Abstract:Aim To determine the effects of API0l34 on en-dothelin (ET)-stimulated proliferation of vascularsmooth muscle cells (VSMC ) and expression ofgrowth factor, such as platelet derived growth factor(PDGF-B ) and basic fibrob1ast growth factor(bFGF), and oncogene c-sis and c-myc.Methods The experimental models of proliferationof cultured porcine aortic smooth muscle cells inducedby ET were established, and 3H-thymidine (3H-TdR ) incorporation, flow cytometry, immunohistochemistry and Northern blot assaies were used.Results API0134 may drop 3H-TdR data in-creased by ET and hold-back VSMC from static phase(G,/G, ) to DNA synthetic phase (S) and mitoticphase (G2/M ). Furthermore, API0134 could re-verse the enhanced expression of antigen PDGF-B,bFGF and reduced expression of oncogene c-sis and c-myc induced by ET.Conclusion API0134 might inhibit DNA synthesisand proliferation of VSMC, related to the mechanismof molecular biology of controlling growth factors andoncogene.

Abstract:Aim Oxidized low density lipoprotein (LDL), verylow density lipoprotein (VLDL ) and high densitylipoprotein(HDL) were occured in vivo in endogenoushypertriglyceridemia (HTG ) human.Methods Human plasma triglycerides (TG), totalcholesterol (TC), high density lipoprotein cholesterol(HDLC), apolipoproteins A l, AⅡ, B₁₀₀, CⅢ, Eand lipoperoxide (LPO) levels in 25 endogenous HTGpatients whose plasma TG levels were>2. 26 mmol/Land TC<6. 21 mmol/L, and in 25 normal age-matchedpersons (TG < 2. 26 mmol/L, TC < 6. 21 mmol/L )were measwred. Blood glucose, TC, TG and HDLCwere performed by enzyme method. Plasmaapolipoprotein were determined by radial immunuodiffusion (RID) assay. The human plasma LDL, VLDLand HDL were isolated by the one step density gradientultra-centrification method. The oxidative modificationof LDL, VLDL and HDL was identified by agarose gelelectrophoresis, absorbance at 234 nm and fluorescenceof thiobarbituric acid reaction substance(TBARS).Results The results showed that plasma TG, LPOand apolipoprotein B₁₀₀ levels in endogenous HTGgroup were significantly higher than that of the controlgroup (p<0. 01). Plasma HDLC and apolipoprotein AⅠ contents in endogenous HTG group were significantly lower than that of the control group (p< 0.01). The electrophoretic mobility of LDL,VLDL andHDL was increased and absorbance at 234 nm andTBARS of LDL, VLDL and HDL in endogenous HTGgroup were significantly higher than that of the controlgroup (p<0. 01).Couclusion The results suggest that not only LDLbut also VLDL and HDL were oxidatively modified invivo of endogenous HTG patients-

Abstract:Aim This work investigated the protective role ofcalcitonin gene-related peptide (CGRP) on balloon de-nuded injury of rat aorta.Methods Per fusion of denuded arterial rings, de-termination of cGMP and endothelin (ET) with re-dioactive analysisResults 2 weeks after denudation, the contents ofplasma and aorta CGRP were increased. ExogenousCGRP (25 μg/kg, iv) could significantly improve therelaxation of denuded aorta to acetyl choline, inducenitric oxide (NO) synthesis (p<0. 01 ), and inhibitthe over proliferation of denuded intima (p<0. 01 ).Couclusion Exogenous CGRP could prevent therestenosis of balloon denided aorta through promtingthe repairment of denuded endothelium, inhibiting theover proliferation of injured intima.

