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Abstract: Philadelphia chromosome (Ph)/BCR‐ABL‐positive (ph+) ALL is the most common genetic abnormality associated with ALL and has been shown to confer the worst prognosis to both children and adults. Increasing evidence has revealed that the dysregulation of prolyl isomerase Pin 1 contributes to multicancer development and progression, including ALL, although the underlying molecular mechanisms remain unclear. Here, we report that the expression of Pin 1 was enhanced in ph+ ALL patient samples and was associated positively with the expression of BCR‐ABL. Genetically or pharmacologically inhibiting Pin 1 expression or activity produces potent therapeutic efficacy against ph+ ALL. We further demonstrated that BCR‐ABL enhances the prolyl isomerase activity of Pin 1 by decreasing the phosphorylated level of Pin 1 at Ser 71 and interacting with DAPK1. The inhibition of BCR‐ABL activity by imatinib in human ph+ ALL cells reduces the prolyl isomerase activity of Pin 1, further suggesting a key role of the newly identified BCR‐ABL‐Pin 1 axis in ph+ ALL progression. Thus, the combined suppression of Pin 1 and BCR‐ABL proteins may be exploited as an additional target therapy for ph+ ALL. [ABSTRACT FROM AUTHOR]

Objective: To analyze the characteristics and prognosis of acute leukemia(AL) with SET-NUP214 fusion gene., Methods: The clinical data of 17 patients over 14 years old newly diagnosed with SET-NUP214 positive AL admitted in Institute of Hematology and Blood Diseases Hospital from August 2017 to May 2021 were analyzed retrospectively., Results: Among the 17 SET-NUP214 positive patients, 13 cases were diagnosed as T-ALL (ETP 3 cases, Pro-T-ALL 6 cases, Pre-T-ALL 3 cases, Medullary-T-ALL 1 case), AML 3 cases (2 cases M5, 1 case M0) and ALAL 1 case. Thirteen patients presented extramedullary infiltration at initial diagnosis. All 17 patients received treatment, and a total of 16 cases achieved complete remission (CR), including 12 cases in patients with T-ALL. The total median OS and RFS time were 23 (3-50) months and 21 (0-48) months, respectively. Eleven patients received allogeneic hematopoietic stem cell transplantation(allo-HSCT), with median OS time of 37.5 (5-50) months and median RFS time of 29.5 (5-48) months. The median OS time of 6 patients in chemotherapy-only group was 10.5 (3-41) months, and median RFS time of 6.5 (3-39) months. The OS and RFS of patients with transplantation group were better than those of chemotherapy-only group ( P =0.038). Among the 4 patients who relapsed or refractory after allo-HSCT, the SET-NUP214 fusion gene did not turn negative before transplantation. While, in the group of 7 patients who have not relapsed after allo-HSCT till now, the SET-NUP214 fusion gene expression of 5 patients turned negative before transplantation and other 2 of them were still positive., Conclusion: The fusion site of SET-NUP214 fusion gene is relatively fixed in AL patients, often accompanied by extramedullary infiltration. The chemotherapy effect of this disease is poor, and allo-HSCT may improve its prognosis.

Objective: To explore the relationship between occurrence of acute graft-versus-host disease (aGVHD) and various immune cell composition in patients with acute myeloid leukemia (AML) after allogeneic hematopoietic stem cell transplantation (allo-HSCT)., Methods: The clinical data of 104 patients with AML undergoing allo-HSCT in our hospital were retrospectively analyzed, and the hematopoietic reconstitution and occurrence of GVHD were analyzed. Flow cytometry was used to detect the proportion of various types of immune cells in the grafts, the number of graft composition in patients with different degrees of aGVHD was calculated and compared, and to analyze the correlation between the severity of aGVHD in AML patients after allo-HSCT and the immune cell components in the graft., Results: There was no significant difference in the time of hematopoietic reconstitution between the high number group of total number of nucleated cells (TNC) and the low number group, while the time of neutrophil and platelet reconstruction in the high number of CD34 group was significantly faster than that in the low number of CD34 group (P＜0.05), and the total hospital stay also tends to be shorten. Compared with patients in 0-Ι aGVHD group, both HLA-matched and HLA-haploidentical transplantation, the infusion amounts of CD3 + cells, CD3 + CD4 + cells, CD3 + CD8 + cells, NK cells and CD14 + monocytes were higher in patients of Ⅱ-Ⅳ aGVHD group， but the difference was not statistically significant ( P >0.05); In addition, in patients with HLA-haploidentical transplantation, the number of CD4 + CD25 + cells in Ⅱ-Ⅳ aGVHD group was significantly lower than that in 0-Ι aGVHD group (P<0.05), and the same trend was also observed in HLA-matched transplanted patients, but the difference was not significant ( P =0.078)., Conclusion: High number of CD34 + cells in the graft is beneficial to hematopoietic reconstitution in AML patients. To a certain degree, high number of CD3 + cells, CD3 + CD4 + cells, CD3 + CD8 + cells, NK cells and CD14 + cells tend to increase the occurrence of aGVHD, but high number of CD4 + CD25 + regulatory T cells is beneficial to reduce the incidence of aGVHD in AML patients.

