OBJECTIVETo investigate the clinical and cytogenetic characteristics of high-level mixed-lineage leukaemia (MLL) gene amplification in patients with acute myeloid leukemia (AML).

METHODSThe clinical and cytogenetic data of 2 AML patients with high-level MLL amplification from January 2010 to August 2016 were analyzed retrospectively.

RESULTSThe two AML cases were in middle-aged population. They were diagnosed as FAB subtype M5b and M2a respectively. Both of them had complex karyotypes with the aberrations of chromosome 11. One case was confirmed as MLL-PTD involving exons 2-9 by RT-PCR and sequencing. The other case without MLL-PTD was further analyzed by CytoScan HD analysis. The CMA results showed partial gain of 11q accompanied with partial loss in 11q, deletion of regions in 3p, 3q, 4q, 5q, 7q, 8q, 10p, 10q, 12p and 18q, as well as gain of 4p.

CONCLUSIONThe co-existence of -5/5q-, -7/7q- and highly complex karyotype may accelerate the poor prognosis. Thus how those cytogenetic abnormalities influencing the disease prognosis need to be further explored.

OBJECTIVETo detect the expression levels of MRE11 and Ku80 mRNA, and telomere length in bone marrow mononuclear cells of aplastic anemia(AA) patients, and to explore their correlation with pathogenesis of aplastic anemia.

METHODSBone marrow mononuclear cells were collected from 40 cases of AA and 20 normal controls for detecting mRNA expression of MRE11 and Ku80 and telomere length by using real-time quantitative polymerase chain reaction (qPCR), then MRE11, Ku80 and telomere length were analyzed for their correlation.

RESULTSAs compared with controls, the expression levels of MRE11 and Ku80 in patients with AA were significantly reduced, and the telomere length in patients with AA was obviously shortened, respectively (P<0. 05). The telomere length was significantly shorter in the persons aged ≥45 years in comparison with the AA patients and normal control younger than 45 years old (P<0.05). For the AA patients older than or equal to 45 years and less than 45 years in comparison with the controls at the same age, the telomere length was significantly shorter(P<0.05). The expression levels of MRE11 and Ku80 didn't correlate with telomere length (P>0.05). The mRNA expression level of MRE11 correlated positively and significantly with that of Ku80 (r=0.863, P<0.05).

CONCLUSIONThe change of telomere length may play an important role in the pathogenesis and progression of aplastic anemia. The lower expression of MRE11 and Ku80 may be involved in the pathogenesis of aplastic anemia.

OBJECTIVETo study the effect of the DA-EPOCH chemotherapy combined with G-CSF and the CTX therapy with G-CSF on mobilizing and collecting the peripheral blood hematopoietic stem cells and the later hematopoietic recovery.

METHODSForty patients accepted mobilization and collection of peripheral blood stem cells(PBSC) after treated by CTX+G-CSF and DA-EPOCH+G-CSF therapy respectively, and were treated by auto-transfusion after BEAM pre-regimen. The mobilization efficacy, adverse effects and hematopoietic recovery after autologous transplantation were analyzed retrospectively.

RESULTSDuring the CTX+G-CSF mobilization, only one patient achieved the white blood cell(WBC) at 0.8×10/L, while the others were with the lowest WBC level above 2.0×10/L. The platelet counts were all normal with the exception of 3 cases at 80×10/L. The median percentage of CD34cells in one period of collection was 0.99(0.35-1.30)%. The median MNC was (3.80±2.05)×10. The cumulative total of mononuclear cell was (5.84±2.48)×10/kg, and the median CD34cell count was 3.84(3.91-6.5)×10/kg. During the DA-EPOCH+G-CSF mobilization, the peripheral WBC count of patients were decreased to the lowest level at (0.2-1.4)×10/L. The platelet counts were all above 40×10/L except for 1 case in which the platelet count was reduced to 8×10/L. The median percentage of CD34cells in one period of collection was 0.85(0.34-1.2)%. The median MNC was (3.68±1.56)×10. The cumulative total of mononuclear cells was (6.01±2.26)×10/kg, and the median CD34cell count was 4.44(2.7-7.10)×10/kg. There were no statistical differences between the 2 groups in the median percentage of CD34cells, the median MNC, the cumulative total of mononuclear cells and the median CD34cell counts (P>0.05). The average acquired time for granulocyte engraftment was 10.00(9.00-11.00) days, and for platelet engraftment was 12.50(11.00-17.25) days, with no statistical difference(P>0.05). No death occurred during the process of transplantation.

