Abstract:Treatment delay is the main reason without effective treatment in the patients with acute coronary syndrome (ACS). It is urgent to solve this problem. Although the study of clinical pathway for ACS has been applied in July 2011. Using clinical pathway is not optimistic. Most of ACS visited grassroots medical institutions for the first time. However, there is no standard diagnosis and treatment, limited guide line and rescue process in the primary hospital. It is difficult to achieve reperfusion for the ACS. Therefore, the new regional cooperative treatment model may be the effective rescue of ACS.

Abstract:Aim To discuss the correlation of the polymorphism of tumor necrosis factor receptor superfamily 1B (TNFRSF1B) in the 196ed gene (T mutation for G) to the inflammation reaction mediated by the macrophage TNF-TNFR2 signaling pathways.Methods Genetic recombination technology was used to bulid eukaryotic expression vector pcDNA6.0-TNFR2196Met and pcDNA6.0-TNFR2196Arg, together with pcDN6.0 being transferred to macrophage by liposome transfection method respectively. In order to establish stable transfection cell lines, blasticidin was used to select them for 4 weeks after transfected 48 h. Digest the macrophages by pancreatic enzyme and divide them into four groups: control group, pcDNA6.0 empty plasmid group, pcDNA6.0-TNFR2196Met group and pcDNA6.0-TNFR2196Arg group. TNFR2, cIAP₁, cIAP₂ mRNA were detected by RT-PCR, the expression of p-JNK, cIAP₁, cIAP₂, TNFR2 and NF-κB were detected by Western blot, the level of sTNFR2, IL-1β and IL-6 in cell supernatant fluid were detected by ELISA.ResultsTNFR2 gene 196Met and 196Arg expression vector were successfully constructed which was verified by enzyme sequencing method. After transfected to marcophages, TNFR2, cIAP₁, cIAP₂, IL-1β, IL-6 and p-JNK expression were increased (P<0.05) in TNFR2196Arg and TNFR2196Met overexpression group compared with the blank control group and empty plasmid group. Each index decreased obviously in mutant TNFR2196Arg cell group compared with TNF2196Met cell group, especially,the reduction of NF-κB activity mediated by TNFR2196Arg was significant. Conclusions Successfully building stable expression TNFR2196Arg and TNFR2196Met vector will lay the foundation for the next inflammation research. The mutation of TNFR2 mediates inflammatory disease through TNF/TNFR2 signaling pathways, which is molecular mechanism of chronic inflammatory disease.

Abstract:Aim To observe the effect of Alisma Decoction on lipid deposition and the expression of LXRα and ABCA1 on macrophagic foam cell formation.Methods Macrophagic foam cell formation was established after induction with oxidized low density liprotein (ox-LDL) (50 mg/L) and Dil-ox-LDL(10 mg/L)for 24 h respectively and then the cells were intervened with 20% Alisma Decoction-containing serum. Oil red O staining and fluorescence microscopy were used to detect the lipid deposition in cells. RT-PCR and Western blot was utilized to detect the expression of LXRα and ABCA1.Results Macrophagic foam cell formation was established sucessfully after the stimulation with ox-LDL(50 mg/L) and Dil-ox-LDL(10 mg/L). Compared with the control group, lipid deposition was increased significantly in the model group, while lipid accumulation was obviously reduced in macrophage cells after treated with Alisma Decoction-containing serum. The expression of LXRα showed a low level in both the control group and model group. However, after intervened with the Alisma Decoction-containing serum,the expression of LXRα increased distinctively in macrophage cells which was pretreated with ox-LDL. As to ABCA1,the model group expressed much more greatly than the control group. And the macrophage cells treated with the ox-LDL and Alisma Decoction--containing serum group had a high level expression ,but it is lower than the model group.Conclusion Macrophagic foam cell formation was sucessfully established after induction with oxidized low density liprotein (ox-LDL) (50 mg/L) and Dil-ox-LDL(10 mg/L). Alisma Decoction can decrease the lipid deposition in the process of macrophagic foam cell formation, and it may depend on the way of upregulating the expression of LXRα.

