Objective To investigate the effects of Basiliximab on alloreactive T lymphocytes precursors, hematopoietic progenitor cells by ex vivo assay and prophylaxis of graft-versus-host disease (GVHD) in mismatched bone marrow transplantation.Methods Two groups were set up: (1) Basiliximab group including 72 leukemia patients subject to haploidentical bone marrow transplantation (BMT) with Basiliximab for prophylaxis of GVHD without ex vivo T-cell depletion. (2) Control group having 15 patients receiving the same regimen before Nov. 2000, without Basiliximab for GVHD prophylaxis. Limiting-dilution method was used to determine the effect of Basiliximab on reactivity of cytotoxic T lymphocyte precursors (CTLP). In the semi-solid hematopoietic culture system, the effect of Basiliximab on the colony proliferation of CFU-GM, BFU-E and CFU-Meg was measured. Results In Basiliximab group, 72 patients established trilineage with full donor hematopoietic reconstitution. Engraftment rate showed no statistical difference between the two groups. The incidence of grade Ⅱ-Ⅳ GVHD was 12.5 % (9 cases) in Basiliximab group and 33.3 % (5 cases) in control group respectively (P

(0.05)).It was noted that the number of CD4~(+) T cells was less significantly throughout the 18 months after BMT in two groups.The time to reach 200 CD4~(+)cells/ ?l was 6 months,and that to reach normal number of CD4~(+)was 18 months.Median time to reach normal CD3~(+) CD8~(+) and CD19~(+) was 9-12 months,and there was no significant difference between two groups.Conclusions The incidence of severe lethal aGVHD and GVHD-related deaths tended to be less in patients with Basiliximab group than un-treated group in haploidentical BMT.It is useful to use Basiliximab to treat sever GVHD.CD4~(+) reconstitution appeared significantly delayed in two groups.CD4~(+) reconstitution is crucial to control post-transplant opportunistic infections and leukemia relapse.Nevertheless,there was no significant difference in immune reconstitution and the incidence of infection and relapse between the two groups.

20?10 9 /L was (18 1?3 0) days in the recipients.The incidences of grade I GVHD and II GVHD were 28 6% and 5 7%, respectively. Furthermore, the percentages of CD34 + and CD4 + cells in G CSF stimulated bone marrow increased, concomitant with a significant decline in CD8 + cell count. These data demonstrated that G CSF mobilization remodeled the density of hematopoietic progenitors and T lymphocyte subsets in the bone marrow, which in turn accelerated hematopoietic reconstitution and minimized the incidence of severe acute GVHD after allogeneic transplantation.Transplantation of marrow harvests from donors who are pretreated with G CSF might be a feasible measure for a successful allogeneic bone marrow transplantation.

Objective To explore the effects of Basiliximab (Simulect) on reducing the incidence of severe acute GVHD in haploidentical bone marrow transplantation (BMT). Methods Nine patients with leukemia received haplotype Allo-BMT from HLA two or three loci mismatched related donor. Most patients were classified as high risk category. The donors of patients were administrated with G-CSF 250 ?g/day for 7 doses prior to marrow harvest. In addition to combination of CsA, MTX, ATG and Mycophenolate mofetil for GVHD prophylaxis, Simulect was administered to prevent severe GVHD. A total 40 mg Simulect was given in two doses of 20 mg each by 30 min intravenous infusion on 2 h before transplantation and day 4 after transplantation.Results All patients were engrafted. 100 % donors hematopoietic cells after transplantation was determined by cytogenetic evidence analysis. None developed the Ⅱ-Ⅳ acute GVHD. Eight patients could be evaluated for chronic GVHD. All experienced chronic GVHD confined to the skin. The median follow-up duration was 14 months (range 12～20 months). One patient died from CMV infection on 3 months and one patient died from disease relapse on 14 months. The remaining 7 patients were survived in disease free situation.Conclusion The use of Simulect in haploidentical bone marrow transplantation is effective on preventing acute severe GVHD and improving disease-free survival.

OBJECTIVE To assess the clinical effectiveness of antibiotic combined therapy for febrile neutropenia as an empirical treatment.METHODS We analyzed bacterial epidemiology form Jan 2001 to Feb 2003 and performed a study in 202 neutropenic febrile patients after chemotherapy or(HSCT).Three groups were divided.In first group(84 cases) carbapenems and vancomycin were used.In second group(78 cases)and in third group(40(cases)) used cephalosporin or quinolone.RESULTS Carbapenems plus vancomycin were with response rate of 93%,and(without) vancomycin were only 66%.Cephalosporin or quinolone was with response rate only of 30%.(CONCLUSIONS) Strong antibiotic with vancomycin is effective for treating patients with neutropenia and fever(under) limited bacterial epidemiology.

