Abstract:Aim The aim of this study was to examine the effect of naturally regulatory T cells on atherosclerosis plaque in apoliprotein(apo)E~(-/-) Mice,through negative regulating FOXP3 gene. Methods Lentivirus-mediated(siRNA) was used and Foxp3high+⁺ CD25+ Treg cells adoptive transfer assays in high fat diet ApoE~(-/-) Mice were done.Its number was identified by FACS.Inflammatory cytokines were determined by ELISA and the area of atherosclerosis plaque was analyzed. Result We found that the number and function of Foxp3+⁺CD25+ Treg cells in mice injected with Foxp3high+⁺ CD25+ Treg cells was significantly high compared with those of mice injected with FOXP3-siRNA lentivirus. Conclusion Naturally regulatory T cells regulated negatively can promote significantly the progression of atherosclerosis plaque in ApoE~(-/-) Mice.

Abstract:Aim To observe the influence of urantide on the expression of C-reactive protein(CRP) in rats with atherosclerosis and investigate its mechanism. Methods Model with atherosclerosis was induced by feeding high fat diet and arterial intimal injury of intraperitoneal injection of vitamin D3,rats were randomly divided into four groups: control group,model group,fluvastatin group and urantide group,and the expression of C-reactive protein in aortic wall were detected by immunological histochemical method.Cultured vascular smooth muscle cells in vitro were randomly divided into four groups: control group,urotensinⅡgroup,fluvastatin group and urantide group,and the expression of C-reactive protein in supernanant were detected by enzyme-linked immunosorbent assay(ELISA). Results In the plaques of thoracic aorta intima and tunica media,the expression of the positive particles of C-reactive protein in the model group was increasing obviously compared with that in the control group and the C-reactive protein expression in each experiment group of urantide was decreasing by fluvastatin.The C-reactive protein expression in the vascular smooth muscle cells supernatant of each urantide concentration group had downward trend,among which the 10-6 mol/L urantide was decreasing most(P<0.01).Conclusions UrotensinⅡ can facilitate the over-expression of C-reactive protein in atherosclerosis.However,the promotion can be inhibited by urotensinⅡ receptor antagonist urantide.The experiment provides new views and experiment basis for the clinic treatment of atherosclerosis with urantide.

Abstract:Aim To evaluate the effect of rosuvastatin on restenosis and the expression of transforming growth factor β1(TGF-β1) in abdominal aorta of rabbits after balloon injury. Methods Two groups of 10 male New Zealand rabbits received either rosuvastatin or no rosuvastatin after balloon injury.We did abdominal aorta angiography and observed the expression of TGF-β1 with immunohistochemistry and reverse transcription polymerase chain reaction(RT-PCR) six weeks later. Results The average stenosis degree(31.41%±7.08%) and the smallest inner diameters(1.74±0.25 mm) were better in rosuvastatin-treated group than that of the model group(55.25%±10.23% and 1.32±0.33).The average absorbency and the area of positive of TGF-β1 were dramatically lower in rosuvastatin-treated group than that of the model group(0.41±0.12 vs 0.57±0.17,31.37%±4.42% vs 40.58%±7.45%);The quantity of amplification products(22.37±4.42) was lower than that of the model group(42.47±5.41),there were significant differences between the two groups(P<0.05). Conclusion Rosuvastatin can improve the restenosis and decrease the expression of TGF-β1 in rabbits after balloon injury.

