Abstract:Aim To study the expression of P21 gene in vascu-lar smooth muscle cells(VSMC)induced by introduc-tion of wild-type P53 gene and explore the mechanism of regulation for cell cycle progression by wild-type P53 gene.Methods A P53 gene recombinant adenovirus vec-tor,AdCMVP53.was transfected into the cultured SMC derived from human umbilical artery.The gene expression of P21 was quantified by reverse transcrip-tion-polymerase chain reaction(RT-PCR).The con-tent of P21 was detected by immunochemical staining.Flow cytometry was used for cell cycle analysis.Results The level of P21 mRNA and protein was increased in VSMC transfected with AdCMVP53.Overexpression of wild-type P53 could arrest the SMC at G0/G1 phase of the cell cycle.Conclusion Wild-type P53 gene could induce the expression of P21 gene,leading to repression of cell cycle progression.

Abstract:Aim To investigate the effects and mechanisms of Gypenosides on the atherosclerotic plaque formation in rabbit with hypercholesterolemia.Methods 23 male New Zealand white rabbits were assigned to four groups:5 rabbits in control group were fed with a normal diet;8 rabbits in hyperc-holesterolemic group were fed with 1% cholesterol diet for 9 weeks;In gypenosides and vitamine E (Vit E)treat groups,5 weeks after fed with 1% cholesterol di-et the animals were fed with 1% cholesterol diet con-taining gypenosides or Vit E for another 4 Weeks.Sudan IV stain was used to show the relative area ofatherosclerotic plaque.Lipid peroxides of aorta were estimated by the thiobarbituric acid method.Nitrite,the matabolic product of nitric oxide,was determined according to a modified Griess' method.The Ca2+.K+ and Na+ content of aorta were measured in means of atom absorption.Results Gypenosides treatment (120 mg/d,4weeks)inhibited formation of atherosclerotic plaque as shown by 37% reduction of plaque areas comparing to 1% cholesterol treatment along.Gypenosides also decreased lipoperoxidation of aorta by 33%.In addi-tion,gypenosides protected nitric oxide release of aorta against hypercholesterolemia damage and reduced Ca2+content of aorta by 50%.However,both hyperc-holesterolemia and gypenosides had no influence on K+and Na+ content of aorta.Vit E showed the same ef-fect as gypenosides did.Conclusion Gypenosides atlenuates the formatiom of atherosclerotic plague and lipoperoxidation of aorta induced by hypercholesterolemin in rabbit.The mecha-nisms of antiatherogenic effects of gypenosides might include its protection on nitric oxide and its inhibition on calcium overload.

Abstract:Aim To investigate the effect of high density lipoprotein (HDL)on the production of nitric oxide (NO)inhibited by oxidized low density lipoprotein (ox-LDL)in vascular smooth muscle cells.Methods The production of NO in cultured human vascular smooth muscle cells was analyzed by measur-ing nitrite in media with GRIESS reagent.The im-munochemical staining was used to observe the level of inducible nitric oxide synthase(iNOS).The content of mRNA for iNOS was determined by reverse tran-scription-polymerase chain reaction(RT-PCR).Results The ox-LDL could decrease the content of iNOS and level of mRNA for iNOS in vascular smooth muscle cells induced by lipopolysaccharide and inter-feron-γ,leading to inhibition of NO production.However,the addition of HDL before incubation with LDL could increase the production of NO and level of iNOS.0. 25g/L HDL could significantly increase the release of NO,and at concentration of 1.5g/L the production of NO was 4.5 limes higher than the con-trol.The effect of HDL was concentration dependent.Conclusions HDL could inhibit the activity of ox-LDL on production of NO in vascular smooth muscle cells.

Abstract:Aim To understand the mechanism of Chinese medicine ligustrazini against thrombosis and the effects of ligustrazini on plasminogen activator inhibitor type-1(PAI-1)expression in normal endothelial cells(EC)and EC exposed to lipopolysaccharide(LPS).Methods Human umbilical umbilical vein endothelial cells (HUVEC)were cultured by trypsin digestion method.PAI-1 protein in HUVEC conditioned medium was measured by sandwich enzyme-linked immunosorbent assay(ELISA),and PAI-1 mRNA expression was de-termined by Northern blot analysis.HUVEC nuclear factor-kappa B(NF-kB)nuclear translocation was as-sessed by electrophoretic mobility shift assay (EMSA).Results LPS treatment of cultured HUVEC result-ed in a significant increase in PAI-1 protein and mRNA expression by these cells.However,when HUVEC were incubated with LPS plus ligustrazini,the upregu-lation of PAI-1 by LPS was abated.Moreover ligus-trazini was able to decrease the basal level of PAI-1protein and mRNA as compared to control.Nuclear extracts prepared from LPS stimulated stimulated HUVEC demonstrated increased binding to the NF-kB oligonu-cleotide as compared to unstimulated cells,but ligus-trazini did not change those binding in the absence or presence of LPS.Conclusion Ligustrazini inhibited both basal and LPS-induced PAI-1 protein and mRNA expression in EC,and the modulation of PAI-1 in HUVEC by ligus-trazini might have other mechanisms rather than NF-kB pathway.

