A 7-Day Decitabine-Included Conditioning Regimen Accelerated Donor Hematopoietic Engraftment while Reduced the Occurrence of Mucositis without Interfering with Prognosis.

INTRODUCTION: Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is the standard and curative treatment strategy for patients with hematologic malignancies. Recently, decitabine-included regimens have been investigated by several studies including ours, which may prevent relapse of primary malignant diseases. METHODS: This study was to retrospectively evaluate a 7-day decitabine-included regimen with reduced dose of idarubicin for patients with hematologic malignancies who underwent allo-HSCT. RESULTS: A total of 84 patients were enrolled, including 24 cases in 7-day and 60 cases in 5-day decitabine groups, respectively. Patients conditioned with 7-day decitabine regimen showed accelerated neutrophil (12.05 ± 1.97 vs. 13.86 ± 3.15; u = 9.309, p < 0.001) and platelet (16.32 ± 6.27 vs. 21.37 ± 8.57; u = 8.887, p < 0.001) engraftment compared with those treated with 5-day decitabine regimen. Patients in the 7-day decitabine group showed a significantly lower incidence rate of total (50.00% [12/24] versus 78.33% [47/60]; χ2 = 6.583, p = 0.010) and grade III or above (4.17% [1/24] vs. 31.67% [19/60]; χ2 = 7.147, p = 0.008) oral mucositis compared to those in the 5-day decitabine group. However, the occurrence of other major complications post-allo-HSCT and outcomes of patients in these two groups were comparable. CONCLUSION: These results demonstrate that this 7-day decitabine-contained new conditioning regimen seems to be feasible and safe for patients with myeloid neoplasms who receive allo-HSCT, and a large-scale prospective study is needed to confirm the findings of this study.

Conditioning Regimen of 5-Day Decitabine Administration for Allogeneic Stem Cell Transplantation in Patients with Myelodysplastic Syndrome and Myeloproliferative Neoplasms.

Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is a potentially curative treatment for patients with myelodysplastic syndromes (MDS) and myelodysplastic/myeloproliferative neoplasms (MDS/MPN). However, post-HSCT relapse remains a major cause of treatment failure. Here we assessed the efficacy of a new conditioning regimen comprising decitabine (Dec), busulfan (Bu), cyclophosphamide (Cy), fludarabine (Flu), and cytarabine (Ara-c) for allo-HSCT in patients with MDS and MDS/MPN. A total of 48 patients were enrolled, including 44 with MDS and 4 with chronic myelomonocytic leukemia (CMML). Patients received Dec 20 mg/m2/day on days -9 to -5, combined with a Bu/Cy/Flu/Ara-c-modified preparative regimen. At a median follow-up of 522 days (range, 15 to 1313 days), the overall survival (OS) was 86%, relapse incidence was 12%, and nonrelapse mortality was 12%. The incidence of severe acute (grade III-IV) graft-versus-host disease (GVHD) was 23% and that of chronic GVHD was 15%. At 2 years, OS was 74% and 86%, respectively for high-risk and very-high-risk patients with MDS. Survival was promising in patients with poor-risk gene mutations, such as TP53 and ASXL1 (88%), and in those with ≥3 gene mutations (79%). Results of immunomonitoring studies revealed that proper natural killer cells made essential contributions to these favorable clinical outcomes. Overall, this new regimen was associated with a low relapse rate, low incidence and severity of GVHD, and satisfactory survival in allo-HSCT recipients with MDS and MDS/MPN.

[Relationship between Early Peak Temperature and Neutropenia Duration in Acute Leukemia Patients after Chemotherapy and Its Mechanism].

OBJECTIVE: To investigate the relationship between early peak body temperature and neutropenia duration and its potential mechanism. METHODS: A total of 111 patients with CR1 phase acute leukemia (AL) with neutropenia infection were enrolled in this study. The relationship between early peak body temperature and neutropenia duration was analyzed retrospectively, and the IL-6 serum level in patients with different peak of body temperature was detected, and the single cell culture system in vitro was established, the incorparation rate of EdU in vivo was detected, and the effect of IL-6 on mouse hematopoietic stem cells /progenitor cells was analyzed. RESULTS: Out of 111 patients with nentropenia, the body temperature <38 °C and the neutropenia duration 9.5±3.69 d were observed in 44 patients, while the body temperature >38 °C and neutropenia duration 7.33±4.20 d were observed in 69 patients, the differences between 2 groups was statistically signficant (P<0.05). The EdU test showed that the number of EdU+ hematopoietic stem cells and progenitor cells increased. The IL-6 level was different in patients with different peaks of initial bady temperature (P<0.05). The results of amimal experiment showed that the IL-6 could promote the proliferation of hematopoietic stem cells/ progenitor cells in vitro and in vivo. CONCLUSION: For patients with neutropenic infection when initial body temperature peak is <38 °C, the probability of neutropenia duration prolonging after chamotherapy increases, which may relate with promotive effect of pro-inflammatory cytokins on prliferation of hematopoietic stem cells/progenitor cells.

