Objective To observe the inhibition effect of selective cyclooxygenase-2 inhibitor (celecoxib) on the experimental choroidal neovascularization (CNV).Methods Thirty 8-10 weeks old healthy male Brown-Norway(BN) rats were randomly divided into the control,laser and celecoxib group,with 10 rats in each group.At the dosage of 50 mg/kg,celecoxib was garaged twice per day.After 7 days,experimental CNV was induced by Krypon laser on laser group and celecoxib group.Fundus fluorescein angiography (FFA) was performed on days 3,7,14,21,30 after laser photocoagulation.On days 21 after photocoagulation,5 rats in each group were sacrificed and the relative thickness of CNV membranes,the expression of COX-2,vascular endothelial growth factor(VEGF) and matrix metalloproteinase-2 (MMP-2) Were studied by histopathologic or immunohistochemistry examination.Results On days 21 after photocoagulation,the incidence of CNV in the celecoxib group is significantly lower than that in the laser group (X~2=7.1068,P=0v0077);the relative thickness of the CNV membranes in the celecoxib group is reduced 41.38% compared to the laser group,the difference is statistically significant (t=16.7600,P=0.0000).COX-2,VEGF and MMP-2 expression in the CNV membrane of celecoxib group were significantly lower than in control group (t=5.7100,5.8400,8.0200;P=0.0000);the COX-2,VEGF and MMP-2 expressions in choroid and retina of control group were weak.Conclusion Prophylactic celecoxib can reduce the expression of VEGF and MMP-2 by inhibiting COX-2,and prevent the CNV induced by laser photocoagulation.

Objective To investigate the expression of Toll like receptor 2 (TLR2) in glioma and its relationship with the malignant biological behavior of glioma, so as to provide a new therapeutic target for glioma immunotherapy. Methods The tumor tissues of 90 glioma patients undergoing surgical excision were collected, of which WHO gradingⅠ-Ⅱgrade 39 cases,Ⅲgrade 24 cases,Ⅳgrade 24 cases. In addition, the normal brain tissues of 12 patients undergoing routine intracranial decompression were selected. The human glioma cell lines U87-MG, U251-MG and human astrocyte cell line HA were cultured. The expression levels of TLR2 mRNA and protein were detected by reverse transcriptase polymerase chain reaction (RT-PCR) and Western Blot test. The correlation between TLR2 protein and different clinical pathological parameters was analyzed. Results The expression levels of TLR2 mRNA and protein in the glioma tissuesⅠ-Ⅱgrade,Ⅲgrade andⅣgrade were significantly higher than those in normal brain tissues (0.27 ± 0.09, 0.57 ± 0.12 and 0.96 ± 0.18 vs. 0.11 ± 0.05; 0.31 ± 0.05, 0.44 ± 0.05 and 0.71 ± 0.09 vs. 0.02 ± 0.01), there were statistical differences (P<0.05). In different grades of glioma tissues, there were statistical differences in the expression levels of TLR2 mRNA and protein (F = 205.9 and 194.9, P<0.05). The expression levels of TLR2 mRNA and protein in human malignant glioma cell lines U87-MG and U251-MG were significantly higher than those in human astrocyte cell line HA (1.000 ± 0.100 and 0.356 ± 0.060 vs. 0.245 ± 0.030, 0.720 ± 0.100 and 1.800 ± 0.150 vs. 0.004 ± 0.000), there were statistical differences (P<0.05). The expression of TLR2 protein was independent of age, sex and location(P>0.05), but was related to tumor diameter and WHO grading (P < 0.01). Conclusions TLR2 in different grade glioma tissues and glioma cell lines are expressed, and its expression level is associated with the malignant degree of glioma; TLR2 protein not only can be used as a biomarker of gliomas and prognosis, but also provide a new target for the treatment of glioma.

A sensitive analytical method based on reversed microemulsion electrokinetic chromatography ( MEEKC) combined with on_line preconcentration technique was developed for the determination of polycyclic aromatic hydrocarbons ( PAHs ) in cosmetics. For six lipophilic PAHs analytes which are difficult to be separated under conventional conditions, three stacking techniques including large volume sample stacking ( LVSS) , dynamic pH junction and sweeping ( LVSS_DypH_sweep ) were combined to realize the efficient preconcentration and separation. Under the optimum conditions, including the microemulsion buffer with the composition of 2. 4%(w/w)SDS_0. 6% (w/w) octane_6. 6% (w/w)n_butyl alcohol_20 mmol/L NaH2PO4 ( pH 2 . 2 ) , HCB injection time of 20 s ( 16 kPa ) and sample injection time of 80 s ( 16 kPa ) , good enrichment effect was reached with the enrichment factors ranged from 25 to 80 , and the PAHs were analyzed successfully within 27 min. The developed method was used to analyze the PAHs in cosmetics. The recoveries ranged from 90 . 6% to 95 . 9%. The RSD values ( n=5 ) were less than 5 . 1%.

