Efficacy of IA regimen followed by FLAG regimen in the treatment of acute myeloid leukaemia / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the efficacy and toxicity of standard-dose IA regimen (idarubicin and cytarabine) as induction therapy followed by FLAG regimen in patients with acute myeloid leukemia (AML), and its influence on peripheral stem cell mobilization.</p><p><b>METHODS</b>A total of 23 previously untreated de novo AML patients were enrolled. Thirteen patients were male, and 10 female, with ages ranging from 14 to 54 (median: 41) years. Cytogenetic analysis was performed for all patients. The IA regimen contained idarubicin (12 mg x m(-2) x d(-1), days 1 to 3) and cytarabine (100 mg x m(-2) x d(-1), days 1 to 7), and the FLAG regimen contained'fludarabine (50 mg/d, days 1 to 5), cytarabine (2 g x m(-2) x d(-1), days 1 to 5) and granulocyte colony-stimulating factor (G-CSF, 300 microg/d, days 0 to 5).</p><p><b>RESULTS</b>After one course of induction therapy, the CR rate was 91.3%. The CR rate for patients with favourable and intermediate prognostic karyotypes was 100% and 91.3%, respectively. Nineteen patients in CR were consolidated with FLAG regimen, of which 6/9 (66.7%) patients were able to mobilize a sufficient number of CD34+ cells and successfully performed autologous stem cell transplantation. Four patients relapsed. The median survival duration was 19.5 months and median disease-free survival was 14 months. Myelosuppression and infections due to neutropenia were the most frequent adverse effects, severe nonhematologic toxicity and the early death were not observed in all patients.</p><p><b>CONCLUSION</b>IA followed by FLAG regimen is effective and well tolerable in AML patients especially in those with favourable and intermediate prognostic karyotypes, and 1 to 2 courses of this therapy shows no influence on peripheral stem cell mobilization and subsequent autologous stem cell transplantation.</p>

Immunophenotypic features in 143 cases of acute promyelocytic leukemia / 中国实验血液学杂志

This study was aimed to investigate the immunophenotypic characteristics of acute promyelocytic leukemia (APL). CD45/Side Scatter (SSC) gating strategy and multiparametric flow cytometry were used to determine immunophenotype of 143 patients with APL. The immunophenotypic features were compared between newly diagnosed APL patients and relapsed APL patients. 42 patients with HLA-DR(-) (non-APL AML, DR(-)AML) were randomly selected as controls. 31 out of 42 AML patients were CD34 negative, and their immunophenotypes were compared with those in newly diagnosed APL patients. The results showed that (1) CD34 and HLA-DR were both negative in 91.9% of newly diagnosed APL, while the positive rate of CD34 and HLA-DR elevated in relapsed cases (3.0% vs 37.5%, 3.9% vs 37.5%). The positive rate of CD34 in HLA-DR(-) AML group was higher than that in newly diagnosed APL group (23.4% vs 3.0%). The positive level of CD34 in newly diagnosed APL group was lower than that in HLA-DR(-) AML group; (2) the positive rate of CD33 in newly diagnosed APL group was higher than that in other groups (97.0% vs 75.0%, 83.3%, 83.9%), as well as the the positive level of CD33 (p < 0.05). (3) no lymphoid antigen other than CD2 was expressed in newly diagnosed APL group. The positive rate of CD7 was 9.5% in DR(-) AML group and 6.5% in CD34(-)/DR(-) AML group, both were higher than those of newly diagnosed APL group (p < 0.05). It is concluded that the immunophenotyping can provide proof to the rapid diagnosis of APL. For those patients with DR(-) AML, it may be helpful to identify APL depending on following features: low or negative CD34 expression, homogeneous and bright expression of CD33, no lymphoid antigens other than CD2, higher SSC.

Standard-dose of idarubicin in combination with continuous infusion of cytarabine as induction therapy in patients with acute myeloid leukaemia / 中国实验血液学杂志

