ABSTRACT
Objective To explore the effect of stem cell factor (SCF), Flt3 Ligand (FL) and Thrombopoietin (TPO) on the expansion of the mononuclear cells derived from umbilical cord blood ex vivo under hypoxic conditions.Method Human fresh CB mononuclear cells were isolated and cultured with different combinations of cytokine for 7 days under normoxic or hypoxic conditions (3 ％ O2).At day 0 and 7 of culture, total nucleated cells (TNC), CD34+ , CD34+ CXCR4+ , CD34+ CD49d+ and CD34+ CD62L+ cells were assayed by flow cytometry, and colony forming unit (CFU) was determined.The experiment was divided into four groups: group A (cells were cultured in single mononuclear cells under normoxic conditions);group B (cells were cultured in single mononuclear cells under hypoxic conditions);group C (cells were cultured with SCF, FL and TPO under normoxic conditions);group D (cells were cultured with SCF, FL and TPO under hypoxic conditions).Result At 7th day of culture, TNC, CD34+ cells and CFU expansion were significantly increased in groups B, C and D, and the expansion effect in group D was superior to that in groups B and C, but there was no significant difference between groups B and C (P＞0.05).At 7th day of culture, the expression levels of CD49d, CD62L and CXCR4 on expanded CD34 + cells were significantly reduced in group A as compared with those at day 0, and those in groups B, C and D were significantly increased as compared with those in group A, those in group D were significantly higher than in group D than in groups B and C (P＜0.05),but there was no significant difference between groups B and C (P＞ 0.05).Conclusion Hypoxic conditions could enhance the expansion and expression of CD49d, CD62L and CXCR4 of mononuclear cells derived from umbilical cord blood with SCF, FL and TPO.
ABSTRACT
Objective To evaluate the effectiveness and side effect of MT regimen (mitoxantrone plus teniposide) in inductive chemotherapy and explore the relationship between the effectiveness and karyotype. Methods 33 patients with acute monocytic leukemia were divided into two groups according to the treatment history or risk status according to cytogenetics MRC criteria. Group A (n=23) and B (n=10) were primary treatment and no remission following one course of DA (daunorubicin plus cytarabine) or HDA (Harringtonine,daunorubicin plus cytarabine) regimen,respectively. According to MRC criteria,group C (n=29) and D (n=4) were intermediate and adverse group. All the cases received two courses MT regimen chemotherapies to induce remission. The results and side effects were analysed. Results The complete remission rate and effective rate in group A and B were 83 % (19/23) and 60 % (6/10),91 % (21/23) and 70 % (7/10) respectively. The complete remission rate and effective rate in group C and D was 83 % (24/29) and 25 % (1/4),88 % (26/29) and 50 % (2/4) respectively. In complex cytogenetic group and 11q23 abnormal without complex cytogenetic group,CR rate was 0 (0/3) and 100 % (4/4). The time point,count of WBC nadir and the duration of WBC were less than 1×109/L is (7±3) day after chemotherapy,(0.4±0.2)×l09/L,(8±5) day. Chemotherapy related mortality was 0. Conclusion MT regimen was highly effective and safe in inducing remission in acute monocytic leukemia,including the cases which achieved no remission following one course of DA or HDA regimen. The effectiveness of MT regimen relates to the cytogenetics. MT regimen may be highly effective in cases with 11q23 abnormal and poor effective in cases with complex cytogenetic.
ABSTRACT
ObjectiveTo investigate the effect of artesunate on the expression of vascular endothlial growth factors(VEGF)and VEGFR in SHI-1 cell line.MethodsEnzyme-linked immunosorbent assay analysis was performed to detect the amount of VEGF in culture supernatants of SHI-1 cell in the condition of artesunate or not. The expression of VEGFR1 and VEGFR2 in SHI-1 cell in the condition of artesunate or not were detected by flow cytometry.ResultsWithout artesunate,the concentration of VEGF in the culture supernatant of SHI-1 cell were (980.3±2.2) pg/ml in 24 h and (982.4±2.3) pg/ml in 48 h. The expression of VEGFRI in SHI-1 cell were (6.40±3.11) ％ in 24 h and (6.45±2.85) ％ in 48 h. The expression of VEGFR2 in SHI-1 cell were (13.90±2.26) ％ in 24 h and (13.95±1.96) ％ in 48 h. With artesunate at 5, 10, 20 ng/ml, the concentration of VEGF in culture supematant of SHI-1 cell were (234.6±1.8)pg/ml, (114.9±1.6)pg/ml, (108.8±1.5) pg/ml in 24 h and (62.3±1.7) pg/ml, (60.9±1.6) pg/ml, (32.7±1.7) pg/ml in 48 h, respectively. The levels of VEGF in SHI-1 cells treated with artesunate at different concentrations decreased significantly (P ＜0.05).There was significant difference between 24 hours group and 48 hours group(P ＜0.05).The expression of VEGFR1 in SHI-1 cell were (4.30±2.21) ％, (4.20±1.37) ％, (3.90±1.86) ％ in 24 h and (3.80±2.87) ％, (3.60±1.73) ％, (3.00±1.82) ％ in 48 h, respectively. The expression of VEGFR1 in SHI-1 cell treated with artesunate at different concentrations were not significantly different (P ＞0.05). No significant difference between 24 hours group and 48 hours group was observed (P ＞0.05). VEGFR2 expression of SHI-1 cell were(4.40±1.15) ％, (3.10±0.68) ％, (1.10±0.72) ％ in 24 h and (3.00±1.68) ％, (2.20±0.93) ％, (0.60±0.92) ％ in 48 h, respectively. The results indicated that the expression of VEGFR2 in SHI-1 cells treated with artesunate at different concentrations reduced significantly (P ＜0.05),but there was no significant difference between 24 h group and 48 h group (P ＞0.05). ConclusionThe concentration of VEGF in SHI-1 cell was high, and artesunate can down-regulate the expression of VEGF and VEGFR2,but the effect of artesunate on the VEGFR1 was not significant.
