Abstract:Atherosclerosis-related diseases is a complex chronic disease.The exact pathogenesis of the disease is still unclear.So far, chemical drugs used to treat the disease in China are all developed outside China.The scientists worked in Traditional Chinese Medicine take its unique advantages.Under the guidance of TCM basic theory, trial and e-rror, a number of significant achievements have been acquired through clinical practice and drug discovery and development.New drugs which have determined curative effect was created for the prevention and treatment of atherosclerotic disease.They have been recognized and accepted on field of medicine, then go to the world.This article provides an overview of the most representative achievements.

Abstract:AimTo explore the effect of Vitamin K2 on vascular calcifiction in old ovariectomized rats model.MethodsThirty-six ten months female SD rats were allocated into three groups randomly: sham operation group, ovariectomized (OVX) group and OVX＋Vitamin K2 (OVX＋VK2) group.Rats in OVX＋VK2 group were administered orally with Vitamin K2 (30 mg(kg·d), 5 times a week) after three weeks of ovariectomy for 12 weeks.The urine and serum of all rats were kept every three weeks.All rats were sacrificed after 18 weeks, uterus and arteries were observed by histochemical method.The contents of MGP in serum and urine were determined by enzyme-linked immunosorbent assay (ELISA).Relative quantification of MGP mRNA expression in arterial was detected by Fluorescent real-time quantitative reverse-transcription polymerase chain reaction.Expression of ucMGP was detected by immunochistochemistry.Aterial calcification was evaluated by Von Kossa staining.The content of arterial calcium was measured.ResultsAfter 18 weeks of ovariectomized, the pathological section of uterus and aortic had differernt variation in different groups.The content of estrogen in serum had no difference before ovariectomized.Before sacrifice, the content of estrogen of sham group had no change, the other groups reduced (p<0.05).The content of MGP in serum and urine had no difference before ovariectomized, the content of MGP in serum and urine decreased in OVX group, the content of MGP in serum and urine decreased and then increased in OVX＋VK2 group.Relative quantification of MGP mRNA expression of aterial in OVX＋VK2 group was significantly lower than sham group (p<0.01), and was higher than that in OVX group (p<0.01).

Abstract:AimTo investigate the molecular mechanism of inflammatory protein expression via transforming growth factor-β1 (TGF-β1)/Smad pathway in ox-LDL-stimulated human umbilical vein endothelial cells (HUVEC).MethodsHUVEC were treated with 0.1 g/L of ox-LDL.TGF-β1, Smad3, VCAM-1 and ICAM-1 protein expression, and Smad3 phosphorylation were detemined by Western blot.Resultsox-LDL obviously increased the expression of VCAM-1 and ICAM-1 in HUVEC.TGF-β1 expression and Smad3 phosphorylation were enhanced in ox-LDL-stimulated HUVEC.Nuclear extract analysis showed that the phosphorylated Smad3 expression was increased in ox-LDL-treated cells.SIS3 (a specific inhibitor of Smad3) inhibited Smad3 phosphorylation in a dose-dependent manner, however, VCAM-1 and ICAM-1 expression were increased in ox-LDL-treated cells.ConclusionsTGF-β1/Smad signaling involved in the regulation of inflammatory protein expression in HUVEC induced by ox-LDL, the inhibition of Smad3 phosphorylation increased the expression of inflammatory protein, suggesting that TGF-β1/Smad3 signaling repressed the inflammatory protein expression in ox-LDL-treated endothelial cells.

Abstract:AimTo investigate the role of PI3K/Akt in vascular smooth muscle cell (VSMC) migration induced by Chlamydia pneumoniae (C.pn) infection.MethodsThe successful infection of rat VSMCs with C.pn was identified by transmission electron microscope; After VSMCs were pretreated with the specific PI3K inhibitor LY294002, wound-healing assay and Transwell assay were performed to observe the changes in the migration ability of VSMCs；The phosphorylation level of Akt was detected by Western blot.ResultsThe typical C.pn elementary bodies were observed in the infected VSMCs under the transmission electron microscope; The migration ability of VSMCs infected with C.pn was enhanced and significantly higher than that of control group at 24 h postinfection in the cell migration assay (p<0.05); Western blot results showed that the phosphorylation level of Akt was up-regulated and also higher than that of control group at 24 h after infection（p<0.05）.The effects of C.pn infection on the VSMC migration and the phosphorylation level of Akt in the VSMCs were significantly inhibited by the specific PI3K inhibitor LY294002.ConclusionC.pn infection may promote VSMC migration via activation of PI3K/Akt.

