BACKGROUND:The CD4+CD25+FOXP3+Treg cells have immunosuppression effect, and it is speculated that these cells may restrain the occurrence of acute graft-versus-host disease. OBJECTIVE:To observe the variety of the CD4+CD25+FOXP3+Treg cells in the peripheral blood from donors before and after granulocyte colony stimulating factor mobilization, and study the relationship between CD4+CD25+FOXP3+Treg cells and acute graft-versus-host disease. METHODS:Ninety patients with malignant blood diseases who undertook al ogeneic hematopoietic stem celltransplantation and their donors were observed. Granulocyte colony stimulating factor 5μg/kg was injected subcutaneously into the donor per 12 hours for 5 days, and the stem cells were col ected before and after mobilization. The ratio of CD4+CD25+FOXP3+Treg cells in the peripheral blood was detected before and after mobilization with flow cytometry, and the ratio of these cells in the stem cellsuspension was measured by the same method. The patients were divided into two groups according to the CD4+CD25+FOXP3+Treg cells ratio:high dosage group (≥5%) and low dosage group (<5%). The incidence of acute graft-versus-host disease was observed in the two groups after transplantation. RESULTS AND CONCLUSION:The ratio of the CD4+CD25+FOXP3+Treg cells in the donor before and after mobilization were 11.3%and 1.5%,respectively, and there was a significant difference (P<0.05). The ratio of the CD4+CD25+FOXP3+Treg cells was 3.4%in the patients with acute graft-versus-host disease, and 15.7%in the patients with no acute graft-versus-host disease, showing a significant difference (P<0.05). After hematopoietic reconstitution, the incidence of acute graft-versus-host disease was 18.4%in the high dosage group and 48.1%in the low dosage group, and there was a significant difference between the two groups (P<0.05). Therefore, the granulocyte colony stimulating factor can lower the ratio of CD4+CD25+FOXP3+Treg cells in the human peripheral blood. The increase in CD4+CD25+FOXP3+Treg cells can restrain the occurrence of acute graft-versus-host disease.

BACKGROUND:The prognosis of malignant hematologic diseases has improved greatly with the application of the hematopoietic stem cell transplantation. Peripheral blood stem cell transplantation (PBSCT) has been used as an alternative to bone marrow transplantation (BMT).OBJECTIVE:To observe curative effect and clinical outcome in 53 patients with hematological malignancy undergoing allogeneic peripheral blood stem cell transplantation (allo-PBSCT) or autologous peripheral blood stem cell transplantation (auto-PBSCT).DESIGN:Randomized controlled study.SETTING:BMT Center, Hematology Department of the First Affiliated Hospital of Zhengzhou University.PARTICIPANTS:From July 2003 to May 2006, 53 patients (33 males and 20 females) with a median age of 37 years underwent PBSCT. Thirty-five patients received allo-PBSCT, including 13 of acute myelocytic leukemia (AML), 7 of acute lymphocytic leukemia (ALL), 10 of chronic myelocytic leukemia (CML), 2 of multiple myeloma (MM), and 3 of myelodysplastic syndrome (MDS). Eighteen patients underwent auto-PBSCT, including 7 AML, 6 ALL, 2 MM, and 3 non-Hodgkin lymphoma (NHL). Thirty-three donors (20 males and 13 females) with a median age of 35 age in the allo-PBSCT were HLA-identical siblings, 2 donors (5.7%) had one mismatch. Sixteen allografts were sex mismatched. Study was authorized by the Ethic committee of the hospital, and all patients had signed an inform consent.METHODS:① PBSC were mobilized with granulocyte colony-stimulating factor (G-CSF) or chemotherapy combined with G-CSF. A median of 6.2×106 CD34+ cells/kg was infused for allo-PBSCT and 3.0×106 CD34+ cells/kg was infused for auto-PBSCT. Amended BU/CY was used as conditioning regimen in allo-PBSCT and MAC was used in auto-PBSCT. Methotrexate (MTX) combined with cyclosporine A (CsA) and mycophenolate mofetil (MMF) was used as graft-versus-host disease (GVHD) prophylaxis. ALG was used in 1 patient with 1 locus mismatched in allo-PBSCT. ② Time to engraftment was calculated from the time of transplantation to neutrophil recovery ≥ 0.5×109 L-1 and platelet recovery ≥ 20×109 L-1, GVHD and relapse were observed until 1 year of follow-up.MAIN OUTCOME MEASURES:① Time to neutrophil and platelet recovery; ② GVHD occurrence after transplantation; ③ outcome of treatment.RESULTS:All the 53 patients were analyzed. ① Engraftment of neutrophils (> 0.5×109 L-1) and platelets (> 20×109 L-1) was achieved at a median of 13 days for neutrophils and 19 days for platelets in auto-PBSCT, and 12 days and 15 days respectively in allo-PBSCT. ② In allo-PBSCT, I-VI acute GVHD occurred in 31.4% cases, and chronic GVHD developed in 71.4% cases. ③ The relapse rate was 38.9% in auto-PBSCT, and 5.7% in allo-PBSCT. CONCLUSION:PBSCT can provide rapid hematopoietic reconstitution. It is an important method to cure the malignant hematologic diseases.

