Abstract:The sera of hypercholesterolemia (HC)from people of Beijing and Guangzhou were incubated with U937 cells respectively for 40 hours,then binding rate between U937 cell and fluorescencelabeled high density lipoprotein(HDL) were detected on cell sorter. The result showed that the binding rate of cells treated with HC from Beijing was significantly higher(2～20 fold) than that from Guangzhou, of which the binding rate of cells was similar to control cells(treated with normal lipid sera). Apolipoprotein E(Apo E)and its receptor pathway did not play an important role in the binding process,HDL3 is major subclass of HDL which bound to U937 cells,These findings indicated that HC from Beijing has a role to stimulate monocyte cell to take lipids, while HC from Guangzhou doesn't. The difference should exist in plasma component between the masses of Beijing and Guangzhou with HC, which may contribute to high incidence of coronary heart disease in Beijing compared with that in Guangzhou.

Abstract:Fluorescein isothiocyanester(FITC)lablled high density lipoprotein-2 keeps the biological activities of native HDL.After human SMMC-7721 hepatocarcinoma cells were incubated with FITCapolipoprotein E-free HDL-2 at 37℃ for 3h, the cellassociated and endocytic fluorescent strength (FS)were 13.5±0.8 and 9.2±0.5(x±s,n=2) respectively. The cell-released trichloroacetic acid (TCA)precipitable FS and TCA-soluble FS were 5.1±0.4and 0.67±0.17 after the cells were incubated further at 37℃ for 2 h. And the cell-released TCA-precipitable FS was 0.41±0.16 at 4℃ for 2h.These results suggest that:①cell-internalized doesn't take lysosomal pathway;②the released of cell-internalized HDL-2-apolipoprotein is a temperature-dependent retroendocytosis.

Abstract:In this study, the effects of dexamethasone(Dex) on the DNA ynthesis and proliferation of vascular smooth muscle cells(VSMC)were studied by 3H-thymidine(3H-TdR) incorporation and counting cell number. With Northern blot analysis and measuring the amount of nitrite, we observed the expression of inducible nitric oxide synthase (iNOS)gene in VSMC of control and treated by tumor necrosis factorα(TNF-α),TNF-α+Dex respectively.It was found that the DNA synthesis and proliferation of VSMCwere obviously inhibited by dexamethasone,the inhibitory rate was proportional to the dose and the duration of dexamethasone treatment.The results ofNorthern biotting and quantity of nitrite in culturemedium showed that dexamethasone obviously inhibited the expression of iNOS gene and nitric oxide (NO)production in VSMC induced with TNF-α. These results indicate it is not by increasing the NO production that dexamethasone inhibits the DNA synthesis and proliferation of VSMC.

Abstract:Aim Observed the expressive quantity and distribution of adhesion related proteins(ARP) in the mononuclear cells among the patients with coronaryheart disease (CHD) and normal people.Methods Display the distribution of CD11b, CD44and Vn on the mononuclear cells by means of immunohistochemistry and morphological metrology methods.Results The positive area of CD11b,CD44 and Vn shows pink or dark pink colour, and display a ring shape, semilunar shape, cap shape, spot-flate shape and granular shape,Quantitative analysis indicate that the Sum and Ar value increased obviously in the CHD.Conclutions The Expression of the ARP is higher in CHD group then in normal. There is a obvious difference among the three kind of proteins in expresive quantity which from high to low is CD44, Vn,CD11b. In CHD group the increase of the three ARP are different in degree, the first is CD11b, the next is Vn and then it is CD44.

Abstract:Aim This study is to investigate the effects of natural very low density lipoprotein (n-VLDL)subfractions (Apo E-poor VLDL and Apo E-rich VLDL)on triglyceride (TG) and cholesteryl ester (CE) accumulation in mouse peritoneal macrophages (MPM).Methods VLDL for subfraction by HeparinSepharose 6B affinity chromatography was obtained from 400 ml of normal human plasma by ultracentrifugation.MPM were incubated with Apo E-poor VDLD or Apo E-rich VLDL at the concentrations of 10～40 mg protein/L DMEM for 24 hours.Results Followed the increased concentrations of VLDL,the accumulation of TG and CE in MPM increased(P<0.05).The TG content in MPM wasmuch higher in Apo E-poor VLDL group than that in Apo E-rich VLDL group (P<0.01). There was little difference in CE content in MPM between both groups (P>0.05).Conclusion These results suggest that both VLDL subfractions can stimulate TG and CE accurnulation in MPM and accelerate the transformation of macrophages into foam cells.

