Abstract:Aim To observe the effect of Klotho gene modified bone marrow mesenchymal stem cells (BMSC) transplantation on myocardial fibrosis in rats with heart failure. Methods Isolation, culture and amplification of rat bone marrow mesenchymal stem cells. Klotho transfection to BMSC and RT-PCR detection of Klotho mRNA expression in BMSC. The rat model of chronic heart failure were randomly divided into normal group, saline group, EGFP-BMSC group and EGFP-Klotho-BMSC group, with 6 rats in each group. The cardiac function was detected by Doppler ultrasound after 28 days of cell transplantation. The survival and distribution of the cells were observed by fluorescence microscope. The pathological changes of myocardial tissue were observed by HE staining. Masson staining was used to detect the content of myocardial collagen deposition and the degree of myocardial fibrosis. Results Klotho combined with BMSC treatment can better inhibit myocardial fibrosis, reduce myocardial interstitial collagen deposition and improve cardiac function. Conclusion Klotho gene combined with bone marrow mesenchymal stem cells could attenuate cardiomyocytes hypertrophy and restrain collagens hyperplasy in the progression of myocardial fibrosis.

Abstract:Aim To investigate the effect of probucol on the differentiation of inflammatory monocyte subsets and its potential mechanism. Methods The primary single cell suspension of spleen was prepared from 8 week old C57BL/6 mice. The differentiation of monocyte subsets was induced by 100 kU/L recombinant interferon-γ, then intervened with 5,0, 100 μmol/L probucol, in order to observe their effects on the differentiation of monocytes. The cells were stained with anti-CD11b-PE and anti-Ly-6C-FITC antibody, and the differentiation of monocyte subsets was detected by flow cytometry, then the data were analyzed by Flowjo software. Using intracellular cytokine staining and 2′,7′-dichlorofluorescein-diacetate probe, the level of reactive oxygen species (ROS) in monocyte subsets was detected by flow cytometry. NADPH oxidase activity in human monocytic cell line THP-1 was detected by using NADPH oxidase test kit. Results In a concentration dependent manner, 5,0, 100 μmol/L probucol inhibited the differentiation of Ly-6C^hi inflammatory monocyte subsets in the spleen primary cells. The level of ROS in Ly-6C＋ inflammatory monocyte subset was 2 times of that in Ly-6C－ inflammatory monocyte subset. At the same time, 5,0, 100 μmol/L probucol inhibited the production of ROS in Ly-6C＋ inflammatory monocyte subset in a concentration dependent manner. 100 μmol/L probucol significantly inhibited the activity of NADPH oxidase in THP-1 cells. Conclusion Probucol may interfere with the activity of NADPH oxidase, thereby inhibiting the differentiation of Ly-6C^hi inflammatory monocyte subset and the production of ROS.

Abstract:Aim To study the mechanism of microRNA-155(miR-155) depending on cholesteryl ester hydrolase(CEH) to influence foam cells formation. Methods Oil red O staining and HPLC were used to analyze cholesteryl ester content, and the positive cells and cholesterol ester content were observed. Western-blot was used to the expression of CEH, ATP binding cassette transporter A1(ABCA1), scavenger receptor AⅠ(SR-A). Results The positive cell ratio and the cholesterol efflux were dramatically decreased by miR-155 mimics(P＜0.05);the positive cell ratio, the cholesterol efflux, the expression of SR-A were significantly decreased(P＜0.05),while the expression of CEH and ABCA1 were significantly increased in miR-155 mimics group respectively(P＜0.05). The expression of SR-A,the positive cell ratio, the cholesterol efflux were significantly increased(P＜0.05), while the expression of CEH and ABCA1 were significantly decreased in microRNA-155+siRNA-CEH group and siRNA-CEH group respectively(P＜0.05). Conclusion The high expression of microRNA-155 can inhibit the formation of foam macrophages,which may be enhanced by CEH affecting the expression of ABCA1 and SR-A.