Abstract:Aim To explore the mechanisms of influenza virus infection in the formation of atherosclerosis from the cellular and molecular levels through investigating amount of intercellular adhesion molecule-1and vascular cell adhesion molecule-1after human umbilical vein endothelial cell was infected by influenza virus. Methods SYBR Green reverse transcription polymerase chain reaction(RT-PCR),flow cytometry and enzyme-linked immunosorbent assay were used to detect the timing expression of intercellular adhesion molecule-1 and vascular cell adhesion molecule-1 of human umbilical vein endothelial cells infected by influenza virus at 0 h,24 h,48 h and 72 h. Results Intercellular adhesion molecule-1 and vascular cell adhesion molecule-1were measured by these three methods after human umbilical vein endothelial cell was infected by influenza virus.The base level of the two inflammatory factors was expressed at a low level at 0 h,and began to increase after influenza virus infection,reached the peak at 24 h.After 48 h,it declined obviously and remained a relatively high level at 72 h.The amount of intercellular adhesion molecule-1 and vascular cell adhesion molecule-1 in the infected groups was higher than that in the control group(P﹤0.05). Conclusion This study showed that influenza virus infection could increase the expression of intercellular adhesion molecule-1 and vascular cell adhesion molecule-1,and then influenza virus infection might lead to dysfunction of vascular endothelial cells and involve in the inflammatory response of atherosclerosis.

Abstract:Aim To study the change of cerebral vascular in hippocampus of vascular dementia rats after hypodermic injection of basic fibroblast growth factor. Methods After the vascular dementia model was reproduced,the 12 model rats were randomly divided into basic fibroblast growth factor(bFGF) treatment group,normal saline treatment group.The 6 rats were in sham group.The vascular dementia rats in therapy group were treated by hypodermic injection of bFGF.After 5 weeks,abilities of learning and memory of three section rats were tested by using the Morris water maze.The changes of vascular endothelial growth factor(VEGF) in rats serum were tested.Immunochemistry staining of factor Ⅷ-related antigen(F8) was used to observe the number of nestin positive neurons in hippocampus. Results The time spent on the platform was 14.3±3.1 s,7.4±2.9 s and 12.6±2.7 s in the sham,bFGF treatment group and normal saline treatment group.The VEGF in ratsserum was 8.14±1.53,6.07±0.18,9.19±0.29.It was more significantly increased in bFGF treatment group than that in normal saline treatment group(P<0.05).The number of the F8 positive neurons in the hippocampus were 105.38+1.45,101.32+1.28,43.03+2.11.It was more significantly increased in bFGF treatment group than that in normal saline treatment group. Conclusions bFGF by hypodermic injection can migrate to hippocampus area and induce hippocampus to create cerebral vascular.bFGF can increase abilities of learning and memory of rats significantly.

Abstract:Aim To investigate the effect of angiotensin Ⅱ on vascular endothelial growth factor expression of early endothelial progenitor cells. Methods Total mononuclear cells(MNCs) were isolated from peripheral blood by Ficoll density gradient centrifugation,and then the cells were plated on fibronectin-coated culture dishes.After 7 days of culture,several groups of attached cells were incubated with angiotensin Ⅱ(to make a series of concentrations: 10-3 mol/L,10-5 mol/L,10-7 mol/L vehicle control for 24 h),angiotensin Ⅱ+valsartan,angiotensinⅡ+PD123319.The cells were observed under inverted microscope,and characterized as adherent cells double positive for DiL DL-uptake and lectin binding by direct fluorescent staining under a laser scanning confocal microscope.The early endothelial progenitor cells were further documented by demonstrating the expression of cell markers with flow cytometry.Enzyme-linked immunospecific assay(ELISA) were used to assess vascular endothelial growth factor expression. Results Our data indicated that angiotensin Ⅱ can significantly increase the vascular endothelial growth factor expression,with a maximum at 10-3 mol/L after 24 hours(P<0.05);These effects can be attenuated by pre-treatment with valsartan but not PD123319. Conclusion It is suggested that angiotensin Ⅱ induces vascular endothelial growth factor protein secretion via the angiotensin Ⅱ receptor-1 but not angiotensin Ⅱ receptor-2.

