BACKGROUND: Statins can block many intracellular signal transductive pathways and suppress the proliferation of various cells by affecting the synthesis of mevalonic acid and the translation following modification of some membrane-connecting proteins.OBJETCIVE: To investigate the influence of simvastatin on the proliferation of lung fibroblasts, the synthesis of collagen and the secretion of matrix metalproteinase-2 (MMP-2).DESIGN:Completely randomized controlled study.SETTING: At Organ Transplanting Research Institute of Fuwai Cardiovasular Diseases Hospital, Peking Union Medical College.MATERIALS: This study was carried out at the Laboratory of Cardiology of Beijing Union Medical College hospital, Peking Union Medical College from June 2004 to October 2004. Lung fibroblasts derived from neonatal SD rats were co-cultured in vitro with different dosage of simvastatin of 0, 1, 5, 10,50 μmol/L, and 50 μmol/L simvastatin + 200 μmol/L mevalonic acid.METHODS: Lung fibroblasts deriving from neonatal SD rat were co-cultured with different dosage of simvastatine in vitro. Methyl thiazolyl tetrazolium colorinetry was used to detect the cell proliferation, and cell immunohistochemical assay was used to determine the collagen synthesis and meanwhile,MMP-2 content in supernatant was examined with enzyme-linked immunosorbent assay.MAIN OUTCOME MEASURES: The proliferation of fibroblasts, the synthesis of collagen and the secretion of MMP-2 due to different dosage of simvastatin intervention and simvastatin combined with mevalonic acid.presenting the expression of type Ⅰ, Ⅲ of collagen in lung fibroblasts and the level of MMP-2 in 5, 10, 50 μmol/L simvastatin group were obviously lower than those of 0 μmol/L simvastatin group (0. 520 ± 0.010, 0. 334 ± 0.011,0.260±0.012, 0.111±0.011; 0.508±0.011, 0.324±0.014, 0.232±0.015, 0. 083 ±0. 015; 0.445 ±0. 017, 0. 305 ±0. 015, 0.216 ±0. 015,0.068±0.012; 0.561±0.013, 0.361 ±0.012, 0.289±0.012, 0.140value, the mean absorbency( A value) in lung fibroblasts and the level of MMP-2 in 50 μmol/L simvastatin + 200 μmol/L mevalonic acid group were obviously higher than that of the 50 μmol/L simvastatin group(0. 567±0.015, 0.354±0.014, 0.283±0.012, 0.138±0.011, t=4.715-10.950, P ＜ 0.01).CONCLUSION: Simvastatin could suppress the fibroblast proliferation and collagen synthesis, attenuate the secretion of MMP-2 and suppress consequently the adhesion and migration of lung fibroblasts; moreover, it has the capability of anti-cell proliferation by affecting the mevalonic acid pathway.

Objective To study the potential of bFGF to promote human bone marrow derived MSCs to differentiate into cardiomyogenic cells, and its effect on proliferation of MSCs. Methods MSCs isolated from adult human bone marrow were cultured in four different systems. Group A: medium with 5-aza. Group B: medium with bFGF. Group C: medium with 5-aza+bFGF. Control group: medium alone. The morphological changes of MSCs were observed. Then immunocytochemistry staining against ?-actin,cTnT,and Connixin43 was performed. The expression of Nkx2.5, GATA-4 and cTnT was detected by semi-quantitative RT-PCR. The proliferation of MSCs in different groups was measured by MTT. Results The MSCs in both group A and C partially differentiated into myogenic cells and expressed proteins of ?-actin,cTnT,and connixin43. In group A and group C, the mRNA level of Nkx2.5,GATA-4 and cTnT was higher than that of control group. In group B, mRNA level of Nkx2.5 and GATA-4 was higher than that of control group. As compared with the control group, cell proliferation was faster in group B than in group A and C. And the proliferation was faster in group C than in group A. Conclusion bFGF can promote the proliferation of MSCs significantly. When combining with 5-aza, bFGF can promote MSCs to differentiate into cardiomyogenic cells more effectively.