CONCLUSIONDA-EPOCH therapy combined with G-CSF can effectively mobilize the peripheral blood hematopoietic stem cells in NHL patients with higher safety and lower price, and proves to be worth recommending in clinical use.

OBJECTIVETo investigate the alteration and clinical significance of IFN-γ, IL-4, IL-17 and TGF-β levels in serum of patients with chronic lymphocytic leukemia treated with FCR.

METHODSForty-seven CLL patients treated with FCR regimen were enrolled in CLL group, meanwhile 30 healthy persons were selected in control group. The serum levels of IFN-γ, IL-4, IL-17 and TGF-β were detected by ELISA in CLL group before and after treatment and in control group, then the differences of IFN-γ, IL-4, IL-17 and TGF-β levels as well as IFN-γ/IL-4 ratio and TGF-β/IL-17 ratio were compared between 2 groups.

RESULTSBefore treatment with PCR regimen, the IL-4, IL-17 and TGF-β levels as well as TGF-β/IL-17 in CLL group were higher than those in control group (P<0.05), while the IFN-γ level and IFN-γ/IL-4 ratio in CLL group were lower than those in control group (P<0.05); after treatment with PCR regimen, the IL-4, IL-17 and TGF-β levels as well as TGF-β/IL-17 ratio all significantly decreased (P<0.05), while IFN-γ level and IFN-γ/IL-4 ratio significantly increased (P<0.05) as compared with those before treatment, moreover, the IL-4 and IL-17 levels as well as TGF-β/ IL-17 and IFN-γ /IL-4 ratio were no significantly different from those in control group (P>0.05), only the IFN-γ and TGF-β levels were significantly diffrent from control group (P<0.05). The analysis of Binet staging (stage A, B, C) showed that along with pregression of Binet stages, the TGF-γ/IL-17 levels as well as the IFN-γ/IL-4 ratio in CLL group negatively correlated with Binet staging (r=-0.53), while the TGF-β/IL-17 ratio positively correlated with Binet staging (r=0.46). The analysis of grouping accoraing to therapentic efficacy fonnd that the IL-4 and IL-17 levels and IFN-γ/IL-4 and TGF-β/IL-17 ratios in CR and PR groups were significantly different before and after treatment (P<0.05), while those in SD and PD groups did not showed statistical difference before and after treatment (P>0.05).

CONCLUSIONAlong with the progression of disease, the IFN-γ/ IL-4 ratio gradually decreases, and the TGF-β / L-17 ratio gradually increases. The treatment with FCR regimen can overcome this tread, therefore dynamically monitoring the chages of IFN-γ/ IL-4 and TGF-β / L-17 ratios may contribute to guide the clinical treatment.

OBJECTIVETo explore the role of survivin and PI3K/AKT pathway in the pathogenesis of psoriasis vulgaris (PV).

METHODSPlaque-like lesions collected from 22 patients with PV in progressive stage and 18 normal control skin specimens were examined using immunohistochemical staining, Western blotting and real-time quantitative PCR for expressions of survivin, PI3K and AKT in the keratinocytes, and their correlation was analyzed. A small interfering RNA (siRNA) was used to knock down AKT in cultured HaCaT cells, and Western blotting was used to detect the changes in the expression of survivin. RESULTS Compared with normal skin, PV lesions showed obviously up-regulated expressions of survivin, PI3K and AKT in the keratinocytes. Survivin expression was positively correlated with PI3K (r=0.4510, P=0.0351) and AKT (r=0.4423, P=0.0393) in the keratinocytes in PV lesions. In cultured HaCaT cells, siRNA-mediated knockdown of AKT caused down-regulation of survivin expression.

CONCLUSIONSurvivin and PI3K/AKT signaling pathway may participate in the occurrence and progression of PV.