[Additional karyotypic abnormalities analysis in patients with hematological malignancies post-allogeneic hematopoietic stem cell transplantation].

OBJECTIVE: To analyze the karyotype stability in hematological malignancies patients before and after allogeneic hematopoietic stem cell transplantation (allo-HSCT) and its prognostic significance of monitoring. METHODS: The karyotypes and clinical data of 21 patients with hematological malignancies at the initial diagnosis and at relapse after allo-HSCT were retrospectively reviewed. Chromosome analysis was performed by standard 24 h-cultured method and R banding. RESULTS: Karyotypes at the initial diagnosis and at relapse after allo-HSCT were different in 11 patients (52.38%), including chromosome 1, 3, 6, 12, 17, 21. Numberical abnormalities and structural chromosomal abnormalities always occurs together. The median survival time of relapse of the patients with karyotype changes was significantly shorter than that of patients without a karyotype change (79 d vs 522 d, P = 0.027), and that of the patients with trisomy 6 was also significantly shorter than that of the patients without trisomy 6 (9 d vs 275 d, P = 0.005). CONCLUSION: Karyotype changes after relapse are associated with the prognosis of allo-HSCT.

[Preliminary analysis of therapeutic efficacy and prognosis of allogeneic hematopoietic stem cell transplantation in patients with advanced chronic myeloid leukemia].

Chronic myeloid leukemia (CML) at advanced and blastic phase is a disease with poor prognosis, for which allogeneic hematopoietic stem cell transplantation (allo-HSCT) is the only treatment choice with curative potential. This study was purposed to investigate the therapeutic efficacy of allo-HSCT and prognosis of advanced CML patients. The 28 cases of CML in accelerated phase or blast crisis received allo-HSCT were analysed retrospectively in terms curative efficacy, basic characteristics before transplant and prognosis, therapeutic strategy before transplant and prognosis, events after transplant and prognosis. The results indicated that 10 out of 28 patients were in complete remission, showing a 3-year overall survival and disease-free survival rate of 34.9% and 35.7% respectively; 18 patients died. Univariate analysis revealed that the clonal evolution and blast amount are baseline risk factor of poor prognosis, and combination of them can be used to predict the outcome of patients; application of imatinib before transplant and achievement of complete hematologic remission could not improve the prognosis; severe aGVHD among post-transplant events was proven to be a negative prognostic factor. It is concluded that for advanced CML patients received allo-HSCT, clonal evolution and blast percentage are prognostic factors, and the pre-transplant use of imatinib did not influence the outcome.

[Risk factors and prognosis of invasive fungal infections in patients with hematological diseases].

OBJECTIVE: To investigate the incidence, risk factors, prognosis and high risk patients of invasive fungal infections (IFI) in patients with hematological diseases. METHODS: : Over 2-week hospitalized patients from January 2007 to December 2008 were retrospectively reviewed. Logistic regression was used to analyze the risk factors of IFI, and recursive partitioning to reveal high risk patients. Incidence of IFI was estimated by cumulative incidence function, and the prognosis by Kaplan-Meier method. RESULTS: A total of 1048 assessable treatment cycles were recorded and 93 cases of IFI were diagnosed, with an incidence of 8.87 per 100 treatment cycles. Multivariate logistic regression revealed the following risk factors: age (OR 1.025, 95% CI 1.010-1.041, P = 0.002), duration of neutropenia (OR 1.028, 95% CI 1.014-1.042, P < 0.0001) and uncontrolled underlying diseases (OR 2.620, 95% CI 1.608-4.268, P = 0.0001). Recursive partitioning found two groups of high risk patients: (1) patients with uncontrolled underlying diseases and neutropenia duration > or = 58 days (7/12, 58.3%), (2) patients with uncontrolled underlying diseases and age > or = 33 years (40/208, 19.2%). At the end of follow-up, 111 cases of IFI were recorded in 451 patients, with a 1-year cumulative incidence of 27.1%. In patients with established IFI, overall survival rate and IFI related mortality rate at 12 weeks after diagnosis were 83.4% and 13.5% respectively. CONCLUSION: Age, duration of neutropenia and uncontrolled underlying diseases are risk factors of IFI; patients with uncontrolled underlying diseases and age > or = 33 years were at high risk of IFI and need major concern. IFI has a better prognosis and a lower related mortality in this study.