Abstract:Aim To investigate the effect of capsaicin on the inhibition of rat vascular smooth muscle cells (VSMC) induced by high sodium. Methods Vascular smooth muscle cells were isolated from thoracic aorta of male Sprague-Dawley rats by tissue adherent method, and the growth curve of vascular smooth muscle cells was made according to MTT assay. The experimental procedures were performed with the fourth generation of vascular smooth muscle cells. Vascular smooth muscle cells were synchronized by serum starvation for 24 h before capsaicin and high sodium treatment. Vascular smooth muscle cells were treated with different concentration of capsaicin (0.1,0.1,1, 10 and 100 μmol/L) for 72 h in high sodium. The effects of different concentration capsaicin and osmotic pressure on the proliferation induced by high sodium medium in vascular smooth muscle cells were evaluated by MTT assay and CCK-8, respectively. The cell cycle of vascular smooth muscle cells was analyzed by flow cytometry technology. The localization and protein expression of PCNA were employed by immunofluorecence staining and Western blot. The mRNA and protein expressions of TRPV1 were detemined by real-time PCR and Western blot. Results MTT results showed that when the concentration of capsaicin was 10 μmol/L, the proliferation of rat vascular smooth muscle cells began to be inhibited, and the inhibitory effect was increased at 100 μmol/L (P＜0.05), so 10 μmol/L capsaicin was selected for subsequent experiments. CCK-8 results showed that the cells in high sodium group proliferated obviously, but there was no difference between mannitol group, normal group and capsaicin group. Capsaicin prevented notably the decrease of cell number in G0/G1 and G2/M stage and increase of cell number of S stage in vascular smooth muscle cells treated with Na＋ (159 mmol/L). PCNA protein level was also decreased by capsaicin. The decrease of mRNA and protein level of TRPV1 stimulated Na＋ (159 mmol/L) was alleviated by capsaicin. Conclusion Capsaicin can inhibit the proliferation of vascular smooth muscle cells induced by Na＋ (159 mmol/L), the mechanism may be related to the up-regulation of TRPV1 expression.

Abstract:Aim To explore the effects and mechanism of ApoAⅠ on TNF-α expression in macrophages and plasma TNF-α levels from atherosclerotic mice. Methods Male ApoE-/- mice were fed with high cholesterol diet for 12 weeks and then randomly divided into control group, ApoAⅠ group and ApoAⅠ+AG490(a specific JAK2 inhibitor)group； each group were separately received treatment with PBS, ApoAⅠ (40 μg/g), ApoAⅠ (40 μg/g)+AG490(4 μg/g) on the third and the first day before sacrifice, 12 h before sacrifice all groups were administered with LPS via intraperitoneal injection, TNF-α levels in plasma were measured by ELISA. THP-1 macrophage-derived foam cells were randomly divided into control group , ApoAⅠ(10 mg/L)group and ApoAⅠ(10 mg/L)+AG490(25 μmol/L) group; all groups incubated with LPS (10 μg/L). TNF-α level in supernate were measured by ELISA and TNF-α mRNA expression were examined by RT-PCR. Results Compared with the control group, ApoAⅠ can significantly decrease TNF-α levels in mice plasma, after administered with AG490 ApoAⅠ-inhibition of TNF-α secretion induced by LPS markedly weakened(P<0.05). In vitro study demonstrated ApoAⅠ can inhibit TNF-α transcription and expression induced by LPS(P<0.01), AG490 abrogated the anti-inflammatory effect of ApoAⅠ(P<0.05). Conclusion ApoAⅠ inhibited TNF-α transcription and expression through JAK2/STAT3 signaling pathway.

