Leukemia is a malignant hyperplasia disease of hematopoietic system. It is characterized that a certain blood cell system proliferates, gets into blood streaming, infiltrates each organism and apparatus and arouse a series of clinical manifestation. In recent years the study of leukemia has discovered that it was correlated with apoptosis of the cells. The roles of XIAP and survivin in apoptosis inhibition have been especially paid attention. It is discovered that they play major roles in the genesis and resistance to chemotherapeutical drug of tumors. We summarize the biological characteristics and the expressions of XIAP and survivin in the children with acute leukemia.

Objective To investigate the relationship between the expressions of proliferating cell nuclear antigen (PCNA), p53 and Bcl-2 protein and clinical chemothreapy, prognosis in bone marrow cells in children with acute leukemia (AL). Methods Immunohistochemical SP method was used to detect the expressions of PCNA, p53 and Bcl-2 in specimens of bone marrow puncture of 59 children with AL and 15 healthy children as control. Results There was a significant difference in the expressions of PCNA, p53 and Bcl-2 proteins between the initially treated patients and healthy subjects, and between the remission patients and non remission ones. There was not a significnat difference in PCNA expression between the refractory patients and healthy subjects, and PCNA expression was related to the chemotherapeutic sensitivity. There was a significant difference in the 6-week remission rate between the patients with and without PCNA expression, but there was no significant difference in the over 3 years survival rate without illness. The expression levels of Bcl-2 and p53 were significantly higher in the refractory patients than those in healthy subjects. The patients with the high expression of p53 and Bcl-2 were resistant to chemotherapy, low in the remission rate and poor in prognosis. Conclusion The AL patients with PCNA expression were higher in remission rate, and PCNA expression was not associated with long-term prognosis. The AL patients with the expression of p53 and Bcl-2 were lower in remission rate, and their expression was associated with long-term prognosis. Both p53 and Bcl-2 protein may serve as a molecular marker to predict chemotherapeutic sensitivity and prognosis. PCNA, p53 and Bcl-2 may be involved in the pathogenesis of child AL by various ways. It is more valuable for predicting prognosis to simultaneously detect the expression of PCNA, p53 and Bcl-2 proteins.

Objective To investigate the expression and clinical significance of Smac and HtrA2 in children with acute leukemia(AL).Methods Bone marrow samples were obtained from 77 children with AL (including 32 newly diagnosed children,33 complete remission children and 12 relapsed children)and the control group of 15 children without malignant blood disease.The expressions of Smac and HtrA2 protein were measured by streptavidin/peroxidase immunoperoxidase technique(SP) in all children.SPSS 13.0 software was applied to analyze the statistical data.Results Protein Smac was detected only in some samples,but HtrA2 was detected in all samples.The levels of Smac and HtrA2 protein in newly diagnosed AL children were both higher than those of the complete remission children (x2 =17.38,F =2.36,all P ＜ 0.05) and normal controls (x2 =12.89,F =5.26,all P ＜ 0.05),there was a statistical significance,but compared with those in the relapsed children,the difference had no statistical significance (x2 =1.18,F =1.57,all P ＞ 0.05).The levels of Smac and HtrA2 protein in complete remission children were both higher than those of the normal controls,and the difference had no statistical sigmficance(x2 =1.20,F =2.23,all P ＞ 0.05).In the newly diagnosed children,the levels of Smac and HtrA2 protein in children with acute lymphocytic leukemia(ALL) were higher than those of the acute myeloid leukemia(AML),but the differences had no statistical significance(x2 =0.113,t =1.024,all P ＞ 0.05).In newly diagnosed AL children,the complete remission(CR) rate of the negative expression of Smac(Smac-,90.9％) and the low expression of HtrA2(HtrA2low,84.6％) in the level of protein were higher than those of the positive expression of Smac(Smac +,47.6％) and the high expression of HtrA2 (HtrA2high,47.4％),and there was statistical significance respectively(x2 =5.772,4.596,all P ＜ 0.05).The CR rate of Smac-HtrA2low group (100％) was higher than that of Smac+ HtrA2high group(30.8％)in the children with AL,and the statistical data were of great significance(x =9.692,P ＜0.01).The protein level of Smac in newly diagnosed AL children was correlatedwith HtrA2 (r =0.979,P ＜ 0.001).Conclusions Pro-apoptotic protein Smac and HtrA2 may be involved in and af-fected each other in the pathogenesis and progression in AL,but levels of Smac and HtrA2 protein may be not correlatedwith the types of AL.In newly diagnosed AL children,the high expression of protein Smac and HtrA2 predicts poorprognosis.