Objective To analyze the differences in biological functions between bone marrow(BM)-derived CD106 + mesenchymal stem cells(MSCs)and the CD106 - subgroup. Methods The MSCs from normal BM were isolated and expanded.The subgroups of CD106 + and CD106 - MSCs were sorted.The cell proliferation and adhesion functions,chemotactic activities,adipogenic and osteogenic potentials,senescence,and senescence protein 21(p21)were detected.The capacity of translocation into nucleus of nuclear factor-kappa B(NF-κB)when stimulated by tumor necrosis factor(TNF-α)was measured. Results The proliferative ability was higher in CD106 + MSCs than that in CD106 - MSCs.In 48 hours,the value of optical density(OD)was significantly higher in CD106 + MSCs than that in CD106 - subgroup(1.004±0.028 vs. 0.659±0.023, t =3.946, P =0.0225).In 72 hours,this phenomenon was even more pronounced(2.574±0.089 vs. 1.590±0.074, t =11.240, P =0.0000).The adhesive capacity of CD106 + MSCs was significantly stronger than that of CD106 - subgroup(0.648±0.018 vs. 0.418±0.023, t =7.869, P =0.0002).Besides,the metastasis ability of CD106 + MSCs were significantly stronger than that of CD106 - subgroup(114.500±4.481 vs. 71.000±4.435, t =6.900, P =0.0005).The CD106 + MSCs had signifcnatly lower proportions of senescent cells.The expression of aging protein p21 in CD106 + MSCs was significantly lower than that in CD106 - MSCs [(17.560±1.421)% vs. (45.800±2.569)%, t =9.618, P =0.0000].Furthermore,there were no visible pigmenting cells after β-galactosidase staining in CD106 + MSCs subgroup.However,in CD106 - MSCs,some colored green cells were detected.The rate of NF-κB translocation into nucleus after stimulated by TNF-α was significantly higher in CD106 + MSCs than CD106 - MSCs [(37.780±3.268)% vs. (7.30±1.25)%, t =8.713, P =0.0001]. Conclusion Bone marrow-derived CD106 + MSCs possess more powerful biological functions than CD106 - MSCs.

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.

Objective: To identify the role of serum cytokine levels prior allogeneic hematopoietic stem cell trans-plantation (allo-HSCT) in the outcome of severe aplastic anemia (SAA) patients received allo-HSCT treatment., Methods: The clinical data of 117 SAA patients received allo-HSCT were enrolled in this study. The overall survival (OS), graft versus host disease (GVHD) incidence and relationship of serum cytokines with OS and major transplantation complications were retrospectively analyzed., Results: The patients enrolled in this study included 78(66.7%) cases received HSCT matched sibling donors (MSD), 12(10.2%) HSCT of unrelated donors (MUD) and 27 cases received HSCT of haploidentical donors (HID). The 5-years OS was 76.0%(95% CI: 64.4-87.5%); aGVHD cumulative incidence was 49.6%(95% CI: 40.4%-58.8%) and cumulative incidence cGVHD was 31.6%(95% CI:23.1%-40.2%). MSD allo-HSCT had a significantly higher 5-years OS as compared with the other donors(82.3%±6.6% vs 61.3%±11.7%, P<0.05). HLA matching, donor's age, cytomegalovirus/ Epstein-Barr virus (CMV/EBV) infection were important factors of affecting occurence of aGVHD. The patients with higher serum IL-6 had reduced platelet recovery time after transplantation (14.6±1.8 vs 18.3±2.6 d)(P=0.050) and higher serum TNF-α level accompanied by a lower incidence of CMV/EBV infection (37.8%±11.1% vs 58.8±16.8%)(P<0.05)., Conclusion: MSD allo-HSCT is the effective treatment for SAA patients. Donor's type remains the strong predictor of survival. The serum levels IL-6 and TNF-α before transplantation associate with platelet recovery and CMV/EBV infection.

Objective: To investigate the effect of microRNA-382 (miR-382) on the biological properties of human umbilical cord-derived mesenchymal stem cells (hUC-MSC)., Methods: The mimics and inhibitor of miR-382 were transfected into hUC-MSC with lipo2000. Inverted microscopy was used to observe the morphology change of hUC-MSC. The proliferation of hUC-MSC was detected by CCK-8. Oil red O and alizarin red staining were applied to assess the adipogenic and osteogenic differentiation of hUC-MSC. Cetylpyridinium chloride was used to the quantitative analysis of osteogenic differentiation. The expression of Runx2 and some cytokines were detected by RT-PCR., Results: miR-382 did not influence the morphology, proliferation and adipogenic differentiation of hUC-MSC miR-382 inhibited the expression of Runx2, thus could inhibit the osteogenesis of hUC-MSC, being confirmed by alizarin red stain; miR-382 could influence the expression of key cytokines secreted from hUC-MSC, such as IL-6, IDO1, G-CSF, M-CSF, GM-CSF., Conclusion: miR-382 decreases the expression of Runx2 and inhibites the osteogenesis of hUC-MSC. In addition, it also affects the expression of some key cytokines secreted from hUC-MSC.

