Objective To observe the influence of different level of hyperhomocysteinemia on mRNA and protein expressions of KV1 .3 ,CaN,NFAT,IL-6 and TNF-αin lymphocytes of patients with acute ST-segment elevation myocardial infarction (STEMI).Methods We selected 90 STEMI patients and divided them into three groups according to the level of plasma homocysteine:the first experimental group (STEMI group,Hcy<1 5μmol/L, n=30),the second experimental group (STEMI with mild Hhcy group,Hcy 15~30μmol/L,n=30)and the third experimental group (STEMI with intermediate Hhcy group,Hcy>30 μmol/L,n=30 ).Another 30 healthy examined people were selected as control group (n=3 0 ).Peripheral lymphocytes were isolated by Ficoll density gradient centrifugation.The Hcy in the plasma was measured with the IMX assays.Real-time quantitative PCR (RT-PCR)was used to detect mRNA expressions of KV1.3,CnAα,NFAT1,IL-6 and TNF-αand Western blot technique was used to detect the expressions of KV1.3,CnAαand NFAT1.Results The mRNA and protein expression levels of KV1.3,CnAαand NFAT1 in each experimental group were significantly higher than those in control group (P<0 .0 5 or P<0 .0 1 ).Multiple comparison in each experimental group showed that compared with that in the first experimental group,the expression level of the second experimental group increased (P<0.05 or P<0.01)and compared with first and second experimental groups,the expression level of the third experimental group increased (P<0.05 or P<0.01).The mRNA expression levels of IL-6 and TNF-αin each experimental group were significantly higher than those in control group (P<0.05 or P<0.01 ).Multiple comparison in each experimental group showed that compared with that in the first experimental group,the expression level of the second experimental group increased (P<0 .0 5 or P<0 .0 1 )and compared with first and second experimental groups,the expression level of the third experimental group increased (P<0.01).Plasma total Hcy levels were positively correlated with mRNA and protein expressions of KV1.3 in all observed groups (r=0.503 P=0.000,r=0.726 P=0.000).Conclusion The higher level of Hcy in plasma,the higher mRNA and protein expression levels of KV1.3,CnAα,NFAT1 and the higher mRNA expression levels of IL-6,TNF-αin the lymphocyte of STEMI patients,which may be one mechanism for Hcy exacerbating the inflammatory reaction of STEMI.

Objective To investigate the influence of atorvastatin in methylation and expression level of Bcl-2 in human umbilical endothelial cells(HUVECs)treated with homocysteine(Hcy)and to expound potential mechanism of atorvastatin resisting arteriosclerosis.Methods After HUVECs were treated with 0, 2, 4, 8, 16, and 32 mmol·L-1 Hcy for 48 h,MTT was used to measure the inhibitory rates of HUVECs and the half inhibitory concentration (IC50 ). According to the experimental results, the HUVECs cultured in vitro were divided into control group (0.00 mmol · L-1 Hcy ), Hcy group (9.00 mmol·L-1 Hcy ), and atorvastatin group (9.00 mmol·L-1 Hcy+1×10-3 mmol·L-1 atorvastatin).After treated for 48 h,flow cytometry was used to detect the apoptotic rate of cells, the mRNA expression of Bcl-2 was analyzed by fluorescence quantitative PCR,the protein expression of Bcl-2 was detected by Western blotting method, and the methylation level of Bcl-2 promoter region was determined by nest touch-down PCR combined with methylation specific PCR (MSP ). Results Compared with control group,the apoptotic rate of HUVECs in Hcy group was increased(P<0.01),the mRNA and protein expression levels of Bcl-2 were significantly decreased(P<0.01),and the Bcl-2 promoter region methylation level was also decreased(P<0.01).Compared with Hcy group,the apoptotic rate of HUVECs in atorvastatin group was decreased(P<0.01),the mRNA and protein expression levels of Bcl-2 gene were increased (P<0.05), and the Bcl-2 promoter region methylation level was also increased (P<0.05). Conclusion Atorvastatin can prevent the apoptosis of HUVECs induced by Hcy through regulating Bcl-2 methylation.

