Abstract:Mitochondrial DNA differs from chromosomal DNA by its endosymbiotic characteristics of being a foreigner to protoeukaryotic cells circular intronless and multitronics. When mtDNA is recognized as a transgene humankind becomes a trans-genie species occurred naturally under strict definition of transgenic technology. With transgenic human assay we are able to study the biological effect of mtDNA-original homologues in the genome of modem eukaryotic cells. This new discipline is termed Mitochromics. Mitochromic analysis allows the assembling of an ancient mtDNA using eight nuclear mtDNA analogues which has an overall consensus rate of 94% with NC-001806. We believe that this assembled ancient mtDNA sequence will have enormous application in evolution and biomedical studies.

Abstract:Aim To study the effect of recombinant human vascular endothelial growth factor (VEGF) on the production of matrix metalloproteinase-2 (MMP-2) protein and activity in human umbilical veins endotheh'al cell line (ECV304). Methods MMP-2 protein was determined by immunohistochemistry and enzyme linked immuno-sorbent assay (ELISA). MMP-2 activity was detected by zymography. Results MMP-2 could be constitutionally secreted by ECV304 cells. Upon VEGF stimulation , a significant upregulation of MMP-2 protein expression was observed at concentration of 50μg/L ( P< 0.05), further elevation at 100μg/L according to ELISA assay. The activity of MMP-2 for gelatin degradation by zymography confirmed the ELBA result. Conclusion VEGF can induce MMP-2 production in ECV304 cells, the responses probably contribute to the angio-genic and vascular permeable potential of the peptide.

Abstract:Aim The effect of zinc deficiency and zinc replenishment on the distribution of elements in rats. Methods SD rats were randomized into zinc-deficient diet (ZD) group, pair-fed (PF) group, zinc normal group (ZN), zinc supplemented (ZS) group and high zinc (HZ) group. ZD group, ZN group, and HZ group were fed up with zinc deficient dietary (content of zinc< 1 mg/kg), zinc-normal dietary (50 mg/kg) and high zinc dietary (150 mg/kg) for 8 weeks respectively. ZS group were fed with zinc deficient dietary for 3 weeks, then changed to be treated with high-zinc dietary for 5 weeks. PF group were fed with zinc normal dietary according to the actual quantity of ZD group at the former day. All of the rats were killed after 8 weeks. The concentration of Zn, Cu, Fe, and Ca in serum were measured with polarography and with atomic absorption spec-troscopy (AAS) in liver, kidney, spleen, and testicle. Results The concentration of Zn, Fe in serum, liver, spleen, kidney, testiele and Cu in kidney, spleen decreased significantly owing to zinc deficiency, while the concentration of triglyceride, cholesterol, high density lipoprotein and low density lipoprotein in serum remained the same. Conclusions Zinc deficiency not only decreased zinc concentration in organism directly, but also affected the metabolism and distribution of elements. However, it didn' t affect serum significantly.

Abstract:Aim To study the influence of hirudin on thrombin-induced proliferation of vascular smooth muscle cells (VSMC) and its regulated mechanism in vitro. Methods VSMCs were isolated from the thoracic aorta of rabbits and cultured 2 - 5 passages were incubated with thrombin (4.0 ku/L), while hirudin (6.0 ku/L) and heparin (6.0 ku/L) were added except for control group. The effect of hirudin on growth of VSMC was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltet-razohum bromide (MTT) assay and flow cytometry. The expression of platelet-drived growth factor (PDGF) and proliferating cell nuclear antigen (PCNA) in VSMCs was evaluated by immunohistochemistry technique and computer image analysis system. Results Hirudin remarkedly inhibited both the thrombin-induced proliferation and the expression of PDGF and PCNA in VSMC (P< 0.05, P< 0.01). Conclusion Hirudin is able to significantly influence the proliferative activity of VSMCs, whichmay be carried out through inhibition of PDGF and PCNA expression.