Abstract:Aim Total Panax Notoginsen saponin(tPNS) is a traditional herb medicine of Chinese. In this study, we have investigated the release of nitric oxide (NO) from cultured porcine aorta endothelial cells (PAEC) affected by the tPNS for understanding the pharmacological mechanism of tPNS.Methods PAEC were cultured with M199 and compared the morphological appearence of tPNS group with the control under the phase contrast microscope after PAEC exposed to conditional medium (CM) for 24 hours. NO amounts in CM were determinated by the methods of Termin and Green.Results PAEC in tPNS groups were grew normally. The cells of control group were constricted and separated from each other, there were a lot of small vesicles in the cytoplasma. The amounts of NO in CM of tPNS groups were ranged from 350±59 to 410±90nmol·L-1, and compared with the control, the P<0.001. This enhance was ceased by pluinq EDTA in CM. The time course expressed that the amount of NO was raised quickly by pluing tPNS and was maintaining a high level. This effect could stop by adding the EDTA in CM too.Conclusions The results of enhancing the release of NO from PAEC and maintaining the growth of PAEC by tPNS were suggested that the tPNS can playa significant role in some diseases such as atherosclerosis.

Abstract:Aim The purpose of this paper is to investigate whether the lipid peroxidation of endothelial cells had any effect on the production of growth factors by them.Methods The bovine aortic endothelial cells were isolated and cultured using an enzyme-dispersal method.and the media conditioned by endothelial cells with lipid peroxidation induced by cumene hydroperoxide (LPECCM) were collected. The mitogenic activity of LPECCM for Swiss 3T3 cells was determined by incorporation of[3H]-thymidine into DNA of the cells.The effect of lipid peroxidation of endothelial cells on their synthesis and secretion of PDGF was performed with a neutralization assay using anti-PDGF antibodles and Northern blot analysis. Results Cumene hydroperoxide induced storage of lipid peroxide in cultured endothelial cells, and there was an obvious increase in the incorporation of[3H] thymidine into DNA in wiss 3T3 cells treated with LPECCM compared with the control.However.the mitogenic activity of LPECCM for Swiss 3T3 cells were not inhibited by anti-PDGF-AA and anti-PDGFBB antibodies. Conversely, a weaker PDGF-B chain mRNA expression in endothelial cells was observed after exposure to cumene hydroperoxide.Conclusions It seems reasonable to believe that lipid peroxidation of endothelial cells induced by cumene hydroperoxide leads to an increased synthesis and secretion of growth factors other than PDGF in cultured bovine aortic endothelial cells.

Abstract:Aim The effects of API0134 on expression of c-sis mRNA and platelet-derived growth factor B chain protein(PDGF-B)in porcine aortic endothelial cells (EC)induced by oxygen free radical (OFR) were studied in this article.Methods Immunohistochemistry,in situ hybridization and computer image analysis methods were used.Results The results showed,that OFR induces csis mRNA transcription and stimulates PDGF-B expression in EC;The inducible effects of OFR can be inhibited by API0134,the inhibitory effects are dose dependent.Conclusion These data suggest that API0134 may play a role in anti-atherosclerosis.

Abstract:The ability of human monocyte (MC)to interact with human umbilical vein endothelial cell (VEC) midiated by oxidized low density lipoprotein (OLDL) was determinated by the use of a micromanipulation technique. The ability of polymorphonuclear neutrophil (PMN) was observed at same condition to show the speciality of OLDL’s effect.The results showed that treatment of VEC with OLDL caused a significant increase in critical separation stress (Sc) MC-VEC.contrast, of PMN-VEC was not increased.Treatment of MC and PMN with OLDL caused a repid increase in Sc of MC-VEC and PMN.VEC,the Sc was maximal within 1 hour of exposure to OLDL and decreased in 24 hours.The results suggest that OLDL can enhance MC-VEC interation specially, this effect indicate that OLDL is important factor for the binding of MC to VEC and the entry of MC into the vessel wall in atherosclerotic lesion.

