Abstract:Aim To establish a new transgenic mice model for determining the function and roles of human scavenger receptor A in atherosclerosis in vivo. Methods Human scavenger receptor A-I minigene-driven tie-1 promoter was constructed by endonuclease digestion and confirmed by sequence analysis. Transgenic mice were produced via microinjection method. PCR, Southern blot were used to screen the positive transgenic mice. Transgenic mice line established by mating with C57BL/6 mice and offspring were identified by PCR so as to research transgenic mice transmission. Northern blot, RT-PCR, immunohistochemical analysis, light and transmission electron microscopy were used to investigate the expression location of human SR-A and lesions of arteries and vascular endothelial cells in transgenic mice. Results The fragment sequence of mouse tie-1 promoter and human SR-AI cDNA are similar to their sequences in Genebank and no ATG before the translation initiation sites of human SR-A by sequence analysis. About 3.4 kb tie-1-promoter-hSR-AI-cDNA-BGH polyA fragment were obtained by AatⅡand XhoⅠdigests. 561 survival embryos injected with purified human SR-A minigene were implanted into the oviducts of 19 ICR pseudopregnant mice.Among the 54 survived pups from 13 foster mothers, 7 founders were identified with PCR and Southern blot analysis. Integration rate of exogenous was 13%. PCR positive rates were 47.8%, 71.3%, 75.0% and 100% in G1, G2, G3 and homozygous transgenic mice, respectively. The results of RT-PCR and immunohistochemical analysis showed human SR-A I specifically expressed on endothelial cells of aorta, liver and renal artery of transgenic mice. Plasma triglyceride level of transgenic mice was significantly higher than the level of non-transgenic mice (P<0.05 vs control). There was no difference in total plasma cholesterol level between transgenic and non-transgenic mice. But in transgenic mice, total plasma cholesterol level was significantly higher of males than of females (P<0.01). SEM of the luminal surface of aorta revealed an irregular, elevated bumpy and swelling surface. There was disruption of the endothelial layer and some blood cell was observed adhering to the surface of them in transgenic mice. TEM of aorta of transgenic mice showed that vesicles, multivesicle bodies and swelling mitochondria filled in plasma of endothelial cells. Vacuolar and mucoid degeneration were observed in the media of aortic ardh sections in transgenic mice with HE staining. Conclusions A transgenic mice model with overexpressed human SR-A on ECs were successfully established in this study. The transgene stable inherited across generation after generation. The high level of plasma lipid, hydropic and mucoid degeneration of arterial wall in transgenic mice may accelerate the development of atherosclerosis. Our studies provide a new transgenic model for investigation of the function and roles of SR-A in atherosclerosis.

Abstract:Aim [WT5”BZ]To study the changes in activity and mRNA expression of nitric oxide synthase (NOS) in different phases of atherosclerotic lesion.[WT5”HZ] Methods [WT5”BZ]24 male New Zeland white rabbits were divided into normal control group(6 rabbits) and hypercholesterol group(18 rabbits). At the time of the 8th, 12th and 16th week, 6 rabbits of hypercholesterol group were sacrificed. Using NADPH-diaphorase histochemistry, we studied the activity of NOS. The NOS mRNA expression in aortas in the 16th week was also studied with in situ hybridization.[WT5”HZ] Results [WT5”BZ]In the 8th week, in cholesterol group, there were three different aspects of NADPH-d stains in plaques, which were doubtful positive, positive and strong positive. In the 12th week, there were extensive NADPH-d moderate positive stains in plaques, particularly at the border of intima and media. In the 16th week, the activity and mRNA expression of NOS in both intima and media decreased accompanying with the forming of necrosis core.[WT5”HZ] Conclusion [WT5”BZ]Changes in activity and expression of NOS in atherosclerotic lesion are dynamic with different locations of cells.