Objective To determine the relationships between maternal serum copper,amniotic copper,lysyl oxidase (LOX) and collagen Ⅲ in pregnant women with premature rupture of membranes (PROM) and without PROM.Methods One hundred women with PROM were enrolled in this study,and divided into 37-42 weeks,34-36~ +6 weeks and 28-33~ +6 weeks according to gestational age. One hundred non-PROM pregnancies matching the same gestational ages were recruited as control group. Copper of maternal serum and amnion in two groups were compared by FAAS method. Amniotic LOX was analyzed by fluorometry. Amniotic collagen Ⅲ was detected by immunohistochemical method and computer image analysis system(absorbance,A). Linear correlation analysis was used to explore the relationships between maternal serum copper,amniotic copper,LOX and collagen Ⅲ. Results (1)For 37-42 weeks pregnant women,serum copper was correlated positively with amniotic copper in two groups, r= 0.82(P0.05),but amniotic LOX and collagen Ⅲ decreased significantly compared with controls, being [(0.53?0.10)?g/g vs (0.75?0.10)?g/g,P

Objective To study the effect of methadone maintenance treatment(MMT) on reducing HIV infection among injecting drug users(IDUs) in Yueyang city of Hunan province.Methods Local IDUs were provided with MMT along with peer education and behavior interventions,and they were tested for HIV antibody to evaluate the effect of MMT after one year treatment.Results Of the 113 IDUs who were negative at the entry,none turned to be positive after one year treatment and the negative partners of 22 IDUs with positive HIV antibody remained negative one year later,and the negative rate of urine test maintained at 76.10%.Conclusion MMT combined with peer education and behavior interventions will decrease the use of heroin,prevent HIV infection among IDUs and reduce their partners' susceptibility to HIV.

Objective In this study, we try to understand the effects of microenvironment on the differentiation of mesenchymal stem cells (MSCs) by coculturing MSCs with mature cardiomyocytes or culturing MSCs in cardiomyocyte-lysate, in this study. Methods MSCs isolated from mature rats were either cocultured with cardiomyocytes isolated from new born rats with the ratio of 1 to 4, or cultured in the medium containing 4-fold cardiomyocyte-lysate obtained by repeated freezing and defrosting of rat myocardial cells. The morphology of MSCs under light microscopy were observed daily for 7 days and immunostaining against cTnT and CD31 was performed on the 7~ th day. MSCs cultured in ordinary medium were observed as the control. Results Both MSCs cocultured with cardiomyocytes and cultured in cardiomyocyte-lysate were differentiated into myogenic cells and expressed cTnT and CD31 at the 7th day of cultivation. The MSCs in the control group did not change in morphology and express cTnT or CD31. Conclusion Both myocardial cell coculturing system and cardiomyocyte-lysate system can be used to induce bone marrow MSCs to differentiate into cardiomyocyte-like cells and endotheliocyte-like cells.

Objective To explore the effect of tert?butylhydroquinone(tBHQ)on ultraviolet B(UVB)?induced oxidative damages in human im?mortalized keratinocytes(HaCaT),and discuss its mechanism. Methods The cultured HaCaT cells were randomly divided into 4 groups:control group(G1),ultraviolet irradiation group(G2),25μmol/L tBHQ pretreatment before ultraviolet irradiation group(G3),and 50μmol/L tBHQ pre?treatment before ultraviolet irradiation group(G4). The content of reactive oxygen species was detected by DCFH?DA method,and the cell prolifera?tion was evaluated by MTT. Western blot was used to measure the protein expression of nuclear factor E2?related factor 2(Nrf2)in both nuclear fac?tions and whole?cell of HaCaT. The mRNA expressions of CAT and SRX were determined by real?time RT?PCR. Results The content of reactive oxygen species in HaCaT cells was increased,and the cell proliferation rate was decreased significantly after ultraviolet irradiation. The pretreatment of 25 and 50μmol/L tBHQ can inhibit the UVB?induced oxidative damage in a dose?dependent manner in HaCaT cells. Compared with G2 group, tBHQ pretreatment could dose?dependently increase the level of Nrf2 protein in nuclear factions and whole?cell of HaCaT,and also the mRNA ex?pressions of CAT and SRX. Conclusion UVB irradiation can induce oxidative stress damages of HaCaT cells. tBHQ may inhibit the UVB?induced oxidative damages through enhancing Nrf2 expressions and nuclear translocation,then activating the transcription of the downstream antioxidant en?zymes CAT and SRX.