Abstract:Aim To determine the effect of heat shock protein-65 (HSP65) on the anti-oxidant and anti-inflammatory properties of high density lipoprotein (HDL) in apolipoprotein E gene knocked-out (ApoE^－/－) mice.Methods Eight-week-old ApoE^－/－ mice were randomly divided into three groups: phosphate buffer as control group, 5 μg HSP65 group and 25 μg HSP65 group. The mice were boosted twice under the same protocol following three and six weeks. Sixteen weeks after the first immunization, serum lipids, paraoxonase-1 (PON-1) activity, myeloperoxidase (MPO) activity, high density lipoprotein inflammatory index (HII), the level of cytokines interleukin-10 (IL-10) and interferon-γ (IFN-γ) were assessed.Results With the increasing dose of HSP65, HDLC level, IL-10 expression and PON-1 activity reduced gradually, whereas HII, MPO activity, and the IFN-γ expression elevated. Compared with control group, markedly decreased HDLC, IL-10 level and PON-1 activity but a significant enhancement of the serum HII, MPO activity, and IFN-γ levels were resulted in 25 μg HSP65 group (all P<0.05).Conclusion HSP65 could induce inflammatory response and impair the anti-oxidant and anti-inflammatory properties of HDL, which was related to the dose of HSP65.

Abstract:Aim To investigate the effects of perinatal hypoxia on carotid artery endothelium-dependent relaxations of 6 week old rabbits and its molecular mechanism.Methods Ten pregnant New Zealand White rabbits were randomly divided into two groups: normoxic control group (21% O₂, n5) and perinatal hypoxia group (12% O₂, n5) during day 10～29 of pregnancy. Pregnant rabbits had spontaneous deliveries, and only one male offspring from each litter was selected randomly and breast-fed for six weeks. Carotid arteries were obtained from 6 week old rabbits, endothelium-dependent relaxation (EDR) and expression of endothelial nitric oxide synthase (eNOS) and heat shock protein 90 (HSP90) of carotid arteries were examined.Results Perinatal hypoxia significantly impaired carotid artery EDR in 6 week old rabbits (P<0.05). Perinatal hypoxia significantly decreased carotid artery HSP90 expression (P<0.05) and eNOS activities (P<0.05) in 6 week old rabbits.Conclusions Perinatal hypoxia can induce functional impairment in vascular endothelium from 6 week old rabbits. The molecular mechanism involved is that perinatal hypoxia decrease HSP90 expression and eNOS activities of rabbit artery.

Abstract:Aim This paper studies the effect of ERK1/2 signal transduction in focal acute brain ischemia-reperfusion.Methods 36 healthy male SD rats were randomly divided into sham operation group and ischemia group. Ischemia group was further divided into brain ischemia-reperfusion with differernt time (6, 12, 24, 48 and 72 h). Suture-occluded method was used to build cerebral middle artery infarction model (MCAO). CD34, Ang-2 and ERK1/2 were detected through immunohistochemical method.Results Ang-2 and ERK1/2 in focal cerebral ischemia-reperfusion rat model had dynamic change, and both reached peak at 24 hour.Conclusion ERK1/2 signal transduction way takes part in the promotion effect of angiogenesis of angiopoietin.

Abstract:Aim To investigate whether the all-trans retinoic acid (ATRA) will perform anti-inflammatory role and therefore to inquire into its possible mechanism of stabilizing plaques, by establishing the animal models ofcarotid artery endometrial injuring in rabbits and observing the expression level of platelet-activating factor (PAF) and leukotriene B4 (LTB4).Methods 36 New Zealand male healthy rabbits were randomly divided into three groups: contrast group (n＝12), high-fat group (n＝12) and treatment group (n＝12), each group then were divided into three subgroups (A, B, C). Contrast group were provided basic feed while the other two groups were given fatty feed. All groups have to be suffered from artery intima air-drying technique after two weeks. Contrast group only had exposure of carotid artery but no damage to the intima others should have injury to artery intima the treatment group were administered ATRA lavage daily from 3 days before the surgery. On the 7d, 14d and 28d, the target vessels were taken out for morphological observation and measuring of intima thickening. Enzyme-linked immunosorbent assay (ELISA) method was used to detect the expression level of PAF and LTB4 in rabbit serum.Results Contrast group did not see endometrial hyperplasia, PAF and LTB4 had normal small expression. High-fat group were of endometrial hyperplasia obviously, target vessels were stenosis even deformation, PAF and LTB4 had quantity expression. The intima hyperplasia in treatment group was lower than high-fat group (P<0.05) as the same as the expression level of PAF and LTB4.Conclusion ATRA can significantly alleviate injury area endometrial proliferation, the mechanism may be related to its inhibition of PAF activation and expression, then reduce the release of inflammatory chemokines LTB4 in order to restrain the inflammatory response ultimately.