Abstract:Aim To ascertain whether vascular smooth muscle cells inhibit vascular smooth muscle cell proliferation by autocrine or paracrine of growth inhibitors. Methods Conditioned medium from rabbit aortic smooth muscle cell was used in this study. The conditioned medium was filtered by cut off M.W. 100 kDa or 10 kDa (Millipore). Smooth muscle cell proliferation was assayed by the measurement of XTT (Boehringer Mannhein). Results Rabbit aortic smooth muscle cell conditioned medium inhibited smooth muscle cell proliferation, 50% smooth muscle cell conditioned medium achieved 45% inhibitory rate of smooth muscle cell proliferation. This inhibition activity substance is dose dependent, 56℃, 30 min heat stable and M.W.less than 10 kDa. Conclusions Rabbit aortic smooth muscle cell could inhibit smooth muscle cell proliferation by autocrine or paracrine growth inhibitors.

Abstract:Aim [WT5”BZ]To investigate the mechanism involved in the apoptosis of human myeloid leukemia U937 cells induced by oxidized low density lipoprotein (ox-LDL).[WT5”HZ] Methods[WT5”BZ] The apoptotic cells were determined by flow cytometry and DNA fragmentation assay. The content of p53, p21 and Bcl-2 protein were analyzed by immunofluoresence staining and flow cytometric analysis. The level of p53,p21 and Bcl-2 mRNA were quantified by reverse transcription-polymerase chain reaction (RT-PCR). [WT5”HZ]Results [WT5”BZ]Ox-LDL induced apoptosis of U937,which is concentration dependent,and upregulate the level of p53 and p21 mRNA and protein as detected by RT-PCR and flow cytometric analysis.However,expression of Bcl-2 was downregulated after treatment with ox-LDL. [WT5”HZ]Conclusion [WT5”BZ]Ox-LDL upregulated the gene expression of p53, p21 and downregulated Bcl-2 expression, which induced apoptosis of U937 cells.

Abstract:Aim [WT5”BZ]To study the influence of thrombin on gene expression of platelet derived growth factor(PDGF) in cultured vascular smooth muscle cells and probe into the mechanism of proliferation of vascular smooth cells induced by thrombin . [WT5”HZ]Methods [WT5”BZ]Take the incoporation of 3 H-thymidine as the target to evaluate vascular smooth muscle cells proliferation; thrombin-induced expression of PDGF-A mRNA in the vascular smooth muscle cells was detected by RT-PCR, while PDGF-B mRNA expression by Dot blot hybridization. [WT5”HZ]Results [WT5”BZ]Thrombin remarkedly stimulated proliferation of SD rat vascular smooth muscle cells in a dose-depedent and a time-dependent manner; PDGF-A and PDGF-B mRNA was detected in quiescent cultured vascular smooth muscle cells in serum-free medium for 48 hours,the expression of PDGF-A mRNA was increased in VSMC after exposure to thrombin for two hours, and reachd a peak levels after 4～6 houres exposure to thrombin. The rising level of PDGF -A mRNA continued for 12 hours,and felt to the base level 24～48 hours afterwards. [WT5”HZ] Conclusion [WT5”BZ] Thrombin remarkedly stimulated proliferation of SD rat vascular smooth muscle cells in a dose-depedent and a time-dependent manner, thrombin-induced vascular smooth muscle cells proliferation was partially mediated by expression of PDGF- A mRNA.

Abstract:Aim To examine the effects of oxidized low density lipoprotein (ox-LDL) and antioxidant on calcium transportation of human vascular endothelial cells, then explore the molecular mechanism of cytotoxicity induced by ox-LDL. Methods Confluent primary cultures of human umbilical vein endothelial cells (hUVEC) were exposed respectively to ox-LDL or ox-LDL and vitamin E in medium. The uptake and release of calcium by hUVEC were measured. The mortality of hUVEC were assessed by the messurement of LDH. Results Ox LDL enhanced the mortality of hUVEC and the uptake of calcium by hUVEC with a dose-dependence manner, while vitamin E inhibited these effects. Conclusions Ox LDL may stimulate the uptake of calcium by hUVEC, leading to hUVEC injury. Vitamin E could protect hUVEC from such cytotoxic effect by inhibitting cellular uptake of calcium.

