Background: Bcr/abl fusion gene plays an important role in diagnosing and treating chronic myelogenous leukemia. Objective: to detect fusion genes: b3a2, b2a2, b3a3, b2a3 and e1a2 in patients with chronic myelogenous leukemia by using Nested RT - PCR technique. Subjects and methods: Peripheral blood samples were analyzed by Nested RT - PCR assay from 30 adult patients. Results: 28/30 patients showed bcr/abl fusions gene; among them 20/30 patients showed b3a2 fusions gene, 5/30 patients showed b2a2 fusions gene, 2/30 patients showed co-expression of the b3a2 and b2a2. 1/30 showed e1a2; 2/30 patients showed negative fusion gene. Count of leukocytes and platelets of patients with b3a2 fusion genes were 311.3 G/l and 597.5 G/l, respectively and of patients with b2a2 fusion genes were 136.7 G/l and 333 G/l, respectively. Conclusion: Most of patients showed b3a2 fusion gene, while remaining showed b2a2 transcripts or the co-expression of the b3a2, only one case showed e1a2 fusion gene, two patients showed negative fusion gene. There was no case which showed b3a3 or b2a3 fusion gene. Nested RT assay should be used to determine bcr/abl fusion genes for patients with chronic myelogenous leukemia\r\n', u'\r\n', u'
Leukemia ; Myelogenous ; Chronic ; BCR-ABL Positive/ blood ; pathology

We describe two cases of pulmonary arterial hypertension (PAH) that occurred under dasatinib treatment and were resolved after dasatinib discontinuation. Two patients with chronic phase chronic myeloid leukemia (CML) were switched to dasatinib therapy because of hematological progress while receiving imatinib. These patients had New York Heart Association (NYHA) functional class II dyspnea with elevated right ventricular systolic pressure (RVSP), which progressed under dasatinib treatment. After dasatinib treatment was discontinued, subjective symptoms were improved to NYHA functional class I and the follow-up transthoracic Doppler echocardiography showed improved RVSP. Treatment with an alternate tyrosine kinase inhibitor was initiated and had been continued without development of dyspnea or elevation of RVSP. This report suggests that dasatinib can cause the reversible PAH, therefore, routine cardiopulmonary evaluation before and during treatment with dasatinib may be needed in CML patients with clinical manifestations.
Blood Pressure ; Dyspnea ; Echocardiography, Doppler ; Follow-Up Studies ; Heart ; Humans ; Hypertension* ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive* ; Protein-Tyrosine Kinases

The objective of this study was to analyze the level of bcr-abl mRNA in peripheral blood (PB) after allogeneic stem cell transplantation (allo-SCT) in chronic myeloid leukemia patients, providing a experimental basis for diagnosing early relapse. bcr-abl mRNA levels in 78 PB and bone marrow (BM) samples from 15 CML patients after allo-SCT were detected by using real-time quantitative PCR. The results indicated that levels of bcr-abl mRNA before transplantation were high (median 29.303%) and decreased greatly (median 0) at the first month after allo-SCT. During the first year after allo-SCT, the patterns of serial bcr-abl transcripts varied in number, but the overall bcr-abl transcript levels significantly decreased at 6 months after allo-SCT. Majority of patients with undetectable or very low levels of bcr-abl mRNA were monitored after 1 year following transplantation. The hematological features of BM and PB in all detected patients remained normal. PB and BM bcr-abl values were not different significantly and had the similar trend of changes. It is concluded that the bcr-abl mRNA levels in CML patients change greatly early after allograft. Serial monitoring measurements for bcr-abl mRNA contribute to understanding the trend of change and effect of transplantation, also can be a guidance for starting therapy. But detectable levels of bcr-abl mRNA during the first 6 months do not indicate relapse. Measurements of bcr-abl mRNA of PB may be more suitable for routine monitoring long-term disease status in CML after allo-HSCT.
Adolescent ; Adult ; Bone Marrow ; metabolism ; Fusion Proteins, bcr-abl ; blood ; metabolism ; Hematopoietic Stem Cell Transplantation ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; metabolism ; therapy ; Neoplasm, Residual ; diagnosis ; RNA, Messenger ; blood ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Young Adult

OBJECTIVETo understand the predictive value of early monitoring BCR-ABL transcripts in patients with chronic myeloid leukemia (CML) after treatment with tyrosine kinase inhibitor (TKI), and to provides the information for early assessment of prognosis and treatment options.

METHODSBCR-ABL transcripts of 53 CML patients before and after TKI treatment were detected by using real-time quantitative RT-PCR. The relationship between BCR-ABL transcripts level after TKI treatment for 3 months and the later molecular response, progression and mutation was analyzed.