Abstract:AimTo introduce a rapid and efficient model of myocardial infarction (MI) without the requirement of ventilation in the mouse and comprehensively describe its details.Methods40 Male BABL/C6 mice were divided into 2 groups: new method MI (15) and new method Sham (25).In the new method, hearts of total 40 mice were manually exposed through a small incision of the left fourth intercostal space without intubation. In group of MI， the left anterior descending coronary artery was ligated.Hearts from group of Sham were without ligation.This novel MI procedure is rapid, with an average procedure time of 60±15 seconds.We compared the difference between MI and Sham post 28days with analysis of echocardiography, morphology and pathology.ResultsSurvival rate of group MI was 80% and group Sham was 100% 28 days after surgery with our method.The echocardiography showed that the heart function of the mice with 28-day post-MI was markedly impaired.The LVIDd and LVIDs were significantly increased from 3.52±0.10 mm and 2.60±0.19 mm to 4.65±0.08 mm and 4.36±0.13 mm (p< 0. 0001)，while LVEF and LVFS were decreased significantly 66.70%±1.41% and 35.90%±1.01% vs 29.70%±1.64% and 14.20%±0.80%，p<0.0001，respectively.Anatomical analysis of hearts with 28-day post-MI showed enlarged left ventricles and thin ventricular walls.

Abstract:AimTo study the effect of lovastatin on expression and activity of matrix metalloproteinase-9 (MMP-9) in a human umbilical vein endothelial cells (HUVECs) line.MethodsCultured HUVECs were stimulated by 20 μg/L tumor necrosis factor-alpha (TNF-α), and then incubated with 0.05-1.35 μmol/L of lovastatin for 0-24 h.The protein and mRNA level were detected by Western blotting and reverse transcription polymerase chain reaction (RT-PCR) respectively, and the activity of MMP-9 was analyzed by gelatin zymography.ResultsMMP-9 expression was induced by TNF-α in HUVECs, lovastatin attenuated both expression and activity of MMP-9 in a time and dose-dependent manner.0.05 μmol/L lovastatin inhibited MMP-9 expression in mRNA level, but not in protein level, but activity of MMP-9 cannot be inhibited by 0.05 μmol/L lovastatin.Other concentration of lovastatin groups can inhibit MMP-9 expression both in mRNA and protein levels, including activity of MMP-9, the maximum inhibitory effect was induced by 1.35 μmol/L lovastatin.ConclusionExpression and activity of MMP-9 can be inhibited by lovastatin in HUVECs.

Abstract:AimTo investigate the regulation of vascular endothelial growth factor (VEGF) expression after cerebral ischemia stroke in atherosclerotic rats model.MethodsRats were randomly divided into control group and model group, and then the latter, established into the atherosclerotic rats model, were randomly divided into five groups: atherosclerosis (As)-sham, As rat model with sustained ischemia (As-SI) 0.5, 6, 12, 24 h.The cerebral tissues were analyzed with the level of VEGF mRNA by reverse transcription polymerase chain reaction (RT-PCR) and the level of VEGF protein by immunohistochemical method.ResultsTo compare each type of rat model groups with control, and compare four-time-points rat model groups with As-sham, the formers were significantly up-regulated (p<0.05), and changed regularly with the four-time points，among which VEGF mRNA and its protein productions had reached the peak in As-SI 6 h group.ConclusionsThe present study showed that atherosclerotic disease had some positive effects on the expression of VEGF gene and its outcomes during cerebral ischemia stoke happened, and changed with the four-time points.Meanwhile, it had provided a theory to evaluate the occurrence, development, treatment and prognosis of cerebral ischemia stroke complicated by atherosclerotic disease.

