mmol/L, 0.05 mmol/L, 0.1 mmol/L and 0.2mmol/L concentration of L-Arg, the velocity of L-Arg transport in HH group was lower than that in NC group, and in HL group higher than in HH group, and in HP group was much higher than that in HH group and in HL group. (3) Light microscopy showed that vessel well area/total area (WA/TA) and media thickness of pulmonary arterioles (PAMT) were much higher in rats of HH group than those in NC group, WA/TA and PAMT in HP group were obviously improved. CONCLUSION: The above results indicated that there existed a functional disturbance in L-Arg transport of pulmonary artery in rats chronically exposed to hypoxia-hypercapnia, and it was obviously enhanced when liposome was used as L-Arg carrier. Thus, it appears that liposome-L-Arg may have clinical perspective in the treatment of chronic hypoxic pulmonary hypertension.

AIM:To explore whether YAP protein is important in induced pluripotent stem cell ( iPSC)-induced cardiovascular progenitor cell and/or vascular smooth muscle differentiation .METHODS:Using episomal vector based reprogramming , we generated human iPSCs from donor fibroblasts .We used both this iPSCs and human H 1 embryonic stem cells to differentiate into vascular smooth muscle cells (VSMCs) through cardiovascular progenitor cells (CVPC).Western blotting, qPCR and immunofluorescence microscopy were used to check the expression of YAP and related genes during this differentiation process .RESULTS:The results showed that iPSCs expressed pluripotent stem cell markers, such as Oct4, Nanog, Sox2, TRA-1-60 and SSEA3, and could form teratoma in SCID mice.YAP was highly expressed in pluripotent stem cells , but dramatically decreased when CVPC differentiation started .YAP gradually increased dur-ing CVPC three-day differentiation.The TAZ and YAP binding partner TEAD1, but not TEAD2 and TEAD4, have similar expression pattern in CVPC differentiation .Immunofluorescence result confirmed that YAP was activated and accumulated in nucleus .Interesting-ly, both YAP and phosphorylated YAP expression decreased to very low level after CVPC differentiated into VSMCs in 7 days.TEAD4 and TAZ also decreased, while TEAD1, TEAD2 and TEAD3 expression did not change during VSMC differentiation .CONCLU-SION:YAP and TEAD1 expression increased during CVPC differentiation , while YAP and TEAD4 expression decreased from CVPC to VSMCs differentiation , which suggested YAP might have different function during diverse cell differentiation .

AIM: To investigate the roles of nitric oxide/L-arginine(NO/L-Arg) pathway and urotensin-Ⅱ(UⅡ) in the development of pulmonary hypertension induced by chronic hypoxia-hypercapnia in rats.METHODS: Forty male Sprague-Dawley rats were randomly divided into four groups(n=10): normal control group(A),hypoxia-hypercapnia+saline group(B),hypoxia-hypercapnia+L-Arg liposome group(C) and hypoxia-hypercapnia+N-nitro-L-arginine methyl ester(L-NAME) group(D).Contents of UⅡ,UⅡ mRNA and receptor of UⅡ(UT) mRNA in pulmonary arterioles were measured with immunohistochemistry analysis and in situ hybridization,respectively.Change of small pulmonary vascular microstructure was also investigated.RESULTS:(1) The mean pulmonary artery pressure(mPAP) and the weight ratio of right ventricle to left ventricle plus septum [RV/(LV+S)] in B and D groups were all higher than those in A group(respectively,P

AIM: To investigate the expression of matrix metalloproteinases(MMPs) in pulmonary arterioles of rats with chronic hypoxia and hypercapnia-induced pulmonary hypertension. METHODS: MMP-2, MMP-9 and MMP-2 mRNA, MMP-9 mRNA were observed in pulmonary arterioles by the techniques of immunohistochemistry and in situ hybridization. RESULTS: ①The mean pulmonary artery pressure (mPAP) and weight ratio of right ventricle to left ventricle and septum (RV/LV+S) of hypoxia-hypercapnia groups were higher than those of normal control group ( P