Objective To investigate the expression and clinical significance of inhibitor of apoptosis protein XIAP in childhood acute leukemia(AL).Methotis The expression of XIAP protein was detected by immunohistochemical assay in 54 children with AL.including 26 newly diagnosed and untreated AL children.23 children in remission and 5 relapsed.The eontrol included 10 children with normal bone marrow.Results The level of XIAP proteins in the bone marrow of newly diagnosed AL children was higher than that in remission and normal controls(P＜0.05),compared with that in relapsed children,with no statistical difference(P＞0.05).The level of XIAP protein in remission was higher than that in normal controls(P＜0.05).The level of XIAP protein in ALL and ANLL in newly diagnosed AL children was compared. with no statistical difference(P＞0.05).Conclusion The higher expression of XIAP may contribute to the pathogenesis and the progress of the childhood AL.

Objective To detect the levels of insulin-like growth factors in children with acute leukemia (AL). Methods A total of 50 previously untreated AL patients were selected, meanwhile 30 healthy children were selected as normal controls. AL children were given regular chemotherapy. All cases were not given the brain radiotherapy. The levels of insulin-like growth factor-1 (IGF-1), free insulin-like growth factor-1 (fIGF-1), insulin-like growth factor binding protein 3 (IGFBP-3) in AL patients before treatment and 6 months after complete remission were measured by enzyme-linked immunosorbent assay (ELISA), and were compared with those in normal controls. Results Before treatment, compared with normal controls, the serum levels of IGF-1, IGFBP-3 in AL patients were lower while the level of fIGF-1 was higher, and the differences were signiifcant (P<0.01). At six months after complete remission, the levels of IGF-1 and fIGF-1 in AL patients were similar to those before treatment, but were signiifcantly different from those in control group (P<0.05);the level of IGFBP-3 was signiifcantly higher than that before treatment (P<0.01), but was similar to that in control group. Before treatment, the level of IGFBP-3 in AL patients was positively correlated with the level of IGF-1 (r=0.777, P<0.01), and negatively correlated with the level of fIGF-1 (r=-0.714, P<0.01). Conclusion Insuline-like growth factors were involved in the pathophysiological process in children with AL.

Objective To study the relationship between DNA methylation and pathogenesis of childhood immune thrombocytopenic purpura (ITP) by examining the expression of DNA methyltransferase 1(Dnmt1) and DNA methyltransferase 3a (Dnmt3a) mRNA in peripheral blood lymphocytes of the children with ITP. Methods Expression of Dnmt 1 and Dnmt3a mRNA in the peripheral blood lymphocytes in 36 children with newly diagnosed ITP and 26 healthy children were detected using RT-PCR. Results Dnmt1 mRNA expression in peripheral blood lymphocytes in children diagnosed with ITP was 3.02±0.49, significantly lower than 4.58±0.52 in the control group (t=11.95, P<0.001). Dnmt3a mRNA expression in peripheral blood lymphocytes in children diagnosed with ITP was 1.49±0.44, signiifcantly lower than 2.41±0.32 in the control group (t=9.12, P<0.001). Conclusions Children with newly diagnosed ITP have lower DNA methylation status in peripheral blood lymphocytes as compared to that in healthy children. The DNA methylation may play an important role in the etiology of acute ITP in children.

