| Objective: Idiopathic thrombocytopenic purpura(ITP) is the most common hemorrhagenic disease in children. While maturity disturbance of MKs and impaired production of platelets have been regarded as the main pathogenesis of ITP, the mechanism remains still unclear. In recent few years, the transcription factor GATA-1 and nuclear factor erythroid 2(NF-E2) have been paid increasing attention for their involvement in megakaryocytic hematopoiesis: the former was thought to play an important role in proliferation and differentiation of megakaryocyte; while the latter was found to be essential for late megakaryocyte maturation and platelet formation. Therefore, a clear understanding of the levels of GATA-1 and NF-E2 on megakaryocytes in bone marrow will help us to elucidate further the mechanism of ITP as well as to explore their influence upon megakaryocytopoiesis and thrombopoiesis.Subjects and Methods: (l)Bone marrow were respectively aspirated from 28 children with AITP and 19 children with CITP before treatment. 9 cases with AITP undergoing high-dose intravenous immunoglobulin (HDIVIgG) treatment were observed to show good recovery. When the patients with the treatment of HDIVIgG showed a rapid, up-to normal platelet increase, bone marrow was aspirated once more immediately. 21 healthy children were normal controls. (2)Purification of the megakaryocytes from bone marrow: 1.5ml~3ml bone marrow was obtained into modified PBS, and MKs were isolated by immunomagenetic beads. (3)Measurement of the levels of GATA-1 and NF-E2 on Mks: the levels of GATA-1 and NF-E2 weredetermined by ABC immune cytochemisty method, the positive cell ratio and average reactive intensity of them were calculated. (4)Statistical assay: the data were dealt with SPSS 10.0 statistical software.Results: 1. GATA-1 positive cell ratio: before treatment in 28 children with AITP, the mean of GATA-1 positive ratio was 82.36 + 3.53%, while it was 76.29 + 3.87% in normal controls. The mean level of positive rate of GATA-1 before and after treatment with HDIVIgG was 81.89±3.95% and 91.89 + 3.95%. It was 96.16 + 2.83% in CITP group. The positive ratio in AITP group before treatment was higher than in normal controls (P<0.05). In CITP group, it was also higher than in normal controls(P<0.05). Moreover, it was much higher in patients with CITP than in patients with AITP before treatment (P<0.05). After treatment with HDIVIgG, the positive ratio increased significantly compared with that before treatment (Ml.339, P<0.01). 2. GATA-1 reactive intensity on MKs: before treatment in 28 children with AITP, the mean of GATA-1 reactive intensity was 1.884±0.1457, while it was 1.726 + 0.1284 in normal controls. Before and after treatment with HDIVIgG, it was 1.877 ±0.1128 and 2.884+ 0.1683 respectively. It was 3.253 + 0.1565 in CITP group. The GATA-1 reactive intensity in AITP group before treatment was higher than in normal controls (P<0.05). In CITP group, it was also higher than in normal controls (P<0.05). Moreover, it was much higher in patients with CITP than in patients with AITP before treatment (P<0.05). After treatment with HDIVIgG, the reactive intensity increased significantly compared with that before treatment (t=24.487, P<0.01). 3. NF-E2 positive cell ratio: before treatment in 28 children with AITP, the mean of NF-E2 positive ratio was 81.82 + 3.59%, while it was 73.90±3.71% in normal controls. The mean level of positive rate of NF-E2 before and after treatment with HDIVIgG was 79.44 + 2.40% and 91.44 ±5.25%. It was 94.68 ±2.36% in CITP group. The positive ratio in AITP group before treatment was higher than in normal controls (P<0.05). In CITP group, it was also higher than in normal controls (P<0.05). Moreover, it was much higher in patients with CITP than in patients with AITP before treatment (P<0.05). After treatment with HDIVIgG, the positive ratio increased significantly compared with that before treatment (t=9.219, P<0.01). 4. NF-E2 reactive intensity on MKs: before treatment in 28 children with AITP, the mean of NF-E2 reacti... |
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