Abstract:AimTo investigate the effects of pioglitazone on the expression of skeletal muscle AdipoR1 in type 2 diabetic rats, and explore the improvement effect and its mechanisms of pioglitazone on type 2 diabetes and insulin resistance.MethodsForty healthy eight-week-old female Sprague Dawley(SD) rats were randomly divided into normal control group(n=10), diabetic group(n=15), and pioglitazone group(n=15).Type 2 diabetes rat model was reproduced by feeding a high-sugar-fat diet followed by an intraperitoneal injection of a low dose of streptozotocin (STZ).Rats in pioglitazone group were laraged with pioglitazone 10 mg/(kg·d), while those in control group and diabetic group received the same amount of normal saline for 12 weeks.After 3 months, blood was taken from femoral vein and serum adiponectin was measured by enzyme-linked immunosorbent assay (ELISA).The structure of skeletal muscle was observed with light microscope and electron microscope.The protein expression of AdipoR1 in skeletal muscle was detected by immunohistochemical staining.ResultsSerum adiponectin (1.01±0.27 mg/L) in T2DM group was significantly decreased compared with normal control group (1.73±0.32 mg/L), but that of pioglitazone group (1.34±0.43 mg/L) was significantly increased compared with T2DM group (p<0.05).Immunohistochemical staining showed that the expression of AdipoR1 in skeletal muscle was the most obvious in normal control group and was attenuated in T2DM group.The expression of AdipoR1 in skeletal muscle of pioglitazone group was more significant than that in T2DM group, but was weaker than that in normal control group.Light microscope and electron microscope showed no obvious abnormalities in skeletal muscle structure.ConclusionsThe levels of serum adiponectin and AdipoR1 in skeletal muscle decline in type 2 dia-betic rats.Pioglitazone can up-regulate the levels of serum adiponectin and AdipoR1 in skeletal muscle thus it can down-regulate the circulating levels of glucose and lipids and improve insulin sensitivity.

Abstract:AimTo investigate the alteration of expression of calcineurin in the abdominal aortas and levels of inflammatory cytokines injured by balloon in rats.MethodsFourty-eight healthy adult male SD rats were randomly divided into two groups: sham group (n=24) and balloon group (n=24).Thirty days after injury, specimens were obtained from rats.HE staining and immunohistochemistry were performed to measure the expression level of CaN in arterial wall.Pathological changes of the arterial wall were observed by light microscope.The serum concentrations of MCP-1 was determined by Elisa.ResultsIntimal hyperplasia was observed in balloon injured rats, the neointima was not uniform in thickness.The thickness of the intimal layers, and the ratio of the intimal and the medial layers in the sham group was obviously lower than that in the balloon group (P<0.05).Immunohistochemistry assay showed that the expression of CaN was significantly increased in balloon group compared with that in the sham group (P<0.05).The serum concentrations of MCP-1 in balloon group is higher than that in sham group (P<0.05).ConclusionsIntimal hyperplasia in rat abdominal aorta following balloon injury, accompanied by the expression of CaN protein and plasma inflammatory cytokines MCP-1 levels were significantly increased，which indicated that CaN signaling pathway and MCP-1 may play an important role in intimal hyperplasia caused by balloon injury.

Abstract:AimTo investigate the biological distribution of 99mTc-anti-cardiac troponin T monoclonal antibody (99mTc-AcTnTMA) in the rats with acute myocardial damage.MethodsPrepared 99mTc-AcTnTMA and built the rat model of acute cardiac muscle damage.Then 60 rats were randomized into the experimental group, the control group and the blank group.There are twenty rats in one group.In the experimental group, 20 rats with acute myocardial damage were injected with 0.3 mCi 99mTc-AcTnTMA and were killed at 2, 4, 8 and 12 h after injection respectively (5 rats).Blood, liver, spleen, kidney, muscle, colon, lung and heart of each rat were taken and the injected dosage (ID%/g) and the ratio of ID%/g for heart-to-lung ratio (HLR) was calculated.In the control group and the blank group, 20 rats with acute myocardial damage were injected with 0.3 mCi 99mTc-N-IgG and 99mTc-AcTnTMA respectively and were killed and followed subsequent procedure in the same way as the experimental group. ResultsThe value of ID%/g and HLR in the experimental group were significantly higher than the other groups, which hinted that acute injury myocardium could uptake specifically the 99mTc-AcTnTMA and the specific uptake was observed with its peak 4 h after injection.Conclusion99mTc-AcTnTMA could be a useful tracer of myocardial imaging to diagnose acute myocardial damage.