Abstract:Aim Study on the metabolic pathway and the interrelation both low density lipoprotein and lipoprotein (a) in the body of the hedgehogs Methods The purified low density lipoprotein and lipoprotein (a) from the human plasma were obtained by sequence ultracentrifugation and sepharose 6B chromatography.They were injected into the body of the hedgehogs by armpit vein under anaesthesia with etherafter they were labelled with Na125 I tyramine-cellobiose, The animals were put to death 3,6,12,24,48,72,96 hours after low density lipoprotein and lipoprotein (a) were injected. The radioactivity in the blood, liver, kidney, spleen and gall were measured.The results were showed by figures.Results Low density lipoprotein in the blood has at 1/2 of 15 hours, while lipoprotein (a) has at 1/2 of 45 hours (Figure 1). The levels of low density lipoprotein in the liver present two peaks which appear in 3 and 72 hours after injecting, and was the lowest in 24 hours after injecting; on the contrary the levels of lipoprotein (a) in the liver keep relatively stable (Figure 2). The same result was seen in the gall that the levels of low density lipoprotein present two peaks,but the second peak move up and the peak was lower.The level of lipoprotein (a) in the gall was same as in the liver (Figure 3). The levels of low density lipoprotein in the kidney same as in the liver. The level of lipoprotein (a) present a peak in 3 hours after injecting, then it goes down and keep reletively stable (Figure 4). The levels of low density lipoprotein in the spleen present single peak which was in 48 hours after injeting. The levels of lipoprotein (a)goes down in 6 to 12 hours and then goes up (Figure 5).Conclusion The results provide that low density lipoprotein was matabolized through two ways; B or B/E receptor and scavenger path. It is possible that lipoprotein (a) was taken up by passive course or was matabolized by other receptor way.

Abstract:White Carneau pigeons(WCP)have previously been sbown to be genetically susceptible to the development of spontaneous atherogenesis. Analysis of levels of total hydroxyproline and isodesmosine in the thoracic aorta and celiac bifurcation of prelesion,six-week-old WCP and Show Racer pigeones (SRP), revealed an increased accumulation of total collagen and cross-liked elastin in the WCP arterial tissue. Using dot blot hybridization, measurements of steady state levels of several mRNA in total RNA extracted from pigeon aortic tissue were determined.While the increased deposition of extracellular matrix proteins was paralleled by significantly greater recovery of mRNA coding for pro α1(1) collagen and elastin, in RNA extracted from WCP aortic tissue, increased recovery of mRNA coding for an intracellular protem, γ-actin were observed in WCP aortic tissue.No differences in steady state levels of mRNA coding for pro α1(1) collagen and elastin were observed in RNA extracted from liver of WCP and SRP.The experiment results indicate that the cell population in WCP aortic tissue exhibits a markedly different phenotype with respect to a capacity for the biosynthesis of extracellular and intracellular proteins.

Abstract:In the study,the effect of sodium Tanshinone Ⅱ a sulfonate (TN Ⅱa) on c-myc expression in vascular smooth muscle (SMC) was explored with in situ hybridization. It was showed that macrophage-derived growth factor (MDGF) could apparently stimulate high gene expression of c-myc. In contrast, TN Ⅱ a could prevent MDGF’s stimulating effect on SMC, decrease the expression level of cmyc. The experiment showed TN Ⅱ a might play an important role on preventing the proliferation of SMC in atherosclerosis.

Abstract:Aim This study evaluated the association of plasma ferritin (PF) with coronary heart disease (CHD)and its risk factor, It has been suggested that total body iron stores are an independent risk factor for CHD.Methods and Results The plasma ferritin, estrodiol (E2), testosterone(T), total cholesterol (TC),triglyceride (TG), high density lipoprotein cholesterol (HDLC), lipoprotein (a), and fasting blood glucose (FBS) were measured in 106 patients with CHD and 73 healthy controls matched with sex, age and body mass index. The results showed that FBS, TG, and lipoprotein (a) levels were significantly higher and HDLC, HDL3C, HDLC/TC and T levels were significantly lower in patients than in controls, whereas no differences in PF concentrations were found between patients and controls, The PF was positively correlated with FBS and TG.Conclusions Our results do not support increasediron store was an independent risk factor for CHD.