Abstract:Aim To investigate the effects of ferulic acid (FA) on the expression of adhesion molecules on activated human umbilical vein endothelial cells (HUVEC) and to elucidate the molecular mechanisms of its anti-atherosclerosis effect. Methods Cultured HUVEC were treated with FA (0.21, 0.41 and 0.62 mmol/L respectively) and stimulated with 10 mg/L li-popolysaccharide (LPS) 30 min later. Expression of E-selectin was detected by flow cytometry (FCM) after 4 h. Expression of E-selectin mRNA was measured by reverse transcription-polymerase chain reaction (RT-PCR). Cultured HUVEC were treated with FA (0.21, 0.41 and 0.62 mmol/L respectively) and stimulated with H₂O₂ (300 μmol/L) 30 min later. After 2 h, expression of P-selectin on HUVEC surface was detected by FCM. Results FA could inhibit E-selectin and P-selectin expression on activated HUVEC. It could also suppress the expression of E-selectin mRNA in activated HUVEC. Conclusions The inhibitive effects of ferulic acid on the expression of adhesion molecules on activated HUVEC may probably contribute to its anti-atherosclerosis effects.

Abstract:Aim To investigate the effect of resveatrol in red wine on the activity of NF-κB and the expressions of protein kinase Co (PKCα) induced by xanthine and xanthine oxidase in cultured rabbit aortic smooth muscle cell (SMC). Methods At first we used 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide metabolism to study the proliferation, then detected activity of nuclear factor-κB (NF-κB) and the protein and mRNA expressions of protein kinase Ca (PKCα) by using electro-phoresis mobility shift assay (EMSA), immunohistochemistry and in situ hybridyzation in cultured rabbit aortic smooth muscle cells (SMC). Results 100 - 200 μmol/L resveatrol (RES), an effective composition in red wine, was confirmed to inhibit proliferation of smooth muscle cells. At same time, the activity of NF-κB as well as the protein and mRNA expressions of protein kinase Ca in rabbit aortic SMC, which were promoted by the oxygen free radicals induced by xanthine and xanthine oxidase, were inhibited. Conclusions Resveatrol may antagonize the increasing of NF-κB activity as well as protein and mRNA expressions of protein kinase Cμ indued by oxygen free radicals in cultured rabbit aortic smooth muscle cells, which may play an important role in preventing active oxygen species leision to atherosclerosis.

Abstract:Aim To study the change of interleukin-10 (IL-10) in focal ischemia tissue of brain and serum in middle cerebral artery occlusion (MCAO) of rats. Methods The middle cerebral artery (MCA) was occluded for different time with modified Zea Longa' s MCAO model. The level of interleukin-10 in focal ischemia tissue of brain and blood serum was measured with double-antibody sandwich enzyme linked immunosorbent assay (ELISA), both in ischemia group and in control group. Results The content of interleukin-10 in focal ischemia tissue of brain was lower at 3 h after ischemia, compared with the control (P> 0.05), and it fell to the lowest degree at 6 h, compared with the control (P< 0.01), then it increased compared with the control (P< 0.05). The concentration of interleukin-10 in serum fell to the lowest degree at 3 h after ischemia compared with the control (p<0.05), and then it increased slowly compared with the control (p< 0.05). Conclusion Interleukin-10 perhaps afford protection to the cerebral ischemia injury.

Abstract:Aim To explore the relationship between serum leptin and essential hypertension, degree of obesity.Methods Serum leptin concentrations were measured by radioimmunoassay. Systolic blood pressure (SBP), diastolic blood pressure (DBF), body mass index (BMI), body fat percentage and fasting blood glucose (FPG) were determined in 115 patients with grade 1 hypertension (51 males, 64 females), 96 patients with grade 2 hypertension (36 males, 60 females) and 47 normo-tensives (23 males, 24 females). The correlation between leptin and other parameters were analyzed. Results Serum leptin concentrations were strongly correlated with BMI and BF percentage in both male and female ( P< 0.001). The concentration level was significantly lower in normotensives group than in grade 1 hypertension group and in grade 2 hypertension group among same gender. The difference in serum leptin concentrations between the three same gender groups disappeared when body fat percentage was corrected by analysis of covariance. Serum leptin concentrations in male and female separately were significantly correlated with SBP, DBP, age, BMI, body fat percentage and FPG (p<0.05-p<0.001). There was a more highly significant correlation between serum leptin concentrations and SBP, DBF in female than in male, but after adjustment for body fat percentage, partial correlation analysis showed that serum leptin concentrations were not correlated with SBP, DBP in both male and female. Logistic regression analysis revealed that serum leptin concentration was not a significant independent predictor of blood pressure in both male and female. Conclusion Serum leptin concentrations are not directly related to essential hypertension in both male and female, but the concentration levels are actually correlated with body fat percentage.

