Abstract:Atherosclerosis is a chronic inflammatory response caused by activation of endothelial cells, lipid deposition, and other relative pathogenic factor. Omega-3 polyunsaturated fatty acids (n-3 PUFA) play an important protective role in atherosclerosis, however, the underlying mechanism remains unclear. n-3 PUFA could produce different biological active eicosanoid metabolites by cyclooxygenase, lipoxygenase and cytochrome P450 oxidase, alternatively, it competes with arachidonic acid for the common metabolic enzymes to regulate the levels of arachidonic acid-derived eicosanoid metabolites. Based on the metabolic perspective, this paper systematically discusses the atherosclerosis protective mechanism that n-3 PUFA exerts by affecting the eicosanoid metabolism profile.

Abstract:Aim To investigate the mechanism of fibroblast growth factor 21 (FGF21) in promoting cholesterol efflux from foam cells. Methods THP-1 derived foam cells were treated with FGF21 at different concentrations (0,0, 0,0, 400 μg/L) for 24 hours and 200 μg/L FGF21 at different time (0,6, 2,4, 48 h), Western blot, laser confocal were used to detect LC3, and MDC staining for autophagosomes analysis, HPLC, oil red O staining for intracellular cholesterol accumulation measurement, and liquid scintillation counting for cholesterol efflux detection. Results After 200 μg/L, 400 μg/L FGF21 and 200 μg/L FGF21 were applied for 24 h and 48 h, the levels of total cholesterol (TC), free cholesterol (FC) and cholesterol ester (CE) in foam cells were significantly reduced, while their cholesterol efflux was significantly enhanced. Mechanism studies found that FGF21 induced the formation of autophagosomes in foam cells, and the conversion rate of microtubule-associated protein Ⅰ light chain3 (LC3-Ⅰ) to microtubule-associated protein Ⅱ light chain3 (LC3-Ⅱ) was significantly increased. However, after autophagy was inhibited with autophagy-related gene 5(ATG5) siRNA or 3-methyladenine (3-MA) or Bafilomyein A1 (BafA1), effect of FGF21 on decreasing TC, FC, CE and lipid accumulation, and increasing cholesterol efflux were reduced. Conclusion FGF21 promotes cholesterol efflux in foam cells by up-regulating autophagy.

Abstract:Aim To investigate whether the recombinant adenovirus with short chain acyl CoA dehydrogenase (SCAD) injected via tail vein can improve the vascular remodeling in spontaneously hypertensive rats (SHR). Methods The experiment was divided into 6 groups:Wistar+NS group, Wistar+GFP group, Wistar+Ad-SCAD group, SHR+NS group, SHR+GFP group and SHR+Ad-SCAD group. After purification of SCAD and GFP packed with adenovirus, the drug was injected by tail vein for 8 weeks. The cardiac function of rats was detected by echocardiography. The blood pressure changes of rats were detected by non-invasive blood pressure meter. HE staining, Sirius red staining, DHE staining, TUNEL staining, EVG staining, were used to observe the phenomenon of vascular remodeling. The expression of related proteins was detected by Western blot, and mRNA expression was detected by RT-PCR. The free fatty acid (FFA), nitric oxide (NO) content and ATP content were observed. Results (1) After SCAD recombinant adenovirus was injected via tail vein, the expression of SCAD protein was significantly upregulated in the aorta of rats, meanwhile, the mRNA level was observably increased, and the activity of SCAD was markedly increased. (2) In the pathological state of rats, the rising of SCAD can lower blood pressure, improve heart function and vascular lumen size, reduce collagen deposition, result a poor production of vascular ROS, consequently, give lower to apoptosis. (3) Under pathological conditions, overexpression of SCAD can reduce FFA content of serum and aorta, increase the ATP level in aorta, activate eNOS phosphorylation, increase NO production in aorta. Conclusion The upregulation of SCAD in aorta of SHR can reverse hypertensive vascular remodeling, which may be related to decreasing FFA content of serum, increasing NO levels, reducing ROS production and eliminating oxidative stress.