Abstract:Aim To study the effects of chronic intrauterine hypoxic on myocardial inducible nitric oxide synthase(iNOS) and plasma endothelin-1(ET-1). Methods Chronic intrauterine hypoxic rats were divided into three groups:hypoxic group,air mimic control group and blank control group.Each group included 10 offspring rats.We monitored the weight of organs and birth weight of offspring,immunohistochemistry was used to examine the expression of myocardial iNOS,enzyme linked immunosorbent assay(ELISA) was used to measure serum ET-1 expression of offspring at 1 day and 6 month old. Results Organ/body weight percentage showed chronic intrauterine hypoxia could cause a disproportionate growth of offspring organs.The expression of myocardial iNOS of the hypoxic group significantly increased compared with that of the air mimic control group and that of the blank control group(P<0.05),but the air mimic control group was not significantly different from that of the blank control group(P>0.05).There was no significant difference of the expression of the serum ET-1 of all groups of offspring at 1 day old(P>0.05).But there was significant difference of the expression of the serum ET-1 of the hypoxic group compared with that of the air mimic control group and that of the blank control group of offspring at 6 month old(P<0.05),while and the air mimic control group was not significantly different from that of the blank control group(P>0.05). Conclusions Intrauterine chromic hypoxia may be a stress factor for development of the cardiovascular system,and it can induce low birth weight and high expression of myocardial iNOS and serum ET-1,which lead to increase the risk of cardiovascular disease.

Abstract:Aim To explore the effects of Naoxintong on the plaques formation of abdominal aorta and C-reactive protein(CRP) in hypercholesterolemic rabbits. Methods 36 male New Zealand White rabbits were equally divided into 3 groups at randomly: balloon injury + high lipid diet(model group),balloon injury + high lipid diet + Naoxintong(Naoxintong group),balloon injury + high lipid diet + simvastatin(simvastatin group).They were all fed for 16 weeks.The CRP concentrations were detected by enzyme-link-immunosorbent assay,and the histopathological changes of the aortas were detected at the end of the study. Results Compared with 0 week,serum total cholesterol(TC),low-density lipoprotein cholesterol(LDLC) and triglyceride(TG) of three groups were higher in the 8th week and 16th week(P<0.01),high-density lipoprotein cholesterol(HDLC) was higher in Naoxintong group and simvastatin group(P<0.05);Compared with model group,there were significant difference in TC,LDLC,HDLC of Naoxintong group.Compared with 0 week,the levels of CRP were significantly increased in three groups(P<0.01),and at the end of the study,the CRP levels were significantly decreased in Naoxintong group and simvastatin group compared with model group(P<0.01).Compared with Naoxintong group,plaques thickness of model group was significantly different(P<0.01). Conclusion Naoxintong decreased the plasma cholesterol and CRP concentrations and resisted the inflammatory function,which may inhibit the formation of plaques of abdominal aortas in cholesterolemic rabbit.

Abstract:Aim To study the effects of triiodothyronine on the Bax and Bcl-2 of immature cadiocytes while given ischemical reperfusion injury. Methods The cadiocytes of neonate rat were culrured in vitro,and the ischemical reperfusion injury of cadiocytes were induced by anoxia-reoxygenation.Triiodothyronine(T3) was administered before anoxia-reoxygenation at a concentration of 1 μg/L,Bcl-2 mRNA and Bax mRNA were detected by polymerase chain reaction(PCR). Results The Bcl-2 were down-regulated and Bax were up-regulated while given T3 shortly before anoxia-reoxygenation.At the same time,the apoptosis of cadiocytes were reduced. Conclusions Giving triiodothyronine before anoxia-reoxygenation would enhance the ablity of immature cadiocytes against ischemical reperfusion injury,and could reduce the apoptosis of cadiocytes.This effect was associated with the expression of Bcl-2 and Bax.

