Abstract:Aim To investigate the effects of basic fibroblast growth factor (bFGF).newborn calf serum (NCS) and heparin on rat vascular smooth muscle cells (VSMC) migration in vitro.Methods Cultured rat VSMC were starved for 24 h and then scraped with a single-edged razor blade. Cell migration in vitro was assayed by measuring the maximal linear distance of cell nucleus from the cut edge with a calibrated ocular micrometer after cells were incubated with different does of NCS or bFGF or heparin for 24 h, 48 h. 72 h.Results The migration distances of VSMC stimu-lated with various concentrations of NCS or bFGF were significantly longer than those of the control group), respectively(p>0. 001 or P<0. 01). But the migration distances of VSMC treated by heparin and NCS were shorter than the control (20% of NCS)dis-tances. All the effects were in a dose-dependent fash-ion.Conclusion NCS and bFGF could stimulate VSMC migration significantly. However, heparin markedly inhibited VSMC migration induced by NCS.

Abstract:Aim To obtain the protein structure deduced from tree shrew (TS) apolipoprotein CI (apo CI) cDNA se-quence.Methods Using Chou's method modified and some related computer softs, not only TS apo CI cDNA sequence cloned in a constructed cDNA library of TS liver tissue was analyzed and compared with those of other species, but also the deduced amino acid (AA) sequence, its secondary structure and hydrophobility of the protein were determined and predicted.Results TS apo CI cDNA sequence consists of 380 nucleotides encoding an 88 AA apo CI precursor (a 26AA signal peptide and a 62 AA mature protein. The function domains of the protein were predicted to locate in two regions of 13~26 AA and 32~57 residues.The secondary structure may contain several kindsof conformations. There is an obvious hydrophobic region at the C-terminal of the amino acid sequence.Conclusion The present results shows TS apo CI is more hydrophobic and may have a stronger ability of binding lipids than human apo CI.

Abstract:Aim To identify possible mechanisms of the sug-gested high atherogenicity of lipoprotein (a) [LP(a)],the susceptibility of LP (a) to Cu2+-induced oxidation and its effects on proliferation of rabbit aortic smooth muscle cells (SMC) were studied and compared with those of low density lipoprotein (LDL).Methods Human Lp(a) and LDL were isolated and purified from normal blood plasma by ultracen-trifugation, sephacryl S-400 gelpermeation chromatog-raphy. The oxidative modification of Lp(a) and LDLwas identified by agarose gel electrophoresis and thio-barbituric acid reacitive substance(TBARS) method.Cell counting and 3H-TdR incorporation were used to observe the mitogenic effect of lipoproteins and oxi-dized lipoproteins on SMC.Results The susceptibility of LP (a) to oxidation was lower than that of LDL. Native Lp(a) and LDL induced slight increase in number of SMC and 3H-TdR incorporation respectively, which was statistically in-significant (P>0. 05); ox-Lp(a) and ox-LDL promot-ed the increase of cell number and DNA synthesis significantly (P<0. 01), and the promoting effects were greater than those of the corresponding native lipoproteins (P<0. 05, P<0. 01).Conclusion Oxidized LP (a) might play a role in atherogenesis through stimulating the proliferaion of SMC.

Abstract:Aim The effects of replacement of amino acid residue Va1156 with Gln or deletion in Gln235 on the capacity of apolipoprotein AI(APo AI) to promote cholesterol efflux from murine peritoneal macrophages.Methods Cultured mouse peritoneal macrophages were used. 3 H-oleate labled acetyl low density lipoprotein (ac-LDL) was employed as an inducer for foam cell formation. After macrophages were cul-tured with Apo AI, 3 H-cholesterol released from cells to the medium was measured by scintillation spectrom-etry.Results The replacement of Va1156→Glu did not influence the capacity of Apo AI to promote cellular cholesterol efflux. However, a deletion in the residue Gln235 significantly reduced the ability of Apo AI to promote cellular cholesterol efflux.Conclusions Amino acid residue Glu235 of ApoAI is important for cellular cholesterol efflux.