Objective To discuss DNA methylation's effect on pathogenesis of pediatric immune thrombocytopenia (ITP)through detecting the expression level of DNA methyltransferases (DNMTs)mRNA in peripheral blood lymphocytes of children with ITP.Methods Two mL peripheral blood was collected from each of 25 children with persistent and chronic ITP and 20 healthy children (the healthy control group)by using aseptic method in the pediatric ward of the First Affiliated Hospital of Xinxiang Medical University from January 2014 to January 2015.First ethylene diamine tetraacetic acid (EDTA) was used as the anticoagulant.Then separate the mononuclear cells,extract RNA and detect expression levels of DNMT1,DNMT3A and DNMT3B mRNA using reverse transcription-polymerase chain reaction (RT-PCR) method.Results (1) The blood platelet (PLT) of children with persistent and chronic ITP was (36.2 ± 19.6) × 109/L,which was obviously lower than the healthy control group(168.8 ±46.8) × 109/L(t =-11.85,P =0.000).(2)The DNMT1 mRNA expression level of children with persistent and chronic ITP was 0.17 ± 0.05,which was obviously lower than the healthy control group (0.27 ± 0.10) (t =-3.912,P =0.001).The DNMT3A mRNA expression level of children with persistent and chronic ITP was 0.20 ± 0.10,which was obviously lower than the healthy control group (0.32 ±0.11) (t =-3.779,P =0.000).The DNMT3B mRNA expression level of children with persistent and chronic ITP was 0.16 ± 0.1 1,which was obviously lower than the healthy control group (0.31 ±0.11) (t =-4.641,P =0.000).(3) There was positive correlation between the expression of DNMT1 and DNMT3B mRNA(r =0.433,P =0.031).There was positive correlation between the expression of DNMT3A and DNMT3B mRNA(r =0.721,P =0.000).Conclusions (1) Children with persistent and chronic ITP have lower expression levels of DNMT1,DNMT3A,DNMT3 B mRNA,which indicates that DNA methylation contributes to the pathogenesis of pediatric persistent and chronic ITP.(2) DNMTs have synergistic effect on DNA methylation of pediatric persistent and chronic ITP.

Objective To analyze the functional changes and the clinical significance of B cell specific mono-clonal murine leukemia virus integration site -1 (Bmi -1 )and Th1 /Th2 cells in children with newly diagnosed im-mune thrombocytopenia(ITP)by testing the mRNA expressions of Bmi -1,helper T cell -related cytokine interferon (IFN)-γand interleukin(IL)-4 in children with newly diagnosed ITP.Methods Thirty -six cases of patients with newly diagnosed ITP in the experimental group came from the inpatient and outpatient children admitted to the Depart-ment of Pediatrics of the First Affiliated Hospital of Xinxiang Medical University from April to December 201 3.In the control group,26 cases of children requiring selective operation were admitted to the Department of Pediatric Surgery during the same period.The mRNA expressions of Bmi -1,IFN -γand IL -4 in the peripheral blood lymphocytes were detected by means of the reverse transcription -polymerase chain reaction(RT -PCR)method,and were analyzed and compared by t test and linear correlation analysis.Results (1 )The mRNA expressions of Bmi -1,IFN -γand IL -4 in peripheral blood lymphocytes in the experimental group were 2.63 ±0.54,3.84 ±0.43 and 1 .44 ±0.39,respec-tively;while the mRNA expressions of Bmi -1,IFN -γand IL -4 in the peripheral blood lymphocytes in the control group were 3.91 ±0.92,2.88 ±0.57 and 1 .87 ±0.34,respectively.The levels of IFN -γof the experimental group were significantly higher than those of the control group (P <0.001 )and the levels of Bmi -1 and IL -4 in the experi-mental group were significantly lower than those in the control group (all P <0.001 ).(2)The mRNA expressions be-tween IFN -γand IL -4 in the peripheral blood lymphocytes in the experimental group were in negative correlation (r =-0.667,P <0.001 ).The mRNA expressions between IL -4 and Bmi -1 in the same group were in a positive correlation (r =0.776,P <0.001 ).There were no correlation in the mRNA expressions between IFN -γand Bmi -1 (r =-0.206,P >0.05).Conclusions Bmi -1 may be involved in the pathogenesis of ITP by regulating Th cell, and Th cell dysfunction may occur in the children with ITP,and the disproportion between Th1 and Th2 may be due to the advantages of Th1 .