Abstract:Aim To estimate the plasma acylation-stimulating protein (ASP) level in coronary heart disease patients, and to investigate the correlation of plasma ASP with blood lipids. Methods 109 subjects were involved in this study, including 62 coronary heart disease patients and 47 health controls. Enzyme linked immunosorbent assay (ELISA) technique was used to determine the plasma ASP concentration. Blood lipid levels were detected by immune turbidimetry. Results The levels of plasma ASP were increased significantly in coronary heart disease group compared with control group (P< 0.05). Body mass index (BMI), systolic blood pressure, diastolic blood pressure, total cholesterol (TC), triglyceride, low density lipoprotein (LDL) and apolipoprotein B₁₀₀ (apo B₁₀₀) of coronary heart disease group are significantly different from the control group ( P< 0.05). Plasma ASP correlated positively with BMI, TC, triglyceride, LDL and apo B₁₀₀ (respectively r = 0.42, P< 0.01; r = 0.36, p<0.01; r = 0.20, p<0.05; r = 0.31, p<0.05; r = 0.45, p<0.01). Conclusion Plasma ASP takes part in the generation of the disorder of lipid metabolism in coronary heart disease. ASP could be used as a new indicator to evaluate the risk to develop cardiovascular disease.

Abstract:Aim To investigate carotid atherosclerosis in patients with old myocardial infarction and related risk factors of cardiovascular disease. Methods The plaque index and carotid intima-media thickness (IMT) were detected by high-resolution ultrasound technique in 38 patients with old myocardial infarction and 32 matched healthy controls. Results The patients with old myocardial infarction had obviously abnormal plaque index (3.47 ± 1.26 vs 0.35 ± 0.86, p<0.01) and IMT (1.04± 0.17 mm vs 0.63±0.25 mm, p<0.01) compared with the healthy controls. The prevalence rates of atherosclerotic plaques were 84.21% (32/38) and 18.75% (6/32), there was significant difference between the two groups ( P< 0.01). Multiple regression analyses showed that IMT was closely associated with age, total cholesterol and the status of hypertension. Conclusions The patients with old myocardial infarction had abnormal plaque index and IMT, there was close relationship between carotid and atherosclerosis. Carotid ultrasounic examination could help to predict the presence of coronary artery disease.

Abstract:Aim To study the predictive value of inflammatory factors such as C-reactive protein (CRP), fibrinogen and tumor necrosis factor-α (TNF-α) in cardiac events of patients with acute coronary syndromes (ACS). Methods Seventy-six cases of ACS patients were tested of their blood concentrations of CRP, fibrinogen and TNF-α on the day of admission and followed for 7-12 months. Results Concentrations of CRP and fibrinogen in ACS patients were significantly raised compared with the controls, while that of TNF-α showed no difference compared with the controls. Patients who developed cardiac events had higher CRP and fibrinogen levels than those who survived free. Multivariate logistic regression analysis demonstrated CRP and fibrinogen were independent risk factors of ACS. Conclusions CRP and fibrinogen could predict cardiac events in ACS patients in short follow-up period, while TNF-α was of no use in predicting cardiac events in ACS patients.

Abstract:Aim To investigate the possible relationship between oxidized low density lipoprotein (ox-LDL) and LDL-immune complexes (IC) level. Methods We developed a "sandwich" ELISA for measuring plasma LDL-IC level using anti-human IgG (Fab) as the capture antibody and quantitating with monoclonalanti-apoB enzyme conjugate. Oxidixed LDL was also detected by a sandwich ELISA method using monoclonal antibodies against ox-LDL as the capture antibody and quantitating with anti-apoB enzyme conjugate. Their levels were studied in 60 patients with coronary heart disease (CHD) and 50 control subjects. Results Plasma concentrations of TG, lipoprotein (a) and apolipoprotein B in CHD patients were all significantly increased whereas HDLC and apolipoprotein AI levels were decreased. Plasma LDL-IC (2.74 ± 0.73 AU vs 1.38 ± 0.78 AU, P<0.001) and ox-LDL (595.5 ±194.8 μg/L vs 440.3 ± 175.0 μg/L, p<0.001) concentrations in the patients with CHD were both significantly higher than those of control. The relationships between LDL-IC ox-LDL levels and other lipid traits in all the studied subjects ( n = 110) were carried out. LDL-IC levels were positively correlated with TC, TG, LDLC, lipoprotein(a) and apolipoprotein B concentrations while negatively correlated with apolipoprotein AI concentrations respectively. Similarly ox-LDL levels were also found positively correlated with TC, LDLC, or apolipoprotein B concentrations respectively. Furthermore a significantly positive relation between ox-LDL and LDL-IC levels was found (r = 0.313, P< 0.005). Conclusions Increased oxidized LDL and LDL immune complexes levels are risk factors for atherosclerosis.