Abstract:Aim To explore the effect of salvia magnesium lithospermate B (MLB) on myocardial ischemia-reperfusion (IR)-induced cells programmed necrosis and its mechanism. Methods Myocardial infarction model was established by occlusion of the left anterior descending coronary artery in rats. After ischemia 1 h and reperfusion for 3 h, myocardial IR injury model was established. Rats were randomly divided into 7 groups:normal control group, sham operation group, IR group, MLB low dose (10 mg/kg)＋IR group, MLB high dose (30 mg/kg)＋IR group, necrostatin-1 (Nec-1; 3 mg/kg)＋IR group and solvent (normal saline)＋IR group (n＝6~8). Myocardial infarct area was detected by triphenyltetrazolium chloride staining. HE staining was used to observe the morphological changes of myocardial tissue. Serum creatine kinase (CK) activity was measured by spectrophotometry. The expression levels of programmed necrosis associated protein receptor interacting protein kinase 1 (RIPK1) and RIPK3 were detected by Western blot. Results High dose of MLB could significantly reduce the myocardial infarct size and CK release, and improve the myocardial tissue structure of IR rats, with the inhibition of RIPK1 and RIPK3 protein expression, which were better than the positive control drug Nec-1. Conclusion MLB has the function of resisting myocardial cell programmed necrosis, and its mechanism is related to the inhibition of RIPK1 and RIPK3 protein expressions.

Abstract:Aim To study the effects of cholesterol (CH) on proliferation and migration of vascular smooth muscle cells (VSMC). Methods VSCMs were treated with 12.5 mg/L, 25.0 mg/L, 50.0 mg/L CH. VSMC activity were analyzed by MTT kit, the percentage of DNA synthesis were detected by EdU kit, the percentage of S phase cell numbers were measured by flow cytometry and VSMC migrations were detected by scratch assay. The expression of miR145 was analyzed by real-time quantitative PCR. Results 12.5 mg/L, 25.0 mg/L, 50.0 mg/L CH can increase VSMC proliferation, migration and the percentage of S phase cell numbers (P＜0.05). 25.0 mg/L, 50.0 mg/L CH promoted DNA synthesis percentage of VSMC, decreased the expression of miR-145 in VSMC (P<0.05). Conclusion Cholesterol increased the percentage of S phase VSMC number, cells proliferation, DNA synthesis and migration, and miR-145 may be involved in these processes.

Abstract:Aim To explore the key members of the miRNAs target to peroxisome proliferator-activated receptor γ(PPARγ), and reveal the clinical significance of which are closely related to in-stent restenosis(ISR). Methods Cytoscape and its plug-in was used to build the miRNAs-PPAR regulatory network to screen out which are the key miRNAs target to PPARγ. The expression of miRNAs in ISR group were detected by Real-time PCR and the clinical recognition ability of miRNAs on ISR patients were evaluated by ROC curve. Results By bioinformatics analysis, miR-27a/b has a strong regulatory effect on PPARγ, while miR-130a/b is able to regulate more genes in the network. Real-time PCR found that miR-27a/b and miR-130a were differentially expressed in the ISR group and no-ISR group (P<0.05). ROC curve analysis showed that miR-27a has a certain clinical recognition ability to ISR (AUC =0.4,5% CI 0.768~1.00, P<0.001). Conclusion MiR-27a/b and miR-130a/b are key members of the miRNAs target to PPARγ, miR-27a/b and miR-130a were differentially expressed in ISR, miR-27a might have certain clinical diagnostic value in patients with ISR.

Abstract:Aim To investigate the effect of intensive dose atorvastatin therapy on B7-H3, B7-H4 in peripheral blood monocytes of patients with unstable angina pectoris undergoing percutaneous coronary intervention (PCI). Methods The patients with unstable angina pectoris were randomly divided into conventional dose group (atorvastatin 20 mg/d, n=40) and intensive dose group (atorvastatin 80 mg/d, n=40), peripheral blood were collected before and 18 h ~ 24 h after PCI. Enzyme-linked immunosorbent assay was used to detect peripheral blood IL-4, IL-10, IFN-γ and sB7-H3, sB7-H4, fluorescence-based quantitive real-time PCR was used to measure B7-H3 and B7-H4 mRNA. Results The levels of IL-10, sB7-H3, sB7-H4 and the expression of B7-H3 and B7-H4 mRNA in the two groups after PCI were higher than those before PCI, and the increase of the intensive dose group was more significant than that of the conventional dose group (P＜0.05). On the contrary, the levels of IL-4 and IFN-γ in the two groups after PCI were decreased, and the decrease of the intensive dose group was more obvious than that of the conventional dose group (P＜0.05). Conclusion Intensive dose atorvastatin may promote the expression of B7-H3 and B7-H4, thereby reducing the immune inflammation in patients with unstable angina pectoris after PCI.

