Objective To investigate the effects of new compound traditional Chinese medicine prepatations Banxiao capsule on the mobilization of endothelial progenitor cells ( EPCs) .Methods After EPCs from mouse bone marrow treated with variable dose of Banxiao capsule, we detected the proliferation by CCK-8 assay, quantified the number of CD34/Flk-1 double positive cells by flow cytometric analysis, evaluated the migratory function of EPCs by transwell chamber assay, and studied the protein level of Bcl-2/Bax by Western blot analysis.Results Compared with untreated group, the proliferation potential of EPCs in Banxiao low dose (20μg/ml) and high dose (200μg/ml) group was increased(P<0.05), the number of CD34/Flk-1 double positive cells were increased (P<0.05), migrative activity of EPCs was increased (P<0.05),the level of Bcl-2 protein were increased, and the level of Bax protein were decreased.Con-clusion Banxiao capsule can enhance proliferation of EPCs through up-regulating of expression of Bcl-2/Bax.

Objective: To investigate the effect of simvastatin on the expression of vascular endothelial growth factor (VEGF) in rabbit aortic atherosclerotic plaques. Methods :Twenty rabbit aortic atherosclerotic models developed by hyper-lipidemic and hypercholesterol feeding were randomly divided into simvastatin treated group (n = 10) and hyperlipidemic diet group (n = 6); another 8 commonly fed rabbits were taken as control group. VEGF protein levels in atherosclerotic plaques were studied by immunohistochemistry. Results: Positive signal of VEGF in simvastatin treated group and hyperlipidemic group were significantly higher than that in control group(P

Objective: To observe the alteration of type-　 Ⅰ collagen protein gene expression after arterial injury and investigate its effect on the development of restenosis. Methods: Firstly, thee xperimental carotid arterial injury rabbit model was constructed. Then, Norther n blot, in situ hybridization and histomorphometric analysis were used to de tect the expression of procollagen mRNA and the accumulation of collagen protein 1,2,4 weeks after injury. Results: Type-　Ⅰ collag en mRNA increased 1 week after injury, peaked 2 weeks later and decreased 4 week s later. The deposition of the collagen protein account for a high percentage o f space in neointima on histomorphometric analysis. Conclusion: Collagen protein may play an important role in the development of neointima and restenosis.

To examine the effect of benapril on atherogenesis, circulating endothelial cells (CEC), and factor vWF. Twenty four healthy male New Zealand white rabbits were randomly divided into 3 groups. In group C (control group) the rabbits were fed normal rabbit diet for ten weeks, in group AS 1% cholesterol diet was fed, and in group B both 1% cholesterol diet and benapril 5mg/day were fed. Fasting blood samples were collected for the determination of serum total cholesterol (TC), high density lipoprotein cholesterol (HDL c), low density lipoprotein cholesterol (LDL c), and triglyceride (TG) on day 0, 5 and 10 weeks after feeding. The levels of plasma CEC were measured by morphometrical count and the levels of plasma vWF were determined by ELISA. After the animals were sacrificed, aortas were obtained, and they were stained with oil red O. The tissue was sectioned for histological examination. Planimetry was performed with a computer system. Endothelial cells were identified by the presence of factor VIII related antigen, which was the specific marker for endothelial cells. The results showed that the levels of serum Tc, TG, and LDL c were increased progressively in group B and group AS, but the levels of vWF in group AS were significantly higher than that in group B at the 5 th and 10 th weeks respectively ( P

Objective To study the effects of adenovirue-mediated p27kipl gene and its protein product overexpression on neointimal proliferation of rabbit carotid artery after balloon injury. Methods After rabbit arterial carotid injuried, injuried arterial segments were immediately infected by LacZ recombinant adenovirues (AdLacZ) and human p27kipl recombinant adenovirues (Adhp27kip1) in vivo, respectively. Western blot, x-gal stainning, HE stainning, immunochemistry and compute image system were used to analyze the expression of exogenous p27kipl gene and its protein product and the effects on neointimal proliferation. Results Comparison to AdLacZ infected or uninfected arterial segments, Adhp27kip1 infected segments overexpressed p27kipl protein, the peak of expression was in 7-14 d, expression lasted more than 4 weeks. After 4 weeks, in uninfected, AdLacZ infected or Adhp27kipl infected segments, the neointimal area was 1.106 mm2, 0.988 mm2 and 0.278 mm2, respecively; the rate of luminal stenosis was 87.07%, 65.40% and 32.14%, respectively. Conclusion Exogenous p27kipl gene and its protein product overexpression in injuried arterial segments could inhibit neointimal proliferation and luminal stenosis significantly after artery injury.