Objective To explore the role of p16 gene methylation in fibroblasts in the occurrence and development of keloid.Methods Skin tissue specimens were resected from the lesions of patients with keloid and normal skin of healthy human controls.Fibroblasts were isolated from these tissue specimens and subjected a primary culture.An immunohistochemical analysis was performed to measure the expression of p16 protein in tissue specimens,real-time fluorescence-based quantitative PCR to determine the mRNA expression level (expressed as 2-△△Ct) of p 16 and DNA methyltransferases (DNMTs) in fibmblasts,and bisulfite sequencing PCR (BSP) to estimate the methylation status of p16 gene in the tissue specimens and primary fibroblasts.Results The keloid fibroblasts (KFbs) showed significandy lower mRNA expression of p16 gene (0.64 ± 0.18 vs.1.92 ± 0.23,t =10.54,P＜ 0.05),but significantly higher mRNA expressions of 3 DNMTs (DNMT1:2.58 ± 0.23 vs.1.13 ± 0.21,t =11.22,P ＜ 0.05; DNMT3A:4.87 ± 0.46 vs.2.38 ± 0.32,t =10.81,P＜ 0.05; DNMT3B:1.57 ± 0.12 vs.0.57 ± 0.16,t =12.45,P＜ 0.05) compared with the normal fibmblasts (NFbs).The DNA methylation rate in the p16 gene promoter region was significantly increased in keloid tissue (1.81％ ± 0.46％) and KFbs (3.15％ ± 0.94％) compared with normal skin tissue (0.90％ ± 0.35％,F =14.23,P＜ 0.01) and NFbs (0.17％ ± 0.29％,F=37.62,P＜ 0.01).Conclusions The methylation and low expression of p16 gene in KFbs may be associated with the uncontrolled growth of keloid,and DNMTs may play a role in the pathogenesis of keloid.

Objective To study the proliferation,collagen production and related gene expression in keloids and normal skin fibroblast.Methods Isolated primary cells of keloid fibroblasts (KFb,n=12) and normal human dermal fibroblasts (NFb,n=12) were identified,the cell viability and proliferating potential and the cell cycle were detected,and the difference on the collagen synthesis between KFb and NFb were compared.The expression of cell cycle-associated genes such as p21,p16,and p27 was dectected by real-time fluorescent quantitative PCR.Results The phase contrast optical microscopy imaging showed that both KFb isolated from keloid tissues and NFb from normal skin tissues possessed classic and similar fibroblast morphology.But there was a significant difference between cell proliferation,Hyp [(2.30±0.10) μg/ml vs.(1.66±0.13) μg/ml,P＜0.05] and collagen levels [(17.19±0.75) μg/ml vs.(12.37±0.94) μg/ml,P＜0.05].Compared with NFb,KFb exhibited more percentage of G2/M phase cells [(5.90±0.62)％ vs.(16.94 ％±1.93)％,P＜0.05]and less percentage of G0/G1 phase cells [(90.24 ±2.27)％ vs.(75.65±1.92)％,P＜0.05].Cell cycle related genes p16,p21 and p27 were low expressed.Collagen type Ⅰ was highly expressed at mRNA levels in KFb than that in NFb [0.84±0.11,1.32±0.2,1.69±0.12,4.33±0.27 in KFb vs.1.43±0.13,2.56±0.26,2.89±0.37,1.40±0.12 in NFb,P＜0.05].Conclusions There are cell dysfunction and abnormal cellular dynamics in keloid fibroblasts.The formation of keloid likely involves aberrant interactions of some genes that affected its development at different extents.

Objective: To explore the relationship between expression of tumor necrosis factor-α( TNF-α) and electrophysiological heterogeneity in isolated heart tissues and isolated rat ventricular myocytes.The arrhythmogenic mechanisms of TNF-αwere further studied.Methods:Langendorff perfused heart tissues models were used to verify the arrhythmogenic effects of TNF-α.The monophasic action potentials( MAPs) of the endocardium and epicardium from the isolated heart tissues were recorded by elec-trophysiological experiments.The isolated rat ventricular myocytes were obtained by enzymatic dissociation.K+currents(Ito,IK1)were recorded by using whole cell patch clamp technique.Results: Compared to the control group, the difference in MAPD between endocardium and epicardium dramatically increased with TNF-α( P<0.05 ) .TNF-αcould cause MAP duration ( MAPD ) prolongation, and a single dose of TNF-αdifferentially affected the MAPs of endocardium and epicardium of isolated heart tissues.Compared to the control group,the K+currents(Ito,IK1)were dose-dependently decreased with TNF-αin rat ventricular myocytes(P<0.05).However, etanercept had no effects on the MAPD in the absence of TNF-α.Conclusion:TNF-α-induced heterogeneity of MAPD between the endo-cardium and epicardium may provide the substrate for the onset of ventricular arrhythmias during acute myocardial infarction.The effect might be associated with TNF-αcontribute to re-entrant ventricular arrhythmias which resulted from decreased K+currents(Ito,IK1).