Abstract:Aim To study the inhibitory effect of locally delivered c-sis proto-oncogene antisense oligodeoxynucleotide (ODN) on aortic smooth muscle cells (ASMC)proliferation and neointimal thickening. following balloon angioplasty in the rabbit aorta at differential time.Methods 60 male New Zealand, White Fabbits were treated with local delivery of c-sis antisense ODN,c-sis sense ODN, or 0. 9% salt solution via balloon catheter following aorta balloon injury, ASMC proliferation and area in neointima was assessed with immunohistochemistry and image analyzer at 3, 7, 14, 28 days after local delivery.Results ASMC proliferation in neointima were reduced 69. 8%. 59 %, 51. 9%, 47. 8% in c-sis antisense ODN-treated group compared to c-sis sense ODNtreated group at 3, 7, 14, 28 d following aortic injury respectively; and the inhibitory rate of neointimal hyperplasia was 48. 9%, 77. 8%, 72. 6%, 70. 8% at the same time respectively.Conclusion The locally deliveried C-sis antisense ODN inhibited ASMC proliferation and neointimal thickening following rabbit aortic injury, the inhibitory effect was the highest at 3 d and 7 d, then reduced slowly. and caused by antisense mechanism.

Abstract:Aim To investigate the effects of tranilast on proliferation and c-myc protooncogene expression in vascular smooth muscle cells(VSMC)Methods In vitro cultured rat VSMCs model was established.3H-thymidine (3H-TdR)incorporation test and, reverse transcription-polymerase chain reaction (RT-PCR) technique were used.Results Both platelet derived growth factor-AB(PDGF-AB)and AngiotensinⅡ(AtⅡ) Were able to induce expression of c-myc protooncogene and in-creased DNA synthesis in VSMCs.Tranilast markedly inhibited the DNA synthesis in VSMCs in-duced by both PDGF-AB and AtⅡ. In the mean while,the drug was able to inhibit the c-myc protoonco-gene expression in VSMCs induced by both PDGF-AB and AtⅡas well.Conclusion Tranilast can inhibit VSMCs prolifera-tion and expression of c-myc protooncogene. The in-hibitoty effect of tranilast on the proliferation of VSM-Cs may be related to the inhibition of c-myc expres-sion.

Abstract:Aim Whether the mutation of lipoprotein lipase (LPL) gene is associated with coronary heart disease (CHD) or not has been yet knon We study rela-tionship between mutation in intro 6(PvuⅡ) and 8region of LPL gene and CHDMethods We analysed 106 patients in myocardial infarction survivors through a case-control study.The technique of lpoymerase chain reaction (PCR)-re-striction fragment lengths polymorphism(RFLP)was useh to analysed the genotype of LPL gene.23 fac-tors maybe involved in CHD was chosed. Simple and multiple non-conditioned logistic regression analysis demostrated that 6factors were association with CHD.Results The risk factors were diabetes, smoke,hypertension,LPL GENOTYPE pvu ⅡP P and haplo-type H+ P-. While Apo A/B ratio was regarded as a protective factor.Conclusion LPL genotype Pvu P-P- or H+P- oth-er than the standard risks was contribuing to CHD.

Abstract:Aim To observe the changes of plasma lipid and apolipoprotein levels in subjects aged over 40 with im-paired glucose tolerance.Methods 53 subjects whose fasting blood sugar(FBS)<6. 11 mmol/L and 2 h postprandial BS≥6. 66mmol/L were selected as the impaired glucose toler-ance (IGT) group: 27 subjects whose FBS<6.11mmol/L and 2 h postprandial BS<6. 66 mmol/L wereselected as control group. Oral glucose tolerance test and insulin release test were performed by routine methods. Plasma glucose and lipids such as triglyceride (TG), total cholesterol (TC) and high densitylipoprotein cholesterol (HDLC) were measured by en-zyme methods. The plasma low density lipoproteincholesterol (LDLC) was determined by calculation.Plasma insulin was measured by radioimmunoassay.Plasma apolipoproteins were measured by radial im-munodiffusion methods.Results The body mass index (BMI), FBS and fasting plasma insulin of the two groups were statistically the same (P>0. 05). 2 h postprandial BS of IGT group was 47.9%, and 2 h postprandial insulin was 69. 8% higher than those of the control group (P<0.05 and P<0.01). The serum levels of TG and TC of IGT group were significantly higher than those of the control group (P<0. 01 and P<0.05). To some extent but not statistically, HDLC was lower,LDLC was higher than those of the control group. The serum apo CII, CIII and E levels of IGT group were statistical-ly higher than those of the control group (P<0. 01, P<0. 001and P<0. 005). To some extent but not statistically, apo B was higher and apo AI was lower than those of the controlgroup.Conclusion Plasma lipid changes of subjects aged over 40 with impaired glucose tolerance are similar todiabetic dyslipoproteinemia and these changes may berelated to the changes of plasma insulin.

