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Naoxintong capsule has beneficial effects for activating blood circulation, dispersing blood stasis and dredging collateral. It is widely used in the treatment of coronary heart disease, angina pectoris, stroke and cardiovascular disease. However, the pharmacodynamic basis and possible mechanism of its preventive effects are not clear. In this study, 10 male and 10 female C57BL/6 mice were used, and were randomly divided into the control group (saline) and Naoxintong group. Adaptively fed for 7 days in common conditions, mice were given Naoxintong capsule or saline for 3 days via intragastric administration. Serum was collected from 6 mice in each group 1 h after the last administration. Serum proteins were prepared to do two-dimensional gel electrophoresis. Then image analysis and mass spectrometry detection were carried out to screen and identify the differentially expressed proteins and make bioinformatics analysis. It was found that 24 differentially expressed proteins between Naoxintong group and control group. Compared with the control group, 12 proteins were increased, and 12 were decreased. The proteins were involved in apoptosis signal pathway and vascular endothelial growth factor signal transduction pathway, in which vasohibin-1 is a negative feedback regulation factor in angiogenesis. Western blot showed that the expression of vasohibin-1 in Naoxintong group was reduced, which is consistent with the result in two-dimensional electrophoresis. Serum proteins expression is different between Naoxintong and control groups. The targets of these differentially expressed proteins include endothelial cells, inflammatory cells and platelets. The changes on proteins showed that Naoxintong capsule may ameliorate coronary heart disease and ischemic cerebrovascular disease, and provide potential biological markers to prevent ischemic disease.
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Objective To investigate the changes of proteomics in serum of Sprague-Dawley(SD) rats after accumulated irradiation with 137Cs γ-rays.Methods A total of thirty mature SD rats were randomly divided into three groups:0.2 Gy group,2 Gy group and healthy control group.Rats were irradiated at a dose rate of 0.336 mGy/min for 10 d and 20 d continuously.Isobaric tags for relative and absolute quantitation (iTRAQ) was used to analyze the different protein expression in serum of irradiated rats.Gene Ontology,KEGG pathway and protein-protein interaction network analysis were conducted using softwares.Results In total,363 protein spots were identified.Twenty nine proteins were differentially expressed in both groups compared with control,of which 10 proteins were up-regulated and 19 proteins were down-regulated.Based on the information of GO categories,these differentially expressed proteins were mainly located in the cytoplasm and membrane concerning the function of binding and catalytic activity.Analysis with the PAJEK software demonstrated that 16 differentially expressed proteins could form a complicated interaction network where glutathione S-transferase P1 (GSTP1),phosphoglycerate kinase 1 (PGK1) and protein disulfide-isomerase (PDI) might be key nodes.Conclusions Accumulated irradiation can induce differentially expressed proteins in serum of irradiated rats.Analysis on functional roles of the screened proteins GSTP1,PGK1 and PDI may provide insight into further mechanistic investigations and underlying molecular biomarkers induced by accumulated irradiation.
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To screen for serum biomarkers for lung squamous carcinoma, two-dimensional gel electrophoresis (2-DE) was performed to separate serum proteins from healthy individuals and stage 1 lung squamous carcinoma(LSC) patients, respectively. PDquest software was used to analyze 2-DE images, and the differential serum protein spots between the healthy individuals and LSC patients were identified by ESI-Q-TOF MS/MS. Then Western blot and immunohistochemistry were used to detect the expression levels of haptoglobin-2(HP-2), one of the differential proteins, in the sera and tumor tissues in the patients with LSC, respectively. 2-DE maps of serum proteins from healthy individuals and stage 1 LSC patients were established. Ten differential serum protein spots were detected, four proteins of which were identified by MS/MS. Western blot showed that the serum level of HP-2 in the LSC patients was significantly higher than that in healthy individuals, but was not associated with LSC staging. Immunohistochemistry showed that the expression level of HP-2 in the LSC tissues was significantly higher than that in the normal bronchial epithelial tissues adjacent to tumors. The results indicated that serum HP-2 protein is a candidate biomarker for LSC, and might be useful for diagnosis of LSC. Up-regulation of HP-2 in the LSC tissues may contribute to the high serum level of HP-2 in the patients.
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Objective:To improve the early diagnosis of hepatocellular carcinoma(HCC),a reversed-phase high-performance liquid chromatography(RP-HPLC)was employed to identify the differential proteins of serum related to HCC. Methods:The sera from 20 healthy volunteers and 8 HCC patients were collected. After the depletion of six serum proteins of the highest abundance with the Multiple Affinity Removal System (MARS) technologies from Agilent,the sera were subjected to RP-HPLC.The chromatographic conditions were: analytical column:ZORBAX 300SB-C18 (250 mm?4.6 mm,5 ?m);column temperature,25℃;the elution was performed with linear gradient elution of A:0.1% trifluoroacetic acid water solution and B:0.09% trifluoroacetic acid ACN solution at a flow rate of 0.75 ml/min. The detection wavelength was 280 nm. The time procedure was:1%B was held 3 min,then the gradient ran from 1 to 100% B in 100 min, then was held for another 10 min,at last,the gradient ran from 100 to 1%B in 5 min and was held for another 5 min. Results:There was a group of differential proteins of strong hydrophobicity in the serum between healthy persons and patients. Conclusion:This method can be useful in detecting protein expression alteration resulting from the carcinogenesis and development of HCC,and newly discovered biomarkers might be an aid in the early diagnosis and therapy of HCC.
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Objective: To establish a two-dimensional polyacrylamide gel electrophoresis(2-DE) technique for serum proteomics.Methods: Various conditions of serum proteomic 2-DE were adjusted and optimized.Results: A steady 2-DE technique was established.Conclusions: Our established 2-DE technique of serum proteins can be effectively applied in serum proteomics of diseases.