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Article in Chinese | WPRIM | ID: wpr-742872


Objective To evaluate the performance of hook effect of five immunoturbidimetric kits for the detection of specific proteins on biochemical analyzers.Methods Five immunoturbidimetric kits with higher market share that came from Beijing BSBE (A), Sichuan maccura (B), Shenzhen Mindray (C), Ningbo Medical System (D) and Beijing Leadman (E) were used to determine six specific proteins.A series of concentration gradient samples were prepared and tested to compare the performance of hook effect from different manufactures′kits when the analytical measurement ranges were known.Results In the five kits, the upper limits of the safe range of antigen excess about ASO, hs-CRP andβ2-MG were relatively higher in B and C.No hook effect occurred at the approximate concentration of 10 000IU/mL, 1 000mg/L and 226mg/L respectively.The highest upper limits for CysC were C and E kits, and both were greater than 112mg/L.The upper limits of the safety range for other manufacturers were more than 700mg/L about RBP except for D.The maximum upper limit of mALB was D.Hook effect did not appear at the concentration of 43 560mg/L approximately.Conclusion Different manufactures′immunoturbidimetric kits have different hook effect performance.The laboratories should verify the hook effect performance before using the kits, and select the most suitable kit to prevent hook effect.

Article in Chinese | WPRIM | ID: wpr-508105


Objective Tumor cells are able to support their malignant proliferation by changing metabolic models .Prostate cells rely much on lipid metabolism in which ATP-citrate lyase ( ACLY) plays a very important role .The aim of this research was to study the effects of downregulated ACLY on the cell proliferation , cycle distribution and apoptosis of androgen-independent prostate cancer cells DUl45. Methods DU145 cells were divided into two groups:the cells in experiment group were transfected with the small interfering RNA-mediated knockdown of ACLY , while the cells in control group were transfected with meaningless small interfering RNA.Cell counting Kit test ( CCK-8 ) was applied to detect the effects of the downregulation of ATP citrate lyase on the proliferation of DU145.Flow cytometry instrument was used to analyze the variation of cell cycle distribution and apoptosis rate between groups .Western blot was used to detect the change of intracellular Caspase-3 protein content. Results Western blot showed favorable effects of ACLY interference .Compared with control group , ACLY protein content significantly decreased in experiment group ( P0.05), while the percentage of G2 cells decreased and the percentage of S cells in-creased with most cell cycle blocking at G 0/G1 stage, which were of significant difference .Meanwhile the expression of apoptosis pro-tein Caspase-3 upregulated significantly . Conclusion ACLY is of vital significance to maintain the malignant proliferation of prostate cancer cells and its downregulation results in the inhibition of cell proliferation and the promotion of cell apoptosis .

Article in Chinese | WPRIM | ID: wpr-355287


<p><b>OBJECTIVE</b>To identify the biomarkers of renal cell cancer (RCC) through urine metabolic analysis.</p><p><b>METHODS</b>Urine samples of 27 RCC patients, 26 patients with other urinary cancers and 26 healthy volunteers were examined with gas chromatography-mass spectrometry (GC-MS). SIMCA-P+ software was used for principal component analysis (PCA) and orthogonal partial least-squares discriminant analysis (OPLS-DA) to screen for the differential metabolites.</p><p><b>RESULTS</b>PCA (R2X=0.846, Q2=0.575) and OPLS-DA (R2X=0.736, R2Y=0.974, Q2Y=0.897) model were established for the RCC patients and control subjects. Fourteen metabolites were selected as the characteristic metabolites, including pentanoic acid, malonic acid, glutaric acid, adipic acid, amino quinoline, quinoline, indole acetic acid, and tryptophan, whose levels in the urine were significantly higher in the RCC patients than in the normal subjects (P<0.01); the RCC patients showed significantly higher urine contents of pentanoic acid, phenylalanine, and 6-methoxy-nitro quinoline than those with other urinary tumors (P<0.01).</p><p><b>CONCLUSION</b>The urine metabolites identified based on GC-MS analysis can distinguish RCC patients from patients with other urinary cancers and healthy subjects, suggesting their potential as diagnostic markers for RCC.</p>

Biomarkers , Urine , Carcinoma, Renal Cell , Urine , Discriminant Analysis , Gas Chromatography-Mass Spectrometry , Humans , Least-Squares Analysis , Metabolome , Metabolomics , Principal Component Analysis , Software
Article in Chinese | WPRIM | ID: wpr-319464


<p><b>OBJECTIVE</b>To investigate AXIN1-related CSRNP1 gene expression and the mechanism of its transcriptional regulation in TGF-β1-induced tumor cells.</p><p><b>METHODS</b>Human lung carcinoma A549 cells or human prostate cancer PC3 cells were treated with TGF-β1 at different doses (0, 20, 40, and 80 ng/ml) or at 20 ng/ml for 0, 8, 12, or 24 h, and the dose and time effect of TGF-β1 on CSRNP1 mRNA expression in the tumor cells were evaluated with real-time RT-PCR. A549 cells were also treated with TGF-β1 and cycloheximide to clarify whether CSRNP1 expression induced by TGF-β1 required de novo protein synthesis. A549 cells transfected with pcDNA3.1, flag-SMAD3, or flag-SMAD3-mu, after serum starvation, were treated with or without TGF-β1 (20 ng/mL) for 24 h, and the overexpression of wild-type SMAD3 and dominant negative SMAD3-mu mutant were confirmed by Western blotting. The effect of SMAD3 or SMAD3-mu overexpression on CSRNP1 mRNA expression was also measured by real-time RT-PCR.</p><p><b>RESULTS</b>In both A549 and PC3 cells, TGF-β1 dose- and time-dependently stimulated CSRNP1 expression, which required de novo protein synthesis in A549 cells. Overexpression of wild-type SMAD3 significantly increased the expression of CSRNP1 mRNA induced by TGF-β1, while overexpression of dominant negative SMAD3 mutant remarkably reduced CSRNP1 mRNA expression in response to TGF-β1 in A549 cells.</p><p><b>CONCLUSION</b>TGF-β1 may contribute to CSRNP1 expression through SMAD3 activation and downstream signaling in tumor cells.</p>

Apoptosis Regulatory Proteins , Genetics , Metabolism , Axin Protein , Genetics , Metabolism , Cell Line, Tumor , Gene Expression Regulation, Neoplastic , Humans , RNA, Messenger , Genetics , Signal Transduction , Smad3 Protein , Genetics , Metabolism , Transfection , Transforming Growth Factor beta1 , Pharmacology
Article in Chinese | WPRIM | ID: wpr-421270


Cultivation of young teachers is an important part of teaching staff training in colleges and universities. To strengthen teaching ability of young teachers in discipline of medicine laboratory is related to the quality of laboratory medicine personnel training.The paper will be based on some problems of young teacher teaching and discuss some measures of young teachers' teaching ability improvement.