RESULTSThe median values of BCR-ABL transcripts in peripheral blood samples from 30 newly diagnosed patients were 43.99%, which was used as a baseline of BCR-ABL transcripts for molecular response evaluation. Of 53 patients, 31 (58.49%) had a BCR-ABL mRNA ≤ 4.40% (reduced more than 1 log) and 22 (41.51%) greater than 4.40% (reduced to less than 1 log) after 3 months of TKI treatment. The former 31 patients had a significantly higher 18-months cumulative incidence of major molecular response (MMR) (90.32% vs 18.18%, P=0.000) and 3-year cumulative incidence of complete molecular response (CMR) (48.39% vs 0, P=0.000) compared with the latter 22 patients. The lower BCR-ABL level was, the earlier MMR reached. The proportion of patients with a mutation in group of BCR-ABL mRNA>4.40% was significantly higher than that of BCR-ABL mRNA ≤ 4.40% (22.73% vs 0, P=0.021). The incidence of progression increased in group of BCR-ABL mRNA>4.40%, but the difference was not statistically significant (P=0.052).

CONCLUSIONIt is important for the prognosis evaluation of the patients to monitor the level of BCR-ABL transcripts at 3 months after TKI treatment, which might help to early optimization of treatment and to improve curative effect of CML patients.

Adult ; Aged ; Female ; Fusion Proteins, bcr-abl ; blood ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; diagnosis ; drug therapy ; Male ; Middle Aged ; Predictive Value of Tests ; Prognosis ; Protein Kinase Inhibitors ; therapeutic use ; RNA, Messenger ; genetics ; Treatment Outcome ; Young Adult

To explore the hematopoietic reconstitution and transplantation-related complications of two units of unrelated umbilical cord blood combined transplantation for the treatment of adult hematologic malignancies, one adult patient with chronic myelogenous leukemia received two units of unrelated umbilical cord blood combined transplantation. The conditioning regimen was busulfan and cyclophosphamide (Bu-Cy). GVHD prophylaxis regimen consisted of mycophenolate mofetil (MMF), cyclosporine A (CsA) and methotrexate (MTX). The patient received total nucleated cells 4.63 x 10(7)/kg with CD34+ cells 8.34 x 10(5)/kg. Engraftment was documented by the analysis of short tandem repeat with polymerase chain reaction (STR-PCR). The results showed that the STR-PCR analysis for peripheral blood at day 31, 46 and 71 after transplantation suggested that one of two units of cord blood were completely engrafted. The ANC > 0.5 x 10(9)/L in the patient occurred at day 23, blood platelet counts > 20 x 10(9)/L at day 33 and > 50 x 10(9)/L at day 47. The Philadelphia chromosome and bcr/abl fusion gene of the patient also turned to negative after engraftment. Acute GVHD grade II occurred at day 13 and cured after treatment. It is concluded that umbilical cord blood can be used in adult hematopoietic stem cell transplantation. Two or more units umbilical cord blood combined transplantation might be the way to solve the problem of the low counts of nucleated cells when be used for adult.
Adult ; Cord Blood Stem Cell Transplantation ; adverse effects ; Female ; Graft vs Host Disease ; therapy ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; blood ; therapy ; Leukocyte Count

Imatinib resistance has become a major clinical problem for chronic myeloid leukemia. The aim of the present study was to investigate the involvement of MEG3, a lncRNA, in imatinib resistance and demonstrate its underlying mechanisms. RNAs were extracted from CML patients’ peripheral blood cells and human leukemic K562 cells, and the expression of MEG3 was measured by RT-qPCR. Cell proliferation and cell apoptosis were evaluated. Western blotting was used to measure the protein expression of several multidrug resistant transporters. Luciferase reporter assay was performed to determine the binding between MEG3 and miR-21. Our results showed that MEG3 was significantly decreased in imatinib-resistant CML patients and imatinib-resistant K562 cells. Overexpression of MEG3 in imatinib-resistant K562 cells markedly decreased cell proliferation, increased cell apoptosis, reversed imatinib resistance, and reduced the expression of MRP1, MDR1, and ABCG2. Interestingly, MEG3 binds to miR-21. MEG3 and miR-21 were negatively correlated in CML patients. In addition, miR-21 mimics reversed the phenotype of MEG3-overexpression in imatinib-resistant K562 cells. Taken together, MEG3 is involved in imatinib resistance in CML and possibly contributes to imatinib resistance through regulating miR-21, and subsequent cell proliferation, apoptosis and expression of multidrug resistant transporters.
Apoptosis ; Blood Cells ; Blotting, Western ; Cell Proliferation ; Drug Resistance ; Humans ; Imatinib Mesylate* ; K562 Cells ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive* ; Luciferases ; Phenotype ; RNA ; RNA, Long Noncoding*