AIM:To investigate the effect of puerarin on pulmonary vessel collagen metabolism in pulmonary hypertension rats induced by chronic hypoxia and hypercapnia. METHODS: Collagen Ⅰ,Ⅲ and their mRNA were observed in pulmonary arterioles by the technique of immunohistochemistry and in situ hybridization. RESULTS: ① Light microscopy showed media thickness of pulmonary arterioles was much higher in HH(hypoxic-hypercapnia) group than that of NC(normal control) group, and, vessel cavity turned more straiter in HH group than that of NC group.However, the damage of pulmonary arterioles in HP(hypoxic-pueratin) group was much slighter than that of HH group. ② The levels of plasma ET-1 and lung homogenates Hyr were much higher in HH group than those of NC group( P 0.05). CONCLUSION: Puerarin inhibited the deposition of collagen and improved pulmonary vessel remodeling.

AIM:To investigate rat Urotensin-II(ra t U-II)-induced vasoconstriction of rat main pulmonary arteries and the role of mitogen-activated protein kinase(MAPK). METHODS: The main pulmon ary artery was dissected from the male Sprague -Dawley rats and artery ring width was 3-4 mm. Concentration-response curves wer e gene rated to rat U-II(0 03 nmol/L-30 nmol/L).Inhibitor of MAPK,PD 98059(0 1 ?mol/ L -10 ?mol/L) were added into the medium after rat U-II(30 nmol/L)induced vasoc onstriction had reached plateau to construct the relaxant concentration-respons e curves and their EC 50 and E max . RESULTS: Rat U-II was a potent vasoconstrictor of isolated rat main pulmonary arteries [EC 50 =7 95?0 4 0, E max =(14 28?6 34)% of the response to 60 mmol/L KCl]; PD 98059 caused c oncentration-dependent relaxations of rat U-II precontracted arteries [EC 50 =5 91?0 45, E max =(81 39?13 65)%]. CONCLUSION: Rat U- II was a potent vasoconstrictor of rat main pulmonary arteries and this response was med iated through MAPK.

Objective Essential polyunsaturated fatty acids(PUFA) can not been synthesized by the body-self.Serum triglycerides n-6 and n-3 PUFAs directly or indirectly reflect the corresponding unsaturated fatty acids intake from meals.This study was to investigate the relationship of serum triglycerides polyunsaturated fatty acid composition with the ratio of n-6 to n-3 PUFA(n-6/n-3 PUFA) and magnetic resonance imaging measured intra-abdominal fat(MRI-IAF) with other body fat parameters of patients with metabolic syndrome.Methods Thirty-six patients with metabolic syndrome and 41 healthy controls were enrolled in this investigation.The relevance of serum triglycerides polyunsaturated fatty acids with MRI-IAF was observed.A stepwise regression analysis was applied to determine which kind of triglycerides polyunsaturated fatty acid could predict MRI-IAF,waist circumference and body mass index (BMI) more potent.Results In the metabolic syndrome group,blood sugar,lipid profiles,blood pressure,visceral fat accumulation-related parameters and serum triglycerides polyunsaturated fatty acid composition was worse than those in the control group.Compared with the healthy controls,n-6/n-3 PUFA was significantly increased(t=8.564,P＜0.05),although C18∶3 n-3,C20∶5 n-3(EPA),C22∶6 n-3(DHA) and n-3PUFA were significantly declined(t=-2.920,-7.034,all P＜0.05) in metabolic syndrome group.The difference of n-6 PUFA showed no statistically significant difference(t=-0.957,-1.494,P＞0.05).n-6/n-3 PUFA,n-3 PUFA,EPA and DHA were associated with MRI-IAF(r=-0.377,0.565,all P＜0.05); n-6/n-3 PUFA was correlated with the waist circumference(r=0.400,P=0.016) and BMI(r=0.357,P=0.033),while n6 PUFA showed no correlation with body fat parameters.N-6/n-3 PUFA was more potent to predict MRIIAF,waist circumference and BMI(adjusted R2=0.102,0.299,all P＜0.05) than other polyunsaturated fatty acids.Conclusions The ratio of n-6/n-3 PUFA and n-3 PUFA may be positively correlated with EPA and DHA could be inversely associated with MRI-IAF and other body fat-related parameters in patients with metabolic syndrome,while n-6 PUFA did not show such a relationship.The ratio of n-6/n-3 PUFA might be more potent to predict MRI-IAF and other body fat-related parameters.