[The effects of donor peripheral blood stem cell infusion in the prevention of relapse in high risk leukemia after haplotype hematopoietic stem cell transplantation].

OBJECTIVE: To explore the preventative effect of donor peripheral blood stem cell (PBSC) infusion mobilized by granulocyte colony-stimulating factor (G-CSF) for the relapsing patients with leukemia after haplotype hematopoietic stem cell transplantation (HSCT), as well as its therapeutic effect and safety. METHODS: G-CSF was given at 5 - 10 µg×kg(-1)×d(-1) to donor and PBSCs were obtained on day 5 and frozen and allotted for storing. PBSC infusion was given to all the 20 patients on day 90 after HSCT, and the second treatment was given to 4 patients on day 30 after the first infusion. The occurrence of graft-versus-host disease (GVHD), relapse rate of high risk leukemia and long-term survival were evaluate after PBSC infusion. RESULTS: A total of 19 patients had acute GVHD after PBSC infusion for a median of 25 (12 - 60) months, 4 of them were ≥ degree III. The cumulative incidence rate was 22.9%, and all of them accepted PBSC infusion twice. Thirteen patients had assessable chronic GVHD, 10 of them were restricted, and no one died of it. Nine patients died of relapse of leukemia. The remaining 11 patients survived leukemia free, including 4 with chronic myelogenous leukemia, 4 with acute myeloid leukemia (AML), 1 with lymphoma-leukemia and 2 with myelodysplastic syndrome-AML (MDS-AML). Kaplan-Meier analysis showed the disease free survival rate of 2-year was 52.5%. CONCLUSION: The prophylactic donor PBSC infusion mobilizing with G-CSF is effective, safe and feasible for the relapse of leukemia.

[Treatment of severe intestinal acute graft-versus-host disease with CD25 monoclonal antibody in haploidentical hematopoietic stem cell transplantation].

OBJECTIVE: To study the feasibility of CD25 monoclonal antibody (basiliximab) in the treatment of severe III-IV intestinal acute graft-versus-host disease (GVHD) following haploidentical hematopoietic stem cell transplantation (HSCT). METHODS: Twenty patients, 13 males and 7 females, who developed III-IV intestinal acute GVHD after haplotypic HSCT between October 2004 and September 2009, were treated with basiliximab (20 mg/d, d1, 4) and prednisolone (1 mg×kg(-1)×d(-1)) from the day of diagnosis. The therapy was repeated in the second week if the intestinal symptoms showed no improvement. The therapeutic effect was analyzed and the adverse reaction and cytomegalovirus (CMV) infection were observed. RESULTS: Ten patients had a complete remission and 5 were in a partial remission. The total effective rate was 75.0%. The clinical symptoms started to lessen in 1-12 days after using basiliximab (average: 7 days). During the 6-64 month follow-up (average: 25 months), 8/10 cases with a complete remission had no acute GVHD relapse, and the other 2 relapsing patients experienced a remission after a re-administration of basiliximab. Nine patients survived with a longest period of 64 months. Four withdrew corticosteroids and the other 5 stayed on a low-dose maintenance corticosteroid regimen. The 2-year disease-free survival was 47.5% by Kaplan-Meier calculation. CONCLUSIONS: Basiliximab is feasible in the treatment of III-IV intestinal acute GVHD after haplotype HSCT. It does not increase the relapse of leukemia or the incidence of infections.

Overexpression of miR­21 is involved in acute monocytic leukemia­associated angiogenesis by targeting IL­12.

Angiogenesis is important in pathophysiological processes, including the pathogenesis of acute monocytic leukemia (AML). MicroRNA­21 (miR­21) is overexpressed and exhibits oncogenic activity in cancer. However, the biological mechanism underlying the effect of miR­21 in AML remains to be fully elucidated. In the present study, the expression levels of miR­21 and vascular endothelial growth factor (VEGF) were determined in 26 patients with AML and 28 healthy individuals. The secretion of VEGF was also measured following the transfection of THP­1 cells with miR­21 mimic or inhibitor. The supernatants of the THP­1 cells, which were transfected with miR­21 mimic, inhibitor or small interfering RNA (si)VEGF, respectively, were used to incubate human umbilical vein endothelial cells (HUVECs), following which tube formation of the HUVECs was measured. miR­21 targets were predicted using a biological target prediction website and confirmed using a luciferase assay. The effects of interleukin (IL)­12 were investigated by examining the tube formation of HUVECs and the secretion of VEGF following recombinant human (rh) IL­12 pretreatment. The results revealed that miR­21 and VEGF expression was significantly increased in the peripheral blood monocytes of the patients, compared with the healthy controls. There was negative correlation between the expression of IL­12 and miR­21 in the serum of patients with AML. Furthermore, supernatant VEGF levels from the miR­21 mimic­transfected THP­1 cells were increased, whereas a decreasing trend was observed in the miR­21 inhibitor group. The angiogenic ability of the HUVECs pretreated with supernatant from the THP­1 cells transfected with miR­21 mimic was higher, and was lower in THP­1 cells co­transfected with miR­21 mimic and siVEGF, compared with the miR­21 mimic only group. A luciferase assay demonstrated that IL­12 was the direct target of miR­21, and the level of IL­12 in the supernatant of THP­1 cells transfected with miR­21 mimic was increased. IL­12 pretreatment increased VEGF expression and angiogenic ability in HUVECs. The inactivation of miR­21 or activation of its target gene may be a potential therapeutic strategy in human AML.

Association of VEGF and VDR gene- gene and gene- smoking interaction on risk of multiple myeloma in Chinese Han population.