Abstract:Aim To investigate the role of DNA methylation of Rap1A in Hcy-induced RAW264.7 cell proliferation. Methods RAW264.7 cells in logarithmic growth phase were divided into control group (Control) and Hcy treated groups with different concentrations (0,0, 0,0, 100 μmol/L Hcy). Cell viability was detected by XTT to screen optimal intervention time and concentration of Hcy after cells were treated for 24 hours. Edu test was used to analyze cell proliferation. The expression levels of Rap1A mRNA and protein were measured by qRT-PCR and Western blot respectively. Methylation-specific PCR (MSP) was used to detect the change of Rap1A promoter region. The changes of Rap1A were examined by immunofluorescence staining. Rap1A interference adenovirus was transfected into RAW264.7 cells and stimulated with Hcy to detect the changes of mRNA and protein levels of Rap1A and cell proliferation. Results Compared with control group, the cell viability was enhanced after cells were treated with different concentration of Hcy, which was the most obvious when cells were treated by 100 μmol/L Hcy (P＜0.01), as well as cell proliferation(P＜0.01). But there was no time and concentration dependence. After Hcy stimulation, the expression of mRNA and protein of Rap1A increased significantly(P＜0.01). The result of MSP revealed that the methylation level of Rap1A promoter region was decreased (P＜0.01). Interfering with the expression of Rap1A can partially reverse the proliferation of cells induced by Hcy (P＜0.01). Conclusion DNA hypomethylation of Rap1A promoter region may play an important role in macrophage proliferation induced by Hcy.

Abstract:Aim To observe the targeting regulation of miR-497-5p on nucleotide-binding oligomerization domain-like receptor protein 1 (NLRP1) and its effect on cholesterol efflux. Methods Bioinformatics and luciferase reporter gene assay were used to verify the targeted binding of miR-497-5p and NLRP1. Human THP-1 monocytes were induced into foam cells after treatment with phorbol ester and oxidized low density lipoprotein. miR-497-5p mimic and miR-497-5p inhibitor were used to treat cells. Real-time fluorescence quantitative PCR and Western blot were used to detect the expressions of NLRP1, cysteinyl aspartate specific proteinase 1 (Caspase-1) and apoptosis-associated speck-like protein containing a Caspase-recruitment domain (ASC). The contents of interleukin-1β (IL-1β) and IL-18 in cell culture medium were determined by enzyme-linked immunosorbent assay. Cholesterol efflux detection was performed by liquid scintillator. Lipid content in foam cells was detected by high performance liquid chromatography. Results The luciferase activity of wild type NLRP1 3′UTR reporter gene was significantly reduced by miR-497-5p mimic. Compared with the control group, the mRNA and protein expression levels of NLRP1, ASC and Caspase-1 in miR-497-5p mimic group were significantly down regulated, and the secretions of IL-1β and IL-18 was significantly reduced. Compared with the control group, miR-497-5p mimic significantly promoted the cellular cholesterol efflux and reduced the contents of total cholesterol, free cholesterol and cholesterol ester in cells. Conclusion miR-497-5p may inhibit the inflammatory response of macrophage-derived foam cells and promote cholesterol efflux by targeting regulation of NLRP1.

Abstract:Aim To study the effect of repetitive transcranial magnetic stimulation (rTMS) on the learning and memory function of rats in the recovery stage of cerebral infarction through insulin-like growth factor-1 receptor (IGF-1R) pathway. Methods Adult male SD rats were randomly divided into control group, model group, rTMS group and rTMS+ADW742 group. The latter three groups were used to establish cerebral infarction model by thread bolt method. Two weeks later, rTMS group was given rTMS intervention, rTMS+ADW742 group was given rTMS and ADW742 intervention for two weeks. Morris water maze was used to detect learning and memory function. TUNEL was used to detect apoptosis rate of hippocampus, Western blot was used to detect the expression of cytochrome-c (Cyt-C), cleaved Caspase-3, IGF-1, IGF-1R in hippocampus. Results Compared with the control group, the escape latency significantly prolonged, the times of crossing the platform and the expression of IGF-1, IGF-1R in the hippocampus significantly reduced, the apoptosis rate and the expression of Cyt-C, cleaved in the hippocampus significantly increased in model group(P＜0.05); Compared with the model group, the escape latency significantly was shortened; the times of crossing the platform and the expression of IGF-1, IGF-1R in the hippocampus were significantly increased; the apoptosis rate and the expression of Cyt-C, cleaved in the hippocampus were significantly reduced in model group(P＜0.05). Compared with model group, the escape latency was significantly prolonged; the times of crossing the platform and the expression of IGF-1, IGF-1R in the hippocampus were significantly reduced; the apoptosis rate and the expression of Cyt-C, cleaved in the hippocampus were significantly increased in rTMS+ADW742 group(P＜0.05). Conclusion rTMS can improve the learning and memory function of rats in the recovery period of cerebral infarction, and activating IGF-1R pathway and inhibiting apoptosis in hippocampus is the related molecular mechanism.

