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Objective To investigate the harmonization of results of Prothrombin time(PT),International Normalized Ratio(INR),activated partial thromboplastin time(APTT),fibrinogen(FIB)and thrombin time(TT)with different coagulation analyzers in different or sanle clinical laboratory.Methods PT,INR,Am,FIB and TT for the same quality control material were detected with 14 different coagulation analyzers,which are distributed in 12 clinical laboratories and classified into A,B and C group.MeaJlwhile,PT,INR,APTT,FIB of 139 samples were detected with two different coagulation aJlalyzers in the same laboratory.Results There was no significant difference for detection of level 3 of both INR and TT among the three group analyzers(P＞0.05),but there was significant difference for other tests (P＜0.05).The comparison between groups showed that there was high percentage(66.7%)of consistency for detection of INR,FIB-C and TT between group B and C.The results of two different coagulation analvzers ( ACL Futura and CA 510)in same laboratory showed that there was no significant difference(P＞0.05)for detection of PT,INR,PT-FIB and FIB-C between them,and there was good eorrelation for them in detecting PT,INR,APTT,PT-FIB and FIB-C(r＞0.975).Analysis of bias showed that the bias of PT,INR,PT-FIB and FIBC between the two different coagulation analyzers was acceptable according to CLIA'88.Conclusion There are good agreement for the results between different coagulation analyzers based upon the similar Drinciple in coagulation analysis.
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Objective To investigate the prevalence of plasmid-mediated quinolone resistance qnr and aac(6')-Ib-cr in Enterobacteriaceac in Chiha.Methods A total of 197 clinical isolates with ciprofloxacin≥0.25μg/ml,cefotaxime≥2.0μg/ml and ceftriaxone≥2.0 μg/ml were screened from the 421 non-repetitive clinical isolates of Enterobac teriaceae(Escherichia coli,Klebsiella pneumoniae,Citrobacter freundii,and Enterobacter cloacae)from the nine teaching hospitals in China.qnrA,qnrB,qnrS and aac(6')-Ib gene were detected by PCR.aac(6')-Ib-cr gene was further identified by the digestion with BtsCI followed by sequencing.Conjugation experiments were done.The MIC of ciprofloxacin and other antibacterial agents in donor strain and acceptor strain were determined by agar dilution.Results Qnr was present in 42%(83/197)of isolares,and among these,17 isolates carried qnrA(9%),46 isolates carried qnrB(23%),24 isolates carried qnrS(12%),2 isolates carried qnrA and qnrB,and 2 isolates carried qnrB and qnrS.aac(6')-Ib was present in 46%(90/197)of isolates,40%(36/90)of which carried the cr variant responsible for low-level ciprofloxacin resistance.18 isolates carried qnr and aac(6')-Ib-cr.Qnr wag present in 66% of Enterobacter cloacae isolates,66% of Klebsiella pneumoniae isolates,63% of Citrobacter freundii isolates,and 6% of Escheriehia coil isolates,respectively,aac(6')-Ib-cr was present in 9% of Enterobacter cloacae isolates,22% of Klebsiella pneumoniae isolates,27% of Citrohacter freundii isolates,and 17% of Escherichia coil isolates,respectively,qnr and aac(6')-Ib-cr were present in 20% (83/421)and 9% (36/421) of all isolates respectively. The 13 transconjugants showed 16 to 125 fold increases in the MICs of ciprofloxacin and 16 to 31 fold increases in the MICs of levofloxacin relative to that of the recipient Conclusion Transferable plasmid-medlated low level quinolone resistance associated with qnr and aac(6')-Ib-cr widely exists in the enterobacteriaceae strains and perhaps this may contribute to the rapid increase of bacterial resistance to quinolones in China.