Secondary antifungal prophylaxis in hematological malignancies in a tertiary medical center.

To investigate the efficacy of secondary antifungal prophylaxis (SAP) in patients with hematological diseases, all medical records within two consecutive years were retrospectively reviewed. In all, 57 patients with a history of invasive fungal infections received 149 cycles of further therapy for their underlying hematological diseases. Logistic regression and recursive partitioning were used to discriminate high risk patients for failure of SAP. After a median follow-up period of 120 (12-1,080) days, 11 cases (7.4/100 cycles) experienced failure of SAP, including 5 cases during allo-SCT and 6 cases during chemotherapy, corresponding to cumulative incidence at 24.5%. Multivariate logistic regression revealed two risk factors for failure of SAP: use of high dose corticosteroid (OR 13.5, 95% CI 3.09-58.6, P = 0.0005) and duration of neutropenia ≥ 14 days (OR 7.47, 95% CI 1.69-32.9, P = 0.008). Recursive partitioning found that patients with these two risk factors were in high risk, with SAP failure rate as high as 50.0%. In conclusion, use of high dose corticosteroid and duration of neutropenia ≥ 14 days were risk factors of SAP failure. Patients with the two risk factors concurrently were in high risk and needed special concern.

[Allogeneic stem cell transplantation for 75 cases of acute myeloid leukemia in complete remission: outcome and prognostic analysis].

This study was purposed to evaluate the outcome of patients with acute myeloid leukemia (AML) who received allogeneic hematopoietic stem cell transplantation (allo-HSCT) in complete remission, and to study the prognostic factors. 75 cases of AML in complete remission receiving allo-HSCT from January 2000 to December 2007 were retrospectively analyzed. Major end points of study included overall survival (OS), disease free survival (DFS), relapse rate and transplantation related mortality (TRM). The results showed that 3-year OS and DFS of the study population reached to 58.4% and 53.9% respectively, and the relapse rate and TRM leaded to 16.9% and 29.9% respectively. Incidence of acute GVHD was 59.6%, with 18.7% II-IV aGVHD. Different prognosis was observed between HSCT recipients of alternative donor and HLA-matched related donor (MRD) (3-year DFS was 34.3% vs 60.0%, p = 0.019), between patients of refractory leukemia and the control (3-year DFS was 35.7% vs 58.2%, p = 0.048), between recipients with and without severe aGVHD (3-year DFS was 35.7% vs 54.4%, p = 0.059). Further analysis revealed significantly high TRM in recipients receiving allo-HSCT of alternative donor (p = 0.033) and high rate of severe aGVHD (p = 0.010). Multivariate analysis revealed three negative prognostic factors: donor availability (alternative vs MRD) (p = 0.049, RR = 2.09, 95%CI 1.01 - 4.36), refractory leukemia (p = 0.038, RR = 2.33, 95%CI 1.05 - 5.20) and severe aGVHD (p = 0.040, RR = 2.33, 95%CI 1.04 - 5.20). It is concluded that allo-HSCT is a choice for the AML case at complete remission and TRM is the major cause of the transplantation failure. Donor availability, refractory leukemia and severe aGVHD are confirmed as risk factors of poor prognosis for allo-HSCT patients with AML in CR.

[Effects of killer immunoglobulin-like receptor and human leukocyte antigen class I ligand on the prognosis of related donor hematopoietic stem cell transplantation].

