ABSTRACT
Objective To establish a candidate reference method for determination of serum glycated albumin based on isotope dilution liquid chromatography /tandem mass spectrometry(ID-LC/MS). Methods Establishment of a method.According to the Japan Society of Clinical Chemistry(JSCC) reference method,the serum albumin was firstly purified by SDS-PAGE and decomposed into amino acid fragments.It selected 2H and 13C respectively as internal standards of lysine and DOF-lysine and chose the lysine and DOF-Lysine as standard material.Then the lysine and DOF-lysine were determined according to peak area after the optimization of analysis conditions.The validity of reference method was verified by the glycated albumin standard material JCCRM611-1 of Japan check medical material institutions.The evaluation of precision depended on the determination of three different concentrations of mixed serum samples for three days.The patients'serum(35 specimens)were determined by this method and a routine test method, and then the correlation of measured value was confirmed.Results The total CV of JCCRM611-1 reference material were 3.2%,2.3% and 2.6%, which had the bias of +2.25%, +2.38%, -1.21% by the deviation values.The intra CV was 0.33%-2.53%,CV for between -run assays was 0.83%-1.32%,total CV ranged from 2.27% to 3.2%.Correlation coefficient of the mass spectrometry and routine enzymatic method is 0.993.Conclusions The ID-LC/MS method for the measurement of serum glycated albumin is precise and accurate.In addition,this method showed great correlation with routine enzymatic method.It can be proposed as a candidate reference method for the determination of serum glycated albumin in China.
ABSTRACT
Diagnosis and treatment of diabetes mellitus depend on accurate monitor of blood glucose of laboratory.Glycated albumin,the short-term indicator of blood glucose control, has special advantage and role in the monitor of diabetes mellitus and has been payed more and more attention.The standardization work of determination of glycated albumin at home and abroad is still in the primary stage at present.The consistency and accuracy of measurement results for glycated albumin is needed to be solved urgently.
ABSTRACT
To construct soluble TNF related apoptosis inducing ligand (TRAIL) expression system and investigate the effect of the expression product on tumor cell. It may provide valuable information for research into the immune system of the finless porpoise. The full-length cDNA of TRAIL (designated fTRAIL) was cloned from the total RNA of the finless porpoises blood using RT-PCR techniques and then the extracellular soluble fragments of fTRAIL (designated fsTRAIL) was ligated into pET43.1a. Recombinant soluble fTRAIL (pET43.1a-fsTRAIL) fused with Nus-his tag was efficiently expressed in Escherichia coli BL21 (DE3) and the Nus-His-fsTRAIL protein was purified. The expression of Nus-His-fsTRAIL was verified by Western blotting. In vitro, the effects of the purified Nus-His-fsTRAIL protein on Jurkat and HeLa cells were etected by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrasodium bromide (MTT) assay, TrypanBlue and Flow Cytometry analysis. The expression system pET43.1a-fsTRAIL was constructed and Nus-His-fsTRAIL protein was expressed successfully. In vitro, the Nus-His-fsTRAIL protein was able to inhibit the proliferation and induce apoptosis of Jurkat and HeLa cells in a dose-dependent manner. The Nus-His-fsTRAIL protein has anti-tumor activity against Jurkat and HeLa cells in vitro.
Subject(s)
Animals , Humans , Apoptosis , Blotting, Western , Cloning, Molecular , DNA, Complementary , Escherichia coli , HeLa Cells , Jurkat Cells , Porpoises , TNF-Related Apoptosis-Inducing LigandABSTRACT
The crude venom of the centipede Scolopendra subspinipes mutilans, injected with Escherichia coli K12D31 for 3-4 days showed broad-spectrum antimicrobial activity against Gram-positive. Gram-negative bacteria and fungi. It showed good antibacterial activity against E. coli K12D31 at different temperatures, pH, and ionic strengths. The crude venom was heated at 100 degrees C for 30 min, centrifuged at 10,000 rpm for 30 min at 4 degrees C and the supernatants were obtained, from which an antibacterial fraction having a molecular mass of 3000-5000 Da, was further separated by ultrafiltration. A homogeneous antibacterial peptide named scolopendrin I, having a molecular mass of 4,498 Da, was isolated using cation-exchange chromatography and two steps of reverse-phase high performance liquid chromatography (RP-HPLC). Scolopendrin I did not show any hemolytic and agglutination activities at the concentration below 30 microM.
Subject(s)
Animals , Anisoles/chemistry , Antimicrobial Cationic Peptides/biosynthesis , Arthropod Venoms/chemistry , Arthropods/chemistry , Indoles/chemistryABSTRACT
Objective To study changes of?-glutamyltransferase(GGT)and its isoenzyme in urine and renal cortex of rats with subacute cadmium exposure.Methods Sixty healthy SD male rats were chosen and divided randomly into control group,middle dose group and high dose group,which were orally dosed daily with feed containing0,5and10mg cadmium per kg for six weeks.The activities of GGT and its isoenzyme in urine and renal cortex of the rats were determined in the3rd and6th week respectively.Re sults During the whole experimental period,the body weights and kidney to body weight ratios of control group,middle dose group and high dose group showed no significant differences.The GGT activities of middle dose group and high dose group increased significantly with the prolongation of exposure to cadmium and the increase of cumulative exposure compared with those in the control group.Unusual bands of GGT isoenzyme in the urine and renal cortex homogenate were found in the3rd week and the incidence of unusual bands of GGT isoenzyme was100percent in the6th week in the cadmium-treated rats of middle dose group and high dose group.Con clusion The GGT activities and the unusual bands of GGT isoen-zymes in the urine and renal cortex could be used as sensitive indexes to identify the renal toxic effects induced by cadmium.