[Effect of Sonic Hedgehog Signal Pathway Inhibitor Jervine on Myelodysplastic Syndromes MUTZ-1 Cells].

OBJECTIVE: To study the effect of SMO inhibitor (Jervine) on proliferation, apoptosis and cell cycle of MDS cell line MUTZ-1, and its mechanism. METHODS: The effect of different concentrations Jervine on proliferation of MUTZ-1 cells was detected by CCK-8 method. Apoptosis and cell cycle of MUTZ-1 cells were detected by flow cytometry. Western blot was used to detect the changes of Shh signaling pathway effecting proteins BCL2 and CyclinD1. The expression levels of Smo and Gli1 gene were detected by real-time fluorescent quantitative polymerase chain reaction (RT-qPCR). RESULTS: Jervine inhibited MUTZ-1 cell proliferation in a concentration dependent manner (24 h, r=-0.977), the apoptosis rate of MUTZ-1 cells increased with the enhancement of concentration of Jervine in MUTZ-1 cells (P＜0.001), the cell proportion of G1 phase increased and the cell number of S phase decreased with enhancement of concentration (P＜0.001). The result of RT-qPCR and Western blot showed that the expression of Smo, Gli1 mRNA and BCL2, CyclinD1 proteins decreased (P＜0.05). CONCLUSION: SMO inhibitor can effectively inhibit the growth of MDS cell line MUTZ-1 improve the cell apoptosis and induce cell cycle arrest. Its action mechanism may be related with dowm-regulating the expression of BCL2 and CyclinD1.

[Expression Level and Clinical Significance of Gli1 in Patients with Myelodysplastic Syndrome].

OBJECTIVE: To study the expression level and clinical significance of Gli1 gene in patients with myelodysplastic syndrome(MDS). METHODS: The positive rate of bone marrow CD34+ cells was detected by flow cytometry in 53 patients with MDS.Magnetic beads were used to separate CD34+ cells. The expression of Gli1 on CD34+ cells was detected by RT-qPCR, 25 patients with iron deficiency anemia were selected as controls. The relationship of Gli1 expression with clinical characteristics were analyzed. RESULTS: The expression of Gli1 in patients with MDS (0.73±1.26) was significantly higher than that in the control group (0.07±0.46) (P＜0.05). The expression of Gli1 significantly correlated with platelet count, chromosome grouping and IPSS risk stratification (P＜0.05). The median overall survival time of patients in high and low expression groups were 7 and 20 months respectively (P＜0.05). Multivariate analysis showed that Gli1 and chromosome grouping were 2 independent poor prognostic factors (P＜0.05). CONCLUSION: The expression of Gli1 is high in MDS. Abnormal expression of Gli1 positively correlates with clinical characteristics and prognosis of patients.Gli1 may be involved in the occurrence and development of MDS.

[Comparison of immunological reconstitution and related complications after HLA-matched and HLA haploidentical peripheral blood hematopoietic stem cell transplantation].

OBJECTIVE: To explore the difference of immune function and relationship with main complications after HLA-matched and HLA haploidentical allogeneic peripheral blood hematopoietic stem cell transplantation (allo-PBSCT). METHODS: Sixty-seven patients undergoing HLA-matched (n = 33) or HLA haploidentical (n = 34) allo-PBSCT during the same time period in our hospital from June 2004 to December 2007 were included in this study. Indirect immunofluorescence assay was employed to detect lymphocyte subsets before transplantation and on month 1, 3, 6, 12 and 18 after transplantation and the lymphocyte subsets of 100 healthy people were used as normal control. The comparison of immunological reconstitution and relationship with main complications was carried out with statistical analysis. RESULTS: (1) Comparison of the 67 patients with normal controls showed that CD(3)(+), CD(4)(+), CD(4)(+)/CD(8)(+) at month 1, CD(4)(+), CD(4)(+)/CD(8)(+) at month 3 and CD(4)(+) at month 6 after PBSCT were lower. CD(8)(+) at month 3 and month 6 were higher. (2) The immune function was not statistically different between HLA haploidentical and HLA-matched allo-PBSCT (P > 0.05). (3) The immune function of patients with and without severe infection was not statistically different (P > 0.05). (4) The immune function of patients with chronic graft-versus-host disease (cGVHD) between HLA haploidentical and HLA-matched allo-PBSCT groups was not statistically different. The immune function of patients without cGVHD in two groups was not statistically different (P > 0.05). (5) The immune function of patients with or without relapse was not statistically different (P > 0.05). CONCLUSIONS: HLA-haploidentical PBSCT conditioning including antithymocyte globulin without in vitro T cell depletion is feasible and safe. The immunological reconstitution, incidence of severe infection, incidence of relapse and treatment-related mortality are not significantly different between HLA-matched and HLA haploidentical allo-PBSCT.

[Non-T-cell depleted HLA haploidentical peripheral blood stem cell transplantation for hematological malignancies: report of 36 cases].

OBJECTIVE: To analyze the clinical outcome of human leukocyte antigen (HLA) haploidentical peripheral blood stem cell transplantation (PBSCT) from related donors for hematological malignancies. METHODS: Thirty-six patients with hematological malignancies, with a median age of 25 (11-48) years, were transplanted with PBSC from an HLA-haploidentical family donors: 7 were 1 locus mismatched and 29 were 2-3 loci mismatched. The recipients received myeloablative conditioning regimen, in combination with different immunosuppressants according to the degree of HLA disparity followed by non-T-cell depleted PBSCT. The median number of CD34+ cells were 11 (4.16-21.00) x 10(6)/kg. RESULTS: All patients achieved sustained, full donor-type engraftment. Fifteen patients (41.7%) developed grade I-II aGVHD. Among 29 patients followed up more than 18 months, 17 (58.6%) developed cGVHD. There was no statistical difference in decrease and recovery of T, B and NK cell subsets after transplantation between HLA haploidentical group and HLA identical PBSCT group. The median follow-up duration was 15 (4 -69) months. Five patients (13.9% ) relapsed. The 2-year probability of leukemia-free survival (LFS) was 82.2%. CONCLUSION: Non-T-cell depleted HLA-haploidentical PBSCT is safe and feasible for patients with hematological malignancies after myeloablative conditioning regimen combined with intensive immunosuppressants.