ABSTRACT
The aim of this study was to detect the expression level of eIF4E gene in patients with non-treated, remission and non-remission/relapse acute myeloid leukemia (AML), and other non-malignant haematologic diseases so as to analyze and reveal the relationship of eIF4E gene expression with AML progression. SYBR Green I RT-PCR was used to assay the expression level of eIF4E mRNA extracted from bone marrow mononuclear cells in 30 patients with AML (6 in M2, 5 in M3, 8 in M4, 10 in M5, 1 in M6) and 20 patients with non-malignant hematologic diseases. The β2-microglubin(β2M) was used as internal reference and the formula 2(-ΔCt)×100% was applied to calculate the expression level of eIF4E gene. The results showed that the eIF4E expression level (7.098 ± 5.544)% in patients with non-treated and non-remitted/relapsed AML was significantly higher than that in patients with remission (0.964 ± 0.312)% (P < 0.01) and non-malignant hematologic diseases (0.248 ± 0.163)% (P < 0.01). There was no difference between latter two group patients, even though the expression level of eIF4E gene in patients with M4 and M5 was higher. As compared with non-malignant hematologic diseases, the expression level of eIF4E gene of patients with remission patients showed no significant difference. It is concluded that the over-expression of eIF4E gene has been found in patients with AML, and its level obviously decreases along with remission of disease, thus the eIF4E gene may be a surveillance parameter for disease progression.
Subject(s)
Adolescent , Adult , Aged , Aged, 80 and over , Child , Female , Humans , Male , Middle Aged , Young Adult , Disease Progression , Eukaryotic Initiation Factor-4E , Genetics , Gene Expression , Leukemia, Myeloid, Acute , Genetics , Pathology , Real-Time Polymerase Chain ReactionABSTRACT
In order to investigate expressions of transcription factor GATA-1 and GATA-2 genes in the bone marrow stromal cells (BMSCs) from patients with leukemia or normal controls, bone marrow stromal cells from 34 normal cases and 42 cases with leukemia were cultured long-term in vitro. Nonadherent cells (bone marrow hematopoietic cells) and amplified adherent cells (BMSC) were collected separately. Expressions of GATA-1 and GATA-2 genes were analyzed by using RT-PCR-ELISA; the semi-quantitative expression levels of GATA genes in the BMSCs from patients with leukemia were compared with normal controls. The results showed that expressions of GATA-1 and GATA-2 genes could be detected in the BMSCs and the bone marrow hematopoietic cells from both normal controls and the cases of leukemia. The expression ratio of GATA-1 in the BMSCs from acute lymphocytic leukemia (ALL) (85.7%) was similar to the normal controls (88.2%), whereas the expression ratios in BMSCs from acute myelocytic leukemia (AML) (55.6%) and chronic myelocytic leukemia (CML) (41.2%) were significant lower than the normal controls (P < 0.05). The rank of expression level of GATA-1 gene in the BMSCs was "ALL>AML>normal>CML". There was no difference in the expression level of GATA-2 gene within the BMSCs from normal controls and patients with leukemia. The ranks of expression levels of GATA-1 and GATA-2 genes in bone marrow hematopoietic cells were "AML>normal>ALL>CML" and "AML>CML>ALL>normal". The dominant expression of GATA-2 gene was found in the BMSCs from AML, CML or normal controls. It is inferred that the expressions of GATA-1 and GATA-2 genes in the BMSCs of normal controls and patients with leukemia may influence the regulation of hematopoiesis in the bone marrow stroma and it is worthy of further study to explore their roles in pathogenesis and development of leukemia.