ABSTRACT
Objective To explore the status of histone acetylation modification and their regulatory effect to hMSH2 gene and hMLH1 gene expression in acute leukemia. Methods Reverse transcription-polymerase chain reaction (RT-PCR) was used to measure the expression of hMSH2 and hMLH1 mRNA, and Western blot was used to measure the expression of histone H3, H4, HDACi, hMSH2 and hMLH1 protein in mononuclear cells of 56 acute leukemia patients and 30 healthy volunteers. The mononuclear cells of 30 acute leukemia patients were treated with histone deacetylase inhibitors trichostatin A (TSA), and measured the expression difference of histone H3, H4, HDAC1, hMSH2 and hMLH1 in the mononuclear cells treated with TSA. Results The protein expression levels of hMSH2, hMLH1, histone H3 and histone H4 in those mononuclear cells of acute leukemia patients were 0.4610±0.1211, 0.4013±0.1143, 0.4103±0.1241 and 0.4251±0.1081, respectively, which were significantly decreased comparing with those of healthy volunteers (0.9461±0. 1841, 0.996±0.2021, 0.8971±0. 1194 and 0.9513±0.1953) (t = 3.341, 3.935, 2.843 and 3.575,respectinely, P ＜0.05). The protein expression levels of HDAC1 (0.8841±0.2018) of acute leukemia patients was significantly increased comparing with those of healthy volunteers (0.5142±0.1340) (t= 2.634, P ＜0.05).After treatment with TSA for 48 hours, the protein expression of hMSH2 was increased nearly 1.5-fold, hMLH1 about 1.6-fold, H3 about 2.9-fold and H4 about 3.4-fold comparing with the negative control groups (P ＜0.05),while the protein expression of HDAC1 were decreased comparing with the negative control groups by 40 %.Conclusion There was an low expression phenomenon of histone acetylation in acute leukemia, and histone acetylation played an important role in regulation of the mismatch repair gene expression in acute leukemia.
ABSTRACT
Objective To evaluate the effectiveness and side effects in sibling and unrelated HLA identical allogeneic hematopoietic stem cell transplantation for multiple myeloma patients under 45-year-old at early stage. Methods Three patients with multiple myeloma ranged from 38 to 44-year-old received two courses of chemotherapies and achieved partial remission. Sibling HLA identical allogeneic hematopoietic stem cell transplantations were underwent in case 1 and 2, and unrelated were in case 3. The conditioning regimens for case 1 and 2 included fludarabine, busulfan plus cyclophosphamide, and of case 3 included modified busulfan, cyclophosphamide plus antithymocyte globulin. Cycloporine A combined with methotrexate were used to prevent GVHD in the case 1 and 2, and methotrexate, mycophenolate and cycloporine A were used in case 3. Results All patients achieved full donor chimerism without graft failure. Grade Ⅱ acute GVHD and extensive chronic GVHD were found in case 1, but not in case 2 and 3. The period of follow-up of case 1, 2 and 3 were 48, 27 and 6 months, respectively, and all of them were alive with no signs of relapse. Conclusion The multiple myeloma patients under 45-year-old underwent sibling and unrelated HLA identical allogeneic hematopoietic stem cell transplantation at early stage after chemotherapy remission have the low treatment-related mortality, high complete remission rate and may prolong long-term survival.