BACKGROUND:For pediatric patients with aplastic anemia in China, it is difficult to find human leucocyte antigen-matched sibling donors that are mostly replaced by parental donors.
OBJECTIVE:To retrospectively analyze the clinical efficacy and safety of parental haploidentical peripheral blood hematopoietic stem cel transplantation in children with relapsed and refractory severe aplastic anemia.
METHODS:Seventeen children with relapsed and refractory severe aplastic anemia who had no matched sibling or unrelated donor and failed to respond to immunosuppressive therapy were subjected to parental haploidentical peripheral blood hematopoietic stem cel transplantation. A conditioning regimen of fludarabine+cyclophosphamide+rabbit anti-human thymocyte immunoglobulin antibody and the triple therapy of methotrexate, cyclosporine A and mycophenolate mofetil were applied to prevent graft-versus-host disease.
RESULTS AND CONCLUSION: (1) Of the 17 children, 16 cases (94%) reached hematopoietic reconstitution, and the median time of neutrophils≥ 0.5×109/L and platelets≥ 20×109/L was 13 (11-15) days and 17 (12-28) days, respectively. (2) Incidence of acute graft-versus-host disease was 47% (8 of 17 cases), including 29% (5/17) of grades I-II and 18% (3/17) of grades III-IV. Incidence of chronic graft-versus-host disease was 41% (7/17). (3) With a median folow-up duration of 268 (43-753) days, the overal survival rate was 70.6% (12/17). Five dead cases (29%) belonged to transplantation-related death, including one case of fungal skin infections, one case of graft-versus-host disease, three cases of severe lung infection. No relapse case was reported. These findings indicate that if there are no matched sibling or unrelated donors and the immunosuppression effect is poor, parental haploidentical peripheral blood hematopoietic stem cel transplantation is a safe and effective salvage treatment for children with relapsed and refractory severe aplastic anemia.

Objective To investigate the effect of postremission consolidation therapy with intermedium-dose cytarabine (MDAC) in elderly patients with acute myelogenous leukemia (AML). Methods Clinical data of 61 elderly AML patients (except M3) in postremission who achieved complete remission (CR) in two period of remission induction program were retrospectively analyzed. Results There were 26 cases in MDAC group and 35 cases in standard-dose cytarabine (SDAC) group. In MDAC group and SDAC group, the relapse free survival (RFS) time were 42.7 months and 16.0 months respectively (P= 0.002), the overall survival (OS) time were 44.6 months and 18.2 months respectively (P= 0.004), and the cumulative relapse frequencies rates were 26.9 % (7/26) and 54.3 % (19/35) respectively (x 2= 4.567, P= 0.033). However, 3 years OS rate of the two groups were 23.1%(6/26) and 8.6%(3/35) (x 2=2.496, P=0.114) , and there was no significant difference in the incidence of adverse reactions between the two groups (all P > 0.05). Conclusion MDAC could improve RFS and OS for the elderly AML patients in postremission who received CR in the early stage, and the incidence of adverse reactions is similar to that of SDAC.