Objective: To investigate the risk factors of intestinal acute graft-versus-host disease (aGVHD) after allogeneic hematopoietic stem cell transplantation (allo-HSCT)., Methods: The clinical data of 534 cases of 533 patients undergoing allo-HSCT during Jan 2004 and Sep 2012 were retrospectively analyzed. The effects of donor-recipient HLA mismatching, recipient age, donor age, donor-recipient sex combination, donor-recipient relationship, HSC source, conditioning regimen with or without total body irradiation (TBI) and HLA loci on intestinal aGVHD with different severity were analyzed by Logistic regression., Results: Intestinal aGVHD occurred in 123(23.0%) cases, with 86(16.1%) cases of stage 1 intestinal aGVHD(16.1%) and 37(6.9%) cases of stage 2 to 4 intestinal aGVHD. Multivariate analysis showed that donor-recipient HLA mismatching (OR=2.519, P=0.002), increasing donor age (OR=1.034, P=0.003), female donor for male recipient (OR=1.855, P=0.007) were risk factors for intestinal aGVHD, HLA-B38 (OR=0.256, P=0.032) was its protective factor. Donor-recipient HLA mismatching (OR=2.799, P=0.011), increasing donor age (OR=1.045, P=0.012), HLA-A1 (OR=4.157, P=0.002), A30 (OR=3.143, P=0.005) were risk factors for stage 2 to 4 intestinal aGVHD., Conclusion: Occurrence of intestinal aGVHD and its severity are associated with donor-recipient HLA mismatching, donor age, donor-recipient sex relationships and some HLA loci.

Objective: To evaluate the efficacy and safety of antifungal prophylaxis of itraconazole in patients with acute myeloid leukemia (AML) to probe the relationship of the antifungal effect and the adverse events with serum concentration., Methods: From April 2009 to May 2011, a total of 310 courses from 112 patients referred to our institute were enrolled in this study; of them, 297 courses were eligible for analysis. Eligible cases were randomized into oral group and injection/oral group according to different chemotherapy of induction and consolidation. Blood samples were collected at different time points for measurements of serum itraconazole levels. The morbidity of IFI and the adverse events were analyzed., Results: The morbidities of IFI in injection/oral and oral groups were 10.1% and 20.9%, respectively (P=0.010). 7 and 9 cases in injection/oral and oral groups, respectively were withdrawn from the study because of adverse events, and the difference between these two groups was of no significance. Serum itraconazole levels of injection/oral and oral groups were 672(299-1097) μg/L and 534(210-936) μg/L, respectively (P<0.01)., Conclusion: Antifungal prophylaxis with itraconazole in AML patients was effective and safe. Prophylactic effect with injection/oral itraconazole was superior to oral itraconazole solution; moreover, prophylactic effect of itraconazole was highly correlated with its serum level.

Objective: This study was aimed to observe the efficacy of autologous stem cell transplantation (ASCT) for adult patients with acute lymphoblastic leukemia (ALL), and investigate related prognostic factors., Methods: A total of 86 adult ALL patients underwent ASCT in Institute of Hematology and Blood Disease Hospital from November 2001 to January 2012 were followed up. Clinical characteristics and outcomes of all patients were retrospectively analyzed. Survival and univariate prognosis were analyzed by the Kaplan-Meier method and multivariate analysis by COX regression model., Results: Outcomes were assessed in 81 cases, including 47 standard-risk and 34 high-risk patients. 1-, 3-, 5-, and 10-year leukemia-free survival (LFS) of standard-risk patients were (82.3±5.7)%, (76.9±6.5)%, (74.1±6.8)%, (67.4±8.9)% respectively,and relapse rates (RR) were as of (13.6±5.2)%, (21.6±6.4)%, (24.5±6.8)%, (31.3±9.0)% respectively. 1-, 3-, 5-, and 10-year LFS of high-risk patients were (55.8±8.9)%, (39.8±9.3)%, (39.8±9.3)%, (39.8±9.3)% respectively, and relapse rates (RR) were (38.8±9.2)%, (56.4±10.0)%, (56.4±10.0)%, (56.4±10.0)% respectively. T-ALL, white blood cell count(WBC) more than 30×109/L when first visited, increased LDH, positive fusion gene of TCR and bone marrow transplantation were the adverse prognostic factors. Multivariate analysis showed bone marrow transplantation was an independent adverse prognostic factor., Conclusion: ASCT was a choice for adult ALL patients when suitable donors were unavailable.

