ABSTRACT
Objective To investigate the influence of bloody cervical cell samples on human papillomavirus(HPV) genotyping detection using flowcytometry fluorescence hybridization kit.Methods According to the concentration of hemoglobin(Hb),cervical cell samples were divided into five groups,including group A with Hb of 0 g/L(10 cases),group B with 0 g/L100 g/L.In each group,50% samples were with initial positive results of HPV genotyping detection.Every sample was detected using sample volume of 50,100,200,500 and 1 000 μL different volumes for testing and analysis results.Results For all samples with initial negative results,five samples with initial positive results in group A and five samples with initial positive results,the detected results of different sample volume were without differences.In the 10 samples,with initial positive results of group C,D and E,there were four,four and nine samples respectively,were with false negative detected results of different sample volumes.Conclusion Bloody cervical cell samples could cause false negative results of HPV genotyping detection using flowcytometry fluorescence hybridization kit.It might be necessary to set optimal sample volume according to the concentration of Hb for ensuring the accuracy of genotyping detection.
ABSTRACT
Objective Developing an internal quality control substance of Ureaplasma urealyticum(UU)‐DNA for real‐time PCR to establish an internal quality control system and preliminary evaluation its clinical value .Methods Internal quality control sub‐stance was prepared by mixing samples which Ct value were 24-25(positive sample) and 32 -33(weak positive sample) ,respec‐tively .At the same time ,selecting samples that test results were negative as negative control .The target value ,standard deviation (s) and coefficient of variation(CV) of internal quality control substance were defined by“instant method”for the first 20 runs and Levey‐Jennings quality control(QC) chart after the first 20 runs .Using the“Westdard” multi‐rule quality control methods to ana‐lyze the detection results .Exporting OPSPecs chart by quality control rules in Unity Real Time (URT ) system and setting up new quality control rules according with OPSPecs chart .Results 131 times of the detection of quality control substance were performed totally .The first 20 runs were defined by“instant method”and later 111 runs were defined by Levey‐Jennings QC chart ,the results were stable of quality control substance and reasonable quality control rules .Conclusion Preparing of internal quality control sub‐stance of UU‐DNA used in real‐time PCR might be easy and stable .So ,the internal quality control substance of UU‐DNA could be worthy for practical application in this PCR laboratory .Design internal quality control rules based OPSPecs chart in molecular de‐tection is very simple and practical .
ABSTRACT
Aim To study the sensitivity of human gastric cancer BGC-823 cells to paclitaxel after trans-fection of SHIP2 ( The SH2 domain containing inositol 5-phosphatase 2 ) cDNA. Methods Apoptotic cells were determined by the propidium iodide method using flow cytometry. The levels of protein and mRNA ex-pression were measured by Western blot analysis and qRT-PCR, respectively. pCMV6-SHIP2 plasmid and empty vector were transiently transfected into BGC-823 cells, respectively. Stable cell lines were established after infecting BGC-823 cell with GV112-Puromycin and GV112-Puromycin-Bim( Bcl-2 interacting mediator of cell death) lentivirus particles. pCMV6-SHIP2 plas-mid was transiently transfected into the stable cell lines. Results BGC-823 cells were relatively insensi-tive to paclitaxel compared with SGC-7901 cells. The apoptotic rate was only (25. 6 ± 1. 6)% after the treat-ment with 0 . 3 μmol · L-1 paclitaxel for 48 h in BGC-823 cells. The expression levels of Bim protein and mRNA in BGC-823 cells treated with paclitaxel at dif-ferent time points were not significantly changed. The expression of Bim protein was increased after transfec-tion of pCMV6-SHIP2 plasmid, and the apoptotic rate was up to ( 50. 8 ± 0 . 9 )% in BGC-823 cells treated with paclitaxel for 48h. The expression of Bim protein was significantly inhibited after infecting with GV112-Puromycin-Bim lentivirus particles. The apoptotic rate of infected BGC-823 cells was only ( 27. 6 ± 1. 6 )%after treatment upon paclitaxel for 48h. Conclusion Overexpression of exogenous SHIP2 can increase the expression of Bim, induce apoptosis and enhance sen-sitivity of BGC-823 cells to paclitaxel.