Abstract:Aim To investigate the plasticity of bone marrow stromal stem cells(MSCs) along endothelial lineage under different culture condition. Methods The human MSCs were isolated by density gradient centrifugation. Fluorescence activated cell sorter (FACS) was used to analyze the phenotypic characteristics of MSCs. Fluorescence Immunocytochemistry was used to observe the vWF and/or Flk-1 protein expression of MSCs after cultured with DMEM medium containing 50 μg/L VEGF, 5 μg/L bFGF, 100 mg/L ECGS or with endothelial cells(ECs) conditioned medium or cocultured with mature rabbit ECs for 5 days. Results FACS analysis indicated that the MSCs did not express CD34 (CD34 expressing cells vs negative control: 4.16%±0.16% vs 4.06%± 0.23 %, n=6, p>0.05), but expressed CD105 and CD166 (CD105 expressing cells vs negative control: 90.20%±2.35% vs 4.06%±0.23%; CD166 expressing cells vs negative control: 82.30%±3.22% vs 4.06%±0.23%, n=6, p<0.05). 33.42% MSCs began to express vWF after cultured in DMEM containing VEGF, bFGF and ECGS. 25.71% MSCs began to express Flk-1, but did not express vWF after cocultured with mature ECs for 5 days. The MSCs did not express Flk-1 or vWF after cultured in ECs conditioned medium. Conclusions Cytokines and mature ECs could induce MSCs to differentiate alone endothelial lineage, while ECs conditioned medium could not.

Abstract:Aim To investigate the effects of low density lipoprotein immune complexes (LDL-IC) on the accumulation of cholesteryl ester (CE) in mouse peritoneal macrophages and on the secretion of nitric oxide (NO) from the macrophages. Methods LDL was isolated from fresh human plasma by ultracentrifugation, and then LDL-IC was prepared by incubating LDL with IgG fraction of a sheep anti-LDL antiserum in vitro. After mouse peritoneal macrophages were incubated with LDL-IC, cellular cholesteryl ester mass was quantified by an enzymatic fluorometric methods and the morphological observation and histochemistry of macrophages were performed. In addition, nitric oxide secreted from macrophages to the medium was measured by nitric acid reductase method. Results The levels of cellular CE in macrophages were increased markedly with LDL-IC dose dependent manner, which was significantly higher than that induced by corresponding level of oxidized LDL(ox-LDL)(p<0.01). Whereas native LDL and apoB-IgG did not show any effect on the level of cellular CE. Morphologically, after exposure to LDL-IC, macrophages exhibited a phenotype typical of lipid-laden foam cell, becoming strongly stained with scarlet. In addition, the content of NO in the medium released from the macrophages were decreased and also showed a LDL-IC dose dependent manner. Conclusion LDL-IC participates in atherosclerotic formation not only by inducing the formation of foam cell, but also by damaging the function of NO of macrophages.

Abstract:Aim To investigate the change rule and correlation of carbon monoxide (CO)/heme oxygenase-1 (HO-1) and nitric oxide (NO)/nitric oxide synthase (NOS) system in atherosclerosis and the influence of the two systems on atherosclerotic progress. Methods The rabbits received 1% cholesterol diet (chol group, n=8)or 1% cholesterol diet plus L-arginine (L-arg group, n=8) or L-NAME (L-NAME group, n=8) by drinking water, or 1% cholesterol diet plus heme-L-lysinate (Heme group, n=8) or ZnPP-IX (ZnPP group, n=8) by injection in abdominal cavity for ten weeks. Results Compared with contrast group, in chol group, aortic NO production and expression of NOS were decreased markedly; however, CO production and HO-1 activity were increased obviously (p<0.01). Compared with chol group (40.2%±8.9%), aortic plaque areas of heme group(26.6%±9.2%) was reduced distinctly while aortic CO production and NOS activity were increased obviously (p<0.01). However, compared with contrast group, HO-1 expression and CO production in heme group were decreased markedly (p<0.01) while they were not different from the chol group (p>0.05). Compared with chol group, aortic NOS activity and NO production of L-arg group were increased obviously and aortic plaques area (28.1%±7.7%) was reduced greatly (p< 0.01 ). HO-1 expression and CO production of L-arg group were increased distinctly compared with contrast group while they were not different from chol group (p>0.05). Compared with chol group, aortic c-myc and c-fos expression of both heme group and L-arg group were reduced markedly while in ZnPP and L-NAME group they were not different from chol group. Conclusions The reciprocal relationship between heme oxygenase/carbon monoxide and nitri coxide synthase/nitrogen monoxide system in atherosclerosis could play the inhibitory role against atherosclerotic lesion.