Abstract:Aim The previous studies have demonstrated that the acetylcholine(Ach)-induced endothelium cells(EC)-dependent relaxation were injuried in diabetic models. The present study was to investigate the effect of Losartan, a specific inhibitor of angiotensin Ⅱ (Ang Ⅱ) receptor 1 (AT 1), on dysfunction of EC-dependent relaxation in diabetic rats induced by streptozotocin(STZ) at different time. Methods The Ach-induced EC-dependent relaxation of rat aortic rings were tested in tissue bath, the concentration of plasma glucose and Ang Ⅱ were measured. Results At 4 weeks, the function of EC-dependent relaxation was not injuried in diabetic group, but the concentration of plasma glucose and Ang Ⅱ were increased significantly compared with control group. The percent of Ach-induced relaxation was declined obviously compared with control group at 8, 16 weeks. Treatment with losartan significantly protected the function of EC-dependent relaxation, the percent of Ach-induced relaxation was obviously increased compared with diabetic group. The concentration of plasma glucose was not changed , but the plasma Ang Ⅱ was increased compared with diabetic group. Conclusion Losartan could protect the function of EC-dependent relaxation in diabetic rats.

Abstract:Aim To investigate the effect of fluvastatin on endothelial, coagulative and fibrinolytic functions in experimental atherosclerotic rabbits. Methods 28 male New Zealand rabbits were divided into four groups randomly: normal group,pathological group, 5-week fluvastatin-treatment group (10 μg/g.d )and 9-week fluvastatin-treatment group (10 μg/g.d ). Plasma 6-Keto-prostaglandin F 1α (6-Keto-PGF 1α ),thronboxane B 2(TXB 2),endothelin (ET), nitric oxide (NO), tissue-plasminogen activator (tPA), plasminogen activator inhibitor (PAI) and fibrinogen (FIB) concentrations were assayed in different intervals during the experiment. Results At the end of ninth week,TXB 2/6-keto-PGF 1α ratio, ET levels and PAI levels in the two treat groups were significantly lower than those in the pathological group (P<0.05); TXB 2 concentration in the pathological group was much higher than that in the normal group (P<0.05). NO and tPA concentrations in the pathological group were significantly lower than those in the normal group (P<0.01). Compared with the pathological group, the concentration of 6-Keto-PGF 1α in two treatment groups had a increasing trend, but the concentration of FIB had a lowering trend. Conclusion Fluvastatin can significantly improve endothelial, coagulative and fibrinolytic functions.

Abstract:Aim [WT5”BZ]To investigate the protective effect of Panax Notoginseng Saponins (PNS) on the cultured aortic smooth muscle cells (SMC). [WT5”HZ]Methods [WT5”BZ]In this study, cell culture technique was used in vitro. The protective effect of PNS on the proliferation of SMC was observed by measuring MTT metabolism and cell numbers and observing cell ultrastructure under TEM. [WT5”HZ]Results[WT5”BZ] The serum hyperlipidemia caused the increases in cell numbers and MTT metabolism of SMC. Under TEM, there are numerous rough endoplasmic reticulum and mitochondria and lipids in the cytoplasm of the cells cultured by serumof hyperlipidemia. The PNS can inhibit the proliferation of SMC dose dependently and can inhibit the proliferation of SMC stimulated by serumof hyperlipidemia significantly. [WT5”HZ]Conclusions[WT5”BZ] The PNS may prevent atherosclerosis and inhibit progression of the atheroslerotic lesions by interfering with the proliferation of arterial SMC.