Abstract:Aim To investigate the effect of miR-499a-5p on hydrogen peroxide(H₂O₂)-induced proliferation and apoptosis of cardiomyocytes H9c2, and to explore its mechanism. Methods Cell viability kit (CCK-8) was used to detect the survival rate of H9c2 cells treated with different concentrations of H₂O₂ (0,0, 0,0 μmol/L) for 6 h, and 400 μmol/L treated H9c2 cells were selected as a model group. The model group cells were divided into miR-NC group, miR-499a-5p group, si-NC group, si-APC group, miR-499a-5p+pcDNA group, miR-499a-5p+pcDNA-APC group. Flow cytometry, western blotting(Western blot) and enzyme linked immunosorbent assay (ELISA) were used to detect the survival rate, apoptosis rate, reactive oxygen species(ROS), superoxide dismutase (SOD), melondialodehyde(MDA) and proliferation-related protein expression proliferating cell nuclear antigen(PCNA), cyclins depend on kinase inhibitors(P21),Bü lymphoblastoma-2(Bcl-2),Xü gene associated with Bcl-2(Bax)of each group. Results H₂O₂(0,0, 0,0 μmol/L) inhibited the survival of H9c2 cells in a concentration-dependent manner with an optimal concentration of 400 μmol/L. The expression of miR-499a-5p was significantly decreased and the expression of APC was significantly increased in the model group. Overexpression of miR-499a-5p and inhibition of APC significantly attenuated H₂O₂-induced proliferation inhibition, apoptosis promotion and oxidation stress of H9c2 cells. miR-499a-5p can also target inhibition of APC. Overexpression of APC reversed the damage of H₂O₂-induced cardiomyocytes by miR-499a-5p.Conclusions miR-499a-5p can regulate the proliferation, apoptosis and oxidative stress of cardiomyocytes induced by H₂O₂. The mechanism is related to the targeted inhibition of APC, which will provide a new target for the treatment of myocardial cell injury induced by oxidative stress.

Abstract:Aim To investigate the relationship between vascular dysfunction and change of functional activities of circulating endothelial progenitor cells (EPC) in hypertensive urgency. Methods Thirteen hypertensive urgency patients(n＝13) and twenty normotension(n＝20) were recruited for the research. First of all, blood samples were collected and circulating EPC were isolated and cultured. The migration, proliferation and adhesion activities of circulating EPC were detected with Transwell method and CCK-8 assay respectively.In addition, the level of granulocyte-macrophage colony stimulating factor (GM-CSF), vascular endothelial growth factor (VEGF) and interleukin-6(IL-6) from both plasma and EPC production were measured by ELISA. The level of nitric oxide (NO) from both plasma and EPC production were determined by nitrate reductase method. Finally, brachial artery-based flow mediated dilatation (FMD) was used to determine vascular endothelial function. Results Compared with the normotension group, the migration, proliferation and adhesion activities of circulating EPC, the level of NO from both plasma and EPC production were dramatically decreased in hypertensive urgency group(P＜0.05). Furthermore, there was a strong correlation between FMD, NO (from both plasma and EPC production) and circulating EPC activities (migration, proliferation and adhesion activities).However, there were no significant differences among the level of GM-CSF, VEGF and IL-6 from both plasma and EPC production in two groups(P＞0.05). Conclusion The functional activities of circulating EPC were declined in hypertensive urgency, which is positively correlated with the level of NO and FMD.