Obejective: To investigate the expression level of suppressor of cytokine signaling SOCS3 mRNA in children's acute lymphoblastic leukemia(ALL); to analyze the relationship between the expresion level of SOCS3 mRNA and disease status and risks of ALL, and to explore the application of SOCS3 mRNA in evaluation of ALL disease status, risk and target therapy.

METHODSThe expression levels of SOCS3 mRNA in bone marrow mononuclear cells from 45 cases of newly diagnosed ALL at initial diganosis and induction remission and 13 normal children as controls were detected by SYBR Green fluorescence quantitative PCR; the correlation of SOCS3 mRNA expression level with risk and clinical characteristics of ALL was analyzed by means of statistical method; the immunofluorescence histochemistry method and laser confocal microscopy were used to detect the sites and expression level of SOCS3 mRNA in bone marrow samples of ALL patients.

RESULTSThe SOCS3 mRNA expression level at initial diagnosis of 45 ALL patients was significantly lower than that of normal controls (P<0.05), and that in induction remission of 45 patients was not significant different from normal controls (P<0.05); the SOCS3 mRNA expression level at initial diagnosis of patients with standasd and high risk was higher than that of patients with low risk (P<0.05). The 45 patients were divided into high expression and low expression groups according to SOCS3 mRNA expression level at initial diagnosis by median method, comparison of clinical characteristics in 2 groups was found that the SOCS3 mRNA high expression group had even more higher leukocyte count in peripheral blood, even more higher LDH level and much more poor prognostic genes; in addition, the high expression group showed more higher risk in comparison between 2 group.

CONCLUSIONThe SOCS3 mRNA expression is down-regulated at initial diagnosis, while recovered to normal level after induction remission of disease, thus the SOCS3 can be used as an indicator for evaluation of disease status. The high expression of SOCS3 mRNA up-regulates the disease risk, therefore the SOCS3 mRNA can be used as a factor for evaluation of ALL risk. The treatment of ALL via regulation of SOCS3 mRNA expression level maybe can become a new way. The regulation of SOCS3 mRNA expression level maybe can become a new way for treatment of ALL.

OBJECTIVETo investigate the efficacy and safety of haploidentical hematopoietic stem cell transplantation(hi-HSCT) combined with bone-marrow derived mesenchymal stem cell (BM-MSC) tranfusion for treatment of children with severe apastic anemia(SAA).

METHODSThe clinical data of 25 children with SAA undergoing hi-HSCT and BM-MSC tranfusion were retrospectively analyzed from August 2014 to July 2016.

RESULTSneutrophil engraftment was achieved in all 25(100%) children, with the median time 12(11-22) days. The median time of platelet engraftment was 21(11-130) days in 23(92%) children. Acute graft-versus-host disease(aGVHD) was observed in 16(64%) cases, including 11 case of grade I and 5 cases of aGVHD grade II-IV, and one of them died of grade IV of skin, gut and liver at day 115; 5 cases of chronic GVHD were found, all of them were limited cGVHD. Cytomegalovirus (CMV) viremia was observed in 23(92%) cases, but no CMV disease was developed after therapy. 3 cases of post-transplant lymphoroliferative disease with 23 EBV viremia positive occurred, all of them were cured after rituximab. Hemorrhagic cystitis appeared in 9 cases with only one case of grade III, 22 children suffered from infection, involving 10 cases in lung and 4 cases in liver, 1 patient was diagnosed as Guillain-Barre syndrome. Autoimmune hemolytic anemia was recorded in 1 patient, 22 children survived during a median following-up time of 14(3-27) months.

CONCLUSIONThe hi-HSCT combined with BM-MSC transfusion for treatment of children with SAA has been confirmed to be safe and feasible.

OBJECTIVETo explore the effects of the shock wave on the capacity of mesenchymal stem cells(MSCs) to proliferate and differentiate into osteoblasts.

METHODSMSCs were isolated from the bone marrow of healthy donors. The human bone marrow MSCs(BM-MSCs) were divided into 3 groups including blank control group,osteoinduced group and shock wave group. The MSCs in blank control group were cultured with common mediam; the MSCs in osteoinduced group were treated with osteogenic agents and cultured; the MSCs in shock wave group were cultured with common medium and stimulated by shock wave. The morphology of MSCs in each groups were observed by micoscopy; the CCK-8 was used to detect the proliferation ability of MSCs; the alkaline phosphatase staining and von Kossa staining were used to evaluale the differentiation potential of MSCs in each groups.

