The present study was to investigate in vitro the rat cardiomyocyte injury with targeting of home-made perfluorocarbon lipid particles with avidin-biotin interaction. Neonatal rat cardiomyocytes were cultured in vitro and divided into two groups: TNF-alpha activated group and non-activated group. Those in the TNF-alpha activated group were exposed to 200 ng/ml TNF-alpha solution for 6 hours and then cardiomyocytes in both groups were pretargeted with biotinylated ICAM-1 monoclonal antibodies, and were exposed to streptavidin, and then to homemade green fluorescently-labeled biotinylated perfluorocarbon lipid particles. Cardiomyocytes nucleus stained with Hoechst. The results were detected with fluorescence microscope. As a result, in TNF-alpha activated group, around blue fluorescent cardiomyocytes nucleus, a great amount of green fluorescent particles were found, while there were few green fluorescent particles in non-TNF activated group. It has been shown that ICAM-1 is expressed in the surface of cardiomyocytes when they are stimulated by TNF-alpha. Perfluorocarbon lipid particles associated with ICAM-1 monoclonal antibodies can be targeted to injured cardiomyocytes by avidin-biotin interaction.
Animals ; Animals, Newborn ; Antibodies, Monoclonal ; metabolism ; Cells, Cultured ; Contrast Media ; Female ; Fluorocarbons ; immunology ; metabolism ; Intercellular Adhesion Molecule-1 ; metabolism ; Lipids ; chemistry ; Male ; Microspheres ; Myocytes, Cardiac ; metabolism ; pathology ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; pharmacology ; Ultrasonography

Objective To establish the diagnostic model based on detection of serum biomarkers in pancreatic cancer (PC) associated diabetes.Methods From June 2013 to July 2014, at Ruijin Hospital, School of Medicine , Shanghai Jiao Tong University , 30 patients diagnosed with PC companied with new onset diabetic mellitus and 30 patients with new onset type 2 diabetic mellitus , were enrolled .Serum samples were examined by liquid chromatography-mass spectrometry ( LC-MS) for metabolomics analysis .Orthogonal partial least square ( OPLS ) was performed for raw data analysis to obtain the differentially expressed metabolites between two groups .The first 15 cases of each group were taken as training samples and the left as validation samples.The model was established using logistic regression via stepwise differentially expressed metabolites and clinical data input in training samples .The diagnostic efficiency of the model was verified in validating samples . Results Ten differentially expressed metabolites were identified in PC companied with new onset diabetic mellitus group and new onset type 2 diabetic mellitus group .The differentially expressed metabolites identified in positive ion mode were 3-ketosphingosine , arachidonoyl dopamine , phosphatidylethanolamine ( 18 :2 ) , ubiquinone-1 and valine .The differentially expressed metabolites identified in negative ion mode were C 16 sphingosine-1-phosphate, keto palmitic acid, isoleucine, N-succinyl-L-diaminopimelic acid and uridine.The diagnostic model was established in training samples:p=e(Xβ)/(1+e(Xβ)), ( Xβ) =-158.975-1.891 (age) +0.309 ( phosphatidylethanolamine 18:2 ) +1.035 ( C16 sphingosine-1-phosphate ) +0.084 (isoleucine) +1.1145 ( N-succinyl-L-diaminopimelic acid ).The area under curve ( AUC) of receiver operating characteristic (ROC) of this model was 0.982 in validation samples, the sensitivity and specificity were both 93.3％.Conclusion Serum metabolomics-based diagnostic approach is a promising method for screening PC from new onset diabetic mellitus .