Abstract:Aim Progressive vascular disease is the leading cause of death in patient with diabetes mellitus. Advanced glycation end products (AGE) modified proteins are found in plasma and atherosclerosis lesion of diabetes mellitus patients. The study was conducted to elucidate the infection of AGE on expression of adhesion molecules E-selectin on human endothelial cell. Methods Human umbilical vein endothelial cell were coincubated in vitro with native human serum albumin(HSA), HSA modified with advanced glycation end products(AGE-HSA). The expression of adhesion molecule E-selectin was determined by immunofluorescen staining and flow cytometer analysis. Results E-selectin was not constitutively expressed on hUVEC. AGE-HSA enhanced the express of E-selectin on hUVEC in a time- and dose- dependent manner. HSA had no effect on the expression of adhesion molecule. Conclusion AGE up-regulates the expression of adhesion molecule on human endothelial cell and this can promote the infiltration of monocytes/macrophages in the intima and the formation of atherosclerosis.

Abstract:Aim To investigate the effect of fibrinogen on proliferation,chemotaxis and migration of vascular smooth muscle cell (VSMC) and to explore the possible effect and mechanism of fibrinogen in the formation of atherosclerosis. Methods Rat aorta smooth muscle cells were primarily cultured. VSMC proliferation was evaluated by BrdU incorporation assay and visual cell counting. VSMC migration and chemotaxis were measured with scrape assay,time-lapse microcinematography and Boyden's chemotaxis assay. Glutinase A expression and activity were assessed by Western blotting and zymography techniques. Results BrdU incorporation assay(r=0.880,p<0.05)and visual cell counting (r=0.860,p<0.05)showed that fibrinogen stimulated the proliferation of VSMC dose-dependently in the range between 0.2 g/L and 3.2 g/L. The cell counts of VSMC after 48 h incubation with the concentrations of fibrinogen as 0.4,0.8,1.6 and 3.2 g/L were significantly higher than control group[(6.32±0.39)×10 7 cells/L,(10.63±0.25)×10 7 cells/L,(12.31±0.44)×10 7 cells/L and (13.59±0.43)×10 7 cells/L vs.(5.63±0.34)×10 7 cells/L,(p<0.01)]. The BrdU incorporation rates of fibrinogen of 0.4, 0.8, 1.6 and 3.2 g/L were also significantly higher than control group (5.2%±2.2%,17.5%±2.0%,21.5%±2.3% and 25.5%±2.5% vs.2.0%±0.8%,p<0.01). Fibrinogen stimulated the chemotaxis of VSMC dose-dependently in the range between 0.2 g/L and 3.2 g/L in Boyden's chemotaxis assay (r=0.957,p<0.01). Fibrinogen had no obvious effect on VSMC migration in scrape assay and on VSMC migration at random direction in Time-lapse microcinematography (p>0.05). Fibrinogen induced the up-regulation of expression and activity of glutinase A. Conclusion By recruiting vascular smooth muscle cells from the media into the intima and promoting them proliferate, fibrinogen may be involved in the pathogenesis of atherosclerosis.

Abstract:Aim To investigate the effect of rapamycin on phenotype of vascular smooth muscle cell(VSMC) and intimal hyperplasia in a rat model of restenosis post balloon injury. Methods Twenty-six healthy adult male rats were randomly divided into four groups: sham group (n=2), balloon group (n=8): received balloon injury only, cellulose group(n=8) received the vehicle solution, and rapamycin group(n=8) intramusclar administration of rapamycin (suspended in solution containing 0.2% sodium carboxymethyl cellulose) started 3 days before injury at a dose of 0.5 mg/kg and continued for 14 days at a dose of 0.25 mg/kg. Phenotype of VSMC was observed with electron microscope. Abdominal aortas were analyzed for intimal area, mean intimal thickness, media area and intimal/media area ratio. Results Electron microscope showed the morphilogic characteristics of VSMC changed from contractile to synthetic after balloon injury. VSMC showed less organelles and more myofilament when exposed to rapamycin. Neointimal area and intimal/media area ratio 14 days after injury showed a 36.4% and 28.9% reduce in the animals receiving rapamycin compared with those receiving cellulose respectively(0.14±0.03 mm 2 vs 0.23 ±0.06 mm 2 and 17.70%±3.37% vs 28.86%±10.25%, respectively, p<0.01). Conclusion Rapamycin can influence the phenotype transform of VSMC, and reduce the degree of neointimal hyperplasia.

Abstract:Aim To observe the effects of captopril late preconditioning on intracellular calcium concentration ([Ca 2+ ]i) in cardiac myocytes of the neonatal rat undergone anoxia-reoxygenation injury and evaluate its delayed protective role. Methods The anoxia-reoxygenation models in cultured ventricular myocytes of neonatal rat were established and divided into 7 groups: normal, anoxia-reoxygenation,preconditioning, captopril,captopril + PKC inhibitor,captopril + NOS inhibitor(L-NAME), captopril + NF-κB inhibitor(PDTC), Using spectrophotometry, the activities of malondialde-hyde (MDA)and superoxide disputase (SOD) were observed. The lactate dehydrogenase (LDH) were observed by biochemical instruments. Using Flou-3 /AM loading and flow cytometry technique, the [Ca 2+ ]i were observed. Results [Ca 2+ ]i was increased in anoxia-reoxygenation group (789±9 nmol/L vs. 414±37 nmol/L, p<0.01). The precondition of captopril and anoxia- reoxygenation resulted in a significant decrease in [Ca 2+ ]i(594±5 nmol/L, 507±32 nmol/L vs. 789±9 nmol/L, p<0.01), at one time [Ca 2+ ]i were lightly increased than normal oxygen condition(p<0.05). Before treatment with captopril, we add PKC blocking agent, PDTC, L-NAME to cell media respectively, captopril late preconditioning relieved calcium overload during anoxia-reoxygenation condition, myocytes [Ca 2+ ]i were measured as 676±32 nmol/L,700±37 nmol/L, 689±11 nmol/L, which were significantly increased compared with captopril group(p<0.05); while compared with anoxia-reoxygenation group were still reduced(p<0.05). Conclusion Captopril late preconditioning in cardiac myocytes is triggered by slightly increasing [Ca 2+ ]i and weakening the calcium overload and lipid peroxidation effect during subsequenced anoxia-reoxygenation injury. It may include the route of PKC, NF-κB, NOS.