The objective of this study was to investigate the efficacy and toxicity of standard-dose idarubicin in combination with continuous infusion of cytarabine as induction therapy in patients with acute myeloid leukemia (AML). A total of 38 AML patients were enrolled, including 30 new diagnosed patients, 8 relapsed and refractory patients. Cytogenetic analysis was performed in all patients, 15 patients had cytogenetic aberrations including 4 complex abnormalities. All patients were treated with standard-dose idarubicin [12 mg/(m(2).d), days 1 to 3] and continuous infusion of cytarabine [100 mg/(m(2).d), days 1 to 7]. The results showed that after one course of induction therapy, the overall response rate was 89.5% (34/38), and 32 out of 38 (84.2%) patients achieved complete remission (CR), including 27 of 30 (90.0%) new diagnosed AML patients, 5 (62.5%) refractory and relapsed AML patients, all 4 patients with complex cytogenetic aberrations achieved cytogenetic CR. Out of 6 relapsed patients 2 showed as extramedullary relapse, 4 showed as bone marrow relapse. The median survival duration was > 22 months and median disease-free survival time was > 16 months. Myelosuppression and infections due to neutropenia were the most frequent adverse effects, severe nonhematologic toxicity and the early death were not observed in the patients. It is concluded that standard-dose of idarubicin combined with continuous infusions of cytarabine as the induction therapy is highly effective and well tolerated approach in patients with AML, this regimen provides an opportune moment for hematopoietic stem cell transplantation.

Allogeneic peripheral blood stem cell transplantation for 75 cases of hematologic malignancies / 中国实验血液学杂志

The aim of this study was to explore the clinical effect and complications of allogeneic peripheral blood stem cell transplantation (allo-PBSCT) in hematologic malignancies through retrospective analysis of 75 patients (42 male, 33 female; aged from 13 to 72 years old) received allo-PBSCT from HLA matched (n=61) or haploidentical donors (n=14). 75 patients included 35 patients with chronic myeloid leukemia (CML), 30 patients with acute myeloid leukemia, 5 patients with severe aplastic anemia, 3 patients with acute lymphocytic leukemia, one patients with multiple myeloma and one patients with paroxysmal nocturnal hemoglobinuria. Conditioning regimens were (1) Cy/TBI or Bu/Cy; (2) Cy/TBI+Ara-C; (3) fludarabine+TBI/or (CTX+ATG). Minimal residual disease has been monitored regularly by PCR and FISH. Patients received cyclosporine A and methotrexate or ATG and anti-CD25 monoclonal antibody and mycophenolate mofetil for graft-versus-host disease (GVHD) prophylaxis. Relapsing patients after transplantation received DLI and/or chemotherapy. Patient with CML were treated with imatinib. The results showed that 74 patients had hematopoietic reconstitution, and eventually converted to full donor chimerism by FISH or PCR-STR. The median time for the initial hematopoietic reconstitution was 15 (5-25) days. 46 out of 75 patients were alive and median duration was 23 (2-61) months. Among 29 dead patients, 9 died of disease relapse, 7 died of III-IV grade of acute GVHD and 7 died of severe infection (2 patients developed interstitial pneumonia). 9 out of 14 patients received haploidentical transplantation were alive, and the time of event-free survival was 30 (6-53) months, the mean survival time of 5 died patients was 7 (2-17) months. 16 patients were infected by cytomegalovirus, 2 of them died of interstitial pneumonia. None of them suffered from veno-occlusive disease in the liver. It is concluded that allo-PBSCT is effective to treat refractory hematologic diseases, and DLI/or chemotherapy should be used in the patients relapsing after transplantation.

Tandem double autologous peripheral blood stem cell transplants for de novo multiple myeloma / 中国实验血液学杂志

The purpose of this study was to evaluate the efficiency and safety of tandem double autologous peripheral blood stem cell transplants (T-APBSCT) for de novo multiple myeloma (MM) patients. The clinical data of 3 patients treated by T-APBSCT after chemotherapy were analyzed retrospectively. The first mobilization regimen was cyclophosphamide (CTX) combined with G-CSF 5 microg/(kg x d) and the conditioning regimen for the transplantation was 180 mg/m(2) melphalan. The second mobilization regimen was CTX and VP16 in combination with G-CSF 5 microg/(kg x d) and the conditioning regimen for the transplantation was 180 mg/m(2) melphalan or 10 Gy total body irradiation plus 140 mg/m(2) melphalan. The interval of two in tandem autotransplants was 31, 15 and 27 weeks. For two in tandem APBSCT in 3 patients, the cell number of mononuclear cells (MNCs) transfused was 4.7 x 10(8), 2.798 x 10(8), 6.08 x 10(8)/kg and 1.67 x 10(8), 2.798 x 10(8), 4.28 x 10(8)/kg, while the dose for CD34(+) cells were 3.25 x 10(6), 9.6 x 10(6), 5.91 x 10(6)/kg and 6.9 x 10(6), 9.6 x 10(6), 5.91 x 10(6)/kg for their first and second transplants respectively. The results showed that all patients gained prompt and sustained hematopoietic reconstitution. In double tandem transplantation for 3 patients the interval of absolute neutrophil count (ANC) >or= 1 x 10(9)/L were at day 12, 0, 10 and 12, 25, 0; while platelet count >or= 20 x 10(9)/L were at day 12, 0, 10, and 11, 25, 20 days. The median follow-up time for 2 T-APBSCT was 44 (range 19 - 58) months. Two patients survived, one of them was in complete remission and other was in a stable PR stage, but one out 3 patients died at 58 months after T-APBSCT. It is concluded that the method of T-APBSCT for de novo multiple myeloma is probably safe and effective.