AIM:To establish the GC fingerprint of Huoxiang Zhengqi Solution.METHODS:The volatile constituents of Huoxiang Zhengqi Solution were analyzed by capillary GC with FID detector using hydrodistillation and hexane extraction under n-hepladecane used as the reference substance.RESULTS:GC fingerprint of Huoxiang Zhengqi Solution,16 common peaks were established on the basis of systematic methodology after 10 batches of samples were tested.Variation in the relative retention time of 16 identified common peaks were within 0.5% range.CONCLUSION:The analytical method for Huoxiang Zhengqi Solution is precise and reliable.The research would be helpful to offer an effective pattern for quality control of Huoxiang Zhengqi Solution.

Objective To explore the role of the improved computer aided design of the digital three -dimensional titanium mesh in the repair of skull,to reduce the incidence of postoperative epidural hematoma and epidural fluid.Methods Retrospective analysis of 93 cases of skull repair using the three -dimensional titanium mesh was conducted.49 cases in the conventional group used the conventional three -dimensional titanium mesh. 44 cases in the observation group were given the improved three -dimensional titanium mesh.The clinical effect of the two groups was observed.Results In the conventional group,postoperative epidural hematoma occurred in 4 cases, 1 case of epidural fluid.All patients were fine in the observation group.The improved three -dimensional titanium mesh could reduce the incidence of epidural hematoma and effusion(χ2 =4.745,P =0.029).The conventional group and the observation group both had one case of infection after operation,there was no significant difference between the two groups(χ2 =0.001,P =1.000).Conclusion The improved three -dimensional titanium mesh can effective-ly reduce the incidence of postoperative complications and improve the curative effect of the operation.

Objective To study the effects of peroxisome proliferators-activated receptor gamma (PPA?r) on the biological characteristics of hepatic stellate cells (HSCs); Methods The activated HSCs were divided into four groups: control group, rosiglitazone group, GW9662 + rosiglitazone group and GW9662 group. The cell proliferation was determined with MTT colorimetric assay, The cell apoptosis was studied with flow cytometry. The expression of PPA?r, Type I collagen were detected by RT-PCR, Western blot and immunocytochemistry. Results MTT of HSCs in rosiglitazone group was (0. 49?0. 06) significantly lower than in control group ( 1. 00?0. 045 ), GW9662 group (0. 89?0. 043 ) , GW9662 + rosiglitazone group (0.78?0.049)(t = 15.59,14.68,8.07, P

Objective To investigate the radiobiological effects of VPA-BSANPs on C6 and U87 glioma cells in vitro.Methods C6 and U87 glioma cells were treated with different concentrations of VPA and VPA-BSANPs for 12 h and 24 h,and MTT assay was used to determine cell viability.C6 and U87 cells were treated with different concentrations of VPA and VPA-BSANPs conbined with X-ray irradiation (0,2,4,6,and 8 Gy),and colony formation assay was used to determine plating efficiency (PE).C6 and U87 glioma cells were treated with different concentrations of VPA and VPA-BSANPs for 12 h,followed by X-ray irradiation (0,4,and 8 Gy),and flow cytometry using Annexin V-FITC/PI staining was used to examine cell apoptosis.Western blot was used to evaluate the effects of VPA and VPA-BSANPs on radiation-induced apoptosis protein expression.One-way ANOVA was used for comparison of means with homogeneity of variance between multiple groups,and the t-test was used for comparison of means between two groups.Results Without irradiation,VPA and VPA-BSANPs had no significant inhibitory effects on the proliferation of C6 and U87 cells (P=0.328,0.920).The PE of cells treated with VPA-BSANPs combined with irradiation was significantly lower than that of cells treated with VPA combined with irradiation (P=0.000).In C6 and U87 cells,VPA-BSANPs combined with irradiation increased the expression of p53 and Bax (P =0.000,0.000 and P =0.010,0.002),but reduced the expression of Bcl-2 (P =0.008,0.000).Active caspase-3 fragments were only found in the cells treated with VPA-BSANPs combined with irradiation and VPA combined with irradiation,but were less in the former cells than in the latter cells (P=0.004).The active fragments of peroxisome proliferator-activated receptor were only found in the cells treated with VPA-BSANPs combined with irradiation.Conclusions VPA-BSANPs can increase the radiosensitivity of C6 and U87 glioma cells in vitro,possibly by promoting the apoptosis of tumor cells induced by radiation.