ABSTRACT
Objective To compare the therapeutic and adverse effects of MT regimen (mitoxantrone plus teniposide) and DA regimen (daunorubicin plus cytarabine) on initial treatment to acute monocytic leukemia. Methods 40 patients with initial treatment to acute monocytic leukemia were randomly divided into MT group(n=23) and DA group(n=17). All patients were treated with MT or DA regimen. The result and adverse effects of the two regimens were compared. Results Complete remission(CR) rate in the first course chemotherapy in MT and DA regimen was 65 % and 18 %, respectively. The total CR rate in MT and DA regimen was 83 % and 47 % respectively. The total effective rate was 92 % and 59 %, respectively. Significant differences were found. Severe myelosuppression occurred in both groups. The counts of wbc nadir and the durations of wbc less than 1×10~9/L were not significantly different in two group. The time points of wbc nadir, the start and end time points of wbc less than 1×10~9/L were significantly later in MT group than in DA group. Conclusion MT regimen is significant better than DA regimen in inducing remission in initial treatment acute monocytic leukemia, and it is a good choice for inducing remission strategy. The degrees of myelosuppression in two groups are similar. But the occurrent time of myelosuppression is later in MT group than in DA group. The great attention should be paid to anti-infection and support therapy at time properly.
ABSTRACT
<p><b>OBJECTIVE</b>To evaluate the application of anti-T-lymphocyte globulin (ATG) based nonmyeloablative but profoundly immunosuppressive regimens followed by donor lymphocyte infusion (DLI) for the treatment of hematologic malignancies.</p><p><b>METHODS</b>The protocol was designed to minimize the intensity of the conditioning regimen to the range of nonmyeloablative therapies based on ATG with low-dose busulfan (Bu) and Cytoxan (CTX) (15 - 19.5 mg/kg, 8 mg/kg and 80 mg/kg, respectively). The patients received the first lymphocytic infusion from HLA-identical sibling donors on days 28 - 30 after transplant, and the first T cell dosage of 10(6)/kg followed by the escalated dosage in the range of (0.5 - 1.5) x 10(8)/kg. The total number of procedures were performed at a median of 4.2 procedures (range of 2 - 8 procedures).</p><p><b>RESULTS</b>Engraftment was documented in all six patients in the form of donor-recipient hematopoietic cells mixed chimera at early-stage posttransplant, which was converted gradually into complete chimera by DLI in four patients. Graft-versus-host disease (GVHD) developed in three of six cases, only one of which was severe. To date, four patients are disease free and alive.</p><p><b>CONCLUSIONS</b>Allogeneic donor stem cell engraftment into host can be achieved by nonmyeloablative conditioning regimen based on ATG. Transient mixed donor-recipient hematopoietic cell mixed with chimeras may be successfully converted into complete chimerism by DLI posttransplant. GVHD remains major clinical concern in our study.</p>
Subject(s)
Adolescent , Adult , Female , Humans , Male , Middle Aged , Antilymphocyte Serum , Therapeutic Uses , Graft vs Host Disease , Hematologic Neoplasms , Therapeutics , Hematopoietic Stem Cell Transplantation , Histocompatibility Testing , Lymphocyte Transfusion , T-Lymphocytes , Allergy and Immunology , Transplantation Chimera , Transplantation ConditioningABSTRACT
Objective To evaluate the effectiveness and side-effects of antithymocyte globulin(ATG) and antilymphocyte globulin(ALG) in nonmyeloablative stem cell on transplantation complication.Methods Fourteen cases of hematologic malignancies and 11 cases of sever aplastic anemia(SAA) were treated with allogenic bone marrow transplantation or cord blood haemopoietic stem cell transplantation based on ATG/ALG.Five patients with malignant hematonosis were received donor lymphocyte infusion(DLI) after transplantation.The protocols for graft-versus-host disease(GVHD) prophylaxis consisted of Cyclosporin A(CSA) and methotrexate(MTX) for malignant hematonosis patients or CSA and methylprednisolone(MP) for patients with SAA.Results Three patients had not evidence of engraftment and died from infection at early stage.Other patients recovered haematopoiesis.The mean time of ANC more than 0.5?10~9/L and Plt more than 20?10~9/L were 12.1(3～29) and 20.1(5～79) days posttransplant respectively.Three patients achieved donor complete chimera(CC).Five malignant patients with transient mixed chimerism(MC) grdually converted into complete chimerism by DLI post transplant.Nineteen patients achieved MC and four of them coverted into donor haematopoietic cell complete chimerism.There was no aGVHD in early stage post transplant.There were 1 patient with Ⅰgrade aGVHD,3 with Ⅱgrade aGVHD,2 with skin local cGVHD and 2 with extensive cGVHD after DLI,respectively.There were 2 patients with bacteria infection complications,4 with virous infection and 5 with fungal infection.All patients complicated with chills and fever in the use of ATG and ALG.Conclusion Treatment with ATG and ALG is safety and tolerant for the patients,and can enhance the engraftment of haemopoietic stem cell and decrease the aGVHD.