Objective To investigate the clinical features and characteristics of coronary artery lesion between Hui and Han nationality young pa?tient with acute myocardial infarction(AMI)who were referred to the affiliated hospital of Ningxia Medical University. Methods A total of 189 con?secutive AMI young patients(age≤44 years)who underwent coronary angiography were retrospectively retrieved from the database.Those patients with AMI were divided into Hui group(46 cases)and Han group(143 cases). The clinical features and results of coronary angiogram were com?pared between the two group. Results Compared with Han group,Hui group are more younger than Han group,high prevalence rate of diabetes, lower smoking history and lower drinking history(P<0.05). Coronary angiography showed the incidence of three?vessel lesions was significant low?er in Han group than in Hui group(P<0.05). Both group showed single vessel was the most common lesion. Conclusion Hui nationality patients with acute myocardial infraction are more younger and are are more prone to suffering from diabetes history、lower smoking history and lower drinking history than Han nationality patients. The coronary artery lesions of Hui nationality patients with acute acute myocardial infraction are more three?branch lesions than Han nationality patients.

Objective To assess under different vessel diameter ,the effect of the aspiration thrombectomy catheter in improving the myocardial reperfusion and clinical prognosis in patients with acute myocardial infarction (AMI)who were undergone primary percutaneous coronary intervention(PCI) .Methods 205 patients with AMI immediate implant stents after thrombus suction ,the TIMI flow grade(myocardial infarction thrombolysis treatment test flow classification ) ,postoperative ecg evolution ,incidence of no-reflow MACE in 30 days and MACE in 6 months were compared between conventional thrombus suction group and suction again group(blood vessels of <3 .0 mm and ≥3 .0 mm) .Results The level 3 blood flow rate ,MACE in 6 months in suction again group with blood vessels of ≥3 .0 mm had improved significantly ,but had no beneficial effects in blood vessels of ≥3 .0 mm .Conclusion In AMI patients treated with primary PCI ,application of aspiration thrombectomy catheter with blood vessels of ≥3 .0 mm may im-prove the flow condition before infarction related blood vessels ,reduce MACE .

Objective To investigate the relationship between plasma homocysteine (Hcy), Kv1.3 channel and cardiac troponinI (cTnI) in patients with acute ST-segment elevation myocardial infarction (STEMI). Methods According to the level of Hcy, 80 STEMI patients were divided into STEMI with Hhcy group (Hcy > 15 μmol/L, n=41) and control group (STEMI group, Hcy≤15μmol/L, n=39). The Hcy, blood lipid and cTnI were detected with automatic biochemistry analyzer, respectively. Peripheral lymphocytes were isolated by ficoll density gradient centrifugation. Real-time PCR was used to detect mRNA expression of Kv1.3, and Western blot assay was used to detect protein expression of Kv1.3. Results cTnI concentrations were obviously higher in STEMI with Hhcy group than those in STEMI group (μg/L:22.997 ± 5.880 vs. 12.881 ± 6.343;P<0.01). Multiple linear regression analysis showed that age, gender, hypertension, diabetes mellitus, smoking, family history, total cholesterol (TC), triglyceride (TG), high density lipoprotein-cholesterol (HDL-C) and low density lipoprotein-cholesterol (LDL-C) had no obvious influence on Hcy (P>0.05). The relative expression levels of Kv1.3 mRNA and protein were significantly higher in STEMI with Hhcy group (1.35±0.14, 0.85±0.12) than those in STEMI group (1.00 ± 0.07, 0.64 ± 0.05, P<0.05). Moreover, there was a positive relation between Hcy level and the mRNA and proteinexpression of Kv1.3 channel (r=0.299, r=0.542, P<0.05). There was a positive relation between protein expression levels of Kv1.3 channel and cTnI (r=0.644, P<0.05). Conclusion Our results support that Hcy could exacerbate the concentration of cTnI through playing an important role in the Kv1.3 mRNA and protein expression in lymphocytes.

Objective This retrospective study is designed to analyze the cardiovascular events of CTO to provide new information on secondary prevention of CHD in patients after hospital discharge. Methods 272 patients with definite diagnosis of CTO were enrolled in this study. Patients were divided into two groups according to whether suffering from cardiovascular events, with 167 patients in group A who had not suffered from the cardiovascular events and 105 patients in group B who had suffered from the cardiovascular events. We com-pared the clinical data, severity of coronary artery lesion, treatment in two groups. Results Between two groups, there was statistic signifi-cance in LDL -C, EF, Gensini scots, the number of coronary artery lesion, the number of chronic total coronary occlusion and PCI success. Logistic regression analysis revealed that Gensini scots was the independent factors for prognosis of CTO. Conclusion Gensini scors was the independent factor for prognosis of CTO.