Abstract:Aim To evaluate the effect of recombinant human vascular endothelial growth factor 165(VEGF165) on progression of atherosclerotic plaque in rabbits. Methods Fifteen rabbits were randomly fed with normal diet or high cholesterol diet. Albumin or VEGF165 was administered by a single-intramuscular injection(2 μg/kg)to rabbits fed with cholesterol diet begining three weeks before therapy. Subsets of rabbits from control group, high cholesterol group and VEGF group underwent perfusion-fixation and harvesting of the thoracic aorta for morphometric and immunohistochemical analysis at 42 days. Results In control group, high cholesterol group and VEGF group, significant difference was shown in comparing mean plaque area (0% vs. 1.81%±0.61% vs. 24.12%±3.58%), plaque circumference (0% vs. 6.05%±1.62% vs. 25.71%±1.97%) and maximal plaque thickness (0 vs. 0.06 mm±0.002 mm vs. 0.16 mm±0.007 mm)respectively. There were significant differences(p<0.05) in neovascularization density (number of CD34-positive cell, 0 vs. 12.35±2.02 vs. 61.15±7.55 cells/ mm 2 ) in control group, high cholesterol group and VEGF group at 42 days. Transmission electron microscopy showed intimal vessels were associated with lesion and the capillary lumen contain lymphocytes. There was positive correlation between CD34 positive area and plaque area (r=0.989, p<0.001)in VEGF group. Conclusions Vascular endothelial growth factor 165 increased the rate and degree of atherosclerotic plaque formation in the thoracic aorta in a cholesterol-fed rabbit model.

Abstract:Aim To investigate the effect of infection with adenoviral vector containing cDNA for human catalase on apoptosis of cultured human vascular smooth muscle cells (VSMC). Methods Human VSMC were infected with catalase -contained recombinant adenovirus (AdCat) in vitro. Catalase protein was detected by Western blot analysis. The VSMC apoptosis was determined by flow cytometry and TUNEL assay. Results The expression of catalase in VSMC infected with AdCat was significantly higher than that in uninfected VSMC. The number of TUNEL- labeled nuclei in AdCat-infected cells was greater than that in AdLacZ-infected or noninfected cells. The percentage of apoptosis cells in VSMC infected with AdCat was higher than that in control group (p<0.001). Conclusion These findings indicate that overexpression of Catalase promoted apoptosis in VSMC. It could be a new target to prevent the restenosis after percutaneous trans luminal coronary angioplasty .

Abstract:Aim To study the influence of polygonatic Rhizome on the blood lipid level and its antiatherogenetic effet in experimental hyperlipoidemic rabbits. Methods Fourty adult Newzil and rabbits were randomly divided into normal control group, hypercholesterol model group, Xiezhikang group and polygonatic Rhizome group. Each group was fed respectively with common forage, high cholesterol forage,high cholesterol forage andding xiezhikang and high cholesterol forage andding polygonatic Rhizome. At week 0, 4 and 8,the level of total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (LDLC)and lipoprotein (a) were determined with enzyme method. At the end of week 8,all rabbits were killed to observe the formation of aorta arteriosclerosis with microscope. Results At the and of week 4(before treatment), the levels of serum TC( 10.5 ±1.6, 11.1±2.0 and 11.2±1.9 mmol/L respectively), LDLC(9.28±1.83, 10.04±2.04 and 10.7±1.89 mmol/L respectively), lipoprotein (a)(649±113, 713±101 and 641±108 mg/L respectively) in hypercholesterol group, xiezhikang group and polygonatic Rhzame group were signifycantly higher than that in the control group(serum TC 1.4±0.3 mmol/L, serum LDLC 1.21±0.19 mmol/L, serum lipoprotein (a)63.3±11.2 mg/L, p<0.01). At the end of week 8(after treatment), the level of serum TC(5.9±2.0 mmol/L), LDLC(4.35±1.64 mmol/L)and lipoprotein (a)(45.3±14.0 mg/L) in polygronatic Rhizome group were signifycantly decreased than that in the and of week 4 (p<0.01). The formation of the aortic foam cells in polygonatic Rhizome group (20%) was significantly lower than that in hypercholesterol group (100%,p<0.01). Conclusion Polygonatic Rhizome has the effect of cholesterl-lowering and anti-atherosclerosis in the experimental rabbits, its mechanism needs future explore.