Abstract:Aim [WT5”BZ]To investigate the antiatherogenetic effect of Chinese medicine angelica injection and its mechanisms. [WT5”HZ]Methods [WT5”BZ]Atherosclerotic rabbit models were duplicated by feeding high lipid forage. Eighteen rabbits were divided randomly into three groups: control group (Control), model group (Model) and angelica group (Angelica). Common forage (control group), high lipid forage (model group) and high lipid forage adding angelica injection (angelica group)were given to each group accordingly. At the end of the tenth week, blood samples were obtained through carotid artery intubation for detection of serum lipids and malonyldialdehyde (MDA). Thoracic aorta were prepared for measurement of plaque area. [WT5”HZ]Results [WT5”BZ](1) Serum lipids level of model group and angelica group were significantly higher than those of control group (P<0.01). Apart from TG which was different (P<0.01), there was no difference of TC, HDLC, and LDLC between model group and angelica group. (2) Plaque area of model group was larger than that of angelica group (P<0.01). (3)There was significant difference of plasma MDA between any two groups (P<0.01). [WT5”HZ]Conclusions [WT5”BZ]Angelica can inhibit atherogenesis. The mechanisms maybe correlate with its decreasing serum TG and antilipoperioxide action.

Abstract:Aim To investigate the relationships between QT dispersion (QTd) and plasma endothelin (ET) levels, insulin-sensitivity, and ventricular arrhythmias in coronary heart disease (CHD). Methods Oral glucose tolerance test was conducted in 27 patients with acute myocardial infarction (AMI), 20 patients with angina pectoris and 20 healthy control subjects (in the fourth week after the onset in AMI patients). Blood samples were analysed for glucose, insulin, C-peptides and ET. Linear and/or stepwise regression analysis were used to assess the correlation between QTd and nine insulin-sensitivity parameters and ET levels. Results In patients, QTd was independently, significantly, positively correlated with ET levels. Nevertheless, fasting glucose was an independent, inversely correlated variable in AMI-recovery-period patients. The first-day ET level and QTd were higher in AMI patients with severe ventricular arrhythmias than without (160.5±23.0 ng/L Vs 119.7±15.2 ng/L,P<0.01 for ET;86±19 ms Vs 69±13 ms,P<0.05 for QTd), and both of them were higher than that in control subjects (44.98±14.74 ng/L,P<0.01). QTd was positively correlated with ET(r=0.94 and 0.84,respectively,P<0.01). Conclusions In CHD, especially in AMI, the increase of plasma ET levels is an important independent factor that causes QTd increasing and might further result in severe ventricular arrhythmias. In the recovery period of AMI, low fasting blood glucose is another independent factor of QTd increase.

Abstract:Aim To educidate the changes of lipoprotein(a) [Lp(a) ] in cerebrospinal fluid (CSF) and serum in patients with cerebral infarction. Methods The CSF and the serum Lp(a) levels were measured with an enzyme-linked immunosorbent assay (ELISA) in 80 patients with acute cerebral infarction and 40 control subjects. [WT5”HZ]Results [WT5”BZ]Lp(a) levels in the CSF and serum of cerebral infarction patients were significanthy higher than that of the controls (234±29 μg/L vs 211±21 μg/L and 0.299±0.028 g/L vs 0.275±0.025 g/L). Lp(a) concentration in the CSF of cerabral infarction patients and controls is not related to that of serum ( P>0.05 ). Conclusion The CSF and the serum Lp(a) levels in patients with cerebral infarction were significantly higher than those in cortrols.

Abstract:Aim To evaluate the effect of apolipoprotein E (apo E) polymorphism on serum lipids and the relationship between apo E polymorphism and coronary stenosis. Methods The polymorphis site of apo E gene were studied using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in 95 cases with coronary artery disease (CAD) and in 46 controls of equivalent age. The serum lipids and apolipoprotein B (apo B) levels were measured in the two groups. Results The frequencres of apo E3/4 genotype and apo E4 allele were significantly higher in 55 cases of CAD with multiple vessel dissease (MVD) and in 40 cases of CAD with single vessel disease (SVD) than in 46 controls (P<0.01), while there were significantly higher in the MVD groups than in the SVD groups (P<0.05). Apo E3/4 genotype had significantly higher serum total cholesterol (TC), LDL-cholesterol (LDLC) and apo B levels and lower HDL-cholesterol (HDLC) level compared to apo E3/3 and apo E₂/3 genotypes in the patients and in the controls. In the patients , apo E3/4 genotype had significantly higher TC, LDLC and apo B levels and lower HDLC levels compared with in the controls . Conclusions It is concluded that apoE polymorphism is involved in cholesterol metabolism , where apo E4 allele is associated with an increased risk of CAD. In CAD cases , an increased frequency of apo E4 allele is associated with an increased severity of coronary stenosis. Therefore it is assumed that there is certain relation between apoE4 allale and severity of coronary stenosis.