Objective: To investigate mesenchymal stem cells (MSCs) immunosuppressive activity in the presence of interferon-gamma (IFN-γ) to reveal synergistic immunomodulatory effects of IFN-γ and MSCs., Methods: ① MSCs were cultured in the presence or absence of IFN-γ（100 ng/ml）, the supernatants were collected for measurements of PGE2、HGF and TGF-β1 by ELISA kits. ② MSCs were cultured in the presence or absence of IFN-γ （100 ng/ml）for 48 h. The cDNA was analysed for the expression of human indoleamine 2, 3-dioxygenase（IDO）mRNA by semiquantitative RT-PCR. ③ Mononuclear cells (MNCs) were extracted from peripheral blood of healthy donors. The T cell proliferation was tested in the co-culture system added with MSCs, recombinant human IFN-γ (100 ng/ml) and anti-IFN-γ mAb (5 μg/ml) by BrdU ELISA kit., Results: ①The immunosuppressive cytokines PGE2、HGF and TGF-β1 were detectable within 24-48 h in the supernatants. Their expressions were significantly up-regulated in the presence of IFN-γ. Concentrations of these cytokines were as of (1715.5±628.6) pg/ml vs (1344.5±709.4) pg/ml (P=0.001);(4031.8±1496.8) pg/ml vs (2452.4±1375.3) pg/ml（P=0.011）;(1753.5±413.8) pg/ml vs (1026.6±450.5) pg/ml（P<0.001）,respectively. ②The expression of IDO mRNA was undetectable when MSCs were cultured alone. In contrast, The IDO mRNA expression was remarkably enhanced in the presence of IFN-γ. ③Bone marrow-derived MSCs remarkably suppressed allogeneic T cell proliferation in vitro. Addition of exogenous IFN-γ had no significant effect on the inhibitory capacity of MSCs, the inhibitory ratios of T cell proliferation were （40.4±10.9）% vs（36.7±7.4）% (P=0.272). By contrast, the inhibitory ratio of T cell proliferation was significantly decreased in the presence of anti-IFN-γ mAb[(40.4±10.9)% vs (23.9±7.6)%,P=0.002]., Conclusion: ①Human MSCs constitutively expressed immunosuppressive concentrations of PGE2, HGF and TGF-β1, and their expressions were significantly up-regulated by IFN-γ. ②IFN-γ-induced expression of IDO on MSCs involved in tryptophan catabolism. ③MSCs notably suppressed allogeneic T cell proliferation in vitro. IFN-γ promoted the immunosuppressive capacity of human MSCs, indicating the synergistic immunomodulatory effect of IFN-γ and MSCs.

Background: The alloreactivity of natural killer cell and certain subsets of T lymphocyte are regulated by the interaction between killer immunoglobulin-like receptors (KIRs) of donor cells and human leukocyte antigen (HLA)-class I molecules on target cells. The interaction has been shown to influence the outcome of allogeneic haematopoietic stem cell transplantation (HSCT). Homozygous C1 or C2 and heterozygous C1/C2 were divided by HLA-Cw typing and they influenced the outcome of HSCT., Objective: The purpose of the study was to analyse the impact of interaction between recipient HLA-Cw and donor KIR on outcome., Methods: The genotypes of recipient HLA-Cw ligands and donor KIRs were correlated with the clinical outcomes of 52 patients who received HLA-matched, sibling donor HSCT for myeloid malignancies., Results: The incidence of chronic graft versus host disease (GVHD) was significantly lower in C1 or C2 homozygotes than in C1/C2 heterozygotes (p = 0.000). Higher overall survival (OS) and disease-free survival (DFS) rates were observed in C1 or C2 homozygotes than in C1/C2 heterozygotes (OS, 81% ± 8% vs 54% ± 10%, p = 0.034; DFS, 81% ± 8% vs 54% ± 10%, p = 0.024). A lower incidence of chronic GVHD and higher OS and DFS were observed in the HLA-KIR mismatched group (chronic GVHD, p = 0.007; OS, 84% ± 7% vs 47% ± 13%, p = 0.003; DFS, 84% ± 7% vs 47% ± 13%, p = 0.002)., Conclusion: The interaction between recipient HLA ligand and donor KIR had a significant impact on the outcome of patients receiving matched sibling HSCT. C1/C2 heterozygotes or HLA-KIR matched patients may benefit from additional intensified therapy with better outcome.

Objective: To explore the impact of IFN-γ + 874 polymorphisms on the outcome in HLA matched sibling HSCT., Methods: We used PCR-sequence-specific primer analysis (PCR-SSP) to analyze the polymorphisms of IFN-γ + 874 T/A in 80 recipient and donor pairs from October 2005 to March 2008., Results: Recipients having donors who possessed IFN-γ + 874 A/A genotype had significantly earlier neutrophil recovery compared with those having donors with non-A/A genotype (15 (11 - 27) days vs 18 (12 - 30) days, P = 0.029). And IFN-γ + 874 A/A in both recipients and donors further facilitated neutrophil recovery compared with others (13 (11 - 25) days and 19 (12 - 31) days, P = 0.019). Besides, IFN-γ + 874 A/A in recipients increased the probability of grade II-IV acute graft versus disease (aGVHD) and cytomegalovirus viraemia compared with IFN-γ + 874 T/A or T/T genotype (20% vs 4% P = 0.041, 43.6% vs 16.0% P = 0.032), which lead to increased 5-year transplant-related mortality (TRM) (33.7% ± 6.8% vs 12.0% ± 6.5%, P = 0.050) and decreased 5-year event free survival (EFS) \[(58.2 ± 6.7)% vs (84.0 ± 7.3)%, P = 0.032\] compared with the latter. IFN-γ + 874 A/A in both recipients and donors also significantly increased the probability of grade II-IV aGVHD and cytomegalovirus viraemia compared with the other (21.7% vs 5.9%, P = 0.050; 45.7% vs 20.6%, P = 0.020), which caused increased 5-year TRM \[(31.6 ± 7.5)% vs (13.6 ± 6.5)%, P = 0.048\] and decreased 5-year EFS \[(56.8 ± 7.3)% vs (79.4 ± 6.9)%, P = 0.037\] compared with the other., Conclusion: In HLA-matched sibling HSCT setting, the presence of IFN-γ + 874 T allele in recipients or in both recipients and donors significantly decreased the risk of grade II-IV aGVHD and CMV infection and increased EFS. While IFN-γ + 874 A/A in donors or in both recipients and donors was associated with shorter duration to neutrophil recovery.