Abstract:Aim To access the effects of leflunomide on experimental atherogensis of rabbits. Methods 24 New Zealand White rabbits were devided into three groups, 8 rabbits were selected as normal-diet control group, 8 rabbits as high-diet control, the others as treatment group administered with leflunomide (5 mg/d). Serum lipids, interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α), atherosclerotic plaque/intima size ratio of aorta and the number of infiltrating macrophage in plaques were detected on the 0, 8 th week. Results Leflunomide had no effect on the lipids compared with control groups, the level of circulating TNF-α, IL-6, plaque/intima size ratio and the number of infiltrating macrophage in the treatment group was lower than that of control groups (all p<0.001). Conclusions LEF could inhibit the inflammation in the lesion of atherosclerosis of rabbits and attenuate atherogensis of rabbits.

Abstract:Aim This study was designed to identify the T-type calcium channel blocker Mibefradil as a cardioplegic agent in terms of its efficacy in immature cardioprotection. Methods By a Langendorff model, 32 hearts of 14- to 21-day-old New Zealand rabbits underwent 90 min of global hypothermic (15℃) ischemia protected with a different single dose of hypothermic (4℃) cardioplegia(group ST:St.Thomas solution; group SV:St.Thomas solution combined with 0.1 μmol/L Verapamil; group SM1:St.Thomas solution combined with 0.1 μmol/L Mibefradil;group SM2:St.Thomas solution combined with 1 μmol/L Mibefradil). The coronary flux, percent recovery of the cardiac function, LDH release, myocardium MDA, and myocardium ionized calcium were compared. Results There are no significant difference in coronary flux between the groups. Verapamil provided significantly the worst postreperfused percentage recoveryof the cardiac function (developed pressure: 0.70±0.23, dp/dt: 0.72±0.23, and -dp/dt: 0.67±0.25) than the other groups (p<0.05). There are no significant difference in the cardiac function between others groups. The higher concentration Mibefradil (1 μmol/L) showed a significant reduction of LDH release (6.6±2.2 IU/L vs 9.3±3.2 IU/L)in the coronary flow, and lower myocardium MDA (1.14±0.24 μmol/g vs 1.64±0.39 μmol/g) and ionized calcium (0.147±0.020 mg/g vs 0.184±0.021 mg/g) than group ST (p<0.05). Verapamil showed a highest LDH release (12.2±2.7 IU/L) in the coronary flow and myocardium ionized calcium (0.225±0.041 mg/g)(p< 0.05 ). Conclusion St.Thomas solution combined with Mibefradil provided better protection during ischemia-reperfusion in the immature rabbit heart than St.Thomas' solution, whereas St.Thomas' solution combined with Verapamil showed the worst protective effects in immature hearts

Abstract:Aim To transfect the endothelial nitric oxide synthase(eNOS) gene into the rats' common carotid arteries injured by catheter, then to observe the influence of the transfection on the stenosis degree of the vessles. Methods 55 male Wastar rats of 3 months age were divided into normal control group (n=10), blank control group (n=15), pcDNA3.1(-) transfection group (n=15) and eNOS trnsfection group (n=15). 45 rats were accepted the intimal catheter surgery of the left common carotid except the normal control group. After operation, pcDNA3.1(-) and pcDNA3.1-eNOS were transfected respectively into pcDNA3.1(-) and eNOS transfection groups by carrier of FuGENE 6. The rats were executed 2 weeks, 1 and 2 month after the operation, the transfected common carotid arteries were took out and stenosis degree of the arteries were measured. Results The eNOS mRNA could be detected in cultured smooth muscle cell and the stenosis degree was significantly decreased in eNOS transfection group (p<0.01). Conclusions eNOS gene could be transfected in vivo into the smooth muscle cell succesefully and the transfecton also could obviously decrease the stenosis degree of the catheter injuried arteries.

Abstract:Aim To investigate the change of cyclooxygenase-2 in hypercholesterolemic rabbits carotid artery injured by balloon and the relationship with intimal hyperplasia. Methods Twenty-five New Zealand male rabbits were studied. They were fed with atherogenic diet. Four weeks later, balloon injury was conducted in right carotid artery respectively. A few rabbits were killed at 6 h, 24 h, 1 W, 2 W and 4 W. Blood samples and right carotid artery were collected in time. Intimal hyperplasia was studied by histological morphology method. The expression of tissue cyclooxygenase-2 mRNA was determined by reverse transcription-polymerase chain reaction. Result Intimal hyperplasia was present at day 7 after balboon injury, became more obvious at day 14 and in progression at day 28. The media area was not changed. The intima/media ratio increased as time went on (0.032±0.004,0.030±0.003, 0.030±0.002, 0.251±0.045, 1.111±0.182 and 1.448±0.216). No change was observed in control group. Cyclooxygenase-2 mRNA expression was trace in the carotid atery of the hypercholesterolemic rabbits without balloon injury (0.5±0.21, 0.5±0.25, 0.6±0.19, 0.6±0.26 and 0.5±0.22). The expression increased significantly after balloon injury as time went on (0.6±0.22, 0.8±0.24, 1.2±0.31, 1.6±0.36 and 1.4±0.32). After balloon injury cyclooxygenase-2 expressed stronger in injury group than in control group. Conclusions Cyclooxygenase-2 expressed stronger in injury group than in control group, which indicated that cyclooxygenase-2 may play an important role in restenosis caused by balloon injury.

