The effects of vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF) on proliferation of bovine coronary artery endothelial cells (BCAEC) and smooth muscle cells (BCASMC) were studied in vitro. BCAEC and BCASMC were isolated and cultured and divided into control group, VEGF (50ng/ml) group and HGF (50ng/ml) group. Cells proliferation was measured using MTT method. The results showed that the OD values of control, VEGF, and HGF group in BCAEC cultures were 0.23?0.02, 0.58?0.10, and 0.42?0.12, respectively, and those in BCASMC were 0.31?0.08, 0.45?0.09, and 0.40?0.11, respectively. The proliferation ratios of BCAEC and BCASMC induced by HGF were 152.2%?33.8% and 45.2%?25.3%, respectively, and that by VEGF were 82.6%?18.7% and 29.0%?20.4%, respectively. The results suggested that HGF could promote proliferation and migration of BCAEC and BCASMC, while VEGF could promote proliferation of BCAEC but not BCASMC. The effect of HGF on BCAEC was stronger than that on BCASMC, and the induction strength of HGF was higher than VEGF.

Objective To explore the image characteristics and clinical application of the optical coherence tomography (OCT) in patients with epiretinal membranes of the macular(ERMM). Methods 38 patients, who were diagnosed as or suspected as ERMM, were examined with OCT before and after operation from November, 2002 to October, 2003 in our hospital. Results Macular epiretinal membranes were visible on OCT as high reflective tissues, which were thin or thick, and contiguous to or anterior to the retinal surface. In most fovea, the depth decreased and the thickness increased. ERMM disappeared after operation. Conclusion OCT can display the macular epiretinal membrane and the pathological changes of macular tissues before and after operation. OCT can provide accurate information on the clinical diagnosis and operative efficacy of ERMM.

Background Studies show that retinal neurodegeneration may precede retinal microvascular changes in diabetes mellitus.The apoptosis of retinal ganglion cells (RGCs) is an early finding in retinal neurodegeneration.Toll-like receptor 4 (TLR4) is proved to be up-regulated in diabetic rats retina.However,the impact of TLR4 on RGCs damage in retinal neurodegeneration is poorly understood.Objective The aim of this study was to investigate the expressing change of TLR4 induced by high glucose in RGCs in order to offer a basis for the prevention diabetic retinal neurodegeneration and the study on targeting drugs.Methods RGCs were isolated and purified from the retinas of SPF SD rats aged postnatal 1-3 days by using papain digestion method and then were identified by immunofluorescence technology to detect the expression of Brn3a,a specific marker of RGCs.The cells were divided into normal control group and 10,20,30 mmol/L glucose groups.The expressions of TLR4 mRNA and protein in the ceils were detected by real-time fluorescence quantitative PCR and Western blot analysis in 24 and 48 hours after addtion of glucose.All procedures performed in studies were in accordance with the Association for National Institutes of Health (NIH) Statement for the Care and Use of Laboratory Animals recommendations.The protocol was approved by the Ethics Committee of Renmin Hospital of Wuhan University.Every effort was made to minimize animal discomfort and stress.Results The normal cells grew well with the shape of near roundness after inoculaton.The cells were gradually enlarged and clustered with obvious axons and dendrites 24 hours after purifying.Brn3a showed the positive expression in cultured cells.At 24 hours and 48 hours after glucose culture,the cell structures were gradually invisible in most cells.The expressions of TLR4 mRNA in the cells were 0.945 ±0.237,1.180±0.193 and 0.827±0.213 at 24 hours and 1.509±0.422,2.433±0.617 and 1.435±0.410 at 48 hours after culture in the 10,20 and 30 mmol/L glucose groups,respectively,which were significantly higher than 0.600±0.099 and 0.724±0.302 in the normal control group (all at P＜0.01).The expressions of TLR4 protein in the cells were 0.442±0.147,0.626±0.128 and 0.330±0.153 at 24 hours and 0.464±0.121,0.930±0.441 and 0.394±0.158 at 48 hours after culture in the 10,20 and 30 mmol/L glucose groups,respectively,which were significantly higher than 0.090±0.050 and 0.094±0.070 in the normal control group (all at P＜0.01).Conclusions A large number of RGCs die in a high-glucose environment in vitro,meanwhile,the expression of TLR4 up-regulates in the cells,indicating that TLR4 maybe participate in the damage of RGCs induced by high glucose.