Abstract:Aim To qualitatively and quantitatively evaluate the influence of plaque calcification on the diagnostic accuracy of dual source CT (DSCT) coronary angiography for coronary stenosis assessment. Methods The retrospective analysis enrolled 224 patients. They were diagnosed as coronary artery disease by DSCT angiography, and then received selective coronary angiography. There were 375 lesions with significant coronary stenosis in 224 patients according to DSCT. Of 375 lesions, 234 lesions were with calcification. The influence of characteristics of lesions on accuracy of DSCT angiography for coronary stenosis assessment was evaluated. These characteristics included calcification degree of lesions, artery external diameter of calcified lesions, length of calcified lesions and location of calcified lesions. Results For mild to moderate calcified plaque and severe calcified plaque, DSCT overestimated coronary stenosis by 6.8% (P＝0.0028) and 18.8% (P＜0.0001) separately, and the overestimation of stenosis was more obvious in severe calcified lesions (P＝0.002). For calcified lesions with artery external diameter＜3 mm and artery external diameter≥3 mm, DSCT overestimated coronary stenosis by 7.2% (P＝0.0026) and 17.1% (P＜0.0001) separately, and the overestimation of stenosis was more obvious in calcified lesions with artery external diameter≥3 mm (P＝0.001). There was no difference in the diagnostic accuracy of DSCT for calcified plaques with different lengths (P＝0.792). Conclusion The lesion calcification makes DSCT angiography overestimate coronary stenosis. This phenomenon is more likely to occur in the severe calcified lesions and calcified lesions with artery external diameter≥3 mm.

Abstract:Aim To evaluate the effect of growth differentiation factor-15 (GDF-15) in predicting no-reflow in patients with acute myocardial infraction (AMI) undergoing percutaneous coronary intervention (PCI). Methods Totally 243 consecutive patients with AMI were enrolled and serum GDF-15 levels were measured within 24 hours. All patients were divided into two groups according to the occurrence of no-reflow during PCI. Results There were 44 patients in no-reflow group (average age was 67.00±13.04 ) and 199 patients in normal-reflow group (average age was 65.54±12.98). The serum GDF-15 levels increased in no-reflow group (1073.43±364.38 ng/L vs 714.10±340.98 ng/L, P<0.001). The area under the receiver operating characteristic curves (AUC) was 0.829 (95%CI:0.766～0.892). Logistic regression showed that GDF-15 (OR:1.3,5% CI:1.001～1.004, P＜0.001) and female (OR:2.6,5%CI:1.358～6.610, P=0.007) were independent risk factors for no-reflow during PCI in AMI patients. Conclusion GDF-15 and female were independent risk factors for no-reflow during PCI in AMI patients.