AIMS: To investigate the association of several single nucleotide polymorphisms (SNPs) within vascular endothelial growth factor (VEGF) and vitamin D receptor (VDR) gene polymorphisms and additional gene- gene and gene- smoking interaction with multiple myeloma (MM) risk in Chinese population. METHODS: Generalized multifactor dimensionality reduction (GMDR) was used to screen the best interaction combination among SNPs and smoking. Logistic regression was performed to investigate association between 6 SNPs within VEGF and VDR gene, additional gene- gene and gene- smoking interaction on MM risk. RESULTS: MM risk is significantly higher in carriers with the rs699947- A allele within VEGF gene than those with CC genotype (CA+ AA versus CC), adjusted OR (95%CI) =1.72 (1.19-2.33), and higher in carriers with rs2228570- T allele within VDR gene than those with CC genotype (CT+ TT versus CC), adjusted OR (95%CI) = 1.68 (1.26-2.17). We also found a significant two-locus model (p=0.0010) involving rs699947 and rs2228570, and a significant two-locus model (p=0.0107) involving rs2228570 andsmoking. Participants with rs699947- CA+AA and rs2228570- CT+TT genotype had the highest MM risk, compared to participants with rs699947- CC and rs2228570- CC genotype, OR (95%CI) = 3.12 (1.82 -4.61). Smokers with rs2228570- CT+TT genotype had the highest MM risk, compared to never- smokers with rs2228570- CC genotype, OR (95%CI) = 3.27 (1.74-4.86). CONCLUSIONS: We found that the A allele of rs699947 within VEGF and T allele of rs2228570 within VDR gene, interaction between rs699947 and rs2228570, rs2228570 andsmoking were all associated with increased MM risk.

Adenovirus co-expressing CD40 ligand and interleukin (IL)-2 contributes to maturation of dendritic cells and production of IL-12.

The aim of the present study was to construct a chimeric adenovirus (Ad)5/F35 co-expressing human CD4O ligand (CD4OL) and interleukin (IL)-2 (Ad5/F35 CD40L-IL-2). The infection efficiency to human monocyte-derived dendritic cells (Mo-DCs), expression of genes, phenotype changes and IL-12 production of Mo-DC by Ad5/F35 CD40L-IL-2 were investigated. CD40L and IL-2 from total RNA extracted from human peripheral blood mononuclear cells (PBMCs) were cloned by reverse transcription-polymerase chain reaction and used to construct Ad5/F35 CD40L-IL-2. The infection efficiency, expression of CD40L, and phenotype changes of Mo-DC infected with Ad5/F35 CD40L-IL-2 were analyzed using flow cytometry. The quantities of IL-2 and IL-12 in the supernatants of Mo-DC following infection of Ad5/F35 CD40L-IL-2 were measured by enzyme-linked immunosorbent assay. The CD40L and IL-2 genes were successfully cloned and the Ad5/F35 CD40L-IL-2 was constructed. Ad5/F35 CD40L-IL-2 efficiently infected Mo-DCs with an infection efficiency of >75%, and the infected Mo-DCs expressed CD40L and secreted IL-2. The expression levels of cluster of differentiation (CD)80, CD86, CD40, and human leukocyte antigen-antigen D related on Mo-DC were moderate; however, CD83 was low prior to infection of Ad5/F35 CD40L-IL-2. Those molecules, particularly CD83, were markedly upregulated 24 h after the infection. Increasing quantities of IL-12 in the supernatants were detected subsequent to infection at different time points in a time-dependent manner. Thus, Ad5/F35 CD40L-IL-2 efficiently infected human Mo-DCs and its products, CD40L and IL-2, were subsequently expressed. In addition, infection with Ad5/F35 CD40L-IL-2 stimulated the maturation of Mo-DC and high levels of IL-12 production.

[Clinical analysis of cytomegalovirus infection after allogeneic hematopoietic stem cell transplantation].

The objective of this study was to explore the incidence and therapeutic efficacy of cytomegalovirus (CMV) infection after allogeneic hematopoietic stem cell transplantation (allo-HSCT). The clinical data of 140 patients undergoing allo-HSCT in our department of hematology from 2010-01 to 2012-01 were retrospectively analyzed. The results showed that the incidence of CMV infection was 4.3% (48/140), the time for the first detection of positive CMV-DNA was at day 45 (33 to 68) after allo-HSCT, and the CMV quantitative range was 1.25×10(3) - 5.5×10(6). There were 2 cases of CMV-related interstitial pneumonia and 5 cases of hemorrhagic bladder inflammation. A total of 65 patients suffered from graft versus host disease (GVHD), in which 32 cases (49.2%) were accompanied with CMV infection, CMV-DNA negative in patients treated with ganciclovir, foscarnet sodium anti-CMV was at day 45 (33 to 68) with the effective rate of 100%. 12 patients with CMV infection were accompanied with transient neutropenia and thrombocytopenia. It is concluded that after allo-HSCT the CMV infection occurs frequently. The patients with GVHD have a higher incidence of CMV infection. Ganciclovir and foscarnet sodium are reliable to be used for treatment of CMV infection with fewer adverse reactions.

[A preliminary report on treatment of myelodysplastic syndrome with cyclosporin a and androgen].

The study was aimed to explore clinical result of cyclosporin A (CsA) and androgens for treatment of myelodysplastic syndrome (MDS) with refractory anemia. Four cases of MDS-RA were treated with CsA and androgens, while the changes of blood counts, bone marrow and chromosome were observed. The results showed that substantial hematological response was observed in all four patients, that their anemia improved and all transfusion-dependent patients achieved transfusion independence. In conclusion, CsA and Adr therapy was well tolerated. CsA and Adr therapy offer an alternative treatment of MDS with RA. The mechanisms of the benifical effect from this therapy remain the subject of an ongoing study.