Abstract:Aim To investigate the effect of a synthetic dsRNA analog polyriboinosinic polyribocytidylic acid(PolyI:C) on quantity and activity of endothelial progenitor cells from human umbilical cord blood in vitro. Methods Mononuclear cells were isolated from human cord blood by Ficoll density gradient centrifugation and then the cells were cultured in EBM-2 medium to differentiate into endothelial progenitor cells.Endothelial progenitor cells were identified by demonstrating the expression of characteristic cell surface marker CD34,CD133 and KDR by reverse transcriptase polymerase chain reaction.CCK-8 reagent kit was used to analyze the effect of PolyI:C on the proliferation of endothelial progenitor cells.Flow cytometry was used to detect the apoptosis caused by PolyI:C.Reverse transcriptase polymerase chain reaction was performed to detect firstly the expression of Toll-like receptors in endothelial progenitor cells,secondly the expression of Toll-like receptor 3 and inflammatory cytokines induced by PolyI:C. Results Under basal condition,except Toll-like receptor 9,Toll-like receptor 1-10 were expressed in endothelial progenitor cells with high expression levels of Toll-like receptor 1,Toll-like receptor 3,Toll-like receptor 4,Toll-like receptor 6 and low expression levels of Toll-like receptor 2,Toll-like receptor 5,Toll-like receptor 7,Toll-like receptor 8,Toll-like receptor 10.Expression of Toll-like receptor 3 mRNA was upregulated by PolyI:C and there was dose-effect relationship in experiment range.Compared with thecontrol group,PolyI:C groups(1 g/L and 10 g/L) significantly inhibited the proliferation of endothelial progenitor cells and the inhibition of PolyI:C(10 g/L) on endothelial progenitor cells proliferation was time-dependent.PolyI:C induced the apoptosis of endothelial progenitor cells in dose-dependent manner.PolyI:C also upregulated the mRNA expression of inflammatory cytokines such as interleukin-1β,interleukin-6,interleukin-8,tumor necrosis factor-α,interferon-β. Conclusion Poly I:C inhibits the proliferation of endothelial progenitor cells via induction of cell apoptosis,simultaneously up regulates the expression of inflammatory cytokines probably by activating Toll-like receptor 3.

Abstract:Aim To investigate some biological characteristics of similarities and differences of endothelial progenitor cells from human peripheral blood,umbilical cord blood and adipose tissue and provide the evidences for efficacious applications of endothelial progenitor cells on different adaptive scopes,ways and methods. Methods Mononuclear cells were isolated from umbilical cord blood and adult peripheral blood using a Ficoll-Hypaque density gradient centrifugation.Adipose tissue-derived stem cells were isolated from the adipose tissue by trypsin digestion.The collected cells were cultured.The cell morphology was observed and the cultured cells were identified at the seventh day.The proliferation and migration of endothelial progenitor cells were measured by MTT and Boyden chamber assay.The adhesion function and vasculogenesis activity in vitro of endothelial progenitor cells were analyzed by adhesion assay and in vitro vasculogenesis kit. Results All differentiation cells were identified as endothelial progenitor cells.Endothelial progenitor cells from umbilical cord blood were revealed superior in proliferation,migration,adhesion and vasculogenesis activity(P<0.05).The proliferation and vasculogenesis activity of endothelial progenitor cells from adipose tissue were more powerful than those of endothelial progenitor cells from adult peripheral blood(P<0.05). Conclusion Endothelial progenitor cells from umbilical cord blood may be more suitable for allogeneic stem cell transplantation.Endothelial progenitor cellsfrom adipose tissue may exhibit potential applications in autologous stem cell transplantation for ischemic disease.