Abstract:Aim To establish a new model of coronary microthrombosis model in rats. Methods A total of 87 male Sprague-Dawley rats were used in this study. Rats were anesthetized with ketamine and devided into sham group (n=6),sodium laurate group (n=54) and microspheres group (n=27). In sodium laurate (SL) group, 200 μg of sodium laurate was injected into the coronary artery while in microspheres (MS) group, different amount of microspheres (42 μm in diameter) were injected into the coronary artery, the sham group was injected saline instead. The thrombus induction and consequent of myocardial dysfunction were examined by histopathological analyses and non-radiactive microspheres. Results One hour after SL injection, there is microthrombus in coronary artery and reaches a peak three hours after the injection (66.4%±18.8%). Microscopic examination of Carstairs Stain sections (n=8) revealed that microthrombi containing fibrin strands obstructed the perforating arteries in the myocardium. Multiple small myocardial infarcts were observed in the heart. Multiple microinfarctlets were observed and a more severe inflammatory reaction was seen in SL group. LVEF was also decreased in SL group which is comparable to the injection of 1000 microspheres. Conclusions Coronary microthrombosis model was successfully induced after injection of SL into the coronary artery. The model is useful for the investigation of the pathophysiology of coronary microthrombosis.

Abstract:Aim To investigate the long-term effects of transplanting mononuclear bone marrow stem cell (MBMC) on repairing necrotic myocardium in chronic heart failure rats. Methods First,to establish chronic heart failure rat animal model. MBMC isolated from the rats were directly injected into the myocardium of transplantation group, control group were injected PBS, and sham-operated rats were underwent the surgery but not transplantation. At 4 th, 8 th and 12 th week after transplantation, myocardium were assessed by immunohistochemistry and electron-microscope. Results In heart specimens slice, by immunofluorescence-microscope, it was confirmed that there were newborn cardiomyocyte and blood vessels at the site of transplantation which nucleolus were marked with fluorescence. Myogenic cell similar with cardiomyocyte,desmin and actin could be detected by immunohistochemistry, proliferating cell nuclear antigen(PCNA) detected the proliferated cell. Immature cardiomyocyte,cell ultrastructure changes and newborn blood vessels which endothelium were not complete could also be detected by electron-microscope. All these matter became more and more with time went on. There were no significant discoveries between control and sham-operation groups, and at different time, there appeared dead cases. Conclusions MBMC transplantation can effectively prevent heart function deteriorated in chronic heart failure, regenerate cardiomyocyte and blood vessels. There are no bad responses such as new ischemia, inflammation and maliganant arrhythmia. MBMC can continue to repair necrotic myocardium. The long-term effect is notable.

Abstract:Aim To search for global protein changes in ischemic preconditioning (IP) using proteomic technology. Methods The SD rats were divided into two groups including IP group and sham groups. The left ventricle tissues of preconditioned and sham groups were directly sampled for proteomic analysis. Results Analysis of two dimensional electrophoresis (2-DE) showed: 704±27 protein spots were seen in Sham ventricle gels and 778±35 protein spots in IP ventricle gels. Differential analysis using the Student’s t-test (p<0.05) showed that the expression of 23 protein spots changed, 12 protein spots were identified by mass spectrometry. Conclusions Myocardial IP resulted in the changes of protein expression profiles in the myocardium. The differential proteins might function as molecular chaperone, decrease free radical and promote the energy metabolism of myocardium to confer cardioprotection.

Abstract:Aim To study the effects of low density lipoprotein -immune complexes (LDL-IC) on the cholesterol metabolism and on the expression of low density lipoprotein receptor (LDLR) in monocyte- derived macrophages. Methods PMA-treated THP-1 cell were used as a model of monocyte-derived macrophages. Quantitative analysis of intracellular cholesterol content was performed by cholesterol enzyme link assays. The expression of LDLR mRNA was detected by reverse transcription polymerase chain reaction(RT-PCR)and LDLR protein was detected by Western blot. Results The intracellular cholesterol level treated with LDL-IC was higher than other groups treated with LDL, IgG-IC and negative control. Both LDLR mRNA and LDLR protein values of macrophages stimulated by LDL-IC were significantly enhanced. Conclusions Incubating with LDL-IC can not only markedly elevate the intracellular cholesterol content but also promote the expression of LDLR mRNA and protein which suggest LDL-IC may be an important factor during the development of atherosclerosis.