Objective To investigate the effects of homocysteine(Hcy) on myocardial injury and its possible mechanisms.Methods The selected H9C2 cardiomyocytes were intervened with various concentrations of Hcy and 4-phenyl butyric acid(4-PBA).The H9C2 cells were divided into the control group,H400 group and H400P2 group.The control group used the common medium,the H400 group was added with 400 μmol/L Hcy,the H400P2 group was added with 2 mmol/L 4-PBBA on the basis of H400 group.The cell livability was detected by using cell counting kit-8 (CCK-8).Apoptosis was evaluated by using the terminal deoxynucleotidyl transferase mediated nick-end labelling(TUNEL) staining.The ERO1α expression was determined by using immunocytochemistry,and the protein expression difference was determined by using Western blot.Results The injury of Hey on H9C2 cardiomyocytes showed a concentration-dependent manner(F=2 039.958,P＜0.01).Compared with the control group,the apoptosis percentages and expression levels of PERK,p-PERK,CHOP and ERO1α in the H400 group were increased(P＜0.01);while which in the H400P2 group were decreased,the difference was statistically significant(P＜0.05).Conclusion Hcy mediates myocardial apoptosis through endoplasmic reticulum stress mechanism.

Aim To analyze the risk factor of the cardiac rupture(CR)in patients with acute ST-segment eleva-tion myocardial infarction(STEMI).Based on this,the nomogram model of acute STEMI patients with CR was estab-lished.Methods Through Ningxia Medical University General Hospital's big data research platform and hospital in-formation system retrieval,5 412 patients with acute STEMI from January 2015 to December 2019 were continuously includ-ed in the study,of which 91 patients with CR were included as CR group;5 321 patients non-combined with CR were in-cluded as non-CR group.LASSO regression,univariate and multivariate Logistic regression were used to analyze the risk factors of CR in patients with acute STEMI,and the CR nomogram predictive model was established.The nomogram mod-el was validated and evaluated by using receiver operating characteristic(ROC)curve,Hosmer-Lemeshow test and clinical decision curve analysis(DCA).Results LASSO regression results showed that age,female,hypertension history,first medical contact time,shock index,Killip grade,white blood cell count,d-dimer,lactic acid,anterior myocardial in-farction,β-blocker administration within 24 hours,angiotensin converting enzyme inhibitor/angiotensin receptor antagonist(ACEI/ARB)administration within 24 hours,emergency percutaneous coronary intervention(PCI)were 13 risk factors of CR(P＜0.05).The screened 13 risk factors were analyzed by univariate and multivariate Logistic regression,the results suggested that age,Killip grade,first medical contact time,white blood cell count,not undergoing emergency PCI and not taking ACEI/ARB drugs within 24 hours were the risk factors of CR in patients with acute STEMI.The acute STEMI with CR nomogram model was established according to the above 6 risk variables.The area under the ROC curve before and after the internal verification of the nomogram model was 0.946(95％CI:0.927～0.961),0.947(95％CI:0.927～0.959),and the sensitivity was 0.957 and 0.904,respectively,the specificity was 0.858 and 0.876,respectively,which indicated that the model had good discrimination degree.The Hosmer-Lemeshow test showed that the deviation between the predicted value and the observed value was not statistically significant(x2=12.70,P=0.122),indicating that the no-mogram model had a good calibration.The DCA curve indicated that the predictive probability threshold of the model was from 0.00 to 0.40,and the clinical net benefit was the highest,indicating that the model had good clinical efficacy.Conclusion The nomogram model established in this study has better distinction,calibration and clinical effectiveness.It can effectively predict the probability of acute STEMI with CR,and provide some help for clinical diagnosis and treat-ment,so as to reduce the incidence of CR.

OBJECTIVETo investigate the characteristics for activated coagulation factor VII(F VIIa) and the R353Q, -323 0/10 bp, HVR4 polymorphisms in the gene in patients with coronary heart disease (CHD) and myocardial infarction from Ningxia Hui and Han populations.

METHODSFour hundred and twenty angiographically proven CHD patients in the Hui population, and 508 healthy blood donors were tested for their plasma levels of coagulation factor VII using recombinant tissue factor method. The coagulation factor VII gene R353Q, -323 0/10 bp and HVR4 genotypes were identified by polymerase chain reaction. In addition, 600 Han patients with CHD and 604 healthy Han control subjects were also investigated.

