Objective: To establish an HPLC method for the determination of three bacteriostatic agents ( methylparaben, eth-ylparaben, propylparaben) in taurine eye drops. Methods:The HPLC method was conducted on a Phenomenex Gemini C18 column ( 250 mm × 4. 6 mm, 5 μm ). The mobile phase was 1％ acetic acid-methanol(40 :60). The flow rate was 1. 0 ml·min-1 and the de-tection wavelength was 254 nm with the injection volume of 20 μl and the column temperature of 25℃. Results:Methylparaben, eth-ylparaben and propylparaben showed good linear relationship (r>0. 999)within the range of 1. 00-19. 94μg·ml-1,2. 01-20. 08μg· ml-1 and 0. 21-10. 46 μg·ml-1,respectively. The average recoveries were 99. 20％-99. 90％, and the RSDs were 1. 34％-1. 54％(n=9). Conclusion:The method is accurate, reproducible and stable without interference, which can be used for the determination of three bacteriostatic agents in taurine eye drops.

Objective Islet transplantation has been an effective method for diabetes mellitus. The quality of donor pancreas is important for successful islet isolation. In this study, we evaluated expanded criteria donor usability based on the warm ischemic time, fatty pancreas and perfusion injury. Methods The marginal pancreases include those from cardiac death donor, fatty pancreas and edema pancreas from perfusion injury. Islets were isolated and purified using a modified University of Minnesota method. Islet yield and purity was determined by Dithizone (DTZ) staining and microscopic examination. Islet viability was assessed by AO/EB staining, and islet function was assessed by static glucose stimulation test. Results In the cardiac death donor group, the islet quality, viability, and in vitro function were similar when the warm ischemic time within 15 min. The quality and viability was decreased when the warm ischemic time beyond 30 min, but the function remained well. With 45 min warm ischemic time, insulin release index was decreased significantly. The islet quality, viability, and in vitro function from severe obesity group and severe edema group were decreased obviously. Conclusion Donor factors play a vital role in pancreas transplant outcomes. We concluded that pancreas severe obesity, severe edema and pancreas from cardiac donors (warm ischemic time ＞30 min) are unsuitable for islet isolation.

Objective To explore the application value of Golgi protein-73 (GP73)and AFP in single and combining form in the diagnosis of primary hepatocelluar carcinoma (PHC).Methods Eighty PHC,65 liver cirrhosis,54 chronic hepatitis patients and 50 controls were selected in the First Afiliated Hospital in Xinjiang Medical University from May to September in 2011,GP73 was detected by ELISA and AFP was measured by clinical chemiluminescence.The sensitivity and specificity of each parameter in single and combining form were evaluated.Results Serum GP73 in PHC group 282.0(163.6-366.7) μg/L,liver cirrhosis group 211.8(107.5-295.7) μg/L,chronic hepatitis group 100.3(61.8-191.3) μg/L and control group 58.3(43.4-83.6) μg/L was tested by Kruskal-Wallis(H =106.6,P ＜0.01).GP73 in PHC group was further compared with liver cirrhosis group,chronic hepatitis group and control group using MannWhitney test,significance was found,(U was 1796.0,826.5,154.0,respectively,all P ＜0.01).In the single form,the sensitivity of GP73 [82.5％ (66/80)] was higher than AFP [66.3％ (53/80),x2 =4.65,P ＜0.05],but the specificity of GP73 [63.3％ (107/169)] was lower than AFP [88.7％ (150/169),x2 =28.91,P ＜0.05].There were 27 AFP negative cases in PHC group,but 22 of them were GP73 positive,making the positive rate of GP73 [81.5％ (22/27)] in PHC patients with AFP negative.There were 14 GP73 negative cases of in PHC group,but 9 of them were AFP positive,making the positive rate of AFP [64.3％ (9/14)] in PHC patients with GP73 negative.In series diagnostic test,the specificity of combining form [95.9％ (162/169)] was higher than AFP [88.7 ％ (150/169),x2 =6.00,P ＜ 0.05] ; in parallel diagnostic test,the sensitivity of combining form [93.8％ (75/80)] was higher than GP73 [82.5％(66/80),x2 =4.84,P ＜0.05].In PHC group,52 patients with HBV infection,10 patients with HCV infection and 18 patients without virus infection,GP73 was 309.5 (170.5-370.5) μg/L,351.0 (274.7-397.9) μg/L and 210.1 (156.8-306.7) μg/L,respectively,no significance was found (H =4.0,P ＞0.05).Conclusion GP73 and AFP have a complementary feature of sensitivity and specificity in the early diagnosis of PHC,some PHC cases with AFP negative can be avoided missing efficiently by parallel diagnostic test.