Abstract:Aim To compare the dynamic changes of neuroendocrine hormone levels on different treatment groups,in order to understand the angiotensin converting enzyme inhibitor and β-receptor blockers' effect on neuroendocrine hormone after valve replacement. Methods 60 patients were randomly divided into A,B,C groups,and were given different drugs after treatment.We took blood samples in the preoperative and different time of postoperation to determine plasma level of angiotensinⅡ,atrial natriuretic polypeptide,aldosterone. Results After valve replacement surgery,plasma angiotensinⅡ,atrial natriuretic polypeptide,aldosterone concentrations at all time measured compared with the preoperative levels had no significant differences in A group.But in B group and C group,compared with the preoperative and postoperative 1 week the plasma atrial natriuretic polypeptide concentrations had no significant change,plasma atrial natriuretic polypeptide concentrations measured in postoperative 2 weeks started to decline,plasma aldosterone concentrations started to decline until 4 weeks after operation;until postoperative 16 weeks the plasma angiotensinⅡ concentrations in B group had a significant decrease;and in C group the plasma angiotensinⅡ concentrations had significant decrease in 8 weeks after operation.Compared with B group and C group,the decreased level of plasma angiotensinⅡ,aldosterone concentrations after operation had no significant difference,but the decreased level of plasma atrial natriuretic polypeptide concentrations in C group was better than B group. Conclusion Patients after mechanical heart valve replacement surgery with routine cardiocinetic,diuretic therapy,and in addition supplemented angiotensin converting enzyme inhibitor and/or β-blocker treatment can effectively reduce the neuroendocrine hormone levels.

Abstract:Aim To investigate the relationship between adiponectin,metabolic abnormalities and cerebral artery atherosclerotic stenosis in patients with acute cerebral infarction complicated by metabolic syndrome(MS). Methods 74 patients with acute ischemic stroke were studied who were hospitalized.All subjects were divided into MS group and non-MS group with the criteria of MS published by international diabetes federation(IDF).All patients underwent magnetic resonance angiography(MRA) to measure the intra-and extra-cranial atherosclerotic stenosis and reveal the incidences of stenosis.The values of plasma adiponectin were detected for all patients too. Results The frequency of cerebral arterial stenosis in patients with acute cerebral infarction in MS group and non-MS group were 85.0% and 64.7%,respectively,and there was statistical significance between the two groups(p=0.039).The patients with acute cerebral infarction complicated by MS showed lower levels of plasma adiponectin and higher values of serum triglyceride and the levels of serum total cholesterol(p<0.05),low-density lipoprotein cholesterol and high-density lipoprotein cholesterol had no significant difference compared with the non-MS group(p>0.05).The result of Logistic regression analysis indicated that MS was associated with atherosclerotic stenosis(Or=3.09,p<0.05)and the presense of plasma adiponectin was not the risk factor for cerebrovascular stenosis(Or=1.272,p>0.05). Conclusion The decrease of the levels of plasma adiponectin induced MS and was the cause of metabolic abnormalities.MS was associated with atherosclerotic stenosis.The values of plasma adiponectin was not significantly associated with atherosclerotic stenosis.

Abstract:Aim To explore the relationship between hepatic lipase gene(LIPC)-250G/A polymorphism and lacunar infarction(LI). Methods A case-control study was performed in 152 patients with LI and 108 healthy controls.The polymerase chain reaction-single nucleotide polymorphism(PCR-SNP) was used to determine the LIPC-250G/A polymorphism. Results The LIPC-250G/A genotype distribution in patients with LI was GG 51.32%,GA 36.84% and AA 11.84%,respectively.Allele frequencies for G and A were 0.697 and 0.303 respectively.The serum high density lipoprotein cholesterol(HDL-C) level of the GA and AA genotypes were significantly higher than the GG genotype(P<0.05).There were no differences in the distribution of genotypes and alleles of LIPC-250G/A Polymorphism between patients and their counterparts(P>0.05). Conclusion The LIPC-250G/A polymorphism results in a beneficial profile of blood lipids,while it may not be associated with LI.