Objective:To detect the levels of cytokines in peripheral blood of patients with chronic immune thrombocytopenia (ITP) and analyze their significance in the clinical prognosis of children with chronic ITP.Methods:Thirty patients with chronic ITP who were treated in the Department of Pediatrics, the First Affiliated Hospital of Xinxiang Medical Univercity from October 2015 to October 2018 were followed and enrolled in the experimental group and 40 healthy children in the same hospital were enrolled in the healthy control group.The levels of interleukin-2(IL-2), interferon-γ(IFN-γ), tumor necrosis factor(TNF), interleukin-4(IL-4), interleukin-6(IL-6), interleukin-10(IL-10) and interleukin-17A(IL-17A) in the experimental group and the healthy control group were detected by flow cytometry (CBA). The relationship between cytokines and prognosis of children with chronic ITP were analyzed.Results:Thirty patients with ITP were enrolled. The expressions of IL-2 and IL-17A in the experimental group before treatment were (7.86±3.90) ng/L and (10.45±12.35) ng/L, while those of IL-2 and IL-17A in the healthy control group were (3.11±2.41) ng/L and (2.97±7.04) ng/L. The levels of IL-2 and IL-17A in the experimental group were significantly higher than those in the healthy control group, and the differences were statistically significant ( t=-7.123, -5.582, all P<0.01). The expressions of IL-4 and IL-10 in the experimental group before treatment were (0.38±0.25) ng/L and (1.80±1.25) ng/L, while those of IL-4 and IL-10 in the healthy control group were (3.08±0.26) ng/L and (4.55±3.44) ng/L. The levels of IL-4 and IL-10 in the experimental group were significantly lower than those in the healthy control group, and the differences were statistically significant ( t=8.400, 5.653, all P<0.01). The expressions of IL-6, TNF and IFN-γ in the experimental group before treatment were (7.30±9.16) ng/L, (4.85±7.60) ng/L and (7.68±20.41) ng/L, while those of IL-6, TNF and IFN-γ in the healthy control group were (5.44±4.18) ng/L, (1.97±0.37) ng/L, (4.81±17.71) ng/L. There was no significant difference between the two groups ( P>0.05), and no significant difference in the levels of cytokines between the patients with chronic ITP before and 12 months after treatment ( P>0.05). Conclusions:The changes of T lymphocyte related cytokines are closely related to the pathogenesis and development of chronic ITP in children. There may be persistent immune dysfunction in children with chronic ITP. Dynamic monitoring of cytokines IL-2, IL-4, IL-10, IL-17A, especially IL-17A, is helpful to judge the prognosis of ITP in children, and may be of guiding significance in evaluating clinical prognosis.

Objectives:To analyze the clinical features of juvenile myelomonocytic leukemia (JMML) and investigate the characteristics of diagnosis and treatment of this disease.Methods:The clinical data of seven children patients with JMML who received treatment in The First Affiliated Hospital of Xinxiang Medical University between April 2015 and February 2020 were retrospectively analyzed. The clinical efficacy of different treatments was analyzed.Results:The median age at diagnosis of JMML was 8 months and 4 days for seven children patients. Fever was the principal cause of treatment, and it was mostly accompanied by hepatosplenomegaly. The median value of peripheral blood leukocyte count was 36.1 × 10 9/L, and it was 4.5 × 10 9/L for mononuclear cell count, 88 g/L for hemoglobin level, and 47 × 10 9/L for platelet count. Myeloid immature cells were found in peripheral blood smears of six patients. Chromosome examination results revealed 7-monomer in one patient, and normal karyotype in six patients. Hemoglobin level was increased in six patients. Gene detection results revealed PTPN11+NF1 mutation in one patient, N-RAS mutation in two patients, and K-RAS mutation in one patient. Three patients gave up treatment, three patients received low-intensity chemotherapy , and these six patients died of complicated infection. One patient received allogeneic hematopoietic stem cell transplantation and the patient survived without any event after 14 months of follow-up. Conclusion:The age of JMML onset is low. JMML has poor clinical specificity. Gene detection is helpful for early diagnosis of JMML. Low-intensity chemotherapy can prolong survival period, but it can not improve prognosis. Infection is the principal cause of death in patients with JMML. Hematopoietic stem cell transplantation is the only possible method to cure the disease.