The study was purposed to investigate the effect of alloreactive natural killer (alloNK) cells on immune reconstitution in murine haploidentical bone marrow transplantation (BMT). The murine model of haploidentical BMT was established by using (C57BL/6×BALB/c)BCF(1)(H-2(d/b)) mouse as the donor, and BALB/c (H-2(d)) mouse as the recipient. Recipient mice were divided into BMT group, non-allo-reactive NK (non-alloNK) cell group and alloNK cell group according to different transfusion. The effect of adding alloNK cells to transfusion was assessed by thymus pathology, the proportion of spleen NK cells, the spleen cell proliferation, the IFN-γ and IL-4 concentrations product at 24 and 48 h of recipient spleen cell culture supernatant at 2 months after BMT. The results showed that there were no obvious difference in thymus tissue among 3 groups under the optical microscope. The proportion of recipient spleen NK cells in non-alloNK group was significantly lower than that in BMT group (P < 0.05). There was no significant difference in proliferation of the recipient spleen cells among 3 groups at 2 months after BMT. The IFN-γ concentration product at 24 and 48 h of recipient spleen cell culture supernatant in alloNK group was significantly lower than that in other 2 groups at 2 months after BMT (P < 0.05). The IL-4 concentration in each group was not significantly different (P > 0.05). It is concluded that alloNK cells do not damage the thymus structure and may induce Th2 immune response in murine haploidentical BMT.

Objective: To evaluate the outcome of allogeneic hematopoietic stem cell transplantation (allo-HSCT) from HLA-matched sibling donor (MSD allo-HSCT) for severe aplastic anemia (SAA)., Methods: The clinical data of 41 SAA patients received MSD allo-HSCT from May. 2003 to Aug. 2011 were analyzed retrospectively. 24 patients were male, 17 were female. Median age was 23 (5 - 43) years old. 28 patients had SAA-I, 9 had SAA-II, and 4 had post-hepatitis aplastic anemia. 17 patients received allogeneic bone marrow (BM) transplantation (allo-BMT), and 24 received allogeneic peripheral blood stem cell (PBSC) transplantation (allo-PBSCT). The conditioning regimens: 20 patients received cyclophosphamide (CY) + anti-thymocyte globulin (ATG) + fludarabine (Flu), 21 received CY + ATG + Flu+ cytarabine (Ara-C) ± busulfan (Bu)/melphalan (Mel). Prophylaxis for graft-versus-host disease (GVHD): 25 patients received cyclosporine (CSA) plus short-term methotrexate (MTX), 16 received tacrolimus (FK506) plus short-term MTX. The median number of infused CD34(+) cells were 3.48 (2.39 - 4.80)×10(6)/kg in allo-BMT and 2.95 (1.27 - 5.98)×10(6)/kg in allo-PBSCT, respectively., Results: Hematopoietic reconstitution was observed in all 41 patients (100%). The median time of neutrophils (ANC) reached to 0.5×10(9)/L and platelets (PLT) reached to 20×10(9)/L were 14 (10 - 23) days and 19 (8 - 38) days, respectively. 12 patients developed acute GVHD (aGVHD), out of which 11 developed grade I-II aGVHD, and one developed grade IV. 2 patients occurred chronic GVHD (cGVHD), out of which one with local cGVHD and the other with extensive. 4 patients occurred graft rejection (GR), all of them recovered haemopoiesis and survived after donor PBSC infusion. 5 patients (12.2%) died, out of which one died of extensive cGVHD, and 4 died of invasive fungal infections (IFI). Median follow-up time was 23 (3 - 79) months. 36 patients survived. 5-year estimated overall survival (OS), disease free survival (DFS), and transplant-related mortality (TRM) was (81.1 ± 9.0)%, (68.4 ± 11.0)%, and (18.9 ± 9.0)%, respectively. Univariate analysis showed that lover OS had significant correlation with receiving PBSCT, occurrence of aGVHD, the number of infused CD34(+) cells no more than 2.5×10(6)/kg, the number of red blood cell (RBC) transfusion before transplant more than 30 U and occurrence of IFI after transplantation (P = 0.034, 0.001, 0.006, 0.000, 0.001, respectively). Occurrence of aGVHD had significant correlation with the disparity between donor and recipient ABO blood groups, the number of PLT transfusion more than 100 U, and the number of RBC transfusion more than 30 U before transplantation, the number of infused CD34(+) cells no more than 2.5× 10(6)/kg (P = 0.019, 0.038, 0.005, 0.005, respectively). The occurrence of GR had significant correlation with the number of PLT transfusion more than 100 U before transplantation (P = 0.038)., Conclusion: MSD allo-HSCT is an effective therapy for patients with SAA. Lower number of blood transfusion before transplantation, use of BMT, more number of infused CD34(+) cells can effectively prevent and treat aGVHD and IFI after transplantation, which may improve the efficacy of MSD allo-HSCT for SAA.