Abstract:Aim To observe the effects of endothelial progenitor cells (EPC) conditional medium on the proliferation of vascular smooth muscle cells (VSMC) induced by angiotensin Ⅱ(AngⅡ).Methods Mononuclear cells were isolated from fresh cord blood by 3% Gelatin and density gradient centrifugation. Isolated mononuclear cells were cultured in EBM-2 medium. Flow cytometry and indirect immunofluorescence revealed that EPC expressed hematopoietic progenitor cell-surface antigens as well as endothelial cell-surface antigens. After preparating early endothelial progenitor cell conditioned medium (E-EPC-CM), late endothelial progenitor cell conditioned medium (L-EPC-CM) and human umbilical vein endothelial cell conditioned medium (HUVEC-CM), the effect of E-EPC-CM, L-EPC-CM and HUVEC-CM on AngⅡ-induced DNA synthesis, total protein expression, cell survival and cycle progress was assessed by BrdU incorporation, total protein content, MTT assays and flow cytometry. Western blot were performed to analyze the effect of E-EPC-CM on the AngⅡ-induced phosphorylations of ERK, JNK, p38 MAPK and the expressions of proto-oncogene c-myc and c-fos.Results The DNA synthesis, total protein contents and cell survival obviously increased in AngⅡ (10^-6mmol/L) induced VSMC for 48 h compared with control group, while pretreatment with E-EPC-CM, L-EPC-CM and HUVEC-CM resulted in significant inhibitions in DNA synthesis, total protein content, and cell survival compared with the control groupThe percentage of S phase cells significantly increased, while the percentage of cells in G₁ phase decreased remarkably compared with control group, pretreatment with E-EPC-CM, L-EPC-CM, or HUVEC-CM led to sustained decrease in the percentage of cells in S phase and sustained increase in the percentage of cells in G₁ phase after 48 h of stimulation with AngⅡ Western Blot showed that AngⅡ time dependently induced phosphorylation of p38, JNK, ERK and significantly increased the expressions of c-myc and c-fos after AngⅡ stimulation in VSMC, while pretreatment with E-EPC-CM showed marked inhibitory effect on the AngⅡ-induced phosphorylations of these MAPK and the expressions of proto-oncogene c-myc and c-fos. E-EPC-CM showed strongest inhibitory effect among all treatment groups.Conclusion EPC may inhibit AngⅡ-induced proliferation and phenotype transformation of VSMC through inactivating MAPKs signaling pathways as well as by reducing the expressions of proto-oncogene c-myc and c-fos.

Abstract:Aim To expore the correlation between plasma visfatin and coronary atherosclerosis in Kazak and Han patients with coronary heart disease.Methods Forty Kazak and forty-five Han patients with coronary heart disease were picked out randmly as test groups aging from forty to seventy-five, then both test groups were divided into two subgroups (mild sub-group and heavy subgroup) using Gensini score respectively. Thirty-five Kazak and forty Han healthy people were picked out as control groups. Plasma visfatin was measured using enzyme-linked immunosorbent assay (ELISA), meanwhile liver function, renal function, blood lipids, blood glucose and fructosamine were measured.Results Kazak and Han patients with coronary heart disease had higher levels of plasma visfatin than control groups (P<0.01) the heavy subgroup had higher levels of plasma visfatin than the mild subgroup in Kazak and Han patients with coronary heart disease there was a positive correlation between the level of plasma visfatin and Gensini score in the two nationalities ( r0.925 and P<0.01 for Kazak； r0.967 and P<0.01 for Han).Conclusions Kazak and Han patients with coronary heart disease had higher levels of visfatin than their control group. There was a positive correlation between the level of plasma visfatin and Gensini score in the two nationalities, indicating that visfatin might be involved in the occurrence and development of coronary atherosclerosis.

Abstract:Aim To investigate the incidence and association of lacunar infarction (LI) with varying degrees leukoaraiosis.Methods A total of 130 cases of leukoaraiosis patients admitted in the department of Neurology, Beijing Military General Hospital from Jun 2012 to Dec 2012 and 130 healthy subjects were enrolled. The degree of leukoaraiosis and number of lacunar infarction were evaluated on MRI.Results The incidence of lacunar infarction in patients with leukoaraiosis was significantly higher than that of control group. The degree of leukoaraiosis was significantly associated with the number of lacunar infarction (r＝0.921, P＝0.000). Logistic regression demonstrated that age, hypertension, and lacunar infarction were the risk factors of the leukoaraiosis.Conclusion Occurrence of lacunar infarction has certain correlation with leukoaraiosis.