Abstract:Aim To investigate the effect of lipopolysaccharide(LPS) on the expression of inducible nitric oxide syn-thetase (iNOS) gene and kinetics of the expression ofiNOS induction in rat heart, aorta and kidney tissues.Methods iNOS mRNA expression was determinedby Northern blot analysis and NOS activities were ob-served by NADPH-diaphorase staining.Results The expression activities of iNOS gene inrat three tissues were undetectable be fore LPS treat-ment. iNOS mRNA in the heart, aorta and kidney be-gan increased at 2 h after LPS treatment, and reacheda peak at 6 h, then decreased gradually and returned tocontrol levels at 24 h. NOS histochemical stainingshowed that there was some expression of constitutivenitric oxide synthetase (cNOS) in rat three tissues un-treated by LPS. NOS activities in the three tissues roseobviously after LPS treatment and still remained thehigher leve1 till 24 h. This result suggested that iNOSis considerably steady in the tissues after it is syn-thetized.Concluslon iNOS mRNA and activities are marked-ly induced by LPS in rat heart, aorta and kidney, andthe kinetics of induction is similar in the three tissues.

Abstract:Aim T0 investigate the effects of phorbol myristateacetate (PMA) and some pathogenic factors in Dia-betes mellitus, such as high glucose (HG), high in-sulin (HI), high proinsulin (HPI) and tumor necrosisfactor-α (TNF-α) on free calcium ion in human umbili-cal vein endothelial cells (hUVEC).Metbods To detect calcium ion concentration byfluoresent probes and con focal microscopy in thisstudy.Results HG,HPI and TNF-Q could elevate intracel-lular free calcium ion concentration ([Ca²⁺ ]i) in hU-VEC obviously while HI did not (p<0. 01 ). Under adynamic observation of ([Ca²⁺ ]i) and intranuclear freecalcium ion concentration ([Ca²⁺ ]n) in a single en-dothelial cell (EC) induced by HG, HPI, TNF-α andPMA. we found an ohvious ascent of [Ca²⁺ ]i and[Ca²⁺]n especially in nucleus area. We also observedthat TNF-α and HI could induce calcium oscillation inhUVEC, and PMA at high dose caused evacuation ofCa²⁺ which was most obvious in nucleus area-Conclusion In vascular complications of diabetesmany pathogenic factors injured vascular endothelialcells by Ca²⁺ signaling and thus led to the damage ofnormal vascular function and structure.

Abstract:Aim Studies were performed to produce specificmonoclonal antibodies to bovine milk lipoprotein lipase(bLPL), to verify the specificity of the monoclonal an-tibodies for bLPL, and to characterize them.Metkods Purified lipoprotein lipase (LPL) frombovine milk-56 kDa was used as an antigen for the pro-duction of anti-LPL antibodies in mice. The spleen wasremoved from the animal having the highest titer of an-tibodies to LPL and the cells were fused with mousemyeloma cells. After cultured with hypoxanthine-aminopterin-thymidine (HAT ) culture medium, thisprocedure thus identified only those hybridomas whichproduced antibodies directed against LPL for super-natants by enzyme linked immunosorbent assay(ELISA). After cloning five monoclonal cell strainswere established, named as 2E5, 2G10, 6D7, 8D2 and7F4. The titriton of monoclonal antibodies (McAb) are5 × 10-2~2 × 10-3 for supernatants by ELISA.Results and Conclusion Contitration study showedMcAb recognized three different epitopes of LPL: 6D7and 8D2 to a close epitop; 2G10 and 7F4 to other closeepitope; 2G10 but recognized a far epitope away fromthe two of others. Western blot analysis of bovine milkwith the McAb gave a single protein band of 56 kDa;2G10 gave two protein bands of 50 kDa and 47 kDa.Dot-blot analysis of bovine milk with the McAb 2E5,8D2 and 6D7 showed specific reaction Western blot andDet-blot analysis of human milk with the McAb, hadno cross-reacted.