Abstract:Aim To investigate the efficacy and Safety of simvastatin on patients with coronary heart disease.Methods The study was a random,open,two-dose control trail to evaluate simvastatin administering (10,20 mg/d) to 102 patients with myocardial infarction or angina, whose total cholesterol (TC) was over 2.68 mmol/L and low density lipoprotein cholesterol (LDLC) was over 2.6 mmol/L. Lipid level was determined after 90 and 180 d. Results The blood concentrations of TC and LDLC were significantly decrease after treating withlO and 20 mg simvastatin 90 d and 180 d (P< 0.05)and the curative effect of 20 mg group was more evident than that of 10 mg group (P< 0.05). The TC target rates of 10 mg and 20 mg after 180 d were 21% and 40% and the LDLC target rates of 10 mg and 20 mg after 180 d were 25% and 44% ( P< 0.05). The level of high density lipoprotein cholesterol was gradually increased; the triglyceride was decreased during the following-up, but there were no significant different. The adverse reaction of simvastatin was mild and relatively rate. Conclusion The results suggested that 10 mg or 20 mg of simvastatin could markedly decrease TC and LDLC concentration and was safety and feasible; the effect of 20 mg-group is superior to 10 mg-group.

Abstract:Aim To observe the effect of warfarin to angina pectoris and asymptomatic myocardical ischemia (AsMI) after percutaneous coronary intervention (PCI). Methods 86 cases with angina pectoris or AsMI after PCI were divided into 2 groups. The control group was normative treatment. The therapy group was add Warfarin except normative treatment. It is compared in relieve or disappear time of angina pectoris and the times of AsMI stroke and total ischemia burden and side effects of warfarin in the two groups. Results The relieve time of angina pectoris in therapy and control groups are 3.28 ± 1.67 d and 4.98± 1.69 d (p<0.01), respectively. The disappear time of angina in the two groups are 8.87 ± 0.60 d and 11.04 ± 3.68 d ( P< 0.01), respectively. The effective rate to treatment of angina pectoris are 90.0% and 73.9% ( P< 0.05), respectively . The AsMI stroke difference of total ischemia burden in the two groups are 659.1 ± 257.6 mm· s and 369.7 ± 297.2 mm· s (P< 0.01), respectively. It is explanation that the therapy group have significant clinical effects in angina pectoris and AsMI after PCI compare with control group. Side effects of warfarin have bleeding but no so serious. Conclusion The studydemostrated that warfarin had effect of important assist therapy to angina pectoris and AsMI after PCI. It is safety as careful use.

Abstract:Aim To evaluate conditions of lipid metabolism in the population of Liuzhou. Methods A cohort of the 7 660 residents (3 894 male cases, 3 766 female cases) of Liuzhou, were selected in cluster sampling, average aged 39.5± 11.0 years. Total cholesterol (TC), low density lipoprotein cholesterol (LDLC), triglyceride (TG), high density lipoprotein cholesterol (HDLC), height, weight, waist and hip circumferences of the 7 660 cases were detected and analyzed. Results The average levels of TC, LDLC, TG of both sexes had trend of increasing with age growing. The detecting rate of high TC, high TG, high LDLC and low HDLC in this group were 20.76%, 26.37%, 17.75% and 15.59% , respectively. The dyslipidemia rate had trend of increasing with age growing. The dyslipidernia rate was obviously increased with overweight and obesity. The serum hpid was obviously affected by the waist circumference reflecting fat to accumulate in abdomen. Conclusion The dyslipidemia rate is higher, and the high TG is the most frequent type of the dyslipidemia in this population. Large waist circumference is the major factor affecting lipid metabolism.