Comparison between efficacy and safety of rituximab plus CHOP regimen and CHOP regimen for treatment of newly diagnosed patients with diffuse large B-cell lymphoma / 中国实验血液学杂志

The aim of this study was to compare the efficacy of rituximab plus CHOP regimen and CHOP regimen on newly diagnosed patients with diffuse large B-cell lymphoma (DLBCL), and analyze their toxicities. A total of 69 patients were enrolled from July 2003 to Dec 2006. The patients were non-randomly were divided into 2 groups: 36 received CHOP alone (CHOP group) and 33 received rituximab plus CHOP (R-CHOP group). The complete response (CR) rates, overall survival (OS) and side events of the 2 groups were compared. The results showed that the CR rate in R-CHOP group was higher than that in CHOP group (69.7% vs 47.2%, p = 0.049); especially in patients of male, Ann Arbor III - IV and IPI 3 - 5 (p = 0.017, p = 0.005 and p = 0.000). The estimated mean OS in R-CHOP group was longer than that in CHOP group (45.7 months vs 35.2 months, p = 0.145), and also in the estimated mean progression free survival (PFS) (38.5 months vs. 24.6 months, p = 0.017). The major adverse events in combination group were infusion-related responses which could be well tolerated in patients, and hematological toxicities which were similar to those in CHOP group. In conclusions, Rituximab increases the therapeutic efficacy of CHOP regimen on newly diagnosed patients with DLBCL, without a clinically significant increase in toxicity.

A case report of misdiagnosed mantle cell lymphoma / 中国实验血液学杂志

Mantle cell lymphoma (MCL) is a rare group of non-Hodgkin's lymphoma (NHL), which is difficult to discriminate from other subtype of small lymphocytic lymphoma in morphologic appearance. In order to enhance the understanding of MCL, a case of MCL first diagnosed as follicular lymphoma (FL) was reported. The clinical and laboratory characteristics of MCL were analyzed and summarized. The results showed that the findings of flow cytometry (FCM) and immunohistochemical staining technique were compatible with the diagnosis of MCL. Cytogenetic analysis can detect multiple types of chromosomal abnormalities, including t (11; 14). In conclusion, MCL is a disease which diagnosis is difficulty confirmed. Interphase fluorescence in situ hybridization, multiplex fluorescence in situ hybridization, FCM and immunohistochemical staining technique play important roles in the diagnosis of MCL.

Anti-CD20 monoclonal antibody for the treatment of refractory autoimmune hemolytic anemia / 中国实验血液学杂志

To observe the therapeutic effect and safety of rituximab (anti-CD20 monoclonal antibody) in the treatment of refractory autoimmune hemolytic anemia (AIHA). One AIHA patient refractory to corticosteroid and splenectomy was treated with rituximab, 375 mg/m(2) weekly for four times. Her hemolytic symptoms, adverse effects, hemoglobin (Hb) concentration and other laboratory data were monitored. The results showed that concentration of lactic dehydrogenase (LDH), total bilirubin (TBIL) and indirect bilirubin (IBIL) began to decrease at 11 days after the first dose of rituximab, and decreased to normal range after 45 days. Concentration of hemoglobin increased up to 95 - 100 g/L. The patient remained disease-free 4 months after treatment. No adverse effect was found during the process of treatment. It is concluded that anti-CD20 monoclonal antibody (rituximab) is both effective and safe for the treatment of refractory autoimmune hemolytic anemia.

Immunophenotyping characteristics of T-cell acute lymphoblastic leukemia / 中国实验血液学杂志

The objective of this study was to investigate the immunophenotypic characteristics of T-cell acute lymphoblastic leukemia (T-ALL). Immunophenotyping was performed in 140 T-ALL patients by flow cytometry using a panel of monoclonal antibodies and CD45/SSC gating. The results showed that the T-lineage-associated antigen expressions were CD7 > CD2 > CD3 > CD5 successively. The positive rate of CD10 was 19.42% in patients. Among 140 cases of T-ALL, 12 (8.57%) was accompanied by B-lineage associated antigen expression. Myeloid antigen expression was identified in 31 out of 136 cases (22.79%). None of them expressed CD14 antigen. The positive rate of CD34 was 31.06%. The positive rate of myeloid antigen expression in CD34(+) T-ALL (36.58%) was significantly higher than that in CD34(-) T-ALL (15.38%) (p < 0.01). The expression of CD3 in child T-ALL was higher than that in adult T-ALL, whereas the expression of CD33 in children was lower than that in adults. It is concluded that immunophenotyping is an important tool for diagnosis of T-ALL. Immunophenotypic characteristics of T-ALL is heterogeneous.