BACKGROUND:Cytokine induced kil er cells therapy as an effective means of adoptive immunotherapy, becomes a new way to treat acute myeloid leukemia. But, the researches about sequential cytokine induced kil er cells therapy after autologous peripheral blood stem celltransplantation in acute myeloid leukemia patients are stil less, which deserve further research.
OBJECTIVE:To observe the clinical efficiency and safety of sequential cytokine induced kil er cells therapy after autologous peripheral blood stem celltransplantation in acute myeloid leukemia M2 patients.
METHODS:Total y 45 patients with low-or intermediate-risk acute myeloid leukemia M2 were recruited in this study. Among them, 19 patients received sequential cytokine induced kil er cells therapy after autologous peripheral blood stem celltransplantation and 26 patients only received autologous peripheral blood stem celltransplantation. The relapse rate, disease-free survival, and overal survival were compared between two groups, and safety of cytokine induced kil er cells therapy was observed.
RESULTS AND CONCLUSION:(1) Compared with the patients only receiving autologous peripheral blood stem celltransplantation, the relapse rate was lower (21.05%vs. 38.46%;P<0.05), and elevated percentages of the disease-free survival and overal survival were observed in the patients receiving sequential cytokine induced kil er cells therapy after autologous peripheral blood stem celltransplantation (P<0.05). (2) The 19 patients who received sequential cytokine induced kil er cells therapy after autologous peripheral blood stem celltransplantation al completed the treatment scheme successful y. Only four patients appeared to have chil s and fever, and no more side effects were observed. These findings suggested that the sequential cytokine induced kil er cells therapy after autologous peripheral blood stem celltransplantation can improve the disease-free survival and overal survival of low-or intermediate-risk acute myeloid leukemia M2 patients without remarkable side effects, which is a safe, effective and feasible way for the treatment of acute myeloid leukemia M2.

Objective To analyze the clinical features and prognosis of CD34-positive and CD34-negative adult patients with acute T-lymphoblastic leukemia (T-ALL),and to explore the value of CD34 expression for prognosis of patients with T-ALL.Methods 75 adult patients diagnosed with T-ALL from January 2012 to July 2015 in the Department of Hematology,the First Affiliated Hospital of Zhengzhou University,were analyzed retrospectively.According to the expression of CD34,the patients were divided into CD34-positive group and CD34-negative group,and then the clinical characteristics and prognosis of both groups were analyzed.Results In 75 patients,CD34-positive group had 24 (32.0 ％) patients and CD34-negative group had 51 (68.0 ％) patients.Between the two groups,there was no significant difference in these factors,such as sex,age,infiltration of liver,spleen and lymph nodes,thrombocytopenia,high white blood cell count,abnormal karyotype,complete remission within 4 weeks and central nervous system leukemia (CNSL).The proportions of patients with hemoglobin (Hb) ＜ 90 g/L and expression of myeloid lineage marker were higher in the CD34-positive group than those in the CD34-negative group (x2 =5.888,P=0.015;x2 =10.758,P =0.001,respectively).There were only 18 patients treated with hematopoietic stem cell transplantation (HSCT),57 patients were not.In patients without HSCT,the median survival time in the CD34-positive group and CD34-negative group was significant different (5 months vs.32 months,x2 =9.172,P =0.002).Conclusions CD34 expression in adult patients with T-ALL appears to be associated with Hb ＜ 90 g/L and the expression of myeloid lineage markers.For the patients without HSCT,CD34 is likely negatively related with the prognosis.

