ABSTRACT
In this study, the rapid resolution liquid chromatography-quadrupole time-of-flight mass spectrometry (RRLC-QTOF/MS) was used to profile the metabolites of urine samples from chronic heart failure (CHF) patients and healthy controls to find the differential metabolites which could provide the scientific evidence to explain the pathogenesis of the disease and supply a better therapy plan.Urine samples from 15 CHF patients (age (62.27±3.14) years) and 15 healthy controls (age (65.41±4.63) years) were analyzed by RRLC-QTOF/MS.After processing the data, the multivariate statistical analysis (principal component analysis, PCA) was performed to find the potential biomarkers.Result showed that urine samples of CHF patients were successfully distinguished from those of healthy controls.Two significantly differentially expressed metabolites, uridine and alanyltryptophan, were found and identified as potential biomarkers.The result showed that the LC-MS based metabolomics approach had good performance to identify potential biomarkers, and the disorder of uracil metabolism and Tryptophan metabolism may play an important role in the mechanism of CHF.
ABSTRACT
An untargeted urinary metabonomics method based on ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry ( UPLC-Q-TOF-MS ) has been established to investigate the mechanism of Schisandra chinensis in treating diabetes and its complications. The urinary biomarkers related to the therapeutic effects of Schisandra chinensis on the diabetes rats were analyzed. In urine, 28 kinds of endogenous metabolites were identified as potential biomarkers, including 13 endogenous metabolites in positive ion mode, 15 endogenous metabolites in negative ion mode, and hippuric acid detected both in positive and negative ion modes. The results revealed that Schisandra chinensis mainly affected the pathways of pentose and glucuronate interconversions, riboflavin metabolism, pantothenate and CoA biosynthesis, arginine and proline metabolism, intestinal bacteria metabolism, ascorbate and aldarate metabolism and tryptophan metabolism in diabetic rats. Combined with biological analysis of these pathways, the therapeutic mechanism of Schisandra chinensis on diabetes and its complications was verified. Based on the biological function of each pathway, the effect of Schisandra chinensis on diabetic nephropathy is stronger. Moreover, it also has the effects of protecting liver, decreasing fat and antioxidant activity.
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Objective To construct a eukaryotic expression vector of decorin(DCN),and observe its expression in CHO cells,in order to provide a basis for further study on the anti-tumor effect of DCN. Methods DCN cDNA was amplified by PCR.The human full-length DCN cDNA ligated into pBluescript was used as template.The fragment was ligated to the expression vector pCDNA3 previously digested with XbaⅠ and EcoR Ⅰ.The ligation mixture was transformed into competent E.coli JM109 cells.Transformants containing inserts were confirmed by restrictive digestion and DNA sequencing.The expression vector was transfected into CHO cells using lipofectamine,and transfected cells were cultivated in DMEM containing G418 (800 mg?L-1) for about 2 months.Immunohistochemistry method was used to detect the expression of DCN protein in stably transfected cells.Results The PCR product was about 1 000 bp.The recombinant expression vector was identified by restrictive digestion and DNA sequencing. DCN protein was detectable in stablely transfected cells.Conclusion The recombinant eukaryotic expression vector pCDNA-DEC is constructed successfully and stablely transfected CHO cells are established.
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Author thinks that it is necessary to improve current clinical surgery teaching work by improving teachers'quality,advocating initiative teaching and professional education,guiding students how to deal with the relationship of doctors and patients,increasing the consciousness of law and self protection and making unified teaching content and target.
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Objective:To investiate biological properties of Mesenchymal stem cell(BMSC) from rat bone marrow in vitro,and their effects in inducing immune tolerance.Methods:BMSCs were obtained from rat bone marrow by density gradient centrifugation and selected by cell attachment.The morphology of BMSCs was observed by electron microscope.The expression of surface molecules of CD34 and CD44 was analyzed by flow cytometry.The cells were injected ex vivo intravenously after in vitro culture and then CD4+ CD25+ Treg ratio were determined.The proliferation of thymic cells was estimated by 3H-TdR incorporation method.Results:BMSCs were fibroblast-like cells.The expression ratio of CD44 in BMSCs was 98%,but expression of CD34 was negative.After administration of BMSCs,the ratio of CD4+CD25+Treg(2.5%?0.69%) from peripheral blood of rats was higher than in normal group(0.8%?0.14%) and PBS control group(1.0%?0.23%),P