Abstract:AimEstablishing model of cerebral ischemia-reperfusion injury and intervening with Danhong injection, then observing morphological changes of cerebral cell in damaged area and Golgi, detecting Golgi Matrix Protein 130 (GM130) expression, and exploring protective mechanism of Danhong injection on cerebral ischemia-reperfusion injury in rats.Methods72 healthy adult SD rats were randomly divided into normal controlled group (6), sham operation group (6), ischemia-reperfusion group (30), and Danhong intervention group (30).According to the reperfusion time after ischemia 2 h, sham operation group and ischemia-reperfusion group were respectively divided into 6 h, 24 h, 48 h, 72 h, and 7 days five subgroups, there were 6 rats in each subgroup.After the success of cerebral ischemia-reperfusion model, the rats in Danhong intervention group were interved with Danhong injection(8ml/kg,QD) by intraperitoneal injection from the recovery of reperfusion until they were executed; At the same time, the rats in ischemia-reperfusion group were treated with the same dose of normal saline by intraperitoneal injection.ResultsMorphology showed cortical neuronal survival number were significantly increased in Danhong intervention 7 d group compared with the other groups, and the damage degree was the lightest; In ischemia-reperfusion injury 7 d group, Golgi morphology took significantly impaired change, lack of network structure, lightly staining, granules decreasing or disappearing, and even some broken, but in Danhong intervention 7 d group, Golgi morphology kept normal.GM130 expression decreased with the prolongation of ischemia-reperfusion time, increased with the prolongation of Danhong injection intervention time.GM130 expression of Danhong intervention 7 d group was the highest compared with the other groups（p<0.05).ConclusionDanhong injection may maintain the Golgi stability through the up-regulation of GM130 expression, and thus play a neuroprotective role.

Abstract:AimExploration of the protective effects of Guanxinshutong (GXST)against Oxidative Stress(OS) injury following ischemia/reperfusion.MethodsThirty male Sprague Dawley rats were randomized into three groups: non-MI/R group (Sham, n=10), MI/R group treated with vehicle (Model, n=10) and MI/R group treated with GXST (Drug, n=10).MI/R was induced by ligation of the left anterior descending coronary artery (LAD) for 30 min, followed by 3 h reperfusion in the model and Drug groups.In the Sham group, the LAD was exposed without occlusion.GXST powder (in the Drug group) or saline (in the Model and Sham groups) were administered via direct gastric gavage from 5 days prior to surgery.Blood samples were collected from the carotid artery after 3 h of reperfusion, to determine the levels of CK-MB, LDH, Tn-T, MDA, SOD, TNOS, iNOS and NO.ResultsGXST significantly decreased levels of CK-MB, LDH, Tn-T, MDA and iNOS; increased levels of SOD, TNOS and NO compared with the model group (all p<0.05).ConclusionsGXST is effective in protecting the myocardium against MI/R injury in rats.Its possible cardioprotective mechanism involves inhibition of the OS injury following MI/R.