Objective: To study the clinical characteristics and antimicrobial resistance of bloodstream infections caused by Gram positive bacteria, so as to provide reference for the rational use of antimicrobial agent., Methods: One hundred and eight patients with bloodstream infections of Gram positive bacteria in our hospital from January 2009 to December 2009 were retrospectively reviewed. The clinical manifestations, pathogen types and antimicrobial susceptibility results of pathogens isolated from bloodstream were analyzed., Results: All patients had fever and 31.89% with rigor, 22.41% of the patients had no local infection lesions, 77.59% had clear infection lesions, including oral infections, respiratory tract infections and soft tissue infections. The pathogen testing showed that 12.82% were staphylococci aureus, 50.42% coagulase-negative staphylococci, 24.8% streptococci, 9.4% enterococci and 2.56% Listeria monocytogenes. Antibiotics resistance of staphylococcus and enterococci in our hospital was severe. The percentage of methicillin-resistant staphylococcus aureus in this investigation was 68.92%. The resistant rates of methicillin-resistant coagulase-negative staphylococci (MRCNS) to the most antimicrobial agents were higher than that methicillin-sensitive coagulase-negative staphylococci. One strain of MRCNS was found resistant to teicoplanin and linezolid, and 1 strain of enterococci resistant to teicoplanin and linezolid., Conclusion: Gram positive bacteria shows serious drug resistance, but still keeps highly sensitive to vancomycin, linezolid, teicoplanin and quinupristin/dalfopristin.

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.

Background: Patients undergoing myeloablative allogeneic haematopoietic stem cell transplantation (HSCT) have a higher incidence of acute kidney injury (AKI). RIFLE is a newly developed classification for AKI that includes three grades of severity - AKI-R, AKI-I, AKI-F., Objective: The purpose of this study was to analyse retrospectively major risk factors for AKI at the time of myeloablative allo-HSCT and to use the RIFLE criteria to predict mortality in myeloablative allo-HSCT., Methods: Renal function was evaluated in 143 patients with allo-HSCT by RIFLE criteria in order to assess the incidence, risk factors and mortality rate of various degrees of AKI., Results: The results of this study showed that patients with hepatic veno-occlusive disease (HVOD) have a higher incidence of AKI-F than those without HVOD (P = 0.002). The incidence of AKI-I and AKI-F in patients with grade III-IV acute graft-versus-host disease (aGVHD) and increased total bilirubin was significantly higher than in those without (P = 0.001, P <0.001). HVOD was an independent risk factor of AKI-F (OR 5.058, 95% CI 1.317-19.424, P = 0.018), and increased total bilirubin was an independent risk factor for AKI-F (OR 5.126, 95% CI 1.403-18.998, P = 0.014). Worsening RIFLE category was associated with increased mortality of the patients in the 100 days post-transplant (P = 0.003). In addition, 180-day survival of patients in different AKI classes was significantly different (P = 0.0095)., Conclusion: AKI is common in patients with myeloablative allo-HSCT and is associated with increased risk of death. The RIFLE criteria appear to be an important tool for stratification of these patients on the basis of death risk.

The objective of this study was to investigate the correlation between the gene polymorphisms of drug metabolizing enzymes and the outcome of the first induction chemotherapy in patients with de novo acute myeloid leukemia (AML). 113 de novo AML patients were enrolled in this study. The genotypes of 11 single nucleotide polymorphisms (SNP) in drug metabolizing enzymes were detected by the SNPstream(®) Genotyping System. The correlation between the distribution of genotypes and the complete remission rate of first induction chemotherapy was analyzed by logical regression. The results showed that patients with variant genotype of CYP2D6 (rs16947) had a lower complete remission (CR) rate, as compared to those with wild type (p = 0.033, OR = 0.32, 95%CI 0.112 - 0.915); meanwhile the patients with variant genotype of GSTO2 (rs156697) had a higher CR rate as compared to those with wild type (p = 0.011, OR = 3.023, 95%CI 1.289 - 7.089). Combined analysis of the above polymorphisms, showed that patients with variant genotype of CYP2D6 and wild genotype of GSTO2 (V + W) had lower CR rates in comparison to patients with wild genotypes of both polymorphisms (p = 0.017, OR = 0.183, 95%CI 0.045 - 0.735). It is concluded that CYP2D6 (rs16947) and GSTO2 (rs156697) polymorphisms are independent factors influencing CR rates of the first induction chemotherapy in de novo AML patients.

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.