Abstract:Aim To investigate the relationship between serum fibroblast growth factor-21 (FGF-21), myeloperoxidase (MPO) levels and prognosis in patients with acute myocardial infarction (AMI) after percutaneous coronary intervention (PCI). Methods 108 AMI patients who received PCI in our hospital from January 2016 to December 2018 were selected as the study subjects. According to whether major adverse cardiovascular events (MACE) occurred during follow-up, patients were divided into two groups:poor prognosis group (25 cases) and good prognosis group (83 cases).The levels of serum FGF-21 and MPO were detected by enzyme-linked immunosorbent assay and colloidal gold immunochromatography respectively. The relationship between FGF-21, MPO and MACE in AMI patients after PCI, and the efficacy of FGF-21, MPO in the diagnosis of MACE were analyzed. Results The levels of serum FGF-21 and MPO in the poor prognosis group were higher than those in the good prognosis group, and the differences were statistically significant (P＜0.05). Area under ROC curve (AUC) of FGF-21＋MPO in the diagnosis of MACE in AMI patients after PCI was 0.860, which was higher than that of FGF-21 and MPO alone in the diagnosis of MACE in AMI patients after PCI. In patients with FGF-21＞140.41 ng/L and FGF-21≤140.41 ng/L, the incidence of MACE was 39.58% and 10.00%, respectively, and the difference was statistically significant (P＜0.001). In patients with MPO＞419.42 μg/L and MPO≤419.42 μg/L, the incidence of MACE was 35.00% and 8.33%, respectively, and the difference was statistically significant (P＝0.001). Cox univariate and multivariate analysis showed that FGF-21 and MPO were closely related to MACE in AMI patients after PCI (all P＜0.05). Conclusion The levels of serum FGF-21 and MPO are related to the prognosis in AMI patients after PCI, and high levels of serum FGF-21 and MPO are closely related to MACE.

Abstract:Aim To investigate the clinical effect and postoperative complications of total thoracoabdominal aortic aneurysm repair (tTAAAR) in patients with thoracoabdominal aortic aneurysm (TAAA). Methods A retrospective analysis was made for 14 patients with TAAA who were treated by open surgery from April 2010 to April 2019 in our hospital, including 11 cases of Crawford Ⅱ and 3 cases of Crawford Ⅲ. There were 12 males and 2 females, 28-54 years old, with an average age for (36.1±7.1) years. 4 patients underwent traditional deep hypothermic cardiopulmonary bypass tTAAAR and 10 patients underwent modified normothermia non-cardiopulmonary bypass tTAAAR. The traditional way was to establish the arteriovenous pathway for extracorporeal bypass under deep hypothermia cardiopulmonary bypass, and the improved way was to establish the descending aorta iliac artery bypass under normothermia non-cardiopulmonary bypass. Results The operation of all the 14 patients were completed. The blocking time of descending aorta was (22.2±9.6) minutes, and the ischemic time of spinal cord was (23.0±7.3) minutes. Early postoperative death occurred in 2 cases, acute renal insufficiency in 4 cases, paraplegia in 3 cases, pulmonary infection in 4 cases, transient brain dysfunction in 4 cases. One patient underwent splenectomy during operation and one patient underwent tracheotomy after operation. Conclusion Open tTAAAR is a relatively safe and effective surgical method, which is currently an effective treatment for some complex TAAA.

Abstract:Aim To investigate the association between the fibrinogen/albumin ratio (FAR) and the severity of coronary artery disease in unstable angina (UA) patients. Methods Retrospective analysis was performed on 140 UA patients. According to the receiver operating characteristic(ROC) curve, patients were divided into two sub-groups based on the best critical value of FAR to predict moderate to severe coronary artery disease(Gensini score>20 scores). The clinical data, laboratory test results and coronary artery diseases of the two groups were compared. Multivariate Logistic regression analysis was used to study the related risk factors of moderate to severe coronary artery disease in UA. Results The best critical value of FAR value to predict moderate to severe coronary artery lesions in UA patients was 0.068 8, when the area under the curve was 0.705 (95%CI:0.613~0.797), the sensitivity was 74.0% and the specificity was 60.0%. There were significant differences in diabetes and smoking history between the two groups (both P＜0.05); The white blood cell count, low density lipoprotein cholesterol, total cholesterol, fasting blood glucose and fibrinogen were higher in the high FAR group than those in the low FAR group, while the levels of high density lipoprotein cholesterol and albumin were lower than those in the low FAR group(all P＜0.05); With the increase of the FAR ratio, the vascular lesions in the single branch gradually decreased, and the vascular lesions in the two and three branches and the Gensini score gradually increased. The differences were statistically significant (all P＜0.05). Spearman correlation analysis indicated that FAR was positively correlated with Gensini score (r＝0.606, P＜0.001). Multivariate Logistic regression showed that FAR≥0.068 8(OR＝7.553, P＝0.016) was an independent risk factor for moderate to severe coronary artery disease in UA patients. Conclusion In UA patients, the FAR value≥0.068 8 has certain value in predicting the severity of coronary artery diseases.