Abstract:Aim To analyze the clinical features of patients with chronic heart failure (CHF) complicated with renal insufficiency, and to explore the risk factors of CHF with renal insufficiency. Methods 385 cases of CHF patients who were treated in department of cardiology of our hospital were collected, including 211 males and 174 females, with an average age of 69.62±8.59 years. Applying method of retrospective control study, according to estimated glomerular filtration rate (eGFR), the CHF patients were divided into renal insufficiency group [eGFR＜60 mL/(min·1.73 m²)] and non renal insufficiency group [eGFR≥60 mL/(min·1.73 m²)]. The general information, heart basic diseases, concomitant disease, contrast agent application history, echocardiographic parameters, renal function, blood lipids, N-terminal pro-B-type natriuretic peptide (NT-proBNP), D-dimer, hemoglobin, urine microalbumin and other laboratory examination indexes were analyzed and compared between the two groups. Multivariate Logistic regression was used to analyze the risk factors of CHF patients with renal insufficiency. Results The incidence of renal insufficiency was about 42.1% in 385 patients with CHF. Univariate analysis showed that the differences of age, NYHA heart function classification, hypertension, anemia, diabetes, atrial fibrillation were statistically significant between renal insufficiency group and non renal insufficiency group (P＜0.05). The levels of serum creatinine, urea nitrogen, uric acid, cystatin C and NT-proBNP in renal insufficiency group were higher than those in non renal insufficiency group, and left ventricular ejection fraction and hemoglobin level were lower than those in non renal insufficiency group (P＜0.05). Multivariate Logistic regression analysis showed that age (OR 1.3,5%CI 1.067-2.231), NYHA heart function classification (OR 1.0,5%CI 1.054-3.212), hypertension (OR 1.7,5%CI 1.175-3.292) and cystatin C (OR 1.9,5%CI 1.027-3.851) were independently associated with renal insufficiency. Conclusion The incidence of renal insufficiency was higher in patients with CHF. The elder age, NYHA heart function Ⅳ classification, hypertension and serum cystatin C were independent risk factors of CHF patients with renal insufficiency.

Abstract:Aim To evaluate the diagnostic value of dual-source computed tomography in detecting coronary artery anomalies. Methods Clinical data of 5153 patients who underwent coronary CTA were retrospectively analyzed to determine the CT features of anomalous origin of coronary artery. Results In consecutive 5153 patients, 141 cases with anomalies origin were identified (2.74%), including 102 cases with trunk anomalous origin (1.98%), 39 cases of anomalous branches (0.76%). Conclusion Dual-source CT could accurately display the anomalous origin of the coronary and direction, provide a reference for clinical diagnosis and treatment.

Abstract:Aim To discuss the display value of the direct conformation of the cervical artery interlayer with the technique of monoenergetic⁺. Methods 43 cases of cervical artery interlayer, confirmed by CT angiography (CTA) and/or digital subtraction angiography (DSA), were collected. The dual energy CT non-enhanced imaging (DENECT) scan obtains data, using monoenergetic⁺ technique. DENECT uses χ² to display the rate of the direct image of the cervical artery. Its sensitivity, specificity, accuracy, positive predictive value and negative predictive value were tested by kappa. Results There were 43 cases confirmed by CTA and/or DSA, with 51 sites of 51 vessels. Among 43 cases of NECT images processed by monoenergetic⁺ technique, 16 cases clearly showed direct signs of the cervical artery interlayer and 27 were not clear. NECT showed significantly better than DSA (P<0.05) for the direct signs of the cervical artery interlayer. CTA showed better than DENECT for the display of vertebral artery interlayer (P<0.05), while DENECT showed no significant difference from CTA for the display of carotid and vertebral carotid artery (P>0.05), which positive predictive value and negative predictive value reached more than 61%, with the accuracy of 66% above. Conclusion monoenergetic⁺ technique provides a new method for emergency screening of artery interlayer.