Objective: To investigate the prognosis and hepatitis B serologic marker changes in patients with HBV infection or with HBV infected donors after allogenic hematopoietic stem cell transplantation (allo-HSCT)., Methods: The clinical outcomes of 79 patients receiving allo-HSCT, including 55 with HBV infection and 24 from HBV infected donors were analyzed retrospectively., Results: (1) HBV infection did not interfere with the clinical outcome of allo-HSCT. (2) In 20 HBsAg(+) patients, 13(65.0%) developed HBV reactivation between 0.5 and 10 months after transplantation, 9(45.0%) developed HBV-related hepatitis. (3) For the 35 HBsAg(-) and HBcAb/HBeAb positive patients, 4 (11.4%) occurred HBV seroconversion, 1 of the 4 complicated with severe chronic graft-versus-host disease (cGVHD). (4) There was a significant difference in HBV reactivation rate between the HBsAg(+) and HBsAg-groups (P < 0.01). The incidence of hepatitis occurred within 100 days after HSCT was high in HBsAg(+) patients (P < 0.05). (5) Clearance of HBsAg was observed in 2 HBsAg(+) patients, both of whom received graft from HBsAb positive donors., Conclusions: Donors or recipients infected with HBV is not considered an absolute contraindication for HSCT, but HBsAg positivity is a high risk factor for HBV reactivation and prophylactic lamivudine treatment may be helpful. For patients with HBcAb/HBeAb positivity, seroconversion can be observed, especially after immunosuppressant withdrawal. Adoptive immunity is effective in clearing HBV in these patients.

This study was purposed to investigate the effect of prostaglandin E2 (PGE2) on proliferation of peripheral blood T lymphocytes, and to evaluate the regulatory role of PGE2 on immunological balance between Th1/Th2 and Tc1/Tc2 lymphocytes. The peripheral blood mononuclear cells (PBMNC) were stimulated by anti-human CD3 monoclonal antibody (mAb) and anti-human CD28 mAb, and were cultured in the presence of different concentration of PGE2 for 120 hours. The proliferation of peripheral blood T lymphocytes was assayed according to the manufacture protocol of BrdU Kit; the IFN-gamma and IL-4 levels in supernatants cultured for 24, 48, 72 and 120 hours were detected by ELISA; the ratios of CD4+IL-4+ T cells/CD4+ IFN-gamma+ T cells and CD8+IL-4+ T cell/CD8+IFN-gamma+ T cells were determined by flow cytometry. The cells cultured without PGE2 were used as control. The results indicated that (1) with the raising of concentration of PGE2, the inhibitory rate of T cell proliferation in vitro significantly increased (p=0.001). There was significant positive correlation between inhibitory rate of T cells and PGE2 concentration (correlation coefficient=0.889, p=0.000). (2) the difference between the IFN-gamma concentrations in supernatant cultured for 120 and 72 hours in test groups had no statistical significance (p=0.917). The IFN-gamma concentration increased continually with prolonging of culture time in control group (p=0.046). The IFN-gamma concentrations produced at different times in test group were significantly lower compared with those in control group (p<0.05). The IL-4 concentrations produced at different time had no significant change in test groups (p=0.400). The IL-4 concentration in 24 hours in control group was significantly higher than that at 48, 72 and 120 hours in control group (p=0.007, 0.003 and 0.002). After cultured for 24 hours the IL-4 concentration in test group was significantly lower than that in control group (p=0.037), but after cultured for 48, 72 and 120 hours, the IL-4 concentration in test group did not show statistical difference in comparison with control group (p>0.05). (3) the proportions of CD4+IFN-gamma+T cells in test group and in control group had no significant difference (p=0.767). The proportion of CD4+IL-4+T cells in test group was slightly higher than that in control group (p=0.051). The ratio of CD4+IL-4+T cells to CD4+IFN-gamma+ T cells in test group was significantly higher than that in control group (p=0.011). The proportions of CD8+IFN-gamma+ T cells in test group and in control group had no statistical difference (p=0.441). The proportion of CD8+IL-4+T cells in test group was significantly higher than that in control group (p=0.015). The ratio of CD8+IL-4+ T cells to CD8+IFN-gamma+ T cells in test group were obviously higher than that in control group(p=0.038). It is concluded that the PGE2 inhibits the proliferation of T lymphocytes in vitro. PGE2 influences the production of IFN-gamma and IL-4, and significantly influences peak appearance of IFN-gamma produced by T lymphocyte. PGE2 can continuously inhibit the production of IFN-gamma, but its continuous effect on IL-4 is no significant. PGE2 enhances the ratio of CD4+IL-4+T lymphocytes to CD4+IFN-gamma+T lymphocytes and the ratio of CD8+IL-4+T lymphocytes to CD8+IFN-gamma+T lymphocytes, and regulates development of T cells toward Th2/Tc2 cells.

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.