OBJECTIVE: To study the genotype distribution and the effects of killer immunoglobulin-like receptors (KIR) and human leukocyte antigen (HLA) class I ligand on related donor hematopoietic stem cell transplantation (HSCT). METHODS: The genotypes of donor/recipient HLA-Cw and donor KIR were determined by polymerase chain reaction-sequence specific primer (PCR-SSP) in 87 cases of related donor HSCT (40 cases were haploidentical HSCT, and the remaining 47 cases were HLA-identical sibling HSCT). RESULTS: All the donors possessed KIR2DL1, 2DL2/L3, 2DL4, 3DL2, and 3DL3, and 96.6% of donors possessed 3DL1. The rate of activating KIRs varied. 97.7% of the recipients expressed C1, while the rates of C2, Bw4, and HLA-A3/A11 were different. In haploidentical HSCT, KIR-HLA-mismatched group included 34 cases and the matched group included 6 cases. HLA-HLA-mismatched group included 31 cases and the matched group included 9 cases. In matched sibling donor HSCT, KIR-HLA-mismatched group included 42 cases and the matched group included 5 cases. KIR-HLA-mismatched group had higher 2-year disease-free survival (DFS) rate compared with KIR-HLA-matched group [ (71.5 +/- 6.5 ) % vs. (50.0 +/- 10.7)%, P < 0.05]. CONCLUSIONS: The rate of activating KIR is lower than inhibitory KIRs. Inhibitory KIR2DL1, 3DL1, and 3DL2 may play key roles in the natural killer cell alloreactivity. The DFS rate is higher in KIR-HLA-mismatched group than in KIR-HLA-matched group in related donor HSCT.

[Expansion and cytokine secretion profile of human valpha24(+) NKT cells from different sources].

This study was purposed to investigate the phenotype, in vitro expansion and cytokine secretion profile of Valpha24(+) NKT cells from cord blood (CB), peripheral blood (PB), and granulocyte colony stimulating factor-mobilized peripheral blood mononuclear cells (G-PBMNCs). Fresh mononuclear cells (MNCs) were isolated by the method of gradient centrifugation and then cultured with alpha-GalCer (100 ng/ml), IL-2 (50 U/ml), IL-15 (50 ng/ml) for 12 days. Valpha24(+) NKT cells were purified by anti-Vbeta11 TCR McAb and goat anti-mouse IgG magnetic beads. The phenotype and purity of Valpha24(+) NKT cells were determined by flow cytometry. Cytokine production was analyzed by ELISA. The results showed that Valpha24(+) NKT cells in CB, PB and G-PBMNCs were expanded by 221.5 (95 - 501), 456.5 (101 - 2207), and 756.38 (82 - 20373)-fold respectively. After stimulation by phorbol-12-myristate-13-acetate (PMA) for 24 hours, IL-4 and IFN-gamma produced by Valpha24(+) NKT cells from CB and PB were 180.33 (144.67 - 2253.48) vs 190.67 (110.07 - 6060.16) ng/ml, 864.33 (401.33 - 3386.67) vs 508.49 (253.82 - 8840.00) ng/ml respectively, with IL-4/IFN-gamma ratio of 0.503 +/- 0.642 vs 0.455 +/- 0.562 respectively. After expansion of Valpha24(+) NKT cells from G-PBMNCs, IL-4 and IFN-gamma produced by Valpha24(+) NKT cells at day 9 and day 12 were 139.08 (7.62 - 606) vs 89.3 (0 - 729.2) ng/ml, 14264.8 (1168 - 18059) vs 14488 (1041 - 18261) ng/ml respectively, with IL-4/IFN-gamma ratio of 0.0531 +/- 0.1081 vs 0.0376 +/- 0.1148 respectively. It is concluded that in presence of IL-2 and IL-15, alpha-GalCer can facilitate the rapid short-term expansion of Valpha24(+) NKT cells from CB, PB, and G-PBMNCs. Valpha24(+) NKT cells from G-PBMNCs show much high potential of expansion in comparison to the counterparts from CB or PB (p < 0.05). The activated Valpha24(+) NKT cells can secrete IFN-gamma and IL-4 in large amounts, with IFN-gamma in particular.

[Ameliorative effect of rat bone marrow mesenchymal stem cell transplantation on infracted heart function].

This study was purposed to investigate the effects of rat marrow mesenchymal stem cell (rMSC) transplantation on left ventricular (LV) function in a rat myocardial infarction model. Myocardial infarction was performed in male Lewis rats by ligating the proximal left coronary artery. Rats were randomly divided into 3 groups: sham operation group (only thoracotomy, n = 8), AMI group (DF12 injection, n = 10), rMSC group (Dil-Labeled rMSC transplantation). At 8 weeks later, the cardiac functions including left ventricular ejection fraction (LVEF), left ventricular end systolic pressure (LVESP), left ventricular end diastolic pressure (LVEDP), +dp/dtmax and -dp/dtmax were evaluated by echocardiography and cardiac catheterization. The presence and differentiation of engrafted cells were assessed. CD31 was detected by immunohistochemical staining to demonstrate neovascular formation. The results indicated that the cultured in vitro rMSC expressed CD90, CD44, CD105, CD54; did not express CD34, CD45, CD31, as compared with AMI group, rMSC group showed a significant increase of LVEF, LVESP, +dp/dtmax, -dp/dtmax and a significant decrease of LVEDP. Immunofluorescence demonstrated that some transplanted rMSCs were positive for myosin, suggesting that small number of transplanted rMSCs differentiated into cardiac-like cells. Immunostaining showed marked augmentation of capillary density in the rMSC group than that of AMI group. It is concluded that transplanted rMSCs can differentiate into cardiac-like cells and rMSC transplantation can improve LV function after myocardial infarction in rats.