Abstract:Aim To investigate the influence of cell-modifiedlow density lipoprotein(cm-LDL) on scavenger recep-tors and low density lipoprotein receptors (LDLR ),meanwhile the effect of prostaglandin E2 (PGE2) wasstudied.Methods The receptor activity was detected bymeans of immunocytochemistry and bitin avidin-en-zyme linked immunosorbent assay(BA-ELISA), usingcultured murine peritoneal macrophages and murineskin fibroblasts as models.Results 1. The scavenger receptor activity onmacrophages increased in cm-LDL group, showingheavy brown granules in the membrane and cytoplasmof enlarged cells, while in PGE2 group, most cellswere mildly stained and in control group stained weak-ly. 2. The binding amount of LDL to LDLR on fi-broblasts showed that it markedly decreased in cm-LDL group (54±13 μg/g cell protein ) compared withwhich in control group (144± 8μg/g cell protein), p<0. 01 ; and significantly enhanced in PGE, group (100± 11μg/g cell protein), versus which in cm-LDLgroup, p<0. 05..Conclusion The findings suggested that cm-LDLstimulated scavenger receptor and inhibited LDLR,PGE2 (20 mg/L) markedly inhibited scavenger recep-tor activity and protected the LDLR against the injuryof oxidized LDL.

Abstract:Aim To investigate the effect of basic fibroblastgrowth factor (bFGF) on acute myocardial infarction(AMI) in rabbits.Methods The hearts of heathy male Japan big-earwhite rabbits were exposed through a left thoracoto-my, and the left anterior descending coronary arterywas ligated with a silk. Animals were divided intocontrol group, intravenously infused normal saline (2ml/kg) nitroglycerin (0. 2 g/kg), bFGF 1. 4 pg/kgand 7. 0 μg/kg, respectively, on operation say and onthe 2nd, 4th, 6th, 8th, and 10th day afer operation.We measured the areas of infarct myocardium with te-trazolium chloride and cardiac function by using physi-ological polygraphy, and determined ATP contents inmyocardium.Results Intravenous infusion of bFGF (1. 4 μg/kgand 7. 0 μg/kg) decreased the infarct size significantly(8. 1%±2. 7% and 8. 2%±2. 1% vs control 16. 5%±3. 2%, p<0. 05), Treatment with bFGF alsoincreased the ATP content in infarct myocardium (2. 9±0. 4 and 3. 2±0. 3 vs control 2. 3±0. 3 μmol/g, p<0. 01) and improved the left ventricular. All the effectof bFGF were better than those of nitroglycerin(p<0. 05).Conclusion bFGF may play a role both in stimu-lating recovery of ischemic myocardium and in improv-ing cardiac function in AMI rabbits.

Abstract:Aim This study was to investigate the aspect ofsmooth muscle cell (SMC) apoptosis and inducible ni-tric oxide synthetase (iNOS) gene expression in theearly periods of rabbit aorta injury model.Metkods The endothelium of rabbit abdominalaorta was injuried by a ballon angioplasty catheter,the presence of apoptotic SMC was demonstrated byterminal deoxynucleotidyl transferase-mediated dUTPnick end labeling (TUNEL). The expression of iNOSmRNA was eveluated by reverse transcription-poly-merase chain reaction (RT-PCR) methods.Results TUNEL analysis showed that the mediaSMC underwent apoptosis after injury. The index ofapoptotic SMC was 6% at 0 hour. It was significantlyincreased t0 15% at 8 hours, and reach a maxium(50%) at 24 hours, and maintained a high level at 3days (32% ) and 5 days (20% ). No significant incre-ment of apoptotic cells was observed in adventitia inthe early periods of artery injury. The expression of i-NOS gene mRNA in artery was significantly increasedafter injury, and was parallel with cell apoptotic in-dex.Couclusion Vascular media SMC of rabbit abdomi-nal aorta underwent apoptotic cell death in the earlyperiods of injury caused by ballon catheter. The in-duction of apoptosis coincided with iNOS mRNA ex-pression.