AIM: To isolate and culture endothelial progenitor cells (EPCs) in blood vessel by in vitro amplification from bone marrow of rabbits to provide cytology basis for the implantation of autologous EPCs in the repair of blood vessel endothelium. METHODS: The experiment was performed at the Department of Cardiology, Changzheng Hospital, Second Military Medical University of Chinese PLA from March 2005 to February 2006. ①Dil labeled acetylated low density lipoprotein, FITC labeled BS-1 lectin, mouse anti-human CD34 antibody, rabbit anti-human FIK-1 antibody, mouse anti-human CD133 monoclonal antibody were purchased from molecular probes company, vector company, Biolegend company, Biolegeng company and R&D company. ②Totally 8 New Zealand rabbits were selected to extract the bone barrow. Mononuclear cell was isolated from bone marrow by density centrifugation. With the inoculated density of 1?106/cm2, it was put in the M199 medium containing vascular endothelial growth factor and Basic fibroblast growth factor (bFGF) for in vitro cultivation for 7 days. Cell growth and reproductive activity were observed. ③EPCs were identified by Dil labeled acetylated low density lipoprotein and FITC labeled BS-1 lectin. The cells showed red fluorescence were cells phagocytized acetylated low density lipoprotein, while those with green gluorescence were cells bind with BS-1, and the cells double stained showed orange fluorescence. ④Expressions of CD133, CD34 and Flk-1 were detected with immunofluorescent staining and flow cytometry. RESULTS: ①Observation of cell morphous: New isolated mononuclear cells were round. After cultivation for 72 hours, adherent cells showed colony-like growth, and the cells were round or irregular, and the caryocinesis was relatively obvious. Till the 7th day after cultivation, cell colony was connected each other, showing fusiform endothelioid cells. ②Reproductive activity of EPCs in blood vessel: At days 2-4, the reproduction of EPCs was rapid, and then became slow gradually. Growth curve showed typical "S" shape. At days 6 and 7, the EPCs grew rapidly. The absorbance (A) reached 0.58?0.15 and 0.62?0.23, respectively. ③Result of EPCs identification by Dil labeled acetylated low density lipoprotein and FITC labeled BS-1 lectin: In kytoplasm of EPCs, red fluorescent concentration bind with acetylated low density lipoprotein appeared, with the positive rate of over 95%. Combined rate with BS-1 lectin reached 100% nearly. Double staining rate reached over 90%. ④Result of EPCs immunohistochemical method and flow cytometry: The cell-surface marker CD133, FlK-1 and CD34 were positive. CONCLUSION: Cell colony with the feature of EPCs can be isolated and cultured from rabbit bone marrow by in vitro amplification method successfully.

Objective To investigate the mechanism of migration of vascular adventitial fibroblasts(AFs) enhanced by advanced glycation end-products(AGEs) and the inhibition effect of candesartan.Methods Isolated vascular adventitial fibroblast of Sprague-Dawley(SD) rats were cultured.Migratory potential was estimated with Transwell chamber in vitro.Expression of receptor for AGEs(RAGE) and phosphorylated mitogen-activated protein kinase(MAPK) of AFs were determined by RT-PCR and Western blotting.Results AGEs modified human serum albumin(AGE-HAS,concentration ranged from 0 to 300mg/L) up-regulated the expression of RAGE at mRNA and protein levels,peaked at a concentration of 200mg/L(P

Objective: To assess the autonomic nervous impairm ent in chronic renal failure and its related factors. Methods: F orty adults were randomly selected including in-patients in the nephrology ward and healthy subjects for routine medical examination. The subjects were classifi ed into 4 groups: normal subjects(NS)，normal renal function，nitremia， uremic patients. The time domain measurements of heart rate variability(HRV) and ambula tory blood pressure were analyzed simultaneously . Results: (1) There were significant differences as compared with normal subjects in the time domain measurements of HRV in uremic group. It decreased significantly when the patient was defined as end stage chronic renal failure. There were no significan t differences between NS，normal renal function group and nitremic group. (2) Ti me domain measurements of HRV was significantly lower(P<0.05) in uremia with renal hypertension than in uremia with normal blood pressure. Conclusio n: (1) Patients with chronic renal failure(HRV) have their cardiac auton omic nervous system impaired conspicuously in the course of uremia. (2) There is a positive correlation between cardiac autonomic nervous system impairment in p atients with CRF and renal function levels. Uremia itself is an independent fact or for the impairment of cardiac autonomic nervous system. (3) Renal hypertensio n with uremia may intensify the impairment of cardiac autonomic nervous system of the patients.

The change of PKC activity and cytosolic free Ca 2+ induced by oxLDL and simvastatin in ASMC were measured by its ability to transfer phosphate from 32P-ATP to lysine-rich histone and flow cytometric analysis loading with the Ca 2+ dye fluo-3/Am, respectively. As a result, oxLDL increased total PKC activity in a dose-dependent manner with phase peaking at 14 min, then decreased slowly and maintained for at least 30 min, while oxLDL induced biphasic [Ca 2+]i responses including rapid initial transient phase and sustained phase. Removal of extracellular Ca 2+did not inhibit the rapid phase of oxLDL-induced rise in [Ca 2+]i, but abolished the sustained phase of [Ca 2+]i in response to oxLDL. Activity of PKC was markedly decreased and simvastatin did not impair the initial peak in response to oxLDL but significantly reduced the subsequent plateau phase when simvastatin was added. The results suggested that oxLDL can induce dynamic changes of signal transduction of PKC and cytosolic free Ca 2+in ASMC. The rapid initial transient phase was due to the mobilization of [Ca 2+]i from intracellular Ca 2+ pool and the sustained phase resulted from the influx of extracellular Ca 2+. The inhibition of PKC activity induced by simvastatin may contribute to the changes of intracellular Ca 2+.

To investigate the clinical significance of the expression of CD40L on peripheral blood monocytes and the changes in serum soluble CD40L in patients with acute coronary syndrome, 16 normal controls and fifty-six patients including 24 with SA (8 patients after PTCA), 20 with UA and 12 with AMI entered in this study. The expression of CD40L on monocytes was analyzed by indirect immonofluorescence flow cytometry and serum sCD40L levels were measured by ELISA. The results showed: (1) The expression of CD40L on monocytes in UA and AMI were higher compared with SA and controls ( P 0.05). (2) Patients with UA and AMI had significantly raised serum levels of sCD40L when compared with patients with SA and controls( P