Clinical analysis of pediatric acute myeloid leukemia with EVI1 gene positive / 中华实用儿科临床杂志

Objective To analyze the clinical features and prognosis of pediatric acute myeloid leukemia (AML) with EVI1 gene positive.Methods The nested RT-PCR was performed to detect the EVI1 expression in pediatric AML patients from Jan.2009 to Dec.2011.The patients with EVI1 were investigated on clinical features,curative effects and prognosis.The differences between EVI1 (+) and EVI1 (-) patients were also analyzed.Results The frequency of EVI1 (+) expression was 15.65％ (13/83 cases)in pediatric AML,with the highest incidence in high-risk patients.EVI1 (+) was obviously associated with some unfavorable molecular genetic changes such as complex karyotype,MLL rearrangement,and monosomy 7.There were significant differences for EVI1 (+) group and EVI1 (-) group in the complete remission rate (45.5％ vs 79.3％,x2 =5.497,P ＜ 0.05) and complete remission(CR) rate after the first chemotherapy (18.2％ vs 63.8％,x2 =7.828,P ＜0.01).Although significant difference in death rate was not observed,EVI1(+) group had significantly higher early-death rate (45.5％ vs 8.6％,P ＜0.01).The EVI1(+) group also had lower 4 years event-free survival (EFS) [(21.2 ± 13.8) ％ vs (50.2 ± 9.1) ％,x2 =4.493,P ＜ 0.05] and lower 4 years overall survival(OS) [(32.4 ± 7.1) ％ vs (60.3 ± 10.9) ％,x2 =4.602,P ＜ 0.05] compared with EVI1 (-) group.But binary Logistic analysis did not identify EVI1 (+) as an independent unfavorable prognostic factor.Conclusions The pediatric AML with positive EVI1 expression had lower CR rate,higher early death rate and lower EFS.Positive EVI1 expression is related with an adverse outcome,but is not an independent poor prognostic factor.

Signal pathways of leukemia cell proliferation induced by ouabain / 中国实验血液学杂志

The aim of this study was to investigate the effects of ouabain and some specific signal pathway inhibitors on growth regulation in various kinds of leukemia cell lines and to explore the role of signal pathways participating in proliferation or apoptosis of leukemia cells induced by ouabain. By using MTT, the survival rates of leukemia cell lines were observed after utilizing ouabain and the specific signal pathway inhibitors. The expressions of Na(+), K(+)-ATPase alpha1 subunit of leukemia cells were evaluated by RT-PCR and Western blot. The results showed that low concentration of ouabain (10 nmol/L) could increase the survival rates of lymphocytic leukemia Jhhan cell line and megakaryocytic leukemia M07e cell line, and could up-regulate the expression of Na(+), K(+)-ATPase alpha1 subunit. Proliferation of these leukemia cells induced by ouabain could be inhibited by PP2 and PD98059 with different extents. It is concluded that Na(+), K(+)-ATPase plays an important role in signal transductions through binding to CTS (ouabain), and they can activate complex signal pathways regulating the growth of leukemia cells. The proliferation effects of cells promoted by ouabain are mediated by activation of Src kinase and ERK1/2 dependent signaling pathway.

Effects of ouabain at different concentrations on growth of leukemia cells / 中国当代儿科杂志

<p><b>OBJECTIVE</b>Cardiotonic steroids (CTS) can bind to Na+, K+ -ATPase to activate complex intracellular signaling cascades regulating the proliferation and apoptosis of cells. The aim of this study was to investigate the effects of ouabain at different concentrations on growth regulation in various kinds of leukemia cell lines and explore the pathogenesis of leukemia, the functions of Na+, K+ -ATPase as a signal transduction conductor and its effects on cell growth.</p><p><b>METHODS</b>Using the MTT assay, the survival rates of leukemia cell lines were observed 6, 12 and 24 hrs after treatment with 1 or 10 nmol/L ouabain. The expression of Na+, K+ -ATPase alpha1 subunit of leukemia cells was detected by Western blot.</p><p><b>RESULTS</b>The MTT results showed that ouabain at 1 nmol/L or 10 nmol/L induced proliferation of lymphocytic leukemia B95 and Jhhan cell lines, as well as megakaryocytic leukemia M07e and Meg01 cell lines. Ouabain at 1 nmol/L or 10 nmol/L increased the expression of Na+, K+ -ATPase alpha1 subunit. There were significant differences in the proliferation and the expression of Na+, K+ -ATPase alpha1 subunit of the leukemia cell lines between the ouabain treatment and the blank control groups 24 hrs after ouabain treatment (P<0.05). The proliferation effect of leukemia cell lines was in a direct proportion with the ouabain concentration and incubation time.</p><p><b>CONCLUSIONS</b>Na+, K+ -ATPase plays an important role in signal transductions. Through binding to ouabain, Na+, K+ -ATPase may regulate proliferation of leukemia cell lines of different origins. Ouabain at 1 nmol/L or 10 nmol/L may induce proliferation of lymphocytic leukemia cell lines (B95, Jhhan) and megakaryocytic leukemia cell lines (M07e, Meg01), and the proliferation effect was in a direct proportion with the concentration and incubation time of ouabain.</p>