Abstract:Aim To investigate the effects of leucine-enkepha-line(L-ENK) on the proliferation, the DNA synthesisand c-fos oncogene expression of vascular smooth mus-cle cell (VSMC).Methods Techniques of cell culture, MTTmethod, 3 H-thymidine incorporation and Dot blot.Results L-ENK (10-5 mol/L) could markedly in-hibit the proliferation, DNA synthesis and c-fos onco-gene expression of VSMC. Naloxone (10-6 mol/L),opioid receptor antagonist, had no effect on the prolif-eration, DNA synthesis and c-fos oncogene expressionof VSMC, but, naloxone could antagonized the in-hibitory action of L-ENK to the proliferation ofVSMC. The inhibitory action of L-ENK to the VSMCcould be antagonized by the noradrenaline (10-5mol/L),adrenoceptor activator.Conclusion L-ENK inhibit the proliferation, DNAsynthesis and c-fos oncogene expression of VSMC byactivation of opioid receptors.

Abstract:Aim Naoluotong (NLT) is a compound Chinesepreparation that to be used in the treatment ofatherosclerosis. In this study, the effect of Naoluo-tong is compared with that of the Gypenosides in clini-cal practice.Methods Thorgh double-blind clinical observation,the patients were divided into Naoluotong group andGypenoside group. A color ultrasonic Doppler blood-now acoustic image system(ultramark-9) was used todetect the atherosclerosis plaque in the bilateralcarotid, measure the cervrical artery intima-media tu-nica thickness(CAIMT) and determine blood lipid, su-peroxide dismutase (SOD), lipid peroxide (LPO) andindexes of blood rheology. Part of the patients also hadtheir blood platelet aggregation rate (PAR) and heartfunction examined- Blood and urine routine and liverand kidney function examinations were conducted so asto find out about the safety of the medicine.Results Among the 57 patients with atherosclerosisplaques in the Naoluotong group, disappearance of theplaques occurred in 15. 8% (9/57), reduced plaquesoccurred in 47. 4K (27/57) and unchanged plaques oc-curred in 29. 8K (17/57) of the patients, the total effective rate being 93K (53/57). The total effective rateobtained in the Naoluotong group in resolving plaqueand reducing the cross section area was much higherthan that in the Gypenoside group (p<0. 01 or 0. 05).It was found that Naoluotong could reduce the thick-ness of the thickened tunica intima-media and had bet-ter effect than Gypenosides in reducing total choles-terol (TC ), triglycerides (TG ) and low densitylipoprotein cholesterol (LDLC) ; raising SOD and re-ducing LPO; and in reducing whole blood viscosity,fibringoen, packed cell volume, blood platelet maxi-mum aggregation rate, etc.Conclusion Naoluotong has better therapeutic effect than the total glycoside in Gypenoside treatingatherosclerosis, in that it can prevent the disease fromprogressing further and may even resolve the plaquesformedas a result of atherosclerosis.

Abstract:Aim In order to study the role of insulin-likegrowth of factor-1 receptor (IGFR) in the growth ofvascular smooth muscle cells(VSMC).Methods Antisense and mismatch of oligodeoxynu-cleotides (ODN) of IGFR were used to study smoothmuscle cell of mouse arteries.Results The incorporation of 3H-thymidine inVSMC handling by antisense of ODN decreased 64%compared with normal control group. The determi-nation of the receptor number in binding experimentdecreases 52% cell handing by as incubated differentconcentration of IGF-l (1 ~ 50 ng) caused a dose de-pendent increase in 3H-thymidine incorporation stimu-lated by 20 μg/L IGF-1 was reduced by 62% in anti-sense treated cells the effect of cells incubation of an-giotensin Ⅱ in antisense handling cells was not al-tered,the receptor quantities didn't reduce.Conclusion This experiment showed the impor-tance of ligand-receptor system to VSMC. The den-sity of receptor p1ays a key role to the growth ofVSMC the decreasing of IGF-1 receptor numbercaused by antisense was due to the decreasing of mR-NA in IGF-1 receptor. This hypothesis has beenproved in the experiment of nudease protection.