Abstract:Aim 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 were collected. Then the endothelial cell-conditioned media (ECCM ) were treated by heparin-binding sepharose, and the mitogenic activity of ECCM for swiss 3T3 cells were determined by incorporation of 3H-thymidine into DNA of the cells. The concentration of endothelin in the media of endothelial cells were analysed by radioimmunoassay.Results Cumene hydroperoxide induced storage oflipid peroxide in cultured endothelial cells, and there was an obvious increase in the incorporation of 3H-thymidine into DNA in Swiss 3T3 cells treated with ECCM compared with the control. However, the mito-genic activity of ECCM for Swiss 3T3 cells were in non heparin-binding parts. Then the concentration of en-dothelin in the media of endothelial cells were treated by cumene hydroperoxide were increased obviously compared with the control.Conclusion It seems reasonable to believe that lipidperoxidation of endothelial cells induced by cumene hy-droperoxide leads to an increased synthesis and secre-tion of growth factors, and the mitogenic activities were due to non heparin-binding parts like as endothe-lin.

Abstract:Aim Calcitonin gene-related peptide (CGRP), a princlpal transmitter in cardiac and vascular sensory nerves, is released from the heart during ischemia,which protected the myocardium against ischemia-reperfusion injury in isolated rat hearts. The purpose of this study was to investigate whether CGRP-induced preconditioning has cardioprotective effects in intact rat hearts.Methods 36 SD rats were at random divided into three groups of control, ischemic preconditioning (IPC) and CGRP. CGRP (10 μg/kg) was given 20min before 30 min ischemia. All animals were .sub-jected to 30 min of regional ischemia followed by 2 h of reperfusion. The preconditioning protocol consisted of 3 cycles of 5 min regional ischemia and 3 cycles of 5min of reperfusion. Infarct size as a percentage of the area at risk was determined with nitro blue tetrazoli-um.Results Both of IPC and CGRP-induced precondi-tioning markedly reduced the incidences of ventricular arrhythmias during ischemia and reperfusion. Both ofIPC (15. 7%, P< 0. 01) and CGRP-induced precondi-tioning (28. 8%, P<0.01) also resulted in a signifi-cant reduction in infarct size when compared to control(54. 0%).Conclusion These results support the hypothesis that CGRP-induced preconditioning has cardoprotective effects in intact rat hearts.

Abstract:Aim To observe the regulatory effect of cytokins on the expression of CD44, CD11b and Vn as well as the regulation change of cytokines to above antigen during coronary heart disease (CHD).Methods The distribution of CD44, CD11b and Vnon the mononuclear ce11s was displayed by means of immunohistochemistry, and followed by quantitative process with Quantimet 52O+image analysis system,Finaly, the data were Processed by factorial analysis of statistics.Results The positive area of CD44, CD11b and Vn shows pink or dark pink color, and display a ring,semilunar, cap, spot-flat or granular shape. The dis-tribution of the three antigens the mononuclear cell surfaces is different. Quantitative analysis indicates that the Sum value increased obviously as a result from the effect of cytokines (P<1. 001) Which showed more significant in patients with coronary heart disease than in control (P<0.001).Conclusion Interleukin-1, tumor necrosis factor and interleukin-6 may up-regulate the expression of the adhesive related proteins (ARPs) and the effect of in-terferon-alpha is less intensive in CHD. Which indi-cate that the sensitivity of CHD patient's ARPs expres-sion to regulatory effect of cytokines is much higher than that of normal people.The present study shows that the cytokines can affect the adhesion of mononu-clear cells to endothelium via the regulation of ARPs expression and then interfere with atherogenesis.