In this study, the underlying antileukemic mechanisms of homoharringtonine (HHT) were investigated. K562 cell line was used to observe the effects of HHT on the induction of apoptosis and on the expression of the specific chimeric protein P210(bcr/abl), as evaluated by flow cytometric annexin V-PI dual labeling technique and Western blot. The results showed that HHT induced K562 cells to apoptotic death at the concentrations of 5 - 20 ng/ml, and some of the cells became necrotic when exposed to a higher concentration. The amount of P210(bcr/abl) oncoprotein was decreased by approximately 70% when the cells were exposed to HHT for 48 hours, however, that of its partner P145(c-abl) proto-oncoprotein was not affected. It is clear from the study that HHT is an inhibitor of P210(bcr/abl) oncoprotein and therefore promotes the apoptosis of CML cells. It could be promising that HHT be used extensively in the chemotherapy of patients with CML.

Objective To investigate early Epstein-Barr virus (EBV) reactivation and the outcome of preemptive therapy after allogeneic hematopoietic stem cell transplantation (allo-HSCT). Methods From January 2007 to January 2009, totally 277 patients after allo-HSCT were studied (haploidentical 116,unrelated 75, matched sibling 86). Conditioning regimens were mainly busulfan (BU) + cyclophosphamide ( CY)/fludarabine(Flu) or total body irradiation (TBI) + CY/Flu. Antihuman thymocyte globulin (ATG)was added in haploidentical and unrelated transplants. Plasma EBV DNA was monitored once to twice weekly in the first 3 months after allo-HSCT with real time quantitative polymerase chain reaction (RQ-PCR). EBV viremia was diagnosed when EBV DNA was more than 5 × 102 copies/ml but without symptoms. Acyclovir (10 mg/kg, intravenous drip, 8 h) was used for preemptive therapy and immnuo-suppressants were decreased if possible. Results Totally 33 patients ( 11.9% ) developed EBV viremia with a median time at day 44 (day 19 to day 84). The incidences of EBV viremia in the transplants from matched sibling,haploidentical, unrelated donors were 0, 15.5%, 20. 0%, respectively. There was no significant difference between haploidentical and unrelated transplants ( P = 0. 09 ), but much less EBV viremia was seen in matched sibling transplant ( P = 0. 001 ). Twenty of 33 patients ( 60. 6% ) had complete response to preemptive therapy. The median time to reach EBV DNA negative in plasma was 11 (4-56) d. The median duration of preemptive therapy was 21 (14-60) d. Both univariate and multivariate analysis indicated that haploidentical and unrelated transplants, acute graft versus host disease (GVHD) were the risk factors for EBV viremia. Two-year overall survival in the patients with EBV viremia was significantly lower than that without EBV viremia (54. 2% vs 72. 1%, P = 0. 006 ). Conclusions Our large clinical study has demonstrated that preemptive therapy with acyclovir that is guided by EBV viremia is effective in majority of the patients with high-risk for EBV reactivation after allo-HSCT, which may further decrease the risk for developing life-threatening EBV disease or post-transplantation lymphoproliferative disorder. Haploidentical and unrelated transplants, acute GVHD are the risk factors for EBV viremia which has negative impact on survival.

ObjectiveTo investigate the efficacy of haploidentical blood and marrow transplantation (haplo-BMT) in the treatment of advanced chronic myeloid leukemia (CML).MethodsFrom November 2002 to October 2007,35 patients with advanced CML received haplo-BMT.Eleven patients achieved the second chronic phase (CP2) after treatment with imatinib or chemotherapy or both before pre-conditioning,but there were 13 cases in accelerated phase (AP) and 11 patients in blast phase (BP) at the time of transplantation.By the last follow-up date October 31,2011,the median follow-up time among living patients was 67 months (range,49 to 100 months).ResultsThe cases of HLA-antigen mismatched between donors and recipients as 1,2,and 3 antigens were 1,12,and 22 respectively.The number of mean mononuclear cells and CD34+ cells was (7.19+ 1.37) × 108/kg and (2.54± 1.50) × 106/kg,respectively.All but one patient achieved durable hematopoietic reconstitution. Hyperacute graft-versus-host disease (GVHD) occurred in 28.6％ (10/35) patients.The cumulative incidence of grade Ⅱ to Ⅳ acute GVHD was 48％.Among 27 patients who survived longer than 100 days after transplant,16 (60 ％) had chronic GVHD.Fiveyear overall survival (OS) rate was 46.2％ and 45.5％ in CML-AP and BP (P =0.97),respectively.Five-year probability of OS rate was 81.8％,30.8％ and 27.3％ in patients with CML-CP2,CML-AP and BP at transplant,respectively.The OS of CML-CP2 was significantly higher than CML-AP and BP at transplant (P＜0.01 ).ConclusionHaplo-BMT is a feasible therapeutic mean for patients with advanced CML who have no matched donors available.It is better to perform haplo-BMT at CML-CP2 other than CML-AP or BP.