Abstract:Aim To explore the relationship between serumγ-glutamyl transferase (GGT) levels and coronary complexity, severity and extent. The interaction of serum GGT with the traditional risk factors of acute coronary syndrome (ACS) was evaluated.Methods 413 consecutive patients with ACS who underwent coronary angiography were enrolled. Baseline serum GGT levels were measured and Gensini score was calculated from the study population. All patients underwent test of GGT, clinical characteristics, blood lipid, brain natriuretic peptide (BNP), blood urea nitrogen, serum creatinine, uric acid, and measuring risk factors.Results Serum GGT levels demonstrated an increase from low Gensini tertile to high tertile. In multivariate analysis, a significant positive correlation was found between logGGT levels and the different number of diseased coronary vessels, diabetes mellitus, total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (LDLC) and high sensitive C-reactive protein (hs-CRP) (P<0.05). TC was the strongest influence on logGGT. Further studying the relations between coronary stenosis severity (Gensini score) and serum GGT as well as the traditional risk factors of ACS, the results showed logGensini had a significant positive correlation with serum GGT, age, ALT, smoking, the different number of diseased coronary vessels (P<0.05).Conclusion Serum GGT involved the development of coronary plaque, serum GGT tested has important clinical values in assessing the severity of coronary artery disease.

Abstract:Aim To investigate the expressions of CD54, CD106 on the surface of endothelial cells after interaction with platelets from the patients with coronary heart disease (CHD).Methods To draw blood from acute coronary syndrome patients (acute coronary syndrome group), patients with stable angina (stable angina group) and healthy people (control group), the platelets were extracted and interacted with endothelial cells. Then the expression of CD54, CD106 on the surface of endothelial cells were tested.Results The expression level of CD54 on the surface of endothelial cells in acute coronary syndrome group were obviously higher than in control group (P<0.01), and higher than in stable angina group (P<0.05) but no difference was found in CD54 between control group and stable angina group. The expression level of CD54 was significantly and positively related with the markers of myocardial necrosis such as initial troponin I and initial creatine kinase-MB (CK-MB). There were no obvious differences of the expression level of CD106 among each group (P>0.05).Conclusions After interaction of endothelial cells with platelets, the expression level of CD54 on the surface of endothelial cells from the patients with acute coronary syndrome was obviously increased, and was associated with the markers of myocardial necrosis. We can infer that CD54 is a very important index which can be used to predict the occurrence and severity of CHD.

Abstract:Aim To analyze the difference of Helicobacter pylori (Hp) infection in five types of atrial fibrillation (AF) and to explore the correlation between Hp infection and the different types of AF.Methods According to the 2010 ESC guidelines, AF was divided into five types: initial AF, paroxysmal AF, persistent AF, long persistent AF and permanent AF. 285 cases of hospitalized AF patients were enrolled with a retrospective analysis from January 1, 2007 to April 12, 2013. All patients were taken the examination of plasma antibody of Hp (IgG), ¹³C urea breath test, blood routine, blood biochemistry, high sensitive C-reactive protein (hs-CRP), left atrial diameter (LAD), urinary microalbumin (UMA), glycosylated hemoglobin and plasma homocysteine (Hcy). We analyzed the difference of Hp infection, hs-CRP level, LAD and other indicators in five types of AF. According to the time of occurrence, AF were divided into two groups: less than one year of AF (Group one: including initial AF, paroxysmal AF and persistent AF) and more than one year of AF (Group two: including long persistent AF and permanent AF). Then, we explored the correlation between Hp infection and AF by the Logistic regression analysis.Results The value of ¹³C urea breath test, hs-CRP and LAD in Group one were significantly different with that in Group two (P<0.05). Hp≥4‰, hs-CRP>5 mg/L and LAD>36 mm were sensitive index of AF patients more than one year.Conclusion The value of ¹³C urea breath test, hs-CRP and LAD in patients with AF sustained over one year were significantly higher than that in patients with AF sustained in one year. Hp≥4‰, hs-CRP>5 mg/L and LAD>36 mm were closely related to AF sustained more than one year.

Abstract:Aim To elucidate the relationship between advanced glycation end products (AGE), arterial stiffness and the degree of coronary artery stenosis in Chinese Han patients.Methods The serum concentration of AGE was measured and the brachial-ankle pulse wave velocity (baPWV) was detected in 105 cases of mild/moderate coronary stenosis, 130 cases of serious coronary stenosis and 150 aged-matched control subjects.Results Serum AGE and baPWV were significantly higher in the mild/moderate coronary stenosis and serious coronary stenosis patients when compared with the control subjects (P<0.001). In Logistic regression analysis, serum AGE and baPWV were independently associated with the degree of coronary artery stenosis (P<0.001). Further analysis showed that AGE was significantly and positively correlated with baPWV (r＝0.692, P<0.001).Conclusions Serum AGE was an independent risk factor of the severity of coronary artery disease and might be involved in the development of coronary artery stenosis through inducing arterial stiffness.