Abstract:Aim [WT5”BZ]To invistgate possible associations between lipoprotein(a) [Lp(a)] and the severity ofcoronary artery diseases. [WT5”HZ]Methods [WT5”BZ]Lipids apoprotein and Lp(a) concentrations in 74 patients with chest pain were measured. Of 74 patients who underwent coronary artery angiography with Judking's technique, twenty six patients were normal (control group), the other forty eight were abnormal (the lumen of coronary artery was occluded by 50% or more). The latter was divided into two group saccording to its number of injured vessels. We used ascoring method of coronary artery diseases to reflect the extent of coronary artery narrowing. [WT5”HZ]Results [WT5”BZ]That serum Lp(a) concentrations have a significant difference between normal group and abnormal group. Multiple stepwise regression analyses revealed that Lp(a) is an independent risk factors for coronary artery. In abnormal group, Lp(a) concentrations increase with the severity of coronary artery lesions, there is no statistical difference between the two groups. [WT5”HZ]Conclusions [WT5”BZ]Lp(a) is an independent risk factor for coronary artery disease and, to some degree, has clinical significance in valuing coronary artery diseases.

Abstract:Aim [WT5”BZ]To investigate the relationship between the clinical effects of defibrase on acute cerebral infarction and the time used, and the effect on the platelet aggregation and fibrinogen in the patient`s blood. [WT5”HZ]Methods [WT5”BZ]Divide the 72 cases of acute cerebral infarction (ACI) confirmed by CT scan into the defibrase group (38 cases) and the Radix Salviae Miltorrhizase (RSM) group (34 cases). The defibrase group were treated with defibrase iv drip of 10 units mixed 250 mL of physiological saline q.d. for 3 days. The RSM group were treated with RSM iv drip of 0.4 g mixed with 250 mL of 5% glucose q.d. for 14 days. The platelet aggregation function and the fibrinogen of the patients` blood were measured both before and after the drug was given. The clinical effects were valved at the end of the 1st week and the 4th week.[WT5”HZ] Results [WT5”BZ]Before the drug was gevin the platelet aggregation rate was 65.6%±5.3% and the fibrinogen was 4.78±0.42 g/L in the defibrase group, after the drug was given, the platelet aggregation rate was 45.8%±4.9% and the fibrinogen was 3.16±0.35 g/L with a significant difference (P<0.001).Before the drug was given the platelet aggregation rate was 60.4%±1.9% in the RSM group, after the drug was given the platelet aggregation rate was 49.0%±10.3% with a significant difference (P<0.01). [WT5”HZ] Conclusions [WT5”BZ] Defibrase is superior to RSM in controlling the platelet aggregation function and decreasing the plasma fibrinogen concentration, which improves the patient`s high viscosity state and prevents the thrombosis.

Abstract:Aim [WT5”BZ]The present study was to compare the clinical efficacy of low dose gemfibrozil with normal dose gemfibrozil administered in hyperlipidemic patients. [WT5”HZ]Methods [WT5”BZ]121 subjects were randomized into two groups: 62 Subjects in low dose gemfribrozil group,59 subjects in normal dose gemfibrozil group as controls. Blood lipid levels were determined prior to lipid lowering treatment and at the end of 4th and 8th week treatment respectively. [WT5”HZ]Results [WT5”BZ] At the end of the 4th week treatment, levels of TC and TG in normal dose gemfibrozil group significantly reduced (P<0.01), and levels of HDLC in normal dose group obviously increased (P<0.01), only levels of TG in low dose group reduced (P<0.05), after treatment compared with the prior to treatmant. At the end of the 8th week treatment, levels of TC and TG significantly reduced and levels of HDLC obviousely increased in both groups. Both of difference were statistically significant (P<0.01). [WT5”HZ]Conclusion [WT5”BZ] Not only was the efficacy of low dose group significantly weaker than that of normal dose group but also the side effects of low dose group was lower