Abstract:Aim To understand whether low density lipopro-teins (LDL).very low density lipoprotein(VLDL),especially oxidized low density lipoprotein(ox-LDL)and oxidized very low density lipoprotein(ox-VLDL)have effects on the expression of monocyte chemoat-tractant protein-1(MCP-1)mRNA and protein in cul-tured calf aortic smooth muscle cells(SMCs).Methods After a 24-hour exprosure to LDL,ox-LDL,VLDL and ox-VLDL respectively.the total RNA in calf aortic SMCs was extracted by using the guanidinium isothiocyanate method.The MCP-1mRNA expression in SMCs was examined by slot blot analysis using a γ-32 p-end-labeled 35 mer oligonu-cleotides probe of MCP-1.Meanwhile,MCP-1 pro-tein content in the SMC conditioned media(SMC-CM)of each group was determined by sandwich ELISA.Results Cultured calf aortic SMCs were able to ex-press MCP-1 mRNA and protein,the lever of MCP-1mRNA in SMCs and the concentration of MCP-1 pro-tein in SMC-CM were increased significantly after a 24hour-exposure to ox-LDL and ox-VLDL,respective-ly,whereas the expression of MCP-1 mRNA and pro-tein was only slightly increased when exposed to LDL and VLDL.Conclusion ox-LDL and ox-VLDL can induce a strong expression of MCP-1 in cultured SMCs.

Abstract:Aim To observe the different effects of oxidized very low density lipoprotein(ox-VLDL)and normal VLDL(n-VLDL)on lipid accumulation in smooth muscle cells(SMCs)and attempt to explore receptor pathway of ox-VLDL uptaken by SMCs.Methods The same concentration of ox-VLDL or n-VLDL was incubated with porcine aortic SMCs for 48 hours,total cholesterol(TC)and triglyceride (TG) of the cells were extracted and determined.SMCs were incubated at 37℃ in RPIM 1640 contain-ing different concentration of FITC-ox-VLDL or FITC-n-VLDL for 4 hours,the cells were washed with phosphate buffered saline and the cell surface bound lipoproteins were determined by fluorospec- trometry.Competition experiments were performed as above.except for adding different concentration of unlabeled lipoproteins into the medium.Results ox-VLDL or n-VLDL(200mg/L)was in-cubated with porcine aortic SMCs for 48 hours,caus-ing a 3.2- and 10.6- fold increment of cellular TG in SMCs respectively(p<0.01).less TG was ac cumulated with ox-VLDL as compared with n-VLDL (P<0.01).Scatchard plot of the data of binding ex-periment showed that the Kd of the binding of ox-VLDL or n-VLDL on SMCs were 48.82mg/L and 37.17mg/L respectively,and the Bmax was 2067 μg ox-VLDL/g cell protein and 2596μg n-VLDL/g cell pro-tein respectively.nLDL competed with FITC-ox-VLDL on binding to SMCs as efficient as ox-VLDL,however,acetyl LDL inhibited only 10%.Conclusion More TG was accumulated in cells ex-posed to n-VLDL(P<0.01).FITC-ox-VLDL and FITC-n-VLDL were taken up by SMCs via saturable mechanism with low affinity.The Kd of the binding of ox-VLDL on SMCs was greater than that of n-VLDL and the Bmax was lower than that of n-VLDL.nLDL competed with FITC-ox-VLDL on binding to SMC as efficient as ox-VLDL,acetyl LDL exhibited low competitive effect,indicating that,ox-VLDL was taken up by SMC via LDL or VLDL receptor.