Objective: To investigate the effect of minimal residual disease (MRD) monitoring by multiparameter flow cytometry (MFC) pre-conditioning on prognosis of acute myeloid leukemia in first complete remission (CR₁-AML) after allogeneic hematopoietic stem cell transplantation (allo-HSCT) , and to explore the value of MRD monitoring by MFC in the prognosis evaluation on allo-HSCT in CR₁-AML. Methods: Between April 2012 and March 2015, consecutive 186 patients with CR₁-AML who underwent allo-HSCT were analyzed retrospectively. MRD in BM before conditioning was detected by eight-color MFC. Any level of residual disease was considered to be MRD positive. Results: ①Of 186 patients, MRD was negative in 151 patients, positive in 35 patients (<1% in 25 patients and 1% to 3% in 10 patients) . ② With the median follow up of 18 (5-41) months, two-year DFS was 80.0% (95%CI 68.5%-92.3%) . Univariate analysis showed that MRD positive patients had lower DFS[62.9% (95%CI 50.6%-75.2%) vs 88.9% (95%CI 76.6%-100.0%) , P<0.001], higher relapse[11.4% (95%CI 4.1%-29.0%) vs 3.3% (95% CI 0.6%-20.9%) , P=0.003] and higher NRM [25.7% (95% CI 8.1%-43.3%) vs 7.9% (95% CI 1.3%-26.5%) , P=0.001] after HSCT compared with that of MRD negative patients. Secondary AML showed lower DFS than primary AML [60.0% (95% CI 42.4%-76.6%) vs 86.0% (95% CI 68.4%-100.0%) , P=0.004]. ③Multivariate analysis indicated that MRD positive pre-HSCT was the independent risk factor on DFS [HR=4.565 (95%CI 2.918-9.482) , P<0.001], relapse [HR=5.854 (95%CI 1.538-22.288) , P=0.010] and NRM [HR=3.379 (95%CI 1.361-8.391) , P=0.009] after allo-HSCT in CR₁-AML. Conclusion MRD positive pre-conditioning was the only negative impact factor for patients with CR₁-AML after allo-HSCT. MRD by MFC can be used to assess the prognosis of CR₁-AML after allo-HSCT.

Objective: To analyze the effect of NCCN (2015) risk stratification on prognosis of patients with acute myeloid leukemia (AML) after allogeneic hematopoietic stem cell transplantation (allo-HSCT) . Methods: Retrospective analysis of 258 patients with AML in CR (186 cases in CR₁, 72 cases in CR₂) who underwent allogeneic HSCT in our hospital between April 2012 and March 2015 according to NCCN (2015) risk stratification. Of them, 63 cases were classified as low risk, 112 cases intermediate risk and 83 cases high risk. Results: ①With the median follow up of 18 (5-41) months, two-year disease free surviva (DFS) in 258 patients was 78.0% (95% CI 60.4%-96.6%) . Two-year DFS in AML after transplantation was 78.6% (95% CI 61.0%-96.2%) in low risk, 76.0% (95% CI 84.0%-93.6%) in intermediate risk and 80.3% (95% CI 62.7%-97.9%) (P=0.886) in high risk groups respectively. ②Univariate analysis showed that DFS has no significant difference in patient age, the median disease course before HSCT, the WBC number at the beginning of the disease, blood routine and chromosomes examination before transplantation, extramedullary disease before transplantation, disease status before transplantation, conditioning regimen, donor type, donor and recipient sex, recipient blood type, transfused MNC number, transfused CD34⁺ cell number and transfused CD3⁺ cell number. DFS was significant lower in primary AML than that in secondary AML (P=0.006) and also lower in MRD positive than that in MRD negative (P=0.003) . The accumulative relapse was significant higher in CR₂ compared to that in CR₁ (P=0.046) . Accumulative non-relapse mortality (NRM) was significanlyt higher in secondary AML compared to that in primary AML (P=0.004) and also higher in MRD positive compared to that in MRD negative (P=0.010) . ③Multivariate analysis showed that MRD positive was the only significant factor in DFS and NRM. Conclusion Allo-HSCT treatment of AML CR patients could achieve a high efficacy, which is similar between CR₁ and CR₂ patients. There is no significant correlation between NCCN (2015) risk stratification and the prognosis of AML patients with allo-HSCT treatment. Pre-conditioning MRD status monitored by multiparameter flow cytometry was the only impact factor on DFS and NRM in allo-HSCT for CR-AML patients.