Objective To investigate the efficacy and influencing factors of allo-HSCT in the treatment of MDS patients with ASXL1+.Methods The second-generation sequencing technique was used to detect 22 gene mutations in 247 newly diagnosed MDS patients in our hospital.The patients were divided into chemotherapy group and transplant group according to treatment style.The differences of OS and PFS between the two groups were compared,and the influencing factors of prognosis of transplant patients were analyzed.Results ASXL1+ was detected in 75 patients(30.36%),with a median mutation ratio of 42.93(18.10,58.39)%,10 received supportive treatment,43 received demethylation therapy or demethylation combined with pre-excitation therapy,and 22 received allo-HSCT.2-year PFS rate and OS rate of transplantation group were significantly higher than that of chemotherapy group(P<0.05).The 2-year OS rate in the low ASXL1 mutation load group(VAF≤42.93%)was significantly higher than that in the high ASXL1 mutation load group(VAF>42.93%)(P<0.05).In the context of allo-HSCT in patients with ASXL1+,2-year OS and PFS rates were significantly reduced in patients with RUNX1+ or ASXL1+(P<0.05);Multivariate analysis showed that high mutation load of ASXL1 or U2AF1+ were independent risk factors for OS in transplant patient(P<0.05).U2AF1+ were the risk factors for PFS(P<0.05).Conclusion allo-HSCT significantly improved the prognosis of patients with ASXL1+ MDS.High ASXL1 mutation load or U2AF1+ were independent risk factors affecting the outcome of allo-HSCT.

To analyze the clinical characteristics, diagnosis, treatment and outcome of patients with thrombotic thrombocytopenic purpura (TTP). The clinical data of 69 adult patients with TTP were retrospectively analyzed. There were 19 males and 50 females with a median age of 42 (18-79) years. PLASMIC score 6-7 was recognized in 82.8% (53/64) patients. The activity of von Willebrand factor-cleaving protease (ADAMTS13), which was detected in 21 patients before treatment, was less than 5% in 17 patients and 5%-10% in 3 patients. All 69 patients were treated with plasma exchange (PEX) and/or fresh frozen plasma infusion (PI), 43 of whom were also given glucocorticoid. In addition to PEX/PI and glucocorticoid, rituximab and/or immunosuppressants were administrated in 20 patients. The median follow-up time was 12 (1-57) months. The remission rate was 69.6%, while the relapse rate was 11.6%. The 2-year overall survival (OS) rate was 69.6%±5.5%. The univariate and multivariate analysis showed that relapsed/refractory disease was an independent risk factor for OS. The 2-year OS rate of relapsed/refractory patients was significantly lower than that of the rest patients (41.5%±9.8% vs. 83.7%±5.6%, P<0.001). Regarding the unfavorable prognosis in relapsed/refractory patients, rituximab and/or immunosuppressants are strongly recommended for sake of improving the overall survival.

Objective To analyze the clinical characteristics and prognostic factors of elderly patients with cytogenetically normal acute myeloid leukemia (CN-AML). Methods A total of 104 initial CN-AML patients were enrolled in this retrospective study. The clinical characteristics were collected and analyzed retrospectively. Factors affecting complete remission (CR) were analyzed by using chi square test. Univariate and multivariate analyses of prognostic factors were performed by using Kaplan-Meier and Cox hazard regression model respectively. Results After the first chemotherapy, 72 of 104 patients were able to be evaluated the efficacy, the CR rate was 38.9%(28/72) and total response rate was 55.6%(40/72). The white cell count<100 × 109/L and NPM1 mutation were related to a higher CR rate [59.4%(38/64) vs. 12.5%(1/8), 83.3%(10/12) vs. 36.4%(8/22), P<0.05]. Among 104 patients, the median overall survival (OS) was 6.9 months. Univariate analysis results demonstrated that age≥70 years, secondary AML, white cell count≥100×109/L, FLT3-ITD mutation, CD7 expression, achieving CR beyond 2 cycles of induction therapy and CCI score≥2 were influence factors on OS. In multivariable analysis, FLT3-ITD mutation (HR=7.61, 95%CI 1.80-32.11, P= 0.006) and achieving CR beyond 2 cycles of induction therapy (HR= 10.11, 95 % CI 2.38-43.03, P=0.002) were independent prognostic factors for OS in elderly patients with CN-AML. Conclusion The prognosis of elderly patients with CN-AML is the result of the combined effect of many factors, FLT3-ITD mutation and achieving CR beyond 2 cycles of induction therapy are independent prognostic factors in elderly patients with CN-AML.