Abstract:Aim To study the changes of volume.function and characteristics of L-arginine(L-Arg)transport of platelet(Pt)in patients with essential hypertension (EH).Methods Nitric oxide(NO)production by platelets was assayed by measuring the accumulation of nitrite in the incubation medium using Greiss reaction accord-ing Green' method.Transport of 3H-L-Arg was determined essentially as described by William.In-flux of 3H-L-Arg was determined by measuring cell-associated radioactivity after an approprite period of in-cubation with 3H-L-Arg.Results in EH patients,the quantity and volume of Pt has no obvious difference with the normal.The maximum platelet aggregation ratio(PAR)was much higher than that in normal(P<0.05)and PAR was correlated with NO production.The production of NO and the ability of L-Arg transport in EH was re-spectively lower than that in normol(P<0.01).Maxi-mum transport velocity(Vmax)in EH was only 79%of normal(P<0.01).While the Michael constant (Km)showed no significant difference between the two groups.Conclusion Above results indicate that there exists a founctional and L-Arg transport disturbance in EH.It appears that the improvement of NO production and L-Arg transport in treatment of hypertension may be potentially important.

Abstract:Aim To analyse the internal relation between the serum lipid levels and apolipoprotein E genotypes in the aged.Methods The serum lipid levels(TC,TG,LDLC and HDLC)and the apolipoprotein E (apo E)geno-types in 106 the aged were determined with enzymolo-gy and PCR-RFLP respectively.Results There were three alleles of apo E and five different genotypes in 106 the aged.The aged with ε4allele showed higher TC,LDLC and lower TG levels as well as the aged with ε2 allele showed lower LDLC and higher TG levels than the aged with ε3 allele.Conclusion The alleles of apo E have a certain ef-fect on the serum lipid levels in the aged,but the ef-fect may not be as obvious and strong in the aged as in adults.

Abstract:Aim To determine cholesterol and cholesteryl ester of U937 monocyte within it's foaming procedure,and to obtained a simple and useful method to identify foamcell.Methods Cholesterol and cholesteryl ester was ex-tracted from U937 monocyte by hexane-isopropanol,and it's triglycerides was hydrolyzed by ethanolic KOH,and it's protein contents was removed by trichloroacetic acid.Father separation of cholesterol and cholesteryl ester was accompolished by high per-formance liquid chromatography(HPLC),using an eluent of isopropanol-n-heptane-acetonitrice and a Gen-Pak FAX column(with a little hydrophobic prop-erty,using for nucleic acid separation)and it's choles-terol peak was identified by L-B(Lieberman burchard)reaction and spectrum index analysis.Results Cholesterol concentration has a high linear relationship with it's peak area within 0.05-1.0g/L,the limits of detection of cholesterol is 4mg/g cell pro-tein(100mg cell protein equivalent to 1×108 cells).The cholestelyl ester inside U937 amount to 62% to total cholesterol after cholesterol-loaded for 48 hour.Conclusion This method is useful and simple to i-dentify foam cell.

Abstract:Aim To study the effect of estrogen(E)or estro-gen combined with progestogen(P)on atherogenesis and the mechanism in ovariectomized rabbits.Methods 40 ovarictomized rabbits were randomized into low E,low EP,high E,high EP and control groups.Each group was fed high cholesterol diet for 14 weeks.Group low E,low EP,high E,high EP were replaced different dose of estrogen or estrogen combined with progestogen.Before the experiment,at the end of the fourth or fourteenth week,the serum lipid,MDA and SOD were determined.At the end of the experiment,the plasma 6-keto-PGF1αTXB2 and ox-LDL levels were assayed.After killing the ani-mals,pathomorphological examination was carried out.Results Compared with the control group,the serum levels of total cholesterol(TC),triglyceride (TG),MDA and plasma ox-LDL,TXB2 concentra-tions were significantly lower(P<0.05~0.01)in the groups of the hormone replacement.While the plas-ma 6-Keto-PGF1αconcentrations were signicantly high-er(P<0.05).The plaque areas percent of the whole aortic intima surface areas in the therapy groups re duced by 57.8% averagely(P<0.01).Conclusion Estrogen could inhibit the aortic atherogenesis of the rabbits,its methanisms might be associated with serum lipid-lowing,inhibition of the lipidoxidation and regulation of the prostaglandin metabolism.Progestogen had no anti-atherosclerotic effect on ovariectomized rabbits.