This study was purposed to establish and identify the model of extramedullary infiltration of CML-NOD/SCID mice. 24 mice were irradiated with 270 cGy of (137)Cs and absorbed dose rate 80 cGy/min, and were randomly divided into test group I, test group II and control group. The mice in test group I and test group II were injected with 5×10(6) and 1×10(7) K562 cells per mouse respectively, the mice in control group were injected with 0.2 ml of normal saline. The general situation and survival time of these mice were monitored, the extramedullary infiltration of leukemia cells was detected by histopathology examination and RT-PCR. The results indicated that at 4 - 8 weeks after injection, all the mice of group I and group II displayed extramedullary infiltration, suggesting that CML/NOD-SCID model was successfully established. It is concluded that the model of extramedullary infiltration of CML/NOD-SCID mice can be established by injection K562 cells into caudal vein, and can be confirmed by histopathologic examination and detection of BCR-ABL fusion gene using RT-PCR.
Animals ; Disease Models, Animal ; Female ; Humans ; Injections, Intravenous ; K562 Cells ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; Male ; Mice ; Mice, Inbred NOD ; Mice, SCID ; Neoplasm Transplantation ; Tail ; blood supply

OBJECTIVETo evaluate the overall efficacy and transplant-related mortality (TRM) of related and unrelated allogeneic peripheral blood hematopoietic stem cell transplantation (allo-PBSCT) in chronic myeloid leukemia (CML) patients conditioned with fludarabine-busulfan (FB) reduced intensity regimen.

METHODSForty-four patients received FB (Flud 30 mgxm(-2)xd(-1) x 5 d, BU 4 mgxkg(-1)xd(-1) x 3 d) conditioning followed by allo-PBSCT. Of them, 29 patients were transplanted with related donor and 15 unrelated donor (URD). All patients received mycophenolate mofetil (MMF), CsA and MTX for acute GVHD (aGVHD) prophylaxis. 5 mg/kg rabbit-antithymocyte globulin (ATG-Fresenius) was incorporated in 15 URD recipients.

RESULTSAll patients were successfully engrafted. The median times to ANC above 0.5 x 10(9)/L in related (RG) and unrelated groups (URG) were 13.7 (9 - 18) d and 13.6 (12 - 17) d, and PLT above 20 x 10(9)/L were 15.3 (9 - 20) d and 14.7 (10 - 26) d, respectively. Two patients in RG. 1 in URG developed graft rejection 5 - 8 months after transplantation. One of the two patients in RG received second transplantation and engrafted. The cumulative incidence of aGVHD and cGVHD were 13.8% (4/29) and 46.4% (13/28) in RG, and were 33.3% (5/15) and 57.1% (8/14) in URG respectively. Two patients in RG relapsed after transplantation, and obtained CR again after donor stem cell infusion (DSI). Median time of follow-up was 34.7 (2 - 73) months. Thirty-four patients were alive and 10 died. The main causes of death were IP, GVHD, graft rejection and infection. The 5-year overall survival (OS) probability was 77.0%, and the disease-free-survival (DFS) was 73.9%, of which, 79.0% and 74.1% were in RG, and 73.3% and 73.3% in URG, respectively.

CONCLUSIONSFludarabine-busulfan based reduced intensity conditioning for allo-PBSCT with either related or unrelated donors is a safe, less toxic and curative approach to CML.

Graft vs Host Disease ; prevention & control ; Hematopoietic Stem Cell Transplantation ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; Peripheral Blood Stem Cell Transplantation ; Transplantation Conditioning

PURPOSE: Tc-99m MDP bone scan was performed to evaluate a generalized bone pain in a 24-year-old male chronic myelogenous leukemia patient who received bone marrow transplantation at 7 months ago. The patient had received large amounts of blood transfusion for managing symptoms related to anemia. Bone scan revealed substantial splenic tracer uptake. Magnetic resonance image and laboratory evidence of hemochromatosis suggests that the presence of large quantities of iron in the spleen of this patient may have been responsible for the splenic uptake of the bone scanning agent. The authors report a case of splenic hemochromatosis incidentally found on Tc-99m MDP bone scan.
Anemia ; Blood Transfusion ; Bone Marrow Transplantation* ; Bone Marrow* ; Hemochromatosis* ; Humans ; Iron ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive* ; Magnetic Resonance Imaging ; Male ; Spleen ; Technetium Tc 99m Medronate* ; Young Adult

Imatinib mesylate (IM) is used to treat a wide range of diseases, including Philadelphia chromosome-positive chronic myeloid leukemia (CML), on account of its high tolerability and low incidence of minor adverse events. Hemorrhage is thought to be a rare complication of IM. Recently, IM has been associated with reduced alpha2-plasmin inhibitor and platelet dysfunction. We report here the case of a 33-year-old female patient with CML who experienced subdural hematoma after an incremental increase in IM dosage due to a loss of complete molecular response. This case indicates that physicians should be alert to this atypical cause of headache in patients taking high-dose IM.
Adult ; Benzamides ; Blood Platelets ; Female ; Headache ; Hematoma, Subdural ; Hemorrhage ; Humans ; Incidence ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; Mesylates ; Philadelphia ; Piperazines ; Pyrimidines ; Imatinib Mesylate