Abstract:To study the possibility of expressionof human lecithin cholesterol acyltransferase (LCAT)in skeletal muscles. Constraction of a plasmid vectorcontaining the human LCAT cDNA: transfer the vec-tor into myogenic cell line C2, and detection of LCATactivity in the culture medium. A expression vector ofhuman LCAT drived by CMV E/P, PCMV-LCAT wasconstructed, which successfully expressed in C2 cells.The LCAT activity was detected in the culture medi-um. It seems possible to chance plasma LCAT level bytransfer of LCAT gene into skeletal muscle.

Abstract:Aim To study the changes of serum lipid,Apolipoprotein and fibrinogen in patients of coronaryheart disease with various extents coronary stenosis.Methods According to the extents of angiographiccoronary maximal stenosis, 29 patients with coronaryheart disease (CHD) were classified (I ) obviousstenosis group (OSG, narrowing in luminal diameter>50% ), (Ⅱ ) mild stenosis) group MS. 16 non-CHDpatients proved by angiography served as controls(CG).Results There were no signifiant difference fortotal cholesterol (TC ) among the three groups.Triglyceride (TG), apolipoprotein (apoA I, apoB),high density lipoprotein cholesterol (HDLC), TC/HDLC, TG/HDLC, fibrinogen (Fg), apoA I / apoBin CHD patients were significantly abnormal as com-pared with CG (p<0.05 or p<0. 01, respectively). While TG, HDLC, apoA I, TC/HDLC, TG/HDLCand Fg in OSG changed significantly as compared withMSG.Conclusion TG, HDLC, apo AI, TC/HDLC,TG/HDLC and Fg related to the extents of coronarystenosis and may be of predictable values in coronarystenosis.

Abstract:Plasma concentration of lipoprotein(a) were measured by enzyme linked immunosorbentassay in 20 patients with coronary heart disease(CHD) and 20 healthy peoples. Plasma activity levelsof tissue type plasminogen activator (tPA) and plas-minogen activator inhibitor (PAI) were determined byspectrophotometric assay. Plasma concentration oflipoprotein (a) and activity of PAI in CHD group weresignificantly higher than the control group, however-plasma activity of tPA in CHD group were significant-ly lower. The plasma concentration of lipoprotein(a) in patients with CHD was not correlated with ac-tivity of tPA, but significantly positive correlated withactivity of PAI. It suggests that elevated plasmaconcentration of lipoprotein (a) is a risk factor forCHD. The results of this study showed also that plas-ma activity of tPA decrease and those of activity PAIof increase in the patients with CHD, which implythat reduced fibrinolytic capacity is related to CHD.High levels of lipoprotein (a) plasma may impair thefibrino1ytic system resueting in thrombose especiallyin the arterial system.

Abstract:Aim To explore the correlation between thechanges of leukocyte deformability (LD), leukocyteadhesion function (LAF), expression of cell adhesionmolecule (CAM) and coronary heart disease (CHD).Methods Leukocyte filteration index (LFI), leuko-cyte adhesion rate (LAR), leukocyte CD18 expressionand serum soluble intercellular adhesion molecu1e-1(sICAM-1 ) were measured in 118 patients and 68 con-trols.Results LFI, LAR, leukocyte CD18 expression andsICAM-1 concentration were significantly higher in pa-tients than those in controls (p<0. 001 ),the increaseof all indicators were more obvious in acute myocardialinfarction(AMI) patients than those in unstable angina(UA) and old myocardial infarction (OMI) patients (p<0. 001). In CHD patients, the LAR and LFI werepositively correlaed to leukocyte CD18 expression andsICAM-1 concentration(p<0.001 ), the LFI was posi-tively correlated to the LAR(p<0. 001 ).Conclusions The decrease of LD and the increase ofLAF, leukocyte CD18 expression andsICAM-1 concentration might participate in the oecu-rance of CHD, and were closely related to the CHD pa-tients conditions.