Abstract:Aim To compare the dilation resistance during stents implantation between different types of non-clacified coronary plaques by retrospectively investigating patients who received 64-slice spiral computed tomography angiography before coronary stents implantation.Methods 116 patients who received 64-slice spiral computed tomography angiography before coronary stents implantation from 2007 to 2012 were selected. Non-calcified coronary plaques were divided into positive remodeling group and non-positive remodeling group based on remodeling index. Clinical features, lipid cores, pre-dilatation balloon size and filling pressure, and stent size and pressure were compared.Results High-sensitivity C-reactive protein (hs-CRP) levels were higher in positive remodeling group. The lipid cores were more commonly observed in positive remodeling group. In patients who have typical symptom, unstable angina was more common in positive remodeling group, and stable angina was more common in non-positive remodeling group. The proportion of administration of pre-dilatation was similar in two groups. The size and filling pressure of pre-dilation ballons were similar in two groups. The size of stents was similar in two groups. Stent release pressure in positive remodeling group was significantly lower than that observed in non-positive remodeling group (1 121.64±213.68 kPa vs 1 232.56±201.95 kPa, P<0.01).Conclusions Compared with non-positive remodeling group, the lipid cores were more common in positive remodeling group, therefore the dilation resistance was lower in positive remodeling group during stents expansion. Preoperative identification of lesion remodeling index was helpful for choosing direct stent implantation rightly.

Abstract:Aim To study renin-angiotensin-aldosterone system (RAAS) hormone levels and CYP11B2－344C/T gene polymorphism in essential hypertension patients with obesity, discuss genetic susceptibility of CYP11B2－344C/T genotype and the correlation between susceptible gene and RAAS levels.Methods 60 cases of patiens with 1～2 grade essential hypertension (30 cases essential hypertension patients with obesity, 30 cases essential hypertension patients without obesity), and 60 people from the Examination Center (30 cases obesity people without hypertension, 30 cases healthy people) were enrolled randomly. CYP11B2－344C/T gene polymorphisms were analyzed using polymerase chain reaction-restriction fragment length polymorphism and agarose gel electrophoresis methods, plasma RAAS concentration were detected by radioimmunoassay.Results Multi-compared with number of cases of TT and TC＋ CC genotype, there were significant differences (P<0.05). And differences in essential hypertension with obesity group was the most significant multi-compared with the number of T and C alleles, significant differences were found (P<0.05), and differences in essential hypertension with obesity group was predominant. Plasma renin, angiotensin Ⅱ, aldosterone concentrations in genotype TT cases were much higher than those of TC and CC genotype (P<0.05), but there was no significant differences between CC and TC genotype cases (P>0.05).Conclusions In CYP11B2－344C/T gene polymorphism, TT genotype may be susceptible genotype of essential hypertension patients without obesity, and T allele may be susceptible one of them. Each component of plasma RAAS hormone levels in the TT genotype cases was significantly higher than the CC and the TC genotype ones.

Abstract:Sphingosine-1-phosphate receptors (S1PR) are important transit point for sphingosine-1-phosphate (S1P) to play roles in cells. Five subtypes of S1PR have been identified so far, which are widely expressed in all systems including cardiovascular system, and play various important biological effects in cardiovascular development, angiogenesis, vasorelaxation, vascular barrier integrity, ischemia/ reperfusion injury and atherosclerosis. This paper makes a review on roles and mechanisms of S1PR in cardiovascular system.

Abstract:Epicardial adipose tissue is a part of visceral tissue deposited around the heart, especially around coronary arteries, which can express many kinds of adipokines participating in the occurrence and development of atherosclerosis. Recently it has been confirmed these adipokine can also cause angiogenesis within the coronary atherosclerotic plaque. These neovessels have very strong permeability, which can improve the condition of local hypoxia new blood vessels, on the other hand, may promote the plaque’s progression and lead to its probability of instability increase. Thus, by studying the molecular mechanism of adipokines promoting angiogenesis, the inhibition of angiogenesis in the plaque to enhance the stability of the plaque can become a new target for therapy of coronary heart disease in the future.