Background Experimental study showed that heparanase (HPA)is overexpressed in choroidal neovascularization,suggesting that it may play a role in the pathogenesis of angiogenesis.To certify HPA inhibitor suppress the formation and development of new blood vessel has an important significance for the treatment of choroidal neovascularization.Objective The aim of this study was to investigate the effect of HPA inhibitor on the proliferation of human umbilical vein vascular endothelial cell (UVEC) and the expression of HPA.Methods Hunan UVEC was primarily cultured and passaged and the third generation cells were used in the experiment.Phosphomannopentaose sulfate (PI-88) solution,a HPA inhibitor,was prepared with endothelial cell medium and the end concentrations were 400.00,200.00,100.00,50.00,25.00,12.50,6.25 mg/L respectively.The cells were treated with PI-88 solutions for 24,48 and 72 hours.The growth and proliferation of human UVEC were analyzed using MTT colorimetric assay at absorbance 570 nm.The expression of HPA in the cells was detected by immunochemistry in 48 hours after addition of PI-88.Results Cultured human UVEC showed the fusiform and polygon in shape.The A570 values of human CVEC were significantly different among various concentrations of PI-88 groups (F=2.721,P=0.053 ) and different action time (F=9.656,P =0.002).When PI-88 was administered for 24 hours,the A570 values of human UVEC were insignificantly altered in comparison with the one without PI-88 culture group (P＞0.05 ).However,in 72 hours after experiment,the A570 values were significantly declined as the PI-88 concentration was ＞50.00 mg/L ( P＜0.05 ).When PI-88 was administered for 48 and 72 hours,the A570 values of human UVEC were significantly higher than those of 24 hours in ＜50.00 mg/L groups (P＜0.01 ),but no statistical differences were seen in ＞100.00 mg/L groups among various time points (P＞0.05 ).HPA was intently expressed in the cytoplasm of human UVEC.However,at 48 hours after addition of ＞25.00 mg/L PI-88,the HPA expression was obviously weaker.Conclusions PI-88 can suppress the growth and proliferation of human UVEC at the dose-and time-dependent manner by downregulating the expression of HPA in the cells.

Objective To develop a high performance liquid chromatography (HPLC) for the determination of fourteen dyes in hair dying formulations.Methods Dyes were separated on a amide bonded C_(16) silica column (250 mm?4.6 mm,5?m),the target analytes were ion-paired prior to HPLC analysis with elution employing methanol-0.025 mol/L phosphate buffer (pH=6.0) containing 1 g/L 1-heptanesulfonic acid sodium salt as mobile phase and detection by a photodiode array detector with the detection wavelength of 230 nm and 280 nm.The flow rate was 1.0 ml/min and the column temperature was 25℃.Results The linear range were 10-500 mg/L,the detection limits were 0.3-2.0 mg/L,the coeffieients of variation were less than 10%(except 2-amino-6-chloro-4-nitrophenol at low concentration) and the recovery rates were 71.3%-118.5%.Conclusion The methods of high performance liquid chromatography introduced in this paper is simple,rapid,accurate and is applicable to the analysis of various hair dyes.

OBJECTIVETo investigate clinical effects of arthroscopy in the treatment of medial meniscal cyst.

METHODSFrom June 2011 to January 2013, 7 patients with medial meniscal cyst were treated with arthroscopy. There were 3 males and 4 females,ranging in age from 27 to 63 years old,with a mean age of (43.93±2.10) years old. The cysts have been discovered for 3 to 30 months,with a mean time of (10.6±1.3) months. All the patients complained of knee pain,especially in the medial joint gap. The Pisani sign, Caklin sign and medial McMurray sign were all positive. Preoperative MRI examination confirmed the diagnosis. Lysholm score changes and clinical efficacy were observed through a six-month follow-up.

RESULTSThe postoperative Lysholm scores were all significantly higher than the preoperative scores. According to Sarimo standard, 6 patients got an excellent result, and 1 good.