Background Researches showed that platelet-derived growth factor (PDGF) modulate the expression of matrix metalloproteinase/tissue inhibitor of metalloproteinase (MMP/TIMP) in cells,but the association of expression of MMP/TIMP in retinal pigment epithelial (RPE) cells and the dose and active time of PDGF is unclear.Objective This study was to observe the effects of PDGF on the expressions of MMP-2,MMP-9 and TIMP-1 in cultured RPE cells in vitro.Methods RPE cell line,ARPE-19,was calculated in vitro,and the cells were divided into 5 groups when they reached 70％-80％ confluence.Different concentrations (0,0.1,1,10,50 mg/L) of PDGF was added into the medium respectively for 36 hours,and the expressing levels of mRNA and protein of MMP-2,MMP-9 and TIMP-1 were detected by reverse transcription PCR (RT-PCR) and Western blot assay.In addition,RPE cells in PDGF group were treated with 10 mg/L PDGF for 24,36,48 hours respectively to detect the expressions of mRNA and protein of MMP-2,MMP-9 and TIMP-1 in the cells and to compare with the control group without PDGF.Results PDGF stimulated proliferation of RPE cells in a dose-and time-dependent manner.As the increase of the PDGF concentrations,the expression values of MMP-2 mRNA and MMP-9 mRNA in RPE cells were gradually elevated,with a statistically significant difference among various groups (MMP-2 mRNA:F=79.304,P=0.000;MMP-9 mRNA:F =8.465,P=0.003),and the expressions of MMP-2 mRNA and MMP-9 mRNA were significantly higher in the 1,10,50 mg/L PDGF groups compared with 0 mg/L PDGF normal control group (all at P＜0.05).Also,the expression values of MMP-2 and MMP-9 proteins in RPE cells were gradually elevated with the increase of PDGF concentrations,showing statistically differences among the groups (MMP-2:F=26.550,P=0.000;MMP-9:F=80.993,P=0.000).Compared with the 0 mg/L PDGF group,MMP-2 and MMP-9 expression levels in the 1,10,50 mg/L PDGF groups were significantly up-regulated (all at P＜ 0.05).However,the expression levels of TIMP-1 mRNA and protein group in the cells were not significantly different among various groups (mRNA:F =0.143,P =0.962 ; protein:F =1.955,P =0.178).The expression levels of M MP-2 mRNA,M MP-9 mRNA in the cells were increased in the PDGF group compared with the control group at different time points (MMP-2 mRNA:Ftime =83.250,P=0.002 ; MMP-9 mRNA:Ftime =6.785,P =0.019).Also,the expression values of MMP-2 and MMP-9proteins in RPE cells were increased in the PDGF group compared with the control group at different time points (MMP-2:Ftime =1 l.185,P =0.041 ; MMP-9:Ftime =968.413,P =0.000).The expression levels of MMP-2 and MMP-9 mRNAs and proteins were significant between the two groups at different time points (all at Pgroup =0.000;all at Ptime＜0.05).While the expression changes of TIMP-1 werc not significant between the two groups and among various time points (all at P＞0.05) Conclusions PDGF up-regulates MMP-2 and MMP-9 expressions in RPE cells in a dose-and time-dependent manner.But,PDGF dose not alter the expression of TIMP-1.These results indicate that PDGF disrupt the balance of MMP/TIMP,which may damage the extracellular matrix and therefore facilitate the migration of RPE cells in the pathogenesis of proliferative vitreoretinopathy.

The Y118A、Y118F、Y118T、S378A、S378T、H310A、H310R mutants of Candida albicans sterol 14?-demethylase (CACYP51) were constructed and heterologously expressed in D12667, the reconstructed strain with the deletion of CYP51 gene of the Y12667. With the strains obtained and microsome enzymes separated, the western blot and the ultraviolet absorption spectrophotometry were used to qualitative and quantitative detect the expressed protein, the GC-MS was used to detect the metabolism activity of the protein. The results showed that, the target protein expressed successfully in the reconstructed strains, with the expression level up to 25% of the total microsome proteins. The results also showed that, the wild type protein had the catalytic activity to its nature substrate. While after alteration the wild gene with Y118A、Y118F、Y118T、S378A、S378T、H310A、H310R by a single base substitution, the catalytic activity of protein markedly decreased respectively. So the wild type and mutation CYP51 were expressed successfully in Saccharomyces cerevisiae and the expression products preserved the activity to metabolism their nature substrate.

Objective:To understand the expression levels of Tim-3,a new proinflammatory factor in the early stages of Echinococcus granulosus infection in mice.Methods: BALB/c mice were infected with E.granulosus.Peritoneal macrophages and spleen cells were collected at 1,5,9 and 13 days post-infection.At different time points ,the levels of Tim-3 in peritoneal macrophages and spleen CD3+lymphocyte subsets were detected by FCM , and the relative expression of TLR 4 mRNA was detected by qRT-PCR.Results:There was no significant difference in the expression levels of Tim-3 of CD3+spleen lymphocyte subsets between E.granulosus group and control group (P>0.05).The expression levels of Tim-3 of spleen macrophages (9,13 days) and peritoneal macrophage (5,9,13 days) were much higher in E.granulosus infected group than those in control group with statistical significance (P<0.05).The numbers of macrophages were no change.Compared with control groups,the relative expression of TLR4 mRNA at 1 day post-infection was statistically higher in E.granulosus infected group ( P<0.05 ).Conclusion:During early stage of E.granulosus infection in mice,the levels of Tim-3 expression are upregulated,while the expression of TLR4 are downregulated,which may inhibit the function of macrophages resulting in host-immunity-defensive-system inhibition and immune tolerance of E.granulosus to host.

PURPOSE: In the present study, we investigated the incidence of cardiotoxicity within 5 years of trastuzumab treatment and evaluated potential risk factors in clinical practice. METHODS: The study cohort included 415 patients diagnosed with early breast cancer (EBC). Cardiotoxicity incidence was evaluated in patients receiving trastuzumab and those who did not. Multivariate Cox proportional hazards regression models were used to estimate hazard ratios and 95% confidence intervals of potential risk factors for trastuzumab-related cardiotoxicity after appropriate adjustments. RESULTS: Incidence of cardiotoxicity in patients treated with trastuzumab was significantly higher than that in controls (23.7% vs. 10.8%, p<0.001). This result was adjusted for factors that might increase the risk of cardiotoxicity, such as history of coronary artery diseases or the use of anthracyclines for more than four cycles. CONCLUSION: Our findings indicated that treatment with trastuzumab was strongly associated with cardiotoxicity in EBC patients.
Anthracyclines ; Breast Neoplasms* ; Cohort Studies ; Coronary Artery Disease ; Humans ; Incidence ; Observational Study* ; Prospective Studies* ; Risk Factors ; Trastuzumab