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Objective To investigate the expression of Vacuolar-H +-ATPase (V-ATPase) and P-glyeoprotein (P-gp) protein in colon carcinoma tissues,the correlation between the expression of V-ATPase and P-gp and their clinicopathological significance.Methods In samples from 80 cases of colon cancer,20 cases of colon adenoma and 10 cases of normal colonic mucosa tissues,the expression of V-ATPase and P-gp protein were detected by immunohistochemical method,their relationship was analyzed,the clinicopathological features and prognosis were evaluated.Results In colon cancer,V-ATPase and P-gp protein expression was 72％ and 80％,higher than that in colon adenomas (40％,35％),and in normal colon mucosa (20％,20％),the difference was statistically significant (P ＜ 0.05).There was a positive correlation between the expression of V-ATPase and that of P-gp (r =0.567,P ＜0.01).V-ATPase and P-gp protein expression in colon cancer was associated with TNM stage,lymph node metastasis and tumor differentiation (P ＜ 0.05).Patients with high V-ATPase expression had lower 5-year survival rate than those with low V-ATPase expression (P =0.023),and 5-year recurrence rate was higher than those with low expression (P =0.024).Conclusions The expression of V-ATPase is up-regulated in colon cancer,there is a positive correlation with colon cancer progress and metastasis,and high V-ATPase protein expression predicts poor prognosis.
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Objective To investigate human plasma glycoproteomie changes related to human immunodeficiency virus (HIV) infection,and to identify glycoproteins with potential anti-HIV activity or anti-HIV drug targets. Methods Plasma proteins with lower abundance were enriched through affinity purification to remove albumin and IgG in clinical samples (HIV-positive patient, n= 10, and healthy controls, n= 20). Proteins were separated by two-dimensional electrophoresis (2-DE) and stained by Pro-Q emerald glycoprotein stain kits. The 2-DE image was analyzed by ImageMaster software to find differential glycoproteins. Furthermore, the depleted HIV-positive and healthy control plasma proteins were digested by PNGase F. Glycoproteins were deglycoliszed, separated by 2-DE and analyzed by ImageMaster software. Differential glycoproteins were identified by liquid chromatography combined with high capacity ion trap mass spectrometry (HCT). Results The pretreatment of HIV-positive plasma prior to 2-DE could efficiently remove the high aboundant albumin and IgG in plasma and improve the detection of proteins with low-abundance. High revolution 2-DE gel images of glycoproteins from HIV positive and healthy control plasma samples were obtained. Glycoproteins were successfully deglycolized through PNGase F treatment. Thirteen differential glycoproteins were identified by liquid chromatography combined with mass spectrometry. These proteins included alphalantitrypsin precursor and serine/threonine-protein kinase N1. Conclusions Potential HIV infection related proteins,such as alphal-antitrypsin precursor are successfully identified. Our study may offer some help to understand the molecular mechanism of HIV infection and select new drug targets for preventing HIV infection.
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Many platelet function tests can be used for monitoring antiplatelet drug for the prevention and treatment of thrombotic diseases.These monitoring tests may be chosen based on different antiplatelet drugs which include aspirin,clopidogrel and GP Ⅱ b/Ⅲ a antagonist.There is difference of the drug resistance which is validated by different platelet function tests,and the relation with clinical adverse events should be farther clarifled.
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Objective To establish a new flow cytometric immuno-bead array assay (FCIBA)to detect several platelet-specific autoantibodies simultaneously in a single serum sample.Methods A series of beads of different red fluorescent intensity were used for coating with different anti-platelet membrane protein monoclonal antibodies(anti-GP Ⅱb/a,anti-GP Ⅰ a/Ⅱ a,anti-GP Ⅳ,anti-GP Ⅰ b/Ⅸ and anti-HLAABC)to detect five platelet-specific autoantibodies in serum. The beads captured platelet antigenautoantiboay complex.Subsequently,different platelet-specific autoantibodies in a patient serum can be detected simultaneously by the flow cytometry.In addition,we evaluated the new FCIBA,and compared its results with modified antigen capture ELISA method(MACE).The new FCIBA was used to detect five platelet-specific autoantibodies in serums of autoimmune thrombocytopenic purpura(AITP)and nonautoimmune thromboeytopenic purpura patients.Results The new FCIBA can be used to detect five plateletspecific autoantibodies simultaneously(Anti-GP Ⅱ b/Ⅲ a,Anti-GP Ⅰ a/Ⅱ a,Anti-GP Ⅳ,Anti-GP Ⅰ b/Ⅸand Anti-HLA-ABC).The coefficient of variation(CV)of intra-repetition are 4.82%,6.09%,5.04%,5.73%and 5.30%,respectively.The dilution test results are in good logarithm linearity which are 0.997 2,0.996 6,0.998 8,0.996 5 and 0.998 2,respectively. The resuhs of the new FCIBA are highly correlated with those with MACE method,and the coefficient correlation were 0.928 9,0.922 4,0.889 4,0.910 0 and 0.913 4,respectively(P＜0.01).51.69%samples of AITP patients show positive for platelet-specific autoantibodies as detected by the new FCIBA.Among the AITP patients,the positivity of specific autoantibodies for anti-GPⅡb/ Ⅲ a,anti-GP Ⅰ a/Ⅱ a,anti-GP Ⅳ,anti-GP Ⅰb/Ⅸ and anti-HLA-ABC were 40.82%,24.45%,19.39%,32.65%and 17.35%.Among the 40 non-autoimmune thrombocytopenic purpura patients,none of platelet-specific autoantibodies in serum samples can be detected.Conclusion A new FCIBA is established successfully to detect five platelet-specific autoantibodies coefficient correlation.