CONCLUSIONArthroscopic treatment of medial meniscal cyst has replaced the traditional method, which could retain the normal meniscus as much as possible and repair the meniscus injury simultaneously, as well as get a good curative effect and a good recovery of knee function. This method is worthy of clinical application.

Adult ; Arthroscopy ; methods ; Cysts ; physiopathology ; surgery ; Female ; Humans ; Male ; Menisci, Tibial ; physiopathology ; surgery ; Middle Aged

Objective To investigate the clinical application value of the laparoscopic intraoperative cholan-giography in laparoscopic cholecystectomy,and summarize the experience.Methods The clinical data of 169 cases of laparoscopic cholecystectomy intraoperative cholangiography were analyzed retrospectively.Results 169 patients were successfully completed,131 cases recovered well and no complications occurred after operation.29 patients showed hyperamylasemia,of which 3 patients had intractable hyperamylasemia,8 patients complicated with secondary acute pancreatitis,with fasting,gastrointestinal decompression,enzyme inhibition(plus somatostatin)and acid,analgesic, anti infection,rehydration treatment after remission.Conclusion Laparoscopic cholecystectomy intraoperative cholan-giography is a safe and reliable diagnostic technique,on the occurrence of biliary residual stones in prevention of post-operative prevention and timely detection of bile duct injury during operation and improves the success rate of repair of bile duct injury has important clinical value;control adaptation of intraoperative cholangiography has important clinical significance of reasonable application license.

Objective To study the damage effect of oxidized low density lipoprotein (OX-LDL)on the endothelial cells and its influence in the expressions of Toll like receptor 4 (TLR-4)and epithelial cell adhesion molecule (EpCAM).Methods The human endothelial cells ECV-304 were cultured in vitro and treated by different concentrations of OX-LDL for 24 h,and the cell vitality was measured by Taipan blue rejection test.The reactive oxygen species (ROS)level,mitochondrial membrane potential (MMP),cell cycle and apoptosis were detected by flow cytometry (FCM).The expression of TLR-4 and EpCAM were also analyzed by FCM.Results Compared with control group,the ROS levels in ECV-304 cells in 25,50,100,and 200 mg·L-1 OX-LDL were increased, but the cell vitalities and MMP were decreased (P < 0.05).Compared with control group,the percentages of S phase cells in 25 and 50 mg·L-1 OX-LDL groups were increased,the G0 G1 phase cells were slightly reduced,and the SubG1 phase cells (apoptotic cells)were increased with the increasing of OX-LDL concentration.Compared with control group,the TLR-4 and EpCAM expression levels in ECV-304 cells in 50 mg·L-1 OX-LDL group at 24 h were significantly increased (P < 0.05 ). Conclusion OX-LDL could increase the ROS level and induce the apoptosis in vascular endothelial cells,while increase the cell vitality and MMP.During the damage process,the expresion levels of TLR-4 and EpCAM are up-regulated.

Using the high efficient copper-adsorbing yeast strain Y17 as absorbing material, the major affect factors including pH, original concentration of Cu2+, cell biomass, adsorption time and temperature were examined, and then the absorbing sites of the Y17 was determined. The results showed that the solution pH was the most dominate factor which affected the biosorption of Cu2+, the other affecting factors were the ini- tial concentration of Cu2+, the cell biomass added, and adsorption time, respectively; the temperature had lit- tle effect on the rate of biosorption. The orthogonal experiment showed that the optimal absorption condition was as follow: the solution pH was 5.0, the absorption time was 40 min, the cell biomass of Y17 added was 5.0 g/L, and the concentration of Cu2+ was 8 mmol/L; the highest adsorbing rate was up to 82.7% at this condition. Based on the results of different pretreatments and the desorption of Cu2+, the cell wall of Y17 was identified as the main place occurring boisorption process, and the -NH2 group, -COOH group on the surface of the yeast cells played an important role on the boisorption process.