Abstract:Aim To investigate whether hyperhomocysteinemia could induce vascular inflammatory response in aorta of rabbit. Methods Twenty New Zealand white rabbits were ramdomly divided into control group and methionine group. After twelve weeks, the level of homocysteine (Hcy) were measured, aortic lesion were obsersved, Interleukin-8 (IL-8) were determined by enzyme-linked immunosorbent assay (ELISA), the expression and activation of nuclear factor-κB (NF-κB) were studied by immunohistochemistry. Results Hyperhomocysteinemia was induced after high methionine diet, aortic endotheliocyte came off and local intima thickened. The production of IL-8 and activation of NF-κB were increased by hyperhomocysteinemia. Conclusions Hyperhomocysteinemia can induce vascular inflammatory response. Inflammatory response is one of potential cellular mechanisms by which hyperhomocysteinemia accelerates atherosclerosis.

Abstract:Aim To observe expressional change of proliferating cell nuclear antigen(PCNA) and cyclin D1 of cardiomyocytes in hypertrophic myocardium induced by pressure overload and investigate the relationship between cardiomyocytes hyperplasia and myocardial hypertrophy. Methods Rat models of pressure overload were produced by aorta constriction(AC). Myocardial mechanics were measured by carotid artery canal connected with pressure transducer and the changes of LV/Bwt and myocardium transverse diameter(TDM) were determined. The expressions of PCNA and cyclin D1 were detected by immunohistochemical method and the ratio of positive cardiomyocytes nuclear and total cardiomyocytes nuclear were then calculated. Result Myocardial mechanics and left ventricular mass index of AC group were significantly higher than that of matchable SO group (p<0.05), and progressively increased in 1, 3, 6 weeks after AC. A significant increase of positive ratios of PCNA and cyclin D1 followed by a gradual decline from it peak in the 1st week to normal in 6th week after AC were observed. Conclusion There were not only the enlarging of cellular size but also the higher expression of proliferation marker in early stage of myocardial hypertrophy. This expression of proliferation marker gradually fall to normal level along with the adaption of heart to pressure overload, while the cardiomyocytes size remained progressive enlargement.

Abstract:Aim To investigate the effects of asymmetric dimethylarginine on the expression of soluble intercellular adhesion molecule-1 (sICAM-1), nitric oxide (NO) and endothelin-1 (ET-1) in human umbilical vein endothelial cells (hUVEC), and observe whether L-arginine (L-arg) of different dosage can antagonize the effects of ADMA. Methods ADMA or ADMA plus L-arg was incubated with hUVEC for 24 h. The levels of sICAM-1, ET-1 and NO in conditioned medium were measured by means of Enzyme-Linked Immunosorbent, Griess and Assay radio-immunosorbent. Results ADMA can increase hUVEC to express sICAM-1, ET-1 and decrease hUVEC to generate No in a concentration-dependent manner (p<0.05, respectively), but these effects were gradually dismissed after giving the increasing dosage of L-arg. Compared with the control group, the group of ADMA 1 μmol/L had no significant difference in the expression of sICAM-1, NO and ET-1. Except this group, the other four ADMA groups all had significant difference compared with the control group (p<0.05, respectively). Among the five ADMA groups, they all had significant difference each other (p<0.05, respectively). When the added L-arg/ADMA ratio is big enough (L-arg/ADMA>100), there are no more improvements of endothelial function. The levels of sICAM-1, NO and ET-1 in conditioned medium were significantly correlated with the levels of ADMA (r=0.943, -0.937, 0.934, p<0.01, respectively). Conclusion ADMA can induce endothelial dysfunction by means of increasing endothelial cells to express sICAM-1, ET-1 and decreasing endothelial cells to generate NO. The degree of endothelial dysfunction is significantly correlated with the levels of ADMA. Giving exogenous L-arginine can antagonize the effects of ADMA. Effective measures to regulate the plasma level of ADMA may be the new aim to improve endothelial function and treat cardiovascular disease.

Abstract:Aim To explore the effects of -477C/T single nucleotide polymorphism (SNP) in promoter region of ATP binding cassette transporter A1 (ABCA1) gene on plasma levels of high density lipoprotein cholesterol (HDLC) and its relationship with coronary heart disease (CHD). Methods -477C/T genotypes in promoter region of ABCA1 gene was detected in 124 CHD individuals and 111 healthy individuals by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), and distribution of the -477C/T genotypes was compared between CHD group and healthy group, and also between different CHD clinical situations. The clinical indexes associated with CHD were also compared among the three genotypes. Results The higher proportion of the TT genotype and T allele was verified in CHD group than that in healthy group (p<0.05 and p<0.01). In CHD group, the higher proportion of the TT genotype and the T allele was further affirmed in ACS group than that in SAP group (p<0.01). The proportion of the TT genotype in multi lesions group was higher than that in single lesion group (p<0.05). The plasma levels of HDLC were differed significantly between the TT genotype and the CC genotype(p<0.001)in CHD group. Conclusions The plasma levels of HDLC may be influenced by the -477C/T SNP in the promoter region of ABCA1 gene in CHD group. The -477C/T SNP in the promoter region of ABCA1 gene may associate with the severity of coronary atherosclerosis and CHD.