Therapeutic effectiveness of the ALL-XH-99 protocol for childhood acute lymphoblastic leukemia / 中国当代儿科杂志

<p><b>OBJECTIVE</b>The ALL-XH-99 protocol for the treatment of childhood acute lymphoblastic leukemia (ALL) has been performed in the Union Hospital for 10 years. This study aimed to evaluate the therapeutic effectiveness of the protocol for childhood ALL and to investigate the prognostic factors for childhood ALL.</p><p><b>METHODS</b>This is a retrospective study. The eligible patients were treated with the ALL-XH-99 protocol. However a minor modification based on the ALL-XH-99 protocol was performed in this study, i.e., the high-risk patients as the low- and moderate-risk patients were not administered with cranial irradiation. Event-free survival (EFS) was evaluated using the Kaplan-Meier method and the differences of the EFS among groups were compared with the log-rank test. Prognostic factors for childhood ALL were investigated by the stepwise Cox proportional hazard model.</p><p><b>RESULTS</b>One hundred fifteen patients were eligible for the ALL-XH-99 protocol clinical study. The 115 patients consisted of 62 low-risk, 12 moderate-risk and 41 high-risk patients. The overall EFS at 5 years in the 115 patients was 69.0 +/- 5.0%. The 5-year-EFS in the low-risk, moderate-risk and high-risk patients was 82.0 +/- 6.0%, 77.0 +/- 15.0% and 43.0 +/- 11.0%, respectively (P <0.01). Relapse occurred in 16 patients (13.9%) in a median time of 17 months. Without administering cranial irradiation to all of the patients, the incidence of CNS leukemia relapse (2/115, 1.7%) was not higher than that previously reported. Multivariate analysis showed that the risk degree of leukemia, the presence of t (9; 22)/bcr/abl fusion gene and leukocyte count were independent adverse prognostic factors for ALL and their hazard ratio was 1.867, 3.397 and 2.236 respectively.</p><p><b>CONCLUSIONS</b>The therapeutic effectiveness of the ALL-XH-99 protocol for childhood ALL is satisfactory, with an EFS rate comparable to that of the developed countries. t (9; 22)/bcr/abl is the most important adverse independent prognostic factor for childhood ALL. Cranial irradiation may be eliminated to reduce late adverse effects in all of ALL patients.</p>

Biological Activity of Cytokine-Induced Killer Cells in vitro / 实用儿科临床杂志

Objective To investigate the biological activity of cytokine-induced killer(CIK)cells in vitro.Methods Lymphocytes isolated from peripheral blood in leukemic children were induced with interferon-?(IFN-?),anti-CD3 monoclonal antibody(CD3McAb)and interleukin-2(IL-2)and co-cultured with dendritic cells(DC)to generate DC-CIK cells.The morphology and immunophenotype of these cells were determined by electron microscopy and flow cytometry,respectively.Cytotoxicity of DC-CIK cells against leukemia cell lines was measured by the methyl thiazolyl tetrazolium (MTT) assay.Interleukin-12(IL-12),tumor necrosis factor-?(TNF-?)levels released by DC-CIK cells were quantified by enzyme-linked immunosorbent assays.Results Induced DC-CIK cells were regular,round and transparent with variable cell volume and cellular aggregation.At the 0th-4th day,its amplification was very slow,and it increased quickly at the 5th-8th day,it reached its peak amplification at the 9th-10th day,at approximately 100-fold.The main effector cells in this population were CD3+CD8+ cells and CD3+CD56+ cells.DC-CIK cells were cytotoxic to B95 cells,K562 cells and HL-60 cells,with the highest cytotoxicity towards B95 cells.The expression levels of IL-12 and TNF-? in supernatant were very high.Conclusions DC-CIK cells induced with cytokines displayed powerful amplification and strongly killing activities in vitro.It suggested that DC-CIK cells induced with cytokines may play killing activities through Th1 pathway in vitro,as a result of high secretion of Th1 cytokines,such as IL-12 and TNF-?.