Abstract:AimTo investigate the level of plasma adiponectin in patients with diabetic nephropathy, and its relationship to vascular endothelial function.MethodsFifty type 2 diabetic patients without clinical macrovascular complications were randomly assigned to normal albuminuria group(NAU), microalbuminuria group(MAU) and macroalbuminuria group(CAU) according to the daily urinary albumin excretion (UAE).Plasma adiponectin, soluble vascular cell adhesion molecule-1(sVCAM-1) and other biochemical indexes were measured.Color Doppler examination was followed to evaluate flow-mediated vasodilation (FMD), nitroglycerin vasodilation (NID), cardiologic parameters and intima-media thickness (IMT).ResultsIn CAU group, plasma concentration of adiponectin was 4-fold and 2-fold higher than that in NAU group and MAU group respectively (13.3±5.0 mg/L vs 3.7±1.6 mg/L vs 5.7±2.2 mg/L, p<0.01 and p<0.05), while the difference between NAU group and MAU group was also significant (p<0.05).The ratio of adiponectin to serum creatinine followed the same trend along three groups (p<0.05).Plasma sVCAM-1 was significantly increased in MAU group and CAU group than that in NAU group (p<0.01 and p<0.05).FMD and NID were signifi-cantly decreased in contrast to the increment of plasma adiponectin (p<0.05).IMT was comparable between three groups (p>0.05).Finally, adiponectin was positively correlated with sVCAM-1, UAE, creatinine, left ventricular posterior wall thickness and IMT（r=0.338, 0.704, 0.470, 0.331, 0.324 respectively，all p<0.05）, while negatively correlated with FMD and NID (r=－0.397 and －0.413 respectively, p<0.01).ConclusionsDiabetic nephropathy is accompanied with hyperadiponectinemia, which is closely related to vascular endothelial dysfunction.Adiponectin may act as an early predictor of vascular endothelial dysfunction in these patients.

Abstract:AimTo compare the advantages and weaknesses of two kinds of noninvasive measurements with cardio-ankle vascular index (CAVI) and intima-media thickness (IMT) in the evaluation of arteriosclerosis.MethodsThe subjects were 351 persons(55～70 years old) who came to our clinics because of arteriosclerosis disease.They were divided into three classes of A,B and C.A class was non-arterioscletotic disease group and B class included diabetes mellitus, hypertension, hyperlipidemia and angina pectoris group and C class included coronary stenosis,myocardial infarction(MI) and cerebral infarction(CI).CAVI and IMT for the subjects were simultaneously measured.The relationship between CAVI and IMT and the duration of diabetes was analyzed in 112 of diabetic patients(21～84 years old).ResultsIn age-matched subjects(55～70 years old ), CAVI was higher in myocardial infarction, coronary artery stenosis, cerebral infarction; diabetes mellitus, angina pectoris, and hypertension was the second; non-arterioscletotic diseases group constituted together a “ladder effect” with three layers of mean values; Each layer presented the meaningful difference(p<0.05 or p<0.01) after exclusion of which p value between DM and MI or CS were appreciably high（p=0.130 and 0.118 respectively）.IMT showed much more uncertainties with mean value, and IMT value was even lower in CI and CS groupsthan that in DM group.The correlation of the duration of diabetes mellitus was stronger with CAVI than with IMT.Significant positive correlations were observed between the CAVI measurement value and the duration of diabetes(r=0.499,p<0.001).However in IMT, there was only a weakly correlations with the duration of diabetes (r=0.195,p=0.038).ConclusionsCAVI is a good indicator of cardio- and cerebro-vascular event, and is more likely reflecting arterioscleosis in a whole body than IMT.

Abstract:AimTo understand the level of plasma adipsin in type 2 diabetes millitus and type 2 diabetes millitus with peripheral arterial disease patients and to discuss the effect of adipsin in the occurrence and development of type 2 diabetes millitus and peripheral arterial disease.Methods50 cases of type 2 diabetes millitus, 50 cases of type 2 diabetes with peripheral arterial disease and 50 cases of normal controls were selected.Detected plasma serum and measured the ankle-brachial index.Detected blood pressure, blood glucose, serum insulin and blood fat.Calculated body mass index and insulin resistant index et al.ResultsThe level of plasma adipsin in type 2 diabetes millitus with peripheral arterial disease group was higher than that of type 2 diabetes millitus group and normal control group.The level of plasma adipsin in type 2 diabetes millitus group was higher than that of normal control group.There was negative correlation between adipsin and ankle-brachial index.ConclusionsThe increase of circulating adipsin may be involved in the me tabolism of type 2 diabetes mellitus and peripheral arterial disease.