Abstract:Aim To explore the relationship between cumulative triglyceride exposure (cumTG) and ankle-brachial pulse wave velocity(BaPWV). Methods A total of 23499 participants was selected from Kailuan Study stroke cohort, hypertensive population and elderly population cohort to compose observation population, and finally 14662 cases were included in the study cohort. According to cumTG, the research subjects were divided into four groups. The correlation between cumTG and BaPWV was analyzed by partial correlation analysis. Multivariate linear regression and multivariate Logistic regression were used to analyze the effects of cumTG on BaPWV. Results With the increase of cumTG, the average BaPWV level and the detection rate of BaPWV≥ 1400 cm/s showed an increasing trend. The correlation analysis showed that cumTG was positively correlated with BaPWV (r=0.512, P<0.05), and cumTG was positively correlated with BaPWV (r=0.322, P<0.05) after adjustment for age and sex. Multivariate linear regression analysis showed an increase in BaPWV of 4.507 cm/s for each increase in cumTG. Logistic regression analysis showed that the cumTG second quartile, the third quartile, and the fourth quartile were significantly associated with BaPWV≥1400 cm/s compared with the cumTG first quartile after adjustment for other confounders (95%CI) which were 1.667(1.505～1.845), 2.384 (2.111～2.691) and 3.287(2.887～3.741), respectively. Conclusion cumTG is positively correlated with BaPWV.cumTG is a risk factor for the increase of pulse wave velocity.

Abstract:The effect of cell inflammation and lipid metabolism on atherosclerosis development was reviewed. Imbalance of lipid metabolism resulted in inflammation. Abnormal modification and location of lipoprotein promoted inflammation. Lipid metabolism disorders induced inflammation by monocytes, M1 / M2 drift, and NLRP-3 inflammasomes.At the same time, inflammation promoted the lipid uptake and accumulation, and inhibited the cholesterol efflux from cell.Inflammation and imbalance of lipid metabolism jointly promoted the developement of atherosclerosis. We discussed the role of caveolae/ caveolin, NF-κB pathway, and PPAR pathway in the interaction and regulation of cell inflammation and lipid metabolism.

Abstract:Adipose tissue lipoprotein lipase (LPL) is regulated by angiopoietin-like protein and influences the storage and energy supply of adipose energy. Myocardial LPL is regulated by peroxisome proliferator-activated receptor and adenosine monophosphate activated protein kinase and influences the uptake of free fatty acids in the absence of cardiac energy supply. Macrophage LPL is regulated by microRNA and inhibits the formation of foam cells. The regulation of LPL in various tissues is influenced by many factors, and the effects are different. In this paper, we reviewed the regulation of LPL in adipose tissue, myocardium and macrophages, and the effects of related proteins on LPL activity, in order to further clarify the role and significance of LPL in various tissues.

Abstract:Atherosclerosis, which takes place in the walls of arteries, is a long-term progressive inflammatory pathological process. The rupture of atherosclerotic plaque and thus thrombosis may lead to acute cardiovascular events which threaten human health and life seriously. Recently, the role of neutrophils in atherosclerosis is attractted much attention.Activated neutrophils release web-like structures known as neutrophil extracellular traps (NETs). NETs affect the pathologic process in the patients with autoimmune diseases, acute lung injury, deep vein thrombosis, etc. A large number of studies indicate that NETs is associated with the formation of atherosclerosis and thrombosis. NETs, which act as predictive factors in coronary artery diseases, predict the major adverse cardiac events and the prognosis. This article is aimed to review the role of neutrophil extracellular traps in the development of atherosclerosis.

Abstract:Simple renal cyst is the most common renal cystic disease. More and more evidence indicates that there is a correlation between simple renal cyst and hypertension. However, the relationship between hypertension and the size, location and number of renal cyst is not clear. The underlying mechanism between simple renal cyst and hypertension is unknown yet. In this review, we will summarize the relation between simple renal cyst and hypertension and possible underlying mechanisms. We aim to identify simple renal cyst as an independent predictor of hypertension and clinical intervention target.

Abstract:Cilostazol is a unique antiplatelet agent that has been clinically available for over two decades. As a phosphodiesterase III inhibitor, it reversibly inhibits platelet aggregation yet also possesses vasodilatory and antiproliferative properties. It has been widely applied to avariety of disease states, including peripheral arterial disease, cerebrovascular disease, and coronary artery disease with percutaneous coronary intervention. In this paper we review recent development of cilostazol regarding its pharmacological action and clinical application.