The study was aimed to compare the efficiency of cytomegalovirus (CMV) quantitative PCR and CMV-pp65 antigen test for detection of CMV infection and their clinical significance in patients received allogeneic hematopoietic stem cell transplantation (HSCT). 84 patients received allogeneic HSCT were enrolled in study. Anticoagulant blood samples were obtained from the recipients before and after transplantation and in the convalescence. CMV quantitative PCR and CMV-pp65 antigen test were performed weekly. The results showed that out of 84 patients, 26 cases were positive (30.95%) by CMV quantitative PCR method. Of the 26 cases, 9 cases were CMV antigenemia and 13 cases were CMV disease, the median positive time was 37.1 (7 - 105) days after HSCT. 22 cases were positive (26.19%) by CMV-pp65 antigen test method, the median positive time was 46.6 (10 - 128) days after HSCT. All the 22 positive cases detected by CMV-pp65 antigen test were also positive by CMV quantitative PCR method. Nevertheless, 4 positive cases detected by CMV quantitative PCR but negative detected by CMV-pp65 antigen test method did not develop CMV disease. The CMV disease was found in the cases either with moderate to high copies of CMV quantitative PCR or moderate to high level CMV antigenemia by CMV-pp65 antigen test method. The clearance median time was 17.5 (11 - 28) days by CMV quantitative PCR method after receiving antiviral therapy and was 10.0 (7 - 21) days by CMV-pp65 antigen detection method. It is concluded that both CMV quantitative PCR and CMV-pp65 antigen test can detect the infection of CMV early and effectively in patients received HSCT. CMV quantitative PCR is more sensitive, and CMV-pp65 is more specific. It can be more effective to guide the antiviral treatment and evaluate its efficacy when combining the two methods.

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.

Objective: To explore the value of circulating galactomannan (GM) screening for early diagnosis and treatment monitoring of invasive aspergillosis (IA)., Methods: Serum samples from 141 IA patients for the detection of GM by Platelia Aspergillus (Bia-Rad) were collected before and after systematic anti-fungal therapy., Results: (1) An increase in the clinical diagnosis rate of IA was obtained on the result of GM detection. The GM positivity appeared (10+/-4.1) (8-15) d before positive sputum culture, while (12.6+/-5.7) (6-22) d before the CT positive image. (2) Among the 62 patients with consecutive serum samples, 50 were success in treatment and 12 died. A progressive decrease of GM level was found in the former group, while the rising antigen titres were found in the latter., Conclusion: Compared with other diagnostic test, GM test has an obvious advantage of higher positivity and earlier result. The anti-fungal effectiveness can be estimated by dynamic detection of serum GM.

The objective of this study was to explore the useful value of circulating galactomannan (GM) for early diagnosis of invasive aspergillosis. All 141 patients were classified as 103 patients of clinical and possible diagnosis, and 38 non-Aspergillus patients. 209 serum samples for the detection of GM by Platelia Aspergillus were collected before anti-fungal vaccine therapy. ELISA method was used in detection of GM. The results showed that (1) the sensitivity of 87.5%, specificity of 81.6%, positive prediction of 66.7% and negative prediction of 93.9% were determined by using cut-off value. According to the result of ELISA, the clinical diagnosed patients was up to 48, while the possible diagnosed patients were 55. (2) Among 62 patients with consecutive examinations of serum samples, 50 patients were successfully diagnosed and treated, while 12 patients died. A progressive reduction of GM level was found in survivors, however, the patients of poor prognosis showed higher antigen titres. It is concluded that GM test has more significance for earlier diagnosis of aspergillosis, the concentration of GM is related to prognosis of disease.

This study was aimed to explore the effect of autologous hematopoietic stem cell transplantation (auto-HSCT) and allogeneic hematopoietic stem cell transplantation (allo-HSCT) in adult patients with acute lymphoblastic leukemia and to analyze the related prognostic factors. Clinical data of 114 ALL patients receiving HSCT, including 70 auto-HSCT and 44 allo-HSCT, were retrospectively analyzed. Disease-free-survival (DFS), relapse-rate (RR) and transplantation-related-mortality (TRM) of patients receiving different HSCT were compared. The results showed that the eight-year OS and DFS in a total of 114 adult ALL patients were (40.89+/-5.27)% and (39.50+/-5.22)% respectively. The three-year DFS of ALL patients who received HSCT in phase CR1 and no CR1 were (47.63+/-5.63)% and (17.65+/-9.25)% (p=0.0034). The two-year DFS of patients who received allo-HSCT and had I/II aGVHD was (62.75+/-12.30)%, and the six-month DFS of patients who had III/IV aGVHD was 0, and the two-year DFS of patients without aGVHD was (29.35+/-9.70)% (p=0.005). The three-year DFS of patients with and without maintenance chemotherapy after transplantation were (55.12+/-7.89)% and (33.33+/-11.11)% respectively, there was significant difference between them (p=0.0499). The five-year DFS between patients received auto-HSCT and allo-HSCT in phase CR1 was not significantly different. The RR of patients received allo-HSCT was lower than that of patients received auto-HSCT, but there was no significant difference between them. The TRM of patients received allo-HSCT was higher than of patients received auto-HSCT (p=0.0313). Expression of myeloid antigen and higher LDH level in diagnosis were poor- prognostic factors. It is concluded that auto-HSCT and allo-HSCT completed in phase CR1 may improve prognosis of the patient with ALL as a method for consolidation chemotherapy, but no significant difference exists between the two HSCTs. Patients receiving allo-HSCT and having I/II aGVHD may achieve higher DFS. The maintaining chemotherapy for patients after auto-HSCT may improve therapeutic effect.