The aim of the present study was to work on the efficiency of differential diagnosis of native hapten-gel diffusion assay (NH-GD) on the background of vaccination with S2 or Rev.1.The conditions of NH-GD assay was firstly optimized,its sensitivity,specificity,repeatability and ability of differential diagnosis were determined respectively,and its test result was compared with that of fluorescence polarization assay (FPA).The results showed NH-GD assay with good specificity and repeatability could differentiate Brucella-infected antibody from vaccinated antibody after vaccination with S2 or 122 days after vaccination with Rev.1.And the result of NH-GD assay was highly consistent with that of FPA,which was simple to operate and needed a few simple equipment.Therefore,NH-GD assay was a good method for sheep brucellosis surveillance in China and especially suitable for application in grass-roots areas.

Objective To evaluate the clinical and pathological characteristics and risk factors of the prognosis of Cardia cancer.Methods Clinical data of 52 cases of cardiac cancer patients who were treated by surgery and follow-up data were collected from June 2009 to August 2011.Factors influencing the prognosis of the patients were analyzed by univariate and multivariate analysis.Results The survival rates in 1,2,3years were 73％,54％,46％.Single factor analysis showed that there was a significant correlation between T stage,N stage,TNM stage,classification Siewert,MSLN and venous invasion.Stepwise Cox proportional hazard model showed that T stage,N stage and MSLN were independent risk factors.Peritoneal and liver were the main way of recurrence and metastasis.Peritoneal metastasis was related to T stage and lymph node invasion,while the liver metastasis was related to MSLN.The survival curves of patients with Kaplan-Meier showed that the survival period of patients with liver metastasis and peritoneal metastasis was significantly shortened.Conclusion Standardization of lymph node cleaning is a key factor affecting prognosis,D2 surgery should be an operation standard in cardia cancer.

Objective To investigate the expression of hypoxia inducible factor-1α(HIF-1α) and vascular endothelial growth factor C(VEGF-C) in colon carcinoma,and their relation with clinical pathological parameters and prognosis. Methods Immunohistochemistry was used to detect the expression of HIF-1α and VEGF-C in 65 tissues of colon carcinoma. Results In all cases,the expression of HIF-1α had a positive correlation with the expression of VEGF-C(r= 0.638,P＜0.01).The expressions of HIF-1α and VEGF-C had significant correlation with clinical stage and nodal metastasis(P＜0.05) ,but had no relationship with sex,age and tumor size(P＞0.05). Conclusion The expression of HIF-1α and VEGF-C was correlated with nodal metastasis, tumorigenesis and prognosis of colon/carcinoma.

In this paper, chemical derivatization-high performance liquid chromatography was used to determine the potential genotoxic impurities chloroacetyl chloride and chloroacetic acid, respectively, in the raw material of azintamide.Derivatization was carried out using 2-nitrophenylhydrazine followed by the determination.Separation was performed on a Thermo Syncronis C18 column (250 mm × 4.6 mm, 5 μm), with mobile phase consisting of 0.1% phosphoric acid in water (A) and acetonitrile(B) by gradient elution, at a flow rate of 1 mL/min.The column temperature was 40 °C and the detection wavelength was 226 nm.The blank solvent, derivatization reagent, and azintamide did not interfere with the peak of the test substance, and the target component was well separated from the others.For impurities chloroacetyl chloride and chloroacetic acid, the limits of detection (LOD) were 7.5 ng/mL and 15 ng/mL respectively. There was a good linear relationship between the integral area and the concentration in the range of 30-300 ng/mL.The sample recovery rate was in the range of 87.37% ~ 109.75%.The two methods established in this study have good specificity, good precision, high sensitivity and simple operation, which can be used for the trace determination of potential genotoxic impurities chloroacetyl chloride and chloroacetic acid in the raw material of azintamide.

OBJECTIVE：To establish a method for the content determination of potential genotoxic impurity maleic hydrazide in azintamide raw material. METHODS ：HPLC-FLD method was adopted. The determination was performed on Thermo Syncronis C18 column with mobile phase consisted of 0.2 mol/L acetic acid-methanol （gradient elution ）. The column temperature was set at 30 ℃，the excitation wavelength was 315 nm and emission wavelength was 389 nm. The flow rate was 1 mL/min，and the sample size was 20 μL. RESULTS：The blank solvent and azintamide did not interfere with the determination of maleic hydrazide. The linear range of maleic hydrazide was 19.5-300 ng/mL（r＝0.999 9）. The limit of detection was 4.5 ng/mL and the limit of quantification was 19.5 ng/mL. The recovery ranged from 98.79％ to 103.76％（RSDs were lower than 3.00％，n＝9）. RSDs of precision and stability （24 h）tests were no more than 5.63％，and those of durability tests were less than 2.00％（n＝6）. Maleic hydrazide was not detected in 3 batches of azinamide raw material. CONCLUSIONS ：The method is specific ，sensitive and accurate. It can be used for the trace determination of maleic hydrazide in azintamide or other matrix.