Abstract:The etiology pathology etc of atherosclerosis (As) have been studied widely for many years. From the traditional medicine viewpoint,the mechanism of stasis exsisted in As. So the Chinese drug recipe of removing stagnancy of vital energy of the liver and invigorating blood circulation was subjucted in treatment of As. From the molecullar level of cytobiology, the effect of Chinese drugs on content of lipid peroxide (LPO) and activity of superoxide dismutase (SOD) were studied in vitro cultured SMC of rabbit aorta. The results showed that content of LPO in SMC were lower in Chinese drugs group than that in the control group (P<0. 01), activity of SOD in SMC were higher in Chinese drugs group than that in the control group (P< 0. 01). This Chinese drug recipe may obviously ruduce the content LPO of SMC, raise the activity of SOD, protect the normal function,metabolism and complete constitution. Thus, the Chinese drug recipe reduce the pathological changes of make it tend to motionless.

Abstract:The C57BL/6J mice were assigned at random to two groups; the experimental group was fed cholesterol-rich diets; the control group received normal mouse chow. Mice receiving normal diets were completely free of atherosclerotic lesions. In mice fed cholesterol-rich diets for 16 weeks, the lesions of the aorta and coronary arteries were observed by laser scanning confocal microscopy (LSCM). The typical atherosclerotic plaques, which consisted of amounts of foam cells. were formed near the aortic valves.Roberts & Theompson remarks were 8. 0 ±1. 2. The atherosclerotic lesions were verey similar pathologically to those in the human. The plasma total and free cholesterol levels increased progressively. The obtained results suggested that an atherosclerotic murine model be successfully established. It may provide an avallable experimental model for further investigation into the molecular mechanism of atherosclerosis and screening of antiatherosclerotic drugs.

Abstract:The levels of serum lipoprotein (a)[Lp(a)], apolipoprotein A1 (Apo A1), apolipoprotein B (Apo B), high-density lipoprotein cholesterol (HDLC), low-density lipoprotein cholesterol (LDLC), total cholesterol (TC), triglyceride(TG),malondialdehyde ( MDA ) and superoxide dismutase (SOD) were measured in 30 old healthy subjects and 44 old patients with coronary heart disease (CHD).Lp (a), Apo B, LDLC and MDA were higher, and Apo A1, HDLC and SOD were lower in CHD patients than in controls (P<0. 05 or P<0. 01). Multiple regression analysis revealed that levels of LDLC correlated signiflcantly with level of MDA (r=0. 5139,P<0.001 ) and level of HDLC correlated significantly with serum SOD activity (r=0. 5702,P<0. 001). The results suggested that hyperlipemia in old CHD patients was closer relationship to lipids peroxidation.

Abstract:Aim To investigate the protective role of n-3 polyunsaturated fatty acids on endothelial function.Methods The endothelium-dependent-relaxation (EDR) experiment is investigated in thoracic aorta ring of rat's hypercholesterolemia. Functional tests of nitric oxide synthase (NOS) and smooth muscle cell were done to illustrate the mechanism.Results Results indicate that, the impaired EDR under hypercholesterolemia is due to the incapable of NOS, n-3 polyunsaturated fatty acid can greatly improves the relaxation through recover the function of NOS.Conclusions One of the mechanism of beneficial role of n-3 polyunsaturated fatty acids on atherosclerosis is its protective function on endothelium.

Abstract:In this study the change of metabolist activity was measured from macrophage to macrophage-derived foam cell by incubating mouse peritoneal macrophages in oxidized Iow density lipoprotein, using tetrazolium dye assay. The resultrevealed that the metabolist activity ascended first,then descended, and the peak was at about 3 hours with macrophage in 5 00 mg/L oxidized low density lipoprotein.

Abstract:In this paper, the method of PicoTagTM amino acid analysis was introduced to measure the level of L-arginine in aortic wall of rabbits. Firstly, the sample (isolated artergal ring of rabbits) was extracted with trichloroacetic acid and water-saturated ether. The crude product was further assayed by PicoTagTM amino acid analysis. The peak area of L-arginine's was corrected by 991 purity analysis disk, the L-arginine levelwas determined by comparing with standrad L-arginine peak area. Three groups of samples was assayed. The statistical L-arginine level of the three group is valuable.