Objective To investigate the effect of up-regulation of tetraspanin-5 (Tspan-5) expression on the metastatic ability of colorectal cancer (CRC) cells. Methods The coding sequence of Tspan-5 mRNA was amplified from total RNA of LoVo cells by RT-PCR and cloned into pEGFP-C1 to construct Tspan-5/pEGFP-C1,a eukaryotic expression vector. The expression of Tspan-5 in LST-R1 cells was up-regulated by gene tranfection. Western blotting was used to detect the expression level of Tspan-5/EGFP fusion protein,and laser scanning confocal microscopy (LSCM) was used to observe the distribution of Tspan-5/EGFP in LST-R1 cells. The changes in adhesion,migration and invasion ability of LST-R1 cells were determined by adhesion,migration and invasion experiment in vitro,respectively. Liver metastasis model of nude mice was used to test the changes in liver metastasis ability of LST-R1 cells in vivo. Results The Tspan-5/EGFP fusion protein could be detected by Western blotting in Tspan-5/pEGFP-C1 transfectants. LSCM showed that Tspan-5/EGFP protein located in cell membrane. Adhesion and migration assays showed that up-regulation of Tspan-5 expression level significantly promoted the adhesion and migration ability of LST-R1 cells on a series of extracellular matrix (ECM) components,including collagen IV,FN,LN and VN (P

Objective To analyze the chemical components of Fructus Aurantii (FA) and Fructus Aurantii Immaturus (FAI).Methods HPLC-ESI-MS with Surveg mass spectrometer was used in the study.Chromatographic column: Symmetry Shield TM RP_ 18 (150 mm?3.9 mm, 5 ?m) (Waters, Milford, MA, USA); mobile phase: (A) water (0.6% HAc, pH=2.5), (B) methanol. Gradient elutions: 20%- 40% B (0-48 min); 40% B (48-54 min); 40%-55% B (54-60 min); 55%-95% B (60-75 min); 95% B (75-85 min); 95%-20% B (85-90 min).Flow rate and wavelength were 0.7 mL/min and 283 nm at room temperature, respectively.Results Four kinds of flavonoids were identified as naringin, neohesperidin, naringenin, and hesperidin, synephrine was also identified in FA and FAI. Furthermore, the contents of them were determined individually.The results showed that the chemical constituents in FA and FAI were the same but the contents were different.Conclusion HPLC-ESI-MS method can be efficiently used to study FA and FAI.

AIM: To establish the quality standard for Zhike Pingchuan Capsule (Radix Panacis Quinquefolii, Semen Pruni Armeniacae, Radix Scutellariae, Radix Glycyrrhizae, Bulbus Fritillariae Cirrhosae, etc.). METHODS: Radix Panacis Quinquefolii, Radix Glycyrrhizae, Bulbus Fritillariae Cirrhosae in Zhike Pingchuan Capsule were identified by TLC and ginsenoside Rb_1 was determined by HPLC. The analysis was carried out on C_~18 column by HPLC. The mobile phase was CH_3CN-H_2O(34∶66). The flow rate was 1.2 mL?min~-1 and the detection wavelength was at 203 nm. The column temperature was at 40.0 ℃ and sensitivity was 0.02 AUFS. RESULTS:The average recovery was 97.20% and RSD was 2.25% and the linear range of ginsenoside Rb_1 was in 0.612-~6.120 ?g. CONCLUSION:This method is simple, rapid with a good reproducibility. This method can be used for the quality control of Zhike Pingchuan Capsule.

In order to identify virulence factors of the pathogen, the outer membrane proteins of virulent and avirulent strains of Riemerella anatipestifer were compared by a proteome analysis. Three protein spots differentially expressed between the two strains were observed by 2-DE gels, and were further analyzed using in gel tryptic digestion and peptide mass fingerprinting. Three proteins were identified. W1 was Hsp20, W2 and W3 were transposon. Although the exact role of these proteins has not been characterized, the exclusive expression in virulent strain may indicate that they play an important role in the pathogenesis of Riemerella anatipestifer infection. Although only two virulence factors identified, it opens a path to the further analysis of virulence factors of Riemerella anatipestifer.