[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.

[CD28 expression on CD4(+);T cells and its relationship with IFN-γ/IL-10 ratio in patients with primary immune thrombocytopenia].

OBJECTIVE: To investigate CD28 expression on CD4(+);T cells and its relationship with IFN-γ/IL-10 ration in patients with primary immune thrombocytopenia (ITP) before and after treatment. METHODS: The expression of CD4(+);CD28(+); was detected by flow cytometry, the levels of IFN-γ and IL-10 were determined by double-antibody sandwich ELISA and platelet count was tested by blood cells automatical counter in peripheral blood of 30 patients with ITP before and after glucocorticoid treatment and 26 cases of normal controls. Then the correlations between the outcomes were analyzed. RESULTS: The expression of CD4(+);CD28(+); of ITP patients before treatment was higher than that of the normal control group (P<0.05), however, after treatment the expression between the two groups had no statistical difference (P>0.05). Compared with the control group, the ITP patients before treatment showed the significantly higher level of IFN-γ and the significantly decreased level of IL-10, and the ratio of IFN-γ/IL-10 was significantly raised (P<0.01); while ITP patients after treatment had no statistically significant difference in the above indexes from the normal controls (P>0.05). Moreover, CD4(+);CD28(+); was positively correlated with IFN-γ/IL-10 ratio (P<0.05), and was negatively correlated with platelet count in ITP patients before treatment (P<0.05). CONCLUSION: Co-stimulatory molecule CD4(+);CD28(+); was closely related with immune disorder of ITP. It maybe played a role in the pathogenesis of ITP through involving Th1 advantage state formation.

[Detection and clinical significance of Th1/Th2 cytokines in patients with idiopathic thrombocytopenic purpura].

AIM: To observe Th1 (IL-2 and IFN-γ) and Th2 (IL-4, IL-10) cytokine changes in patients with idiopathic thrombocytopenic purpura (ITP) and explore the correlation between ITP development and Th1/Th2. METHODS: A total of 30 patients with ITP and 26 healthy volunteers were enrolled in this study. Serum levels of IL-2, IFN-γ, IL-4, and IL-10 were detected by enzyme-linked immunosorbent assay (ELISA) before and after treatment. RESULTS: The serum levels of IFN-γ and IL-2 in patients with ITP before the glucocorticoid treatment were significantly higher than those after the treatment and in the control groups (P<0.05), while the serum levels of IL-4 and IL-10 in patients with ITP before treatment were significantly lower than those after the treatment and in the control group (P<0.05). CONCLUSION: Th1/Th2 imbalance plays a prominent role in the development of ITP, and glucocorticoid treatment helps to restore the Th1/Th2 in the patients with ITP.

[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.

Retinoic acid in treating acute promyelocytic leukemia with hyperleukocytosis and its therapeutic strategy / 中国实验血液学杂志

In order to investigate the occurrence of hyperleukocytosis in treating acute promyelocytic leukemia (APL) patients with all trans retinoic acid (ATRA) and to explore the influence of the level of leucocyte on curative effect of ATRA, the APL patients were divided into three different groups according to the count of leucocyte in peripheral blood. Patients with WBC count less than 30x10(9)/L were administered with ATRA alone (the first group), patients with WBC count more than 30x10(9)/L were administered with ATRA alone (the second group) and patients with WBC count more than 30x10(9)/L were treated with ATRA+cytotoxic drugs (the third group). The results showed that hyperleukocytosis were found in 23 out of 39 patients (58.97%). Total remission rates in the second group and in the third group were 91.3%. The remission rates in the first, second and third groups were 100%, 87.5% and 90.9%, respectively. It is concluded that the ATRA in combination with cytotoxic drugs can efficiently control the occurrence of hyperleukocytosis during ATRA-treating APL and reduce the early mortality.

Early prediction of efficaciousness after first course of chemotherapy in de novo acute leukemia / 中国实验血液学杂志

To achieve the complete remission (CR), acute leukemia (AL) patient must get through the period of myeloid ablation after chemotherapy that the white blood cell (WBC) count in peripheral blood decreases rapidly. To observe the relationship of WBC count with therapeutic effectiveness after chemotherapy in previously untreated AL, eighty cases of previously untreated acute leukemia who took the first induction chemotherapy course were analyzed. Blood routine was carried out 2 to 3 times every week while the bone marrow pictures on 12th and 20th day after chemotherapy were observed. 80 patients were divided into 3 groups based on the lowest value of WBC count after chemotherapy: <or= 0.4 x 10(9)/L group, (0.4 - 0.9) x 10(9)/L group and > 0.9 x 10(9)/L group. The results showed that the complete remission rates after the first course of chemotherapy were 60 vs 55.6 vs 27.3% respectively in the three groups which WBC counts were <or= 0.4 x 10(9)/L, (0.4 - 0.9) x 10(9)/L and > 0.9 x 10(9)/L. The total efficaciousness rates were 90 vs 92.6 vs 66.7 respectively in the three groups. The efficaciousness rate of the first two groups significantly differed from the third group (P < 0.01). However, no significant difference was shown between the first and second groups. It is concluded that the leukocyte count after the first course of chemotherapy may be an early indicator to predict the efficaciousness of chemotherapy.