RESULTSThe results of CCK-8 revealed the shock wave could promote cell proliferation as compared with blank control group. The results of alkaline phosphatase and Von Kossa staining showed that the shock wave displayed a stronger ability to promote the human BMMSC differentiation into osteoblasts cells in comparison with the osteoinduced group. The blank control group was weakly positively stainined.

CONCLUSIONThe shock wave treatment can promote proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells.

OBJECTIVETo evaluate safety and efficacy of autologous peripheral blood hematopoietic stem cell transplantation (auto-PBHSCT) with IBu precondition regimen consisting of high-dose idarubicin (IDA) and busulphan (Bu) for treatment of patients with low and intermediate risk acute myeloid leuekmia (AML).

METHODSA total of 11 patients with AML (5 low and 6 intermediate risk patients) treated with auto-PBHSCT with IBu precondition regimen (IDA 20 mg/m, continuous i.v. from d-13 to d-11, Bu 0.8 mg/kg/q6h i.v. for 2h, from d-5 to d-2) from March 2011 to July 2014 were analyzed retrospectively. Adverse effects and transplantation-related mortality (TRM) were evaluated. Kaplan-Meier analysis was performed to calculate the overall survival (OS), disease-free survival (DFS) and cumulative relapse rate (RR). Cox regression was performed for univariate analysis for DFS.

RESULTSAmong the 11 patients, 10 patients obtained hematopoietic reconstitution, 1 patient died during transplantation, thus the TRM was 9.1%. The adverse effects were well tolerated. With median follow-up of 31.6 (8.7-52.5) months, 7 patients (63.3%) were alive, including 6 patients (54.5%) in continuous complete remission (CR). Median OS and DFS were not reached. The 3-year OS, DFS and RR were (57.7±16.3)%, (52.5±17.6)% and 47.5%, respectively. Univartiate analysis indicated that the age, sex, interval between diagnosis and transplantation, white blood cell count at diagnosis, risk-grouping (low or intermediate risk), disease status before transplantation (CR1 or CR2), and count of mononuclear cells for infusion all can not influence DFS(P>0.05, respectively).

CONCLUSIONThe treatment of auto-PBHSCT with IBu precondition regimen for low to intermediate risk AML patients is safe and effective.

OBJECTIVETo investigate the effect of postischemic treatment with endomorphin-1 (EM-1) against myocardial ischemia/reperfusion (IR) injury in rats and on extracellular signal regulated kinase 1/2 (Erk1/2)-dependent signaling pathway.

METHODSSprague-Dawley rats were randomly divided into 5 groups, namely the sham-operated group, IR group, EM-1 post-treatment group (EM50 group), EM-1 post-treatment group with PD98059 treatment (EM50+PD group), and PD98059 post-treatment group (PD group). The hemodynamic indexes of the rats were recorded. After reperfusion, CK-MB, LDH, CTnI, MDA, IL-6, TNF-α, and SOD activities or contents were measured, the infarct size was determined, and the expression levels of Erk1/2, P-Erk1/2 and cleaved caspase-3 were detected using Western blotting.

RESULTSCompared with the sham group, the IR group showed significantly decreased heart rate and mean arterial pressure (P<0.05), which were increased obviously by EM-1 post-treatment (P<0.05). EM-1 post-treatment also resulted in significantly decreased LDH, CK-MB, CTnI, MDA, IL-6, and TNF-α activities or contents (P<0.05), increased SOD activity (P<0.05), reduced the infarct size (P<0.05), and increased the expression level of P-Erk protein (P<0.05). Compared with EM50 group, EM50+PD group showed significantly decreased heart rate and mean arterial pressure (P<0.05), increased LDH, CK-MB, CTnI, MDA, IL-6, and TNF-α activities or contents (P<0.05), decreased SOD activity, increased infarct size (P<0.05), and lowered expression of P-Erk protein (P<0.05).

CONCLUSIONPostischemic treatment with EM-1 protects the heart against IR injury by improving the cardiac function, inhibiting inflammation, and inhibiting oxidative stress and myocardial apoptosis, and Erk1/2 signaling pathway may be involved in this process.