Subject(s)
Adolescent , Adult , Aged , Child , Child, Preschool , Female , Humans , Male , Middle Aged , Bone Marrow Cells , Metabolism , Enzyme-Linked Immunosorbent Assay , GATA1 Transcription Factor , Genetics , GATA2 Transcription Factor , Genetics , Gene Expression Regulation, Leukemic , Leukemia , Blood , Pathology , Reverse Transcriptase Polymerase Chain Reaction , Stromal Cells , MetabolismABSTRACT
Objective To study the incidence,risk factors,clinical outcome,management and prevention of pure red cell aplasia (PRCA) following major ABO-incompatible allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods Forty-two patients underwent major ABO-incompa- tible allo-HSCT,including major ABO-mismatch in 33 patients,major plus minor ABO-mismatch in 9 patients,and 27 recipients with blood group O.Thirteen patients underwent bone marrow transplan- tation,25 peripheral blood stem cell transplantation,and 4 cord blood transplantation.Six patients re- ceived donor-type plasma replacement before transplantation.Cyclosporine A (CsA) and methotrexate (MTX) were used for prophylaxis of graft-versus-host disease (GVHD).Results All 42 patients had sustained engraftment.PRCA occurred in 11/42 patients (26.2%).All the 11 cases of PRCA were in blood group O recipients of grafts from blood group A donor (n=9) or blood group B donor (n= 2);6 patients with blood group O who received donor-type plasma exchange before transplantation did not develop PRCA.PRCA resolved spontaneously in 8 cases with transfusion support.Two patients were treated by donor-type plasma exchange,resulting in the decrease of isoagglutinin titer,followed by complete recovery of erythropoiesis.One patient responded to rituximab and achieved complete re- mission of symptoms of PRCA.Univariate analysis revealed that the most significant risk factors asso- ciated with PRCA were blood group O recipient,blood group A donor,blood group O recipient of graft from blood group A donor;only blood group O/A in recipient/donor pair was identified as being significantly associated with the occurrence of PRCA by multivariate analysis (RR 10.999,95% CI 1.975-61.258,P
ABSTRACT
In order to investigate expression of SCL (stem cell leukemia) gene in bone marrow stromal cells (BMSC) and bone marrow cells from patients with leukemia and normal individuals, bone marrow mononuclear cells from AML (18 cases), CML (17 cases), ALL (7 cases) and normal individuals (33 cases) were cultured long-term in vitro. Nonadherent cells (hematopoietic cells) and amplified adherent cells (BMSC) were collected respectively. RT-PCR-ELISA assay was then performed to detect expression of SCL gene. The expression ratio of SCL gene were analyzed and its expression level was normalized by beta(2)M gene acting as an internal reference for the purpose of semi-quantitative analysis. The results indicated that the expression ratio of SCL gene was lower in BMSC from AML (27.8%) and CML (11.8%) than that in normal controls (69.7%, P < 0.05), while was higher in the nonadherent cells from CML (64.3%) than that in its corresponding BMSC (P < 0.05). Semi-quantitative analysis showed that SCL gene expression level in nonadherent cells from AML was higher than that in its corresponding BMSC (P < 0.05). In conclusion, the low-level expression state of SCL gene in BMSC from patients with AML and CML may be involved in the abnormal regulation of hematopoiesis in myelocytic leukemia.
Subject(s)
Adolescent , Adult , Child , Child, Preschool , Female , Humans , Infant , Male , Basic Helix-Loop-Helix Transcription Factors , Bone Marrow Cells , Metabolism , DNA-Binding Proteins , Genetics , Gene Expression , Leukemia , Metabolism , Proto-Oncogene Proteins , Genetics , Stromal Cells , Metabolism , T-Cell Acute Lymphocytic Leukemia Protein 1 , Transcription Factors , GeneticsABSTRACT
Six out of 20 patients undergoing a major ABO-incompatible allogeneic stem cell transplantation (allo-HSCT) developed pure red cell aplasia (PRCA), which did not show any effects on granulocyte and platelet engraftment, and incidence of grade II-IV aGVHD. All the 6 cases of PRCA were in blood group O recipients of grafts from blood group A donors (n = 5) or blood group B donor (n = 1), suggesting that donor/recipient pair (A/O) is associated with a high risk of PRCA after major ABO-incompatible allo-HSCT. Erythroid engraftment occurred spontaneously in four cases without specific intervention other than the RBC transfusion, which coincided with the decrease of isoagglutinin titers below 8, and the remaining 2 patients with prolonged erythroid aplasia( > 300 days) despite therapy with erythropoietin (EPO) were successfully treated by plasma exchange with donor-type plasma replacement. Cyclosporine did not appear to have played any role in causing PRCA in our patients, however, the occurrence of GVHD may facilitate the recovery of erythropoiesis.
Subject(s)
Adolescent , Adult , Female , Humans , Male , Middle Aged , ABO Blood-Group System , Cyclosporine , Therapeutic Uses , Erythrocytes , Pathology , Erythropoiesis , Graft vs Host Disease , Hematopoietic Stem Cell Transplantation , Red-Cell Aplasia, Pure , Transplantation, Homologous , Treatment OutcomeABSTRACT
Abstract We investigated the effects of IL-3, IL-6 and IL-4, alone and in combination with G-CSF and GM-CSF on hematopoietic reconstitution following murine allogenic bone marrow transplantation. The most effective combination for increasing the circulating absolute neutrophil count (ANC) above the control value at day 6 posttransplant was the combination of IL-3 and GM-CSF and showed a significant synergistic effect. When used alone, both IL-6 and G-CSF raised platelet count at day 6 after transplantation (312?10~(9)/L, P=0.02 and 309?10~(9)/L, P=0.01 respectively) compared to control mice(216?109/L). In combination, only IL-3 with G-CSF significantly increased the day 6 platelet count (328?10~(9)/L). We found that neutrophil recovery at day 7 after transplantation was significantly suppressed by IL-4 in a dose-dependent fashion and that the recovery of nentrophils accelerated by IL-3, and G-CSF was also significantly suppressed by IL-4.