Abstract:Aim To investigate the predictive value of miR-150 for major adverse cardiovascular events (MACE) after percutaneous coronary intervention (PCI) in patients with acute ST segment elevation myocardial infarction (STEMI). Methods 130 patients admitted to our hospital from January 2016 to January 2019 who were diagnosed with STEMI and emergency PCI were selected. The patients were divided into MACE group (n＝36) and non-MACE group (n＝94) according to whether MACE occurred within 6 months after PCI. The expression level of miR-150 and general clinical data were compared between two groups. Risk factors of MACE in acute STEMI patients 6 months after PCI were analyzed by Logistic regression. Spearman correlation analysis was used to analyze the correlation between miR-150 and various risk factors. Receiver operating characteristic curve (ROC) was used to analyze the predictive value of miR-150 for MACE within 6 months after PCI in patients with acute STEMI. Results There were no significant difference between the two groups in gender, age, comorbidities (diabetes, hypertension, coronary heart disease, hyperlipidemia), infarction site, postoperative medication, emergency PCI time, high density lipoprotein cholesterol, low density lipoprotein cholesterol, total cholesterol, triglyceride and hemoglobin (P＞0.05). Heart rate (HR), systolic blood pressure, diastolic blood pressure, platelet, C-reactive protein (CRP), creatine kinase-MB (CK-MB), neutrophil lymphocyte ratio (NLR) in MACE group were significantly higher than those in non-MACE group, and left ventricular ejection fraction (LVEF) and miR-150 levels were significantly lower than those in non-MACE group (P＝0.000). Multivariate Logistic regression analysis showed that age, hypertension history, HR, LVEF, CRP, CK-MB, NLR, miR-150 were all independent risk factors for MACE within 6 months after PCI in patients with acute STEMI. The results of correlation analysis showed that miR-150 was negatively correlated with hypertension history, HR, CRP, CK-MB, NLR, and positively correlated with age and LVEF (P＝0.000). ROC curve showed that the cut-off point of MACE diagnosed by miR-150 within 6 months after PCI in acute STEMI patients was 0.23, and the area under curve was 0.905 (95%CI 0.871-0.939). Conclusion Low level of miR-150 is an independent risk factor of MACE within 6 months after PCI in patients with acute STEMI, and the detection of miR-150 level can help to evaluate the prognosis of patients with acute STEMI.

Abstract:Atherosclerosis (As) is a complicated chronic disease characterized by lipid deposition in the blood vessel wall, involving inflammatory and proliferative cascade of major functional cells, including smooth muscle, endothelial cells and immune cells. At present, the cognitive mechanism of atherosclerosis and the treatment of atherosclerotic cardiovascular disease (ASCVD) have been substantially improved, but the mortality rate and economic burden are still high. Long non coding RNAs(lncRNAs) is a transcription product that does not have the function of encoding proteins. It is widely involved in the regulation of cell biological functions such as cell development, proliferation, differentiation and apoptosis. It has been extensively studied as an important biomarker and therapeutic target in research fields such as tumor diseases and nervous system diseases. At present, a large amount of evidence indicates that lncRNAs also play an important role in the regulation pathway of atherosclerosis, which not only can further understand atherosclerosis, but also provide new directions for developing new diagnostic markers and treatments.

Abstract:The pathological characteristic of coronary heart disease (CHD)is rupture or erosion of atherosclerotic plaques of coronary artery which induces platelet aggregation, thrombosis, coronary artery spasm,microvascular embolism.They cause acute or subacute hypoxia or ischemia of myocardium. Studies show that coronary arterial endothelial cells can differentiate from venous endothelial cells. This review illustrates the classification of angiogenesis , the process of differentiation and clinical significance of coronary venous endothelial cells to arterial endothelial cells and the exploration of potential therapeutic targets. It aimed at providing reference for the future coronary angiogenesis.

Abstract:MicroRNA (miRNA) is a kind of evolutionarily highly conserved noncoding small molecule single stranded RNA (about 18-24 nucleotides), which negatively regulates gene expression at posttranscriptional level. Vulnerable plaques refer to plaques that are easy to form thrombus or may rapidly develop into criminal lesions in the process of atherosclerosis. Research shows that miRNA is involved in almost all steps of atherosclerosis, including endothelial cell and vascular smooth muscle cell injury and dysfunction, monocyte infiltration and platelet dysfunction, and plays a beneficial or harmful role. MiRNA will also become a new field to further study the mechanism of bidirectional regulation of vulnerable plaque by traditional Chinese medicine.