Abstract:Aim To investigate the condition of very low density lipoprotein (VLDL) receptor mRNA expression by smooth muscle cells (SMC).Methods Rabbit aortic SMC was cultured by digestion method. Total RNA from cultured SMC with or without interleukin-1β) (lL-13β) was extracted by the acidguanldinium thiocyanate-phenol-chloroform method. followed by mRNA isolation using an oligo (dT)-cellulose column. The expression of specific VLDL receptor mRNA in SMC was determined by Northern blot analysis.Results The VLDL receptor mRNA in cultured rabbit aortic SMC can be detected by Northern blot analysis. After cultured SMC exposure to 1 μg/L of lL, 1β for 15 h, the level of specific VLDL receptor mRNA increased by two fold as much as unstimulated cells.Conclusion Cultured rabbit aortic SMC was able to express VLDL receptor mRNA, and IL-1β could up-regulate the expression of VLDL, receptor mRNA in the cultured SMC. It suggested that SMC might take in lipoproteins through VLDL, receptor, and so became foam cells. IL-1β might be concerned with pathogen-esis of atherosclerosis by its effect on SMC VLDL re-ceptor mRNA expression.

Abstract:Aim To investigate the Protective effects of SK-827 on brain ischemia,/reperfusion (I/R )injury in rabbits.Methods Plasma lactate dehydrogenase (LDH) ac-tivity. malondialdehyde (MDA) content, lactate con-tent and brain ATP, MDA 1evel were assayed in I/Rgroup and SK-827 treated group.Results In SK-827 group, LDH activity, lactate content and MDA content in plasma and brain de-creased, ATP level in brain increased and encephalede-ma improved significantly as compared with I/Rgroup.Conclusion SK-827 might have protective effect onbrain I/R injury in rabbits.

Abstract:To study the correlation between the activity of Na+-K+ -ATPase on erythrocyte membrane and arteriosclerotic cerebral infarction. Activity of Na+-K+ -ATPase on erythrocyte membrane was determined in 37 cases of arteriosclerotic cerebral infarction (ACI)and 25 normal control subjects. Activity of Na+ -K+-ATPase on erythrocyte membrane in ACI was obviously lower than that of control (P<0. 001), and the activity of Na+ -K+ -ATPase on erythrocyte membrane was negative correlated with infarct size on brain CT scans in ACI(r= -0. 58, P<0. 01).It suggested that the reduced activity of Na+ -K +-ATPase on erythrocyte membrane may have some ef-fect in the development of ACI.

Abstract:Aim To investigate possible oxidative injuries topatients during hemodialysis process with differentmembranes.Method The levels of malondialdehyde(MDA)andsuperoxide dismutase (SOD) in both plasma and redblood cell, gultathione peroxidase (GPX) in whole blood samples of maintenance hemodialyzed patientswere exanlined before and after dialysis with cupo-phane, polysulphone and hemophan membranes,and were compared with those of health controls.Result A significant increase of MDA level and adecrease of SOD activity after dialysis with cupophane membrane, while no significant differences were ob-served with polysulphone and hemophan membranes.Conclusion The use of polysulphone and hemophan membranes for long term hemodialysis is benefit to Prevention and cure any syndrome caused by the in-crease of peroxide radicals.

Abstract:Aim To analyze the lipid profile changes in chronicrenal failure patients.Methods Forty cases of chronic renal failure patients were studied. Triglyceride (TG), total cholesterol (TC), high density lipoprotein cholesterol (HDLC), low density lipoprotein cholesterol (LDLC).apolipoprotein AI (Apo AI) and apolipoprotein B (ApoB) were assessed.Results The values of TG, Apo B were higher andvalues of HDLC, Apo AI, Apo AI/Apo B were lowerin chronic renal failure patients compared with control group. The values of TC and LDLC, however, were no different from that values in the control group.Conclusion Lipids abnormorlities were present in some chronic renal failure patients.

Abstract:In order to extract genomic DNA from frozen blood clots,a new exclusive method was esteblished,Long time application of it indicates that this method is easy, stable and expeditious, About 30 μg DNA is obtained from 1 cm3 clo,It is very suitable to pre-pare genomic DNA from large samples of blood gotten in epidemic fields survey,