Abstract:Aim To investigate the changes of transcranial doppler (TCD) in mind-body relaxing therapy (MBRT) of patients with cerebral arteriosclerosis (CAS). Methods 60 CAS patients were randomly divided into mind-body relaxing therapy with routine treatment group (n=30) and routine treatment group (n=30) for the treatment of 8 weeks. The clinical effects were assessed by TCD. Results Compared with before treatment, Vm, Vs, and Vd of base artery (BA) and left middle cerebral artery (LMCA )were significantly increased, RI and PI were significantly decreased in mind-body relaxing therapy with routine treatment group after treatments of 8 weeks (p<0.05 or p<0.01); the significant rate and total effctive rate of TCD frequency spectrum in mind-body relaxing therapy with routine treatment group were higher than routine treatment group (p< 0.05 ). Conclusion The mind-body relaxing therapy can significantly improve the blood flow velocity, reduce the resistance of cerebral blood vessel, and increase therapeutic effect of the patients with CAS. Mind-body relaxing therapy with routine treatment group has better effect in treatment of CAS than routine treatment group.

Abstract:Aim To detect the expression and distribution of macrophage inflammatory protein-1α (MIP-1α)in atherosclerotic tunica intima of artery from type 2 diabetics and non-diabetics in order to try to find out the effect of MIP-1α on the formation and development of atherosclerosis in type 2 diabetics. Methods We used the immunohistochemistry method to detect the expression and distribution of MIP-1α in atherosclerotic tunica intima of artery from 13 type 2 diabetics, 15 non-diabetics and normal tunica intima of artery from 7 healthy people, respectively. Then computer image analyzer was used in relative quantu analysis. Results MIP-1α was absent in normal arteries with average gray value of 234.27±8.04. There was different expression of MIP-1α in different-stage tunica intima of artery from non-diabetics with atherosclerosis: With the progression of atherosclerosis, the immunoreactivity of MIP-1α increased with average gray value of 168.40±7.69 in fatty steaks and 173.67±6.56 in fibrous plaques. MIP-1α was mainly in endothelial cells and foam cells during fatty steaks, while MIP-1α was mainly in foam cells and there was little expression in endothelial cells during fibrous plaques. Average gray value of MIP-1αin fatty steaks from type 2 diabetics was 132.73±6.01. There was a significant difference between average gray value in fatty steaks from non-diabetics and average gray value in fatty steaks from type 2 diabetics (p<0.05). Average gray value of MIP-1α in fibrous plaques from type 2 diabetics was 138.68±9.41. There was a significant difference between average gray value in fibrous plaques from non-diabetics and average gray value in fibrous plaques from type 2 diabetics (p<0.05). There were strong expression of MIP-1αin endothelial cells and foam cells during two stages of type 2 diabetics. Conclusions MIP-1α was absent in normal arter-

Abstract:Aim To investigate the endothelial function in patients with acute coronary syndrome (ACS) and the effect of percutaneous coronary intervention (PCI) therapy on endothelial function. Methods Endothelium- dependent flow-mediated dilation (FMD)and endothelium-independent nitroglycein-mediated dilation (NMD)of brachial artery were assessed by high-resolution color Doppler ultrosound. One hundred and twenty patients with ACS and thirty control subjects were enrolled in the study. First, the groups were matched for age, sex and other risk factors, people with hypertension and diabetes were excluded. FMD and NMD of two groups were compared. Second, basline FMD and NMD of all patients were measured. Then, patients were divided into PCI and medical groups. FMD and NMD for basline and three months later of two groups were compared. Results Compared with healthy controls, ACS patients had lower FMD (8.29±5.11 vs 10.64±3.82, P=0.029), but equal NMD (20.37±9.29 vs 18.41±5.83, P=0.226); For PCI group, FMD of basline and three months later was (7.86±5.51, 5.26±7.20), respectively (P=0.037); while of medical group, FMD of basline and three months later was (7.91± 4.52 , 7.14±6.99), respectively (P=0.401), No significant difference was found in NMD between two groups, before and after therapy (p>0.05). Conclusions The endothelial function was impaired in ACS patients; The treatment of PCI augment endothelial dysfunction.

Abstract:Aim To assess the relationship between hypertension and aortic dissection. Methods The study consisted of three groups: aortic dissection group (n=19), hypertension group (n=42) and control group (n=41). The structure and function of thoracic aorta were evaluated by multiplane transesophageal echocardiography. Results Compared with control subjects, the systolic and diastolic diameter, intima-media thickness(IMT), stiffness of thoracic aorta were increased and the compliance of thoracic aorta were decreased significantly (p<0.05～0.001) in patients with hypertension and aortic dissection. These changes were aggravated with increasing the grade of hypertension. The systolic and diastolic diameters of thoracic aorta were dialated significantly in patients with aortic dissection than that of in patients with hypertension (p<0.01), and the intima-media thickness, compliance and stiffness were not statistically diffferent between two groups(p>0.05). Conclusions The damaged structure and function of thoracic aorta are the pathological foundation of aortic dissection. The hypertension is the initial cause of these pathological changes.

