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AIM:To determine the biological feature of circulating endothelial cells (CECs) in acute promye-locytic leukemia ( APL) patients before and after treatment , and to analyze the relationship between CECs and the clinical characteristics .METHODS: The CECs were sorted from peripheral blood by magnetic-activated cell sorting and then counted by 3-color flow cytometry.The cells were identified by immunofluorescence staining for the expression of CD 146, CD31, CD144, VEGFR-2, CD45 and CD133.The CECs were cultured in vitro, and the tube formation and colony-forming rate were determined .RESULTS:Increased quantity of CECs was observed in CD 34 positive group and group with WBC >10 ×109/L (P<0.05).The quantity of CECs had a significant difference among low risk , medium risk and high risk groups (P<0.05).The positive rate of CD133 and quantity of CECs significantly reduced in 32 APL patients when they gain complete remission after treatment (P<0.05).The amount of tube formation and colony-forming rate were significant-ly reduced after treatment (P<0.05).The ratio of CECs quantity from APL patients after treatment to that before treatment had a negative correlation with arsenic concentration in urine on day 7 during As2O3 treatment (P<0.05).CONCLU-SION:Accurately counting CECs may be helpful for evaluating prognosis and designing treatment strategy .
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Objective To establish a HPLC/MS/MS method for the determinati on of gentiopicroside in human urine.Methods The urine sample was treated by solid-phase extraction with internal standard of caffeine;The RESCEK C8 colum n(150mm?2.1mm,5 ?m) was used as the analytical column with a mobilep hase consisting of methanol-10mmol? L-1 NH4AC buffer(pH=6.5)-acetonit rile(50∶40∶10,V/V),the flow rate was 0.2 mL? min-1;A triple quadruple tandem mass spectrometer was used as the detector,Electrospray ioniza tion source was applied and operated in positiveion mode.Gentiopicroside and caffeine were detected by monitoring the ion transition of m/z 374.1→ 195.2 and m/z 195.2→ 138.2 respectively.Results The linear range was 30～ 9000 ng?mL-1(r=0.9980) for gentiopicroside in human urine.The recovery was 91.10% ～ 9 6.21 %,The absolute recovery was 100.52% ～ 103.83%,The within-day and between-day precisions were less than 10 %.Conclusion The method is proved to be sensitive,accurate,rapid,specific.
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Objective:To establish a method for determination of diisopropylamide dichloroacetate(DADA) concentrations in urine.Methods:GC-HS was applied to quantitative analysis.Using Agilent DB-5MS capillary column(30m?0.25mm),the column temperature was ascended from 40℃ to 200℃ according to program.Initial temperature was 40℃ and maintained 2 mins,then temperature rose to 80℃ at 5℃/min rate and maintained 2 mins,finally temperature rose to 200℃ at 20℃/min rate and maintained 8 mins.The injector temperature was 200℃.The detector temperature was 300℃.Nitrogen was used as carrier gas,and the flow rate was 1 ml/min.The flow rate of hydrogen was 35 ml/min.The flow rate of air was 350 ml/min.An internal standard was secondary butanol.Results:The calibration curve was linear within 8～800?g/ml.The recovery rate of the extraction were 88.75%～98.06%.The recovery rate were 97.36%～103.25%。Within-day and between-day RSD were 2.95%～6.62% and 2.45%～5.61% respectively.Detectable concentration limited was 2?g/mL.Conclusion:The GC-HS method is simple,reliable,and suitable for urine DADA concentration analysis.