Expression of Ki-67 and Bcl-2 in adults and children with acute lymphoblastic leukemia and its clinical significance / 中国实验血液学杂志

To evaluate the expressions of proliferative antigen Ki-67 and apoptosis-antagonizing protein Bcl-2 as well as their clinical significance, immunohistochemistry staining with SAP was used to detect Ki-67 antigen and Bcl-2 protein in 18 cases of children with acute lymphoblastic leukemia (ALL) and 43 cases of adults with ALL. The results showed that the levels of Ki-67 and Bcl-2 expression in children with ALL were lower than that in adults, but only Bcl-2 expression had significant difference. Both in children and in adults, the levels of Ki-67 expression in T-ALL and My(+) ALL were higher than that in B-ALL and null-ALL. The highest complete remission rate (CR) was seen in the group with lower expression of both indexes (Ki-67 and Bcl-2). The lowest CR rate was seen in the group with higher expression of both indexes. It is concluded that the levels of Ki-67 and Bcl-2 expression in children and adults with ALL were closely related with the subtype of ALL and chemotherapeutic effects.

Expressions of ki-67 and bcl-2 in 29 cases of chronic lymphocytic leukemia and their clinical significance / 中国实验血液学杂志

To investigate the expressions of proliferative antigen Ki-67 and apoptosis-antagonizing protein Bcl-2 in patients with chronic lymphocytic leukemia (CLL) and their clinical significance, immunohistochemistry method was used for detection of Ki-67 and Bcl-2 in bone marrow or peripheral blood of 29 patients with CLL. The results showed that the level of Ki-67 expression in advanced stage of CLL was higher than that in early stage of CLL and there was significant difference between these stages of CLL, but there was no significant difference between expression levels of Bcl-2 in two stages. The survival time in the group with Ki-67 expression < or = 8% was longer than that in the group of Ki-67 > 8%, and there was no significant difference of survival time between high and low groups in terms of Bcl-2 expression. It is concluded that detection of Ki-67 antigen and Bcl-2 protein for CLL patients can reflect the status of proliferation activity and apoptosis suppression of leukemia cells in patients; the level of Ki-67 expression closely correlate with the Binet stage and prognosis of CLL.

Monitoring of bcr/abl fusion gene by interphase-dual-color and dual-fusion fluorescence in situ hybridization in CML after allo-HSCT / 中国实验血液学杂志

This study was aimed to investigate the sensitivity and clinical application value of interphase-dual-color and dual-fusion fluorescence in situ hybridization (DD-FISH). The minimal residual disease (MRD) in 19 patients with chronic myelogenous leukemia (CML) after allogeneic hematopoietic stem cell transplantation (allo-HSCT) was detected by DD-FISH, and the detected results were compared with those of conventional cytogenetics (CC) and reverse transcription-polymerase chain reaction (RT-PCR). The samples were collected from bone marrow or peripheral blood or smears of bone marrow. The results indicated that 14 out of 19 patients achieved and maintained continuous complete molecular remission after transplantation. In these patients, CC assay displayed normal donor karyotype, result of RT-PCR was negative, complete donor chimerism was detected after 2 months of transplantation, result of DD-FISH was negative, average time of the follow-up survey was 11.25 months, MRD did not increase. Results of CC and RT-PCR in 1 patient showed negative, while FISH of sex chromosome showed mixed chimerism, result of DD-FISH was positive, MRD did not increase, no therapy was given for this patient, clinical state of patient was stable. Three patients with hematological relapse demonstrated obvious increase of MRD detected by DD-FISH and sex FISH, result of RT-PCR was found positive in them, but the abnormal result of CC was observed only in 1 patient. After donor lymphocyte infusion and imatinib mesylate treatment, these 3 patients achieved cytogenetic remission again, results of DD-FISH, CC and PCR were negative in them. DD-FISH, CC and PCR in bone marrow and peripheral blood from one patient with extramedullary relapse revealed negative results, and the complete chimerism was found in this patient. It is concluded that interphase-dual-color and dual-fusion fluorescence in situ hybridization is a more reliably sensitive and practicable method for monitoring MRD in patients with CML after allo-HSCT, and can be used in detection of chromosome sample and blood or bone marrow smears. Dynamic detection of bcr/abl fusion gene level by FISH may predict disease changes and guide individual therapy.