Abstract:Aim To observe the plaque morphological characteristics and oxidized low density lipoprotei (ox-LDL) expression in the carotid arteries of brain ischemia autopsy cases. Methods Serial sections of the carotid arteries of the 8 cases with cerebral ischemia were taken and the morphology of the plaques were observed microscopically. Twenty stable and unstable plaques were randomly selected to do the immunohistochemical staining for ox-LDL. Computer image analysis system was used to measur the thickness of fibrous cap and the ox-LDL content of the stable and unstable plaques. Results 26(21.5%)of the 121 carotid artery plaques in the 8 cases were unstable plaques;while 95 plaques belong to stable plaques(78.5%)(p<0.01). The thickness of the fibrous cap in unstable and stable plques were 0.2±0.1 mm and 0.5±0.2 mm respectively(p<0.01). The immunohistochemical staining for ox-LDL were positive in both unstable and stable plaques. But the postive area was significantly larger in unstable plaque than that in stable plaques (19.4±3.3 and 14.3±2.5 respectively, p<0.01). Conclusion the carotid arteries consist of unstable and stable plaques. The unstable plaque had a relatively thinner fibrous cap than that of stable plaque and the ox-LDL postivity area was larger in unstable plaque than in stable plaque. This indicates that ox-LDL may bear a close correlation with plaque stability.

Abstract:Aim To investigate whether individuals with essential hypertension have increased cytokines, specifically interleukin-1β(IL-1β) and tumor necrosis factor-α(TNF-α), secretion in peripheral blood mononuclear cell (PBMC). Methods In a cross-sectional study involving 12 apparently healthy control and 33 patients with essential hypertension, concentrations of IL-1β and TNF-αwere compared in PBMC culture supernatant. PBMC were isolated by gradient centrifugation. IL-1βand TNF-αconcentrations in supernatant from PBMC were measured by enzyme-linked immunosorbent assay(ELISA). Results There is no difference in the spontaneous secretion of IL-1β(182±66 ng/L vs 154±63 ng/L, p<0.05) and TNF-α(142±74 ng/L vs 168±61 ng/L )by PBMC between normal control and hypertensive patients. After stimulation by lipopolysaccharide, both IL-1β(2 179±660 ng/L vs 182±66 ng/L, p<0.05; 154±63 ng/L vs 2 854±631 ng/L) and TNF-α(442±134 ng/L vs 142±74 ng/L; 500±176 ng/L vs 168±61 ng/L, p<0.05) secretion by PBMC increased dramatically compared with the spontaneous secretion. After stimulation by lipopolysaccharide, patients had increased IL-1β(2 179±660 ng/L vs 2 854±631 ng/L; p< 0.05 ) secretion by PBMC compared with controls. Conclusions These data suggest that PBMC in patients with essential hypertension are preactived and inflammation mechanism may be involved in the pathogenesis of hypertensive disease.

Abstract:Aim To detect the peripheral venous blood' concentration of interleukin-18 in patients with coronary heart disease and angina pectoris and evaluate its clinical correlation with angina pectoris. Methods Eighty hosipitalized patients were enrolled in this research. Forty cases were unstable angina pectoris, twenty cases were stable angina pectoris, twenty cases were normal controls. All patients were performed with coronary artery angiography and Doppler echocardiography. Interleukin-18 was measured with ELISA methods. Results The concentrations of interleukin-18 in patients with coronary heart disease and angina pectoris were much higher than controls', patients with coronary heart disease and left ventricular ejection fraction≤50% had higher IL-18 concentrations than those >50%, but there were no difference between unstable angina pectoris and stable angina pectoris. Conclusions The concentrtion of Interleukin-18 was increased with coronary arteriosclerosis and the decrease of left ventricular ejection fraction, but not related to the stability of atherosclerosis plaques. Interleukin-18 perhaps had important values in diagnosing coronary artery diseases.

Abstract:Aim To evulate the value of 12-lead holter in the diagnosis of coronary artery diseases. Method 60 cases were assessed by 12-lead holter and coronary angiography. Postive standard of 12-lead holter was ST-segment horizontal or downward sloping depression of greater than 1 mm (0.1 mV) at least 60 ms after the J point 80 ms in near two leads. Postive standard of coronary angiography was stenosis of coronary artery diameter reduction more than 50 percent than near normal vessel. Results ST-segment horizontal or downward sloping depression of 56 branches stenosis in 32 cases reaches postive standard,postive rate is 69.64% (p<0.05). In 35 branches of stenosis diameter is lower than 50%,only 9 cases ST-segment horizontal or downward sloping depression reaches postive standard, false postive rate is 25.7%. Conclusions 12-lead holter has an important value in diagnosis coronary artery disease,espically in aging and severe coronary artery disease.

Abstract:Aim To investigate the association of the blood von Willebrand factor (vWF) and C-reactive protein (CRP) of diabetes. Methods Blood specimens of 93 new diagnotic diabetic patients whose blood glucose level was more than 10 mmol/L were collected. The vWF and CRP were assayed by ELISA. Results The level of vWF and CRP of diabete group increased significantly than the control normal group, there were not any associative relationship in the level of blood vWF and CRP. Conclusions The elevated concentration of plasma vWF and serum CRP showed that the new diagnotic diabetic patients had the endothelia cell dysfunction and chronic systemic inflammation, vWF and CRP were the independent promoting factor of atherosclerosis.