[Differentiation of bone marrow derived from mesenchymal stem cells into cardiomyocyte-like cells induced by co-culture with rat myocardial cells].

The study was purposed to investigate the differentiation ability of mesenchymal stem cells (MSCs) into myocardial cells in vitro. Rat bone marrow-derived MSCs were labeled and co-cultured with neonatal rat cardiomyocytes (CM) for 5 - 7 days. The expression of cell surface antigens was detected by flow cytometry, and the expression of muscle-specific marker myosin and troponin T in labeled cells was detected by immunofluorescence. The results showed that in vitro cultured MSCs expressed CD90, CD44, CD105, CD54, not expressed CD34, CD45, CD31. After co-cultured with neonatal rat CM, labeled MSCs differentiated into cardiomyocyte-like cells expressing myosin and troponin T. It is concluded that MSCs can differentiate into cardiomyocyte-like cells when co-cultured with neonatal myocardial cells in vitro. In co-culture of two kind of cells in ratio of four to one showed obvious efficacy differentiating MSCs into CMs.

[Impact of donor KIR2DS5 genotype on outcome following haploidentical hematopoietic stem cell transplantation].

The aim of this study was to evaluate the impact of donor killer cell immunoglobulin-like receptor (KIR) and recipient HLA genotypes on outcome following haploidentical hematopoietic stem cell transplantation (HSCT). 26 patients with hematologic diseases received non T-cell-depleted (TCD) in vitro transplant from haploidentical donor. Donor/recipient HLA and donor KIR genotypes were determined by polymerase chain reaction-sequence-specific primer (PCR-SSP). Donor/recipient KIR/HLA subgroup was assessed by donors KIR and recipients HLA-Bw4, Cw1 group and Cw2 group alleles. Hematopoietic reconstitution, incidence of graft versus host disease (GVHD), disease-free survival (DFS), infection and transplant-related mortality (TRM) were analyzed between every two groups. The influence of donor activating KIR on outcome following haploidentical HSCT also has been studied. The results showed that hematopoietic reconstitution, incidence of GVHD, DFS, infection and TRM were not significantly different between every two groups (p>0.05). There were 4 cases of severe GVHD in C2 mismatched group. The donor activating KIR2DS5 positive group had higher 2-year DFS compared with the negative group [(85.7+/-13.2)% vs (31.2+/-12.8)%, p<0.05]. It is concluded that KIR/HLA genotypes between donor and recipient influence the outcome following haploidentical HSCT. Donor activating KIR2DS5 may improve DFS in non TCD haploidentical HSCT.

[Allogeneic hematopoietic stem cell transplantation for 16 patients with aplastic anemia].

OBJECTIVE: To evaluate the efficacy of allogeneic hematopoietic stem cell transplantation (allo-HSCT) for aplastic anemia (AA). METHODS: Twelve patients with severe AA (SAA) and 4 with chronic AA (CAA) received allo- HSCT. The effectiveness and complication were analyzed retrospectively. RESULTS: Hematopoiesis reconstitution was achieved 14 patients (87.50%). The median time of neutrophils reached to 0.5 x 10(9)/L and platelets reached to 20 x 10(9)/L were 14 (11 - 16) and 14 (10 - 33) days, respectively. Six cases developed grade I - II acute graft-versus-host disease (aGVHD), chronic local GVHD occurred in 2 patients. Graft rejection occurred in 3 cases. Thirteen cases survived with a median of 10 (0.5 - 84) months at the end of follow-up. Three cases died of un-engraftment, graft rejection (GR) and interstitial pneumonia (IP) each. CONCLUSION: Allo-HSCT is an effective therapy for patients with AA. Enhancing immunosuppressive treatment for conditioning and GVHD prophylaxis may reduce the incidence of GR and GVHD.