Abstract:Aim To analyze the lipid disorders in type II dia-betic patients accompanied with coronary heart disease (CHD)and the important roles of lipid disorder in the development of CHD.Methods The study included 171 cases of healthy control subjects,230 cases of type II diabetic pa-tients,64 cases of CHD patients,and 61 cases of type II diabetic patients accompanied with CHD.All the subjects were examined the serum levels of triglyceride (TG).total cholesterol(TC),low density lipopro-tein-cholesterol(LDLC),high density lipoprotein-cholesterol(HDLC),HDL2C,HDL3C,apolipoprotein A,apolipoprotein B,and lipoprotein(a).Results The serum levels of TG,TC,LDLC,and Apolipoprotein B were significantly higher in type II diabetic patients than in the control subjects.The lipoprotein(a)level seemed to be higher in type II dia-betic patients than in the control subjects.The levels of TG,TC,LDLC,and lipoprotein(a)were higher in CHD patients than in the control subjects.which were similar to the type II diabetic patients.The levels of TG,apolipoprotein B were higher in type II diabetic patients accompanied with CHD than in diabetic or CHD patients separately.The level of HDL3C was lower in type II diabetic patients accompanied with CHD than in diabetic patients.The results of logistic regression showed that the increased levels of TG,lipoprotein(a),and TC were the high risk lipid factors for CHD.Conclusion There existed more severe lipids disor-ders in type II diabetic patients accompanied with CHD.The lipids disorders in type II diabetic pa-tients were atherogenic.The high risk lipid profile for CHD was the increased levels of TG,lipoprotein (a),and TC.

Abstract:Aim To study the abnormalities of lipoprotein and apoliprotein in patients with acute myocardial infarc-tion(AMI),unstable angina pectoris(UAP)and stable angina pectoris(SAP).Methods The subjects studied were 21 cases with AMI,27 cases with UAP,35 cases with SAP and 50cases as controls.The serum lipids and lipoprotein and apolipoprotein of each fasting serum were exam-ined simultaneously.Results 1.In AMI.UAP and SAP groups the serum levels of Lp(a)(360±364mg/L,297±301mg/L and 232±200mg/L respectively),Apo B100(0.89±0.28g/L,0.90±0.24g/L and 0.84±0.25g/L re-spectively)and Apo E(43.4±8.3mg/L,42.8±10.6mg/L and 42.6±8.1mg/L respectively)were signifi-cantly higher than that in healthy control group(Lp (a)124±110mg/L,Apo B1000.62±0.16g/L and Apo E 34.5±8.0mg/L);while in AMI,UAP and SAP groups.serum levels of HDLC(1.12±0.29mmol/L,1.18±0.25mmol/L and 1.25±0.28mmol/L respectively).HDL2C(0.35±0.14mmol/L,0.37±0.12mmol/L and 0.37±0.11 mmol/L respectively)and Apo AI(1.09±0.23g/L,1.09±0.24g/L and 1.13±0.23g/L respectively)were signifi-cantly lower than that in healthy control group(HDLC 1.405±0.26mmol/L,HDL2C 0.49±0.26mmol/L and ApoAI 1.25±0.21g/L).2.The ratio of LDLC/HDLC was higher in AMI group than that in SAP group.3.In serum lipid abnormality detectable rat in AMI,UAP and SAP groups,the increased serum Apo B100(53.13% to 57.89%),Apo E(43.48% to 63.64%)were the highest;and the next was the decreased HDLC(39.39% to 62.96%).In serum lipid abnormality pattern in AMI,UAP and SAP groups the increased LP(a)(37.97%)was the most frequent one.Conclusion Increased Lp(a).ApoB100.ApoE and decreased HDLC play an important role in coronary heart disease(CHD)and increased ratio of LDLC/HDLC is a predictor of CHD.

Abstract:Aim To ivestigate the association of lipid metabolism disturbance with haemostatic activity and fibrinolytic activity.Methods Serum lipids,some plasma haemostatic and fibrinolytic parameters were determined in 61 pa-tients with hyperlipidemia(hypertriglyceridemia in 45and mixed hyperlipidemia in 16)and compaerd with those in 18 healthy controls.Results The plasma activities of type-1 plasmino-gen activitor inhibitor(PAI-1),coagulation factor VII (F VII)and X(F X)were significantly higher,tissue type plasminogen activitor(tPA)was significantly lower in hyperlipidemic patients when compared with those in healthy controls(P<0.05~0.001). Fur-thermore serum triglyceride level was positively corre-lated with plasma activities of PAI-1,F VII and F X(P <0.05~0.001),and negatively correlated with tPA activity(P<0.01).Serum cholesterol level was also positively correlated with plasma fibrinogen level and PAI-1 activity(P<0.05).Conclusion Hyperlipidemia,especially hyper-triglyceridemia may increase coagulation activity and decrease fibrinolytic activity,benefiting the formation of thrombosis.Therefore,hyperlipidemia is an im portant risk factor for atherosclerosis.