it was 4% (3/75). GP73-positive rate in HCC was higher than that of the normal liver tissues (χ2=73.60, P<0.05). sCLU-positive rate in HCC was also higher than that of the normal liver tissues (χ2=207.94, P<0.05). GP73 expression was positively correlated with sCLU expression in HCC (r=0.405, P<0.05). GP73 and sCLU were associated with clinicopathological features including tumor differentiation, TNM stage and vascular invasion (P<0.05); GP73 and sCLU had no correlation with the patient’s gender, age, HBsAg, cirrhosis, AFP value, portal vein thrombosis and tumor numbers (P>0.05). GP73 was associated with survival but not sCLU. Conclusion:GP73 and sCLU have higher positive rates in HCC and GP73 is positively correlated with sCLU. The expression of GP73 and sCLU are probably closely related with the invasion of HCC, which can help evaluate the prognosis of the patients.

Objective: To obtain polycystin-1 intracellular region. Methods: cDNA of polycystin-1 intracellular region was generated by PCR and then cloned into pProEX Hta, which was prokaryotic expression vector. After verified by sequencing, the recombinant was transformed into E.coli host to express and purify the fusion protein by affinity chromatography. Results: 660 bp cDNA of polycystin-1 intracellular region and 2.6×104 fusion protein were obtained. Conclusion: The fusion protein containing polycystin-1 intracellular region is obtained and is helpful for preparing anti-polycystin-1 monoclonal antibody.

Objective: To obtain polycystin-1 intracellular region. Methods: cDNA of polycystin-1 intracellular region was generated by PCR and then cloned into pProEX Hta, which was prokaryotic expression vector. After verified by sequencing, the recombinant was transformed into E.coli host to express and purify the fusion protein by affinity chromatography. Results: 660 bp cDNA of polycystin-1 intracellular region and 2.6×104 fusion protein were obtained. Conclusion: The fusion protein containing polycystin-1 intracellular region is obtained and is helpful for preparing anti-polycystin-1 monoclonal antibody.

Nasal spray can treat local diseases such as rhinitis, it also plays an important role in the treatment of systemic diseases including vaccine immunity. As a drug-device combination product, spray pattern is often used as the quality indicator of nasal spray to ensure its quality, plume geometry can not only be combined with the spray pattern to evaluate the performance of the nasal spray, but also can predict the deposition of the nasal spray in the nasal cavity. This article systematically reviews the definition and measurement methods of the spray pattern and plume geometry of nasal spray and their correlation with nasal deposition behavior. The measurement parameters of spray pattern and plume geometry are defined. The influence of formulation, device and trigger parameters on spray pattern and plume geometry is clarified. The correlation between various parameters and nasal deposition is analyzed. The measurement parameters are classified according to the size and shape of the spray. Plume angle is closely related to the deposition of drugs in the nasal cavity. Jet-like plume with a smaller plume angle can increase the navigation ability of the nasal spray in the curved anatomical structure of the nasal cavity, which is conducive to increase drug deposition. This makes it possible to increase deposition of the nasal spray in the nasal cavity via appropriately increasing viscosity and thixotropy of the nasal spray formulation. This review provides the theoretical basis for the high quality nasal spray product development.

Objective To study the effect of bromocriptine(BRC) on the activation of T lymphocyte stimulated by phytohemagglutinin(PHA).Methods After CD4+ T cell line Jurkat E6-1 cells were stimulated by PHA,prolactin(PRL) and BRC,respectively,the expression of linker for activation of T cells(LAT) and zeta-chain T cell receptor associated protein kinase 70 000(ZAP-70) mRNA of T lymphocytes were checked by RT-PCR.The expression of PRL mRNA of T lymphocytes was detected by Real time PCR.The expression of CD25(cluster of differentiation) as a marker of early activation on the surface of T lymphocytes was detected by flow cytometry,and the activation of nuclear factor-?B(NF-?B) was detected by luciferase reporter system.Results 1.BRC inhibited the expression of ZAP-70 as the common signal molecules both in the T lymphocyte activation pathway and PRL-prolactin-prolactin receptor(PRLR) signal transduction pathway,and decreased the expression of PRL mRNA produced by activation T lymphocytes.2.BRC enhanced the expression of LAT mRNA as another important signal molecular on the T lymphocytes and CD25 on the surface of the T lymphocytes.3.The activation of NF-?B of T lymphocytes was decreased.Conclusions BRC might inhibit the activation of T lymphocytes by inhibiting the expression of ZAP-70,the common signal molecules between T lymphocytes activation and PRL-PRL pathway,and PRL mRNA,the like-T lymphocyte growth factor.