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Objective To study and develop the whole blood red cell lysing reagents in order to replace the commercial kit and reduce the cost.Methods Flow cytometry was used to compare the home-made red cell lysing reagents with the commercial kit on the separation of white cells into three groups,on the ratio of lymphocyte subsets and on the mean fluorescence intensity(MFI)of lymphocyte subsets.Results Compared with the commercial kit,the home-made lysing reagents had no significant difference on the separation of white cells into three groups,on the ratio of lymphocyte subsets and on the MFI of T lymphocytes,B lymphocytes,Helper-Inducer T-lymphocytes and Suppressor-Cytotoxic T-lymphocytes.Conclusions The home-made lysing reagents had similar effects as the commercial kit on the separation of white cells,on the ratio of lymphocyte subsets and on the MFI of T lymphocytes,B lymphocytes,Helper-Inducer T-Lymphocytes and Suppressor-Cytotoxic T-lymphocytes,but the cost is much lower.
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Objective To quantitate platelet GPIIb/IIIa occupancy and to evaluate the performance of the method, and investigate GPIIb/IIIa occupancy for the patients with leukemia.Methods GPIIb/IIIa occupancy was quantified by flow cytometry (FCM) and the method was evaluated according to guidelines published by NCCLS and ICSH; meanwhile,GPIIb/IIIa occupancy for 13 healthy donors and 16 patients with acute leukemia was investigated.Results The results demonstrated coefficients of variation (CV) for within-batch, between-batch and overall imprecision were
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<p><b>OBJECTIVE</b>To quantitatively analyze apoptotic and secondary necrotic cells under apoptosis conditions.</p><p><b>METHODS</b>The cells of Burkitt lymphoma (BL) cell line Raji were incubated with 1.0 micromol/L dexamethasone (DEX) for 2, 4 and 8 h respectively, then stained with Annexin V-FITC (fluorescein isothiocyanate conjugated) which was used to detect the exposed phosphatidylserine (PS) on the epimembrane resulting from a loss of phospholipid asymmetry in the early stage of apoptosis, and also stained with propidium iodide (PI) which allows analysis of secondary necrotic cells related with cell membrane and DNA damage that probably representlate stage of apoptosis, then apoptotic cells were quantified by flow cytometry (FCM). Furthermore, Annexin+ /PI- and Annexin+ /PI+ cells were sorted by fluoresence-activated cell sorter (FACS), and identified by electron microscopy (EM) and DNA gel electrophoresis.</p><p><b>RESULTS</b>The percentage of apoptotic cells was found to increase with the incubation time (r = 0.97). This method was sensitive with low detection limit (0.02%), and was reproducible with low coefficient variance (CV) (4.2%). Meanwhile, the Annexin+ /PI- and Annexin+ /PI+ cells were identified as apoptotic and necrotic cells under EM, and DNA extracted from the Annexin+ /PI- cells was characterized by "ladder pattern".</p><p><b>CONCLUSIONS</b>Annexin-V assay is a specific, sensitive, accurate, reproductive and quantitative method for analyzing apoptotic cells.</p>
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Humanos , Anexina A5 , Apoptose , Linfoma de Burkitt , Patologia , Dano ao DNA , Necrose , Fosfatidilserinas , Metabolismo , Propídio , Células Tumorais CultivadasRESUMO
Cortical neurons of mouse embryo were cultured for 7 days.The infection group was co cultured with HTNV(A9 strain) and the control group with devitalized HTNV(A9 strain) for 4h.Both groups were divided randomly into 9 subgroups acccording to different time points.The expression of fos gene was stained with immunohistochemistry.The resulfs showed that cell nuclei of neurons in the infection group displaying a purple blue color.At o,1,2,3,and 4h after infection,the expression rates of fos positive cells were 50 0%( P
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Objective To set up a calibration system for automated hematology analyzers with fresh blood in clinical laboratory.Methods Fresh blood assigned by a traceable measurement system was used to calibrate nine hematology analyzers, and compared the bias before and after calibration.Results In the parameters to be calibrated for the hematology analyzers, there was about 55.6% (25/45) over allowable bias before calibration but 15.6% (7/45) after calibration with fresh blood. Among the results of bias over allowable upper limit were mostly existed in 3-part differential hematology analyzers, and mainly focused on WBC and PLT.Conclusion It is available to calibrate different hematology analyzers with fresh blood in a clinical laboratory.