RESULTS(1) The plasma F VIIa levels was significantly higher in patients with CHD and myocardial infarction than that in healthy control subjects and angor pectoris (P<0.01) in both Hui and Han populations. (2) There were significant differences in the distribution of genotypes and allelic frequencies of the R353Q between myocardial infarction and angor pectoris disease in the Hui population (P<0.05). So was the -323 0/10 bp locus in both the Hui and Han population. (3) The F VIIa level was significantly higher in individuals with RR genotype than those of Q allele carriers in the Hui population.

CONCLUSIONThere are polymorphisms of the F VII gene R353Q, -323 0/10 bp and HVR4 in the Hui and Han populations. The Q allele might be a protective factor against myocardial infarction in the Hui, and the plasma F VIIa level may be influenced by the R353Q polymorphism of the F VII gene. The 10 allele may be a protective factor against myocardial infarction in both the Hui and Han populations.

Aged ; Asian Continental Ancestry Group ; ethnology ; genetics ; Case-Control Studies ; Factor VII ; genetics ; metabolism ; Female ; Humans ; Male ; Middle Aged ; Myocardial Infarction ; ethnology ; genetics ; metabolism ; Polymorphism, Genetic

BACKGROUND:There is an internal relationship between hyperhomocysteinemia and vascular calcification.However,the pathogenesis of hyperhomocysteinemia promoting vascular calcification is still unclear. OBJECTIVE:To investigate the role of bone morphogenetic protein-2 in hyperhomocysteinemia-induced vascular calcification. METHODS:Human carotid wax samples were divided into a calcified group(n=29)and a non-calcified group(n=13)according to the presence or absence of calcified plaque.Sixteen ApoE-/-mice were randomly divided into a control group and a hyperhomocysteinemia group,with 8 mice in each group.Bone morphogenetic protein-2 vector was used to transfect rat thoracic artery smooth muscle A7r5 cells,and gradient concentration of homocysteine(50,100,200,and 400 μmol/L)was utilized to treat A7r5 cells.Calcification was detected by alizarin red staining and hematoxylin-eosin staining.The interaction of bone morphogenetic protein 2 with Runt-related transcription factor 2 was detected by immunofluorescence,and the expressions of bone morphogenetic protein 2,Runt-related transcription factor 2,and α-smooth muscle actin were detected by immunohistochemistry and western blot assay. RESULTS AND CONCLUSION:(1)Human carotid artery tissue staining revealed that compared with the non-calcification group,inflammatory cells increased and calcification positive rate increased in the calcification group(P<0.05).Compared with the non-calcification group,the expressions of bone morphogenetic protein-2 and Runt-related transcription factor 2 were up-regulated,and the expression of α-smooth muscle actin was decreased in the calcification group(all P<0.05).(2)The staining of mouse arterial specimens exhibited that,the positive rate of calcified area in the hyperhomocysteinemia group was significantly higher than that in the control group(P<0.05);serum homocysteine level in the hyperhomocysteinemia group was significantly higher than that in the control group(P<0.05).Compared with the control group,the expressions of bone morphogenetic protein-2 and Runt-related transcription factor 2 were up-regulated,and the expression of α-smooth muscle actin was decreased in the hyperhomocysteinemia group(all P<0.05).(3)A7r5 cell culture analysis demonstrated that with the increase of homocysteine concentration gradient,the degree of calcification,the content of bone morphogenetic protein-2 and Runt-related transcription factor 2 protein in A7r5 cells increased(P<0.05),and the content of α-smooth muscle actin protein decreased(P<0.05).(4)The A7r5 cell culture analysis of overexpressed bone morphogenetic protein 2 showed that the calcification degree of the overexpressed bone morphogenetic protein 2 group was increased compared with the corresponding control group,the β-sodium glycerophosphate group,and the homocysteine group.RUNt-related transcription factor 2 expression up-regulated(P<0.05)and α-smooth muscle actin expression down-regulated(P<0.05).(5)The expression of bone morphogenetic protein 2 increased in A7r5 cells cultured with homocysteine in calcified medium,and the expression of Runt-related transcription factor 2 increased with the increase of bone morphogenetic protein 2 expression.(6)The results confirm that bone morphogenetic protein-2 is a key target gene in the regulation of smooth muscle cell phenotypic transformation resulting in vascular calcification by hyperhomocysteinemia.Targeted regulation of bone morphogenetic protein-2 reduces hyperhomocysteinemia-induced vascular calcification.