Molecular cytogenetic characteristics of chronic lymphocytic leukemia / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To explore the molecular cytogenetic characteristics in patients with chronic lymphocytic leukemia (CLL).</p><p><b>METHODS</b>Interphase fluorescence in situ hybridization (FISH) was used to detect trisomy 12, deletion of 13q14 and 17p13 in 60 patients with CLL.</p><p><b>RESULTS</b>Out of the 60 patients, 41 (68.3%) had at least one kind of molecular cytogenetic aberrations. Two (3.3%) had two kinds of abnormalities. Trisomy 12 was found in 12 (20.0%) cases, 13q14 deletion in 24 (40.0%) cases and 17p13 deletion in 5 (11.7%) cases. The number of trisomy 12 cells ranged from 4.0% to 34.0%, 13q14 deletion ranged from 22.0% to 93.0% and 17p13 deletion ranged from 6.0% to 68.0%. There was no significant difference among each Binet stages.</p><p><b>CONCLUSION</b>FISH is a more rapid, accurate and sensitive technique in analysis of chromosome aberrations in CLL. FISH may provide accurate information of molecular cytogenetics for CLL.</p>

JWA gene in regulating committed differentiation of HL-60 cells induced by ATRA, Ara-C and TPA / 中国实验血液学杂志

The study was aimed to explore the role of gene JWA, a novel retinoic acid responsible and cytoskeleton associate gene, in regulating committed differentiation of HL-60 cell and the molecular mechanism in the course of differentiation and apoptosis of leukemic cells. By using FCM, the changes of CD13, CD14, CD15, CD11b and cell cycles were detected in HL-60 cells treated with ATRA (10(-6) mol/L), Ara-C (10 ng/ml) and TPA (10(-8) mol/L) respectively. The samples were determined by semi-quantitative reverse transcript-polymerase chain reaction (RT-PCR) and Western blot for the expression of JWA, Bcl-2, HSP27 and HSP70 at day 0, 2, 4, 6, 8. The results showed that HL-60 cells committedly differentiated into granulocyte-, monocyte-, macrophage-like cells. As a result, JWA was up-regulated in a time-dependent manner, while Bcl-2 was down- regulated at the same time. In ATRA and TPA group, the change of HSP70 had positive correlation with JWA, and negative correlation with Bcl-2. The expression of HSP27 was not detected. Contrast to the cells from APL patient, the expression of JWA need not be activated by ATRA in advance. In this study, we also exposed HL-60 cells in higher dose of Ara-C (20 ng/ml), and JWA expression underwent opposite trend comparing with in lower dose of Ara-C (10 ng/ml). It is concluded that JWA may play double important roles in regulating ATRA and TPA-induced differentiation and apoptosis in leukemic cells. The JWA expression had a negative correlation between induction and cytotoxic response. The difference of JWA expressions between HL-60 cell and ANLL patient cells would be involved in different leukemia pathogenesis.

Expression of P27(Kip1) and its clinical significance in acute and chronic leukemia / 中国实验血液学杂志

To evaluate the expression of cyclin dependent kinase inhibitor P27(Kip1) in leukemia and to investigate its clinical significance, the P27(Kip1) protein in bone marrow or peripheral blood samples from 82 cases of leukemia was measured by Western blot and enhanced chemoluminescence (ECL). The results showed that the expression of P27(Kip1) protein in ALL was higher than that in ANLL (P = 0.033) and also that in CML (P = 0.008). P27(Kip1) expression in CLL was higher than that in CML too (P = 0.017). In acute leukemia, the effective rate (CR and PR) of initial chemical therapy in the group of P27(Kip1) > 0.655 was higher than that in the group of P27(Kip1) < or = 0.655, P = 0.041. For ANLL and ALL patients, the survival time in the group of P27(Kip1) > 0.655 was longer than that in the group of P27(Kip1) < or = 0.655, P = 0.0065. There were similar statistical significance for ANLL and ALL patients, P = 0.0271 and P = 0.0266 respectively. There was a negative correlation between chromosomal abnormalities and P27(Kip1) expression in ALL patients (r = -0.775, P = 0.04). The expression of P27(Kip1) protein appeared nothing to do with sex, age, white blood cell number, blast cell number in peripheral blood, serum LDH or uric acid. In conclusion, the expression level of P27(Kip1) protein is in relation to the effect of initial chemical therapy and survival time, so that the lower P27(Kip1) expression may associated with poor prognosis in acute leukemia.