Evaluation Of Erythroblast Macrophage Protein Expression In Mononlcear Cells And Hematopoietic Function Of Bone Marrow

PartⅠEstablishiment and evaluation of Quantitative Measurement of EMP mRNA detectionObjective:To establish fluorescence real-time quantitative reverse transcriptase polymerase chain reaction (FQ-RT-PCR) for the of EMP mRNA detection.Methods:A pair of primers and a TaqMan probe of EMP gene were designed using primer 2.0 software. Quantitative measurement of EMP mRNA was established and evaluated. EMP sequence was amplified by conventional PCR and purified PCR products were cloned into pGEM-TEasy vector as external standard and the method was evalued.Results:The sensitivity was 5copies/ml, the linear range was 7～7×10 copies/ml, the coefficient variation was 2.80～3.15% and 3.45～4.50% in intra and inter assay. The PCR product was specific for EMP sequence.Conclusions:The quantitative method for EMP mRNA detection was established. FQ-RT-PCR for EMP mRNA was a sensitive, specific, rapid and efficient method. PartⅡErythroblast macrophage protein expression in mononlcear cells and hematopoietic function of bone MarrowObjective:To detect the expression of EMP mRNA and EMP protein in patients with hematologic disease after hematopoietic stem cell transplantation (HSCT).Results:Five experimental groups were designed for observing the change in bone marrow, including two control groups, the first group (10 donors) and the second group (46 patients before HSCT), and three groups after HSCT (at the point of 30,90 and 180 days). EMP protein of bone marrow was detected in group 3, group 4 and group5 but not in group 1 and group 2 by western blot analysis. Mean erythroblastic island quantity, EMP mRNA level of bone marrow in group3, group 4 and group5 was significantly higher than that of the two control groups respectively.Conclusions:These results indicated that the expression of EMP could indicate the hematopoietic status and may have an important function for bone marrow hematopoiesis.