Objective To explore the establishment of a rat model of acute radiation-induced liver injury and sig-nificance of the dynamic changes of TGF-β1 expression.Methods Forty healthy 6-week old male SD rats were randomly divided into model group (n=30) and control group (n=10).The right liver of rats in the model group was given a single dose of 25 Gy 6 MV X-ray irradiation.Histopathological examination using HE staining and transmission electron microsco-py were conducted to observe the liver pathological changes in rats at 3, 5, and 10 days after irradiation, serum TGF-β1 was detected, and relevant indicators of liver function ( ALT, AST, ALP) were determined.Statistical analysis was per-formed using SPSS 17.0 software.Results At 3, 5 and 10 days after irradiation, early pathological changes in the liver cells were observed by electron microscopy, the expression of TGF-β1 was gradually increased with the time prolongation, and significant differences were found between the model group and the control group at different time points (P<0.05). The light microscopic observation of liver tissues did not show significant differences between the control group and model group.The liver ALT, AST, ALP at different time points did not show significant differences between the two groups ( P>0.05).Conclusion Electron microscopy can be used to evaluate the early changes of radiation-induced liver injury, pri-or to the alterations visible by routine light microscopy.TGF-β1 can be used to predict the degree of radiation-induced liver injury, and may be used as a sensitive serum cytokine in predicting the degree of radiation-induced acute liver injury.

OBJECTIVETo investigate the expression profiles of serum Golgi protein-73 (GP73) in liver cirrhosis and primary hepatic carcinoma (PHC) and determine its clinical value for differential diagnosis.

METHODSSerum protein expressions of GP73 and alpha-fetoprotein (AFP) were detected by enzyme-linked immunosorbent assay and chemiluminescence assay, respectively, in patients with PHC (n=80), liver cirrhosis (n=65), and healthy controls (n=50). Inter-group changes were assessed by Kruskal-Wallis test, and significance of these differences was assessed by Mann-Whitney test. A receiver operating characteristic (ROC) curve was plotted to evaluate the diagnostic efficiency and determine the cut-off values for GP73 and AFP. Sensitivity and specificity were compared by the Chi-squared test. Correlation between serum GP73 expression and clinical parameters was determined by Spearman's rank correlation analysis.

RESULTSThe PHC group showed significantly higher serum GP73 (282.0 mug/L) than the liver cirrhosis group (211.8 mug/L) and control group (58.3 mug/L) (H = 93.30, P less than 0.01). For differential diagnosis of PHC and liver cirrhosis, the cut-off value was 318.1 mug/L for GP73 and 13.4 mug/L for AFP. Sensitivity of GP73 was lower than AFP (45% (36/80) vs. 65% (52/80); X2 = 8.02, P less than 0.05). Specificity of GP73 was lower than AFP but no significance was found (83.1% (54/65) vs. 87.7% (57/65); X2=0.27, P more than 0.05). The areas under the ROC curves were not significantly different between GP73 and AFP (0.65 (95% confidence interval (CI): 0.54~0.72) vs. 0.75 (95% CI: 0.67~0.83); Z = 1.88, P more than 0.05). The area under the ROC curves increased but not significantly (0.80 (95% CI: 0.73~0.88) vs. 0.75 (95% CI: 0.67~0.83); Z=2.61, P more than 0.05). Serum GP73 was correlated with liver cirrhosis (r=0.27), vascular invasion (r=0.29), and TNM staging (r=0.27) (all P less than 0.05), but not with sex (r=0.13), age (r=0.10), enhanced AFP (> 13.4 mug/L; r=0.03), tumor size (r=0.18), or distant metastasis (r=0.04), all P less than 0.05.

CONCLUSIONSerum GP73 and AFP have comparable diagnostic efficiency, but the sensitivity of AFP is superior for differential diagnosis of liver cirrhosis and primary hepatic carcinoma. Elevated serum GP73 may be correlated with liver tumor load and aggressiveness.

Adult ; Aged ; Carcinoma, Hepatocellular ; diagnosis ; Case-Control Studies ; Diagnosis, Differential ; Female ; Humans ; Liver Cirrhosis ; diagnosis ; Liver Neoplasms ; diagnosis ; Male ; Membrane Proteins ; blood ; Middle Aged ; Sensitivity and Specificity ; Transcriptome ; alpha-Fetoproteins ; metabolism