Objective To investigate the potential change of cellular cholesterol efflux and the functional changes of ABCA1 on macrophages from diabetes animals. Methods High energy diet was given to golden hamster to make animal model of hyperlipidemia. Diabetes was induced by a single intraperitoneal injection of STZ (30 mg/kg). Macrophages were isolated and incubated with apoA-I or 8-br-cAMP in vitro. Cellular cholesterol content was measureal before and after treatment by HPLC. Results Intracellular cholesterol content of diabetic animals was higher than that in high energy diet and normal controls. Expression of ABCA1 mRNA was upregulated in diabetic group after incubation with apoA-I and cAMP. When macrophages were incubated with apoA-I, cholesterol efflux increased with the prolongation of incubation time.The amount of intracellular cholesterol efflux in 3 groups of animals showed the following relationship: diabetic animals exceed high energy diet fed animals, and the latter exceeds normal animals. Conclusion Intracellular lipid efflux depends mainly on the presence of extracellular apoA1 as well as membrane ABCA1 protein. The deposition of cholesterol in the macrophage of diabetic and insulin resistant animals may be related to the characteristic changes of quantity and function of apoA1 in these animals.

Objective To investigate the relationship between inflammatory factors, coronary artery dilation, and their clinical significance. Methods The cases undergone coronary angiography in our hospital last year were collected and divided into three groups: the first one included 11 patients whose angiography showed coronary artery dilation, the second group included 35 cases of atherosclerosis, and the third includes 24 cases with normal angiography. sES, MMP9 and TIMP1 were measured by ELISA method. Results Patients with coronary artery dilation were found to have significantly higher sES and MMP-9 level in comparison with atherosclerosis group and normal group[(153.7?152.7)ng/L,(90.1?54.2)ng/L,(76.5?37.2)ng/L, respectively](P

Objective To observe the expression and location of TF and TFPI in femoral artery atherosclerotic plaque.Methods We detected the expressions and locations of TF and TFPI in femoral artery atherosclerotic plaque by immunohistochemical and double-stain immunohistochemical method.We detected TF mRNA and TFPI mRNA expressions in atherosclerotic plaque by RT-PCR,with the normal umbilical artery as a control.ResultsThe normal umbilical artery contained little TF,TFPI and their mRNA in the adventitia.A great deal of TF,TFPI and their mRNA were found in the tunica intima of the femoral artery atherosclerotic plaque.Conclusion Expression of TF,TFPI and their mRNA of all types of cells and stroma in the proliferative tunica intima.

Objective To explain the effects of C-reactive protein(CRP) on lectin-like oxidized low density lipoprotein receptor-1 expression on THP-1 derived macrophages and the related signal transduction pathways.MethodsTHP-1 cells were differentiated into macrophages with the stimulation of PMA.THP-1 derived macrophages were incubated with CRP and co-incubated with inhibitors of NF-?B、AP-1 and MARK signal transduction pathways.The expression of LOX-1 antigen and mRNA was analyzed by ELISA and RT-PCR.Results CRP stimulated the expression of LOX-1 antigen and mRNA on macrophages in a dose-dependent manner.NF-?B inhibitor BAY11-7085 suppressed the inducible effects of CRP on LOX-1 expression.Conclusion CRP increased LOX-1 expression on THP-1 derived macrophages at transcription and post-transcription levels.The NF-?B signal transduction pathway may be involved in such process.