Abstract:Using acoustic quantification(AQ)techonology to observe influence of QTd on left ven-tricular function in 51 cases of myocardiol infarction patients.Using 24 hours dynamic electrocardiogram (DCG)to detect cardiac arrhythmias in 43/51 cases of myocardiol infarction patients,and compared with 30cases of normal control subjects.The results showed that QTd in myocardial infarction(MI)patients was signifficantly longer than that in normal group(P<0.01),and the left ventricular systolic and diastolic function were also demaged compared with normal group.QTd in MI accompanying cardiac arrhythmia group was signifficantly longer than that in MI accom-panying sinus rhythm group(P<0.01),and left ven-tricular systolic and diastolic function in MI accompa-nying cardiac arrhythmia group were demaged com-pared with MI accompanying sinus rhythm group.It is concluded that QTd in MI patients was longer,and it may have relationship with left ventricular dysfunc-tion and the incdence of cardiac arrhythmia in myocar-diol infarction patients.

Abstract:Aim To explore the providing leads of coronary artery collateral vessels in patients with coronary atherosclerotic heart disease on predicting prognosis and guiding treatment.Methods 68 patients with total occlusion of coro-nary arteries through coronary arteriograph were divid-ed into good development of collateral vessels group and poor development of collateral vessels group.Compared their ECG ST-T changes,motion stress testing.QRS scores,ventricular wall motion scores,ejection fraction,left ventricular end diastolic pressure and cardiac index between the two groups.Results Compared with the poor development of collateral vessels group,in the good development of collateral vessels group the ECG ST-T changes and left ventricular end diastolic pressure were obvious less(P <0.01),QRS scores and ventricular wall motion scores were lower(P<0.01),ejection fraction and cardiac index were higher(P<0.01),their positive rate of motion stress testing was not significant differ-ence(P<0.05).myocardium has not infarcted in 4patients of the good development of collateral vessels group,treadmill stress testing was negative in three of four cases.Conclusion Observing collateral vessels during coronary arteriograph can provide valuable leads in pa-tients with coronary atherosclerotic heart disease on predicting prognosis and guiding treatment.

Abstract:Aim To study the effects of benazepril and ni-trendipine on renal hemodynamics and urinary β2-mi-croglobulin(β2-MG)in the hypertensive patients.Methods A randomized-cross trial comparison of benazepril and nitrendipine in the treatment of 58 hy-pertensive patients divided into two groups for 4 weeks was preformed.The renal hemodynamics and urinary β2-MG were determined in all patients before and after treatment.The urinary β2-MG was measured with ra-dioimmunoassay and renal hemodynamics was detected by means of radionuclide.Results Benazepril not only increased the effective renal plasma flow,but also significantly reduced the filtration fraction and urinary β2-MG(P also<0.01)after treatment.There was no change on the renal hemodynamics and β2-MG after the treatment of ni-trendipine.Conclusion The data indicate that benazepril could improve the renal function better than nitrendipine in the hypertensive patients.

Abstract:Aim To establish a immunocytochemistry method of detection to thrombin receptor(TR).Methods TR activated by thrombin can release a peptide that contain 41 amino acids(TR1-41)from N-terminal of TR.The TR1-41 was synthesized accord-ing to the amino acids sequence of TR,then mice were immunized utilizing the TR1-41,4 cell strains produc-ing TR-McAb were gotten.Preparations of atheroscle-rotic human left ventricular myocardium of four died patients were cut out.The preparations were made in-to paraffin sections.Results A immunocytochemistry method-alkaline phosphatase antialkaline phosphatase(APAAP)to de-tect the thrombin receptor was established and the TR of myocardium were detected successfully.Conclusion The method established can detect TR expression of tissue and organ.

Abstract:Aim To study the clinical practical value of region-al cerebral blood flow(rCBF)spect for amaurosis fu-gax(AF).Methods 20 normal persons and 42 AF Patients were examined by rCBF spect.and compared with X-ray CT at the same time.followed up for 24(18-32)months and checked again by rCBF spect.Results The T/NT of visual area in occipital lobe of AF Patients' rCBF spect was more lower than con-trol group.the sensitivity and accurate late of AF patients' rCBF spect was more higher than CT,the AF patients' prognosis of normal rCBF spect was best and the rCBF spect had close relation with clinical prognosis observation(90.5%).Conclusion rCBF spect played an important clinical role on diagnosing,pledicting prognosis and following up with AF patients.