Abstract:AimTo study the relationship between plasma level of interleukin-8 (IL-8) and the polymorphism of IL-8 single nucleotide with the susceptibility to coronary heart disease (CHD).MethodsPolymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) was used to detect genotype of IL-8 －251A/T in CHD cases and control subjects.Plasma levels of IL-8 were detected by enzyme-linked immunosorbent assay.ResultsCompared with that of control subjects, plasma levels of IL-8 of CHD cases were significantly increased (p<0.05).The overall distribution of IL-8 －251A/T genotype and allelotype in CHD patients and controls were not significantly different (p>0.05).However, a statistically significant lower frequency of AA genotypes was observed in acute coronary syndrome patients compared to other patients (OR=0.43, 95%CI was 0.2～0.97, P=0.04).ConclusionsThe results suggest that plasma level of IL-8 may co-relate with the susceptibility of CHD, －251A/T single nucleotide polymorphism is not associated with the susceptibility of CHD, but the genetic diversity may influence the clinical manifestation of CHD.The AA genotype may reduce the risk of acute coronary syndrome.

Abstract:AimTo investigate the association of the G894T,T786C polymorphism of the endothelial nitric synthase (eNOS) gene with essential hypertension (EH) in the Han and Kazakh populations in Xinjiang.MethodsBased on the case-control study,346 patients with essential hypertension(EH group)and 385 healthy people (NT group)in Han Chinese population, and 363 patients with essential hypertension(EH group)and 370 healthy people (NT group)in the Kazakh population were selected from Tacheng Region of China by random cluster sample.After physical examination and questionnaire, blood samples were selected, the G894T,T786C polymorphisms of eNOS gene were detected with SNaPshot system.Genotype frequency, allele frequency, linkage disequilibrium (LD)and haplotypes were calculated and compar-ed.ResultsOverweight, obesity, triglycerides disorder, age 51 and over are risk factors of EH in Han and Kazakh populations.Distribution of allele frequencies G894T, T786C polymorphism of eNOS gene, the total population in EH and NT group are significantly different between two ethnic (P<0.05).G894T and T786C polymorphism do not have strong LD.It constituted four kinds of haplotypes of GT (75.3% and 79.6%), GC (10.8% and 10.5%), TT (5.7% and 9.8%), TC (8.2% and 0.1%) in Han and Kazakh populations.The maximum distribution of haplotypes frequencies was GT in two ethnic, and the minimum are TC (Han), TT (Kazakh).The formation of haplotypes (GT, TT, TC) frequency was significantly different between two ethnic (P<0.05).ConclusionsThe polymorphism of eNOS gene G894T, T786C may not be associated with the essential hypertension in the Han and Kazakh populations in XinJiang.

Abstract:Calcium (Ca2+) entry into non-excitable cells is mainly carried by store-operated calcium channels (SOCC).The best characterized SOCC current is the Ca2+ release-activated Ca2+ (ICRAC) current.Recently, store-operated calcium entry (SOCE) mediated by the STIM, Orai and TRPC involved in a variety of physiological and pathological processes.This paper describes the molecular elements of store-operated calcium inux complex (SOCIC), their interaction, as well as the role of SOCE in the pathogenesis of pulmonary artery hypertension (PAH), which aimed at providing new research directions about prevention and treatment of PAH.

Abstract:GALNT2 is a key member of polypeptide:N-acetylgalactosaminyltransferase family (GalNAc-Ts), and recent GWAS based epidemiological study reveal that its polymorphism can interrupt HDLC and TG levels in blood, and is significantly associated with cardiovascular disease.GALNT2 participates in the first step of O-linked glycosylation and is regarded as a cardiovascular disease susceptible gene recently.Therefore, its function and genetic difference as well as the relationship with cardiovascular disease will become a hot research point in the future.This review will focus on the recent advances in research of its biological characteristics, functions, gene polymorphisms and relationship with lipids levels and cardiovascular disease.

Abstract:Cardiovascular calcification happens early and seriously in uremia, of which cardiac valve calcification is relatively common.It can cause not only valve stenosis and (or) incompetence but also arrhythmia, ventricular hypertrophy, cardiac insufficiency, even sudden death, etc.Researches show that many factors were involved in cardiac valve calcification in uremia, such as calcium phosphate metabolism imbalance, parathyroid hormone, vitamin D, inflammation, fetuin-A, etc.Cardiac valve calcification is the imbalance of the results of calcification promoters and inhibition.Its mechnism in recent years is reviewed in this paper.