Abstract:Aim To investigate the effects of various antihypertensive drugs on pulse wave velocity(PWV) and study the correlation of effects with high sensitivity C-reactive protein(hs-CRP) and von Willebrand factor(vWF) in patients with isolated systolic hypertension. Methods 147 patients with isolated systolic hypertension were enrolled in this study and these patients' blood pressure fulfiled the criteria of systolic blood pressure(SBP) ≥140 mmHg and diastolic blood pressure(DBP) < 90 mmHg.These patients were devided radomly into nifedipine GITS group,benazeprilat group and valsartan group and were prescribed nifedipine GITS 30 mg/d,benazeprilat 10 mg/d and valsartan 150 mg/d respectively.Blood pressure,brachial-ankle pulse wave velocity(Ba-PWV),the levels of hs-CRP and vWF were measured at baseline and 2 weeks after treatment. Results Compared with nifedipine GITS group and benazeprilat group,Ba-PWV significantly decreased in valsartan group after 2 weeks(P<0.01),the level of Ba-PWV in nifedipine GITS group,benazeprilat group and valsartan group were-413.3±107.3 cm/s,-563.3±100.3 cm/s and-717.3±147.3 cm/s respectively.Compared with the other two groups,the levels of hs-CRP and vWF significantly decreased in valsar tan group(P<0.05),and the SBP,DBP and PP significantly decreased in nifedipine GITS group compared with the other two groups(P<0.05).Correlative study indicates the change of Ba-PWV in valsartan group was positively correlated with the change of hs-CRP and vWF(r=0.96,p=0.02;r=0.67,p=0.01),but not with the change of SBP,DBP and PP(P>0.05).And the change of Ba-PWV in the nifedipine GITS group was positively correlated with the change of SBP,DBP and PP(r=0.45,p=0.03;r=-0.27,p=0.02;r=0.75,p=0.00),but not with the change of hs-CRP and vWF(P>0.05).Conclusions Nifedipine GITS,benazeprilat and valsartan all can decrease Ba-PWV.However decreasing Ba-PWV and stiffness of vessel in alsartan group mainly originates from suppressesing inflammation and improving endothelial function,but the decrease of Ba-PWV in nifedipine GITS group mainly originates from the decrease of stretching pressure exerted by the blood against the vessel walls combined with expanding of artery.

Abstract:Aim To provide a basis for the primary prevention of diabetes,hyperlipemia and hyperuricemia by comparing the difference on blood glucose,blood lipids and serum uric acid levels in community groups and regular physical examination groups,and the prevalence rates of hyperglycemia,hyperlipemia and hyperuricemia. Methods The difference on fasting blood glucose,blood lipids(including total cholesterol(TC),triglyceride(TG),high density lipoprotein cholesterol(HDLC),low density lipoprotein cholesterol(LDLC) and serum uric acid levels were compared in community groups and regular physical examination groups of Taiyuan in our hospital.The total number of these two groups were 9960. Results Fasting blood glucose,bood lipids and serum uric acid had no difference between community groups and regular physical examination groups.LDLC and uric acid between the two groups were significantly different.The incidence of diabetes and hyperlipemia in community groups were significantly higher than that in regular physical examination groups.The incidence of hyperuricemia had no difference between the two groups.The incidence of these illnesses had sex difference.The incidence of diabetes,hyperlipemia and hyperuricemia in women changed along with age. Conclusions Regular physical examination is helpful to diagnose the diabetes and hyperlipemia.The incidence of hyperlipemia and hyperuricemia is influenced by age.The old age women have high risk for hyperuricemia.

Abstract:A mass of evidence has showed that atherosclerosis is a chronic inflammatory disease,and monocytes play a crucial role in the pathogenesis of atherosclerotic plaques.It is found that in recent studies monocytes in human peripheral blood are heterogeneous.There is a growing awareness that distinct functional monocyte subsets play different roles in several inflammatory and immunological diseases,like atherosclerosis,rheumatoid arthritis,Kawasaki disease.This review aims to make a brief summary for the phenotype and function of monocytes as well as discussion about their correlation to atherosclerosis in mice and humans.

Abstract:CD4⁺ and CD8⁺ T lymphocytes is significant during the development of atherosclerosis,and in recent years a growing number of studies have shown that the CD4⁺ and CD8⁺ T lymphocytes are involved in the immune and inflammatory responses of atherosclerosis which receives the attention of scholars home and abroad,and also becomes the challenge in the field of cardiovascular disease and immunology.In this paper,CD4⁺ and CD8⁺ T lymphocytes and atherosclerosis correlation research progress are reviewed.