Abstract:Aim [WT5”BZ]To study whether serum non-high density lipoprotein cholesterol (nHDLC) level is related to coronary heart disease (CHD). [WT5”HZ]Methods [WT5”BZ]The triglyceride, total cholesterol and high density lipoprotein cholesterol in the serum of 183 patients with coronary heart disease with hyperlipoproteinemia and 194 healthy individuals were tested separately. The low density lipoprotein cholesterol and the very low density lipoprotein cholesterol were calculated according to Friedewald`s formula LDLC = TC - (HDLC + 0.458 TG) and the non-HDL Ccontent was calculated with Frost method. [WT5”HZ]Results [WT5”BZ]There was significant difference in serum low density lipoprotein cholesterol (LDLC) level and nHDLC level between the CHD with Ⅱa and Ⅱ_b patient group and control group (P<0.001). There was no significant difference in serum LDLC level between the CHD with Ⅳ patient group and control group (P>0.05), while serum nHDLC level was significantly higher in CHD with Ⅳ patient group than control group (P<0.001). [WT5”HZ]Conclusions [WT5”BZ] This article provides examples of the utility of non-HDL cholesterol concentrations in clinical medicine.

Abstract:Aim To evaluate the spiral CT manifestations and its diagnostic efficacy in the aortic dissection. Methods Twenty-three patients underwent 25 spiral CT examinations totally. The spiral CT features were classified into specific and non- specific spectrums for analysis. Results Specific signs emerged as true and false double aortic lumen in 22 cases (95.7%),the intimal flap 22 (95.7%). Non-specific signs include irregular aortic dilatation in 23 cases all (100%),thickening and calcification of the aortic wall 21(91.3%),intraluminal thrombus 4 (17.4%). The diagnostic accuracy of spiral-CT achived 95.7%. Conclusion Spiral CT is a valuable tool in the assessment and diagnosis of aortic dissections. were from eighteen to twenty five years old without orthodontic history Chengdu were randomly collected in. All teeth must be free of visible malform, aberration, and fully erupted. Impressions were taken, and measurements were made, parallel to the occlusal plane at the greatest mesiodistal width of each tooth on dental cast with electronic caliper (0 01 mm). Each tooth was measured twice and the average of the two readings was taken as the dimension for that individual tooth. Mesiodistal crown diameters were statistically analyzed with t test and linear regression, using the SAS (Statistic Analyses System) computer package. Results: It was found that mesiodistal crown diameters had significantly statistical difference in gender. Hence the mesiodistal crown diameters were assessed and investigated separately for the male and the female. It was also found that the total mesiodistal crown diameter of lower incisors was closely correlated with the total mesiodistal crown diameter of canines and premolars. The correlation coefficients between the independent variable and dependent variables were 0 599 for upper canines and premolars and 0 654 for lower canines and premolars in the males and 0 586 and 0 635, respectively, in the females. The P were found to be less than 0 001 for both arches. The correlation coefficients had significantly statistical difference. Conclusion: The equations predicting overall mesiodistal crown diameters of upper and lower unerupted canines and premolars for male and female Chinese people were established respectively: Maxillary (y=10 87+0 51 x (male)), y=11 88+0 46 x(female), Mandibular (y=10 36+0 50 x(male), y=10 03+0 49 x(female)), y(total mesiodistal crown diameter of canines and premolars), and x(total mesiodistal crown diameter of lower incisors).\;Key words:\ mesiodistal diameter\ \ space analyses\ \ linear regression\ \ predicting equation