Imatinib mesylate, a specific inhibitor of BCR-ABL tyrosine kinase, has been mainly used for the management of chronic myeloid leukemia (CML). Common side effects of imatinib include hematologic toxicity, liver dysfunction, edema, fluid retention, gastrointestinal symptoms, and skin rashes among others. We experienced a patient with imatinib-associated diarrhea, who was successfully treated with hangeshashinto. The patient was 61 year-old female, who first consulted our clinic in June, 2005. Her complaint was frequent diarrhea(4 or 5 times a day) which occurred after the introduction of imatinib. Four weeks after hangeshashinto was administered, her bowel movement improved from watery diarrhea to loose stool twice a day. Eight weeks later, her symptom disappeared. It is generally accepted that imatinib should be administered continuously to obtain a complete clinical remission of CML. From the viewpoint of eastern and western medicine integration, it is significant that a Kampo medicine contributed to continuous therapy with imatinib.
IMATINIB ; Diarrhea ; BCR Gene ; week ; Treated with

OBJECTIVE: To analyze the significance of various abnormal signal patterns appreared in CML and B-ALL patients by using BCR/ABL/ASS1 tricolor dual-fusion probe, and to explore its application value in detecting BCR/ABL fusion gene and ASS1 gene deletion. METHODS: 50 newly diagnosed CML patients and 50 newly diagnosed B-ALL patients were detected by fluorescence in situ hybridization (FISH) with BCR/ABL/ASS1 tricolor dual-fusion probe. Meanwhile, karyotype analysis was performed on all the patients using the 24 hours short-term culture and R-banding. RESULTS: Among the 50 CML patients, Ph was found in 49 cases, 5 normal interphase karyotype was observed in 1 case. FISH detection showed that BCR/ABL fusion gene existed in all patients (100%), while the positive signal pathway showed that 1R1G2B2F was observed in 39 cases (78%), 2R1G2B1F in 2 cases (4%) and 1R1G2B1F in 6 cases (12%), simultaneous existence of 1R1G1B1F and 1R1G2B3F in 1 case (2%), 2R1G1B1F in 1 case (2%) 1R1G3B3F in 1 case (2%). FISH detection also showed that the karyotype of 6 case at ASS1 gene deletion (1R1G1B1F) all were simple t (9; 22) translocation, and other abnormalities not were observed. Among 50 cases of B-ALL, Ph was found in 13 cases, the numerical aberration and structural aberration of non t (9; 22) in 16 cases, normal karyotype in 20 cases, absence of mitotic phase in 1 case. FISH detection showed that 16 cases (32%) had BCR/ABL fusion gene including 13 cases (26%) of 1R1G2B2F, 1 case (2%) of stimultaneous exitance of 1R1G2B2F and 1R1G3B3F 1 case (2%) of 2R1G1B1F, 1 case (2%) of 1R1G3B2F. FISH detection also showed that 3 cases had BCR/ABL fusion gene, including 1 case with ASS1 gene deletion (2R1G1B1F), 1 case with classical t (9; 22) translocation (1R1G2B2F) and 1 case with BCR/ABL fusion gene and increase of ASS1 gene copy (1R1G3B3F). CONCLUSION Tricolor dual-fusion FISH probe for detecting BCR/ABL fusion gene and ASS1 gene deletion is simple, rapid, sensitive and stable. It can detect various forms of molecular fusion and avoid the false positive results due to coincidental overlap of signals generated by D-FISH probe and ES-FISH probe. In addition, this detection method not only can directly observe the presence or absence of ASS1 gene deletion, but also improve the reliability of the positive results of newly diagnosed BCR/ABL fusion gene and accuracy of monitoring results of minimal residual disease for the subsequent visit.
Fusion Proteins, bcr-abl ; genetics ; Gene Deletion ; Humans ; In Situ Hybridization, Fluorescence ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; Reproducibility of Results

Sixty patients with chronic myeloid leukemia (CML) underwent bone marrow trephine biopsy at presentation. All the biopsies were decalcified, paraffin-embedded and stained with H&E, Gomori's reticulin and Masson Trichrome. A detailed study of the histology including the morphology and topographic distribution of megakaryocytes was done. 55 patients presented in chronic phase. Of these there were 37 cases (67%) of CML-granulocytic (CML-G) type and 18 cases (33%) of CML-granulocytic megakaryocytic (CML -GM) type. Five cases presented in blast crisis. 73% of CML-G had low-grade fibrosis while 83% of CML-GM had high-grade fibrosis. This was statistically significant. On follow-up 25% of CML-G went into blast crisis while all the CML-GM patients remained stable to date. Bone marrow biopsy is a useful investigation in patients of CML at diagnosis as it provides prognostic information. Evaluation of megakaryopoiesis, grading of fibrosis and localization of blasts are possible on a trephine biopsy.
BCR Gene ; biopsy characteristics ; upper case gee ; Myeloid Leukemia, Chronic ; Fibrosis

OBJECTIVETo compare allelic frequencies of 15 short tandem repeat (STR) loci (D8S1179, D21S11, D7S820, CSF1PO, D3S1358, TH01, D13S317, D16S539, D2S1338, D19S433, vWA, TPOX, D18S51, D5S818 and FGA) between chronic myeloid leukemia (CML) patients and non-related healthy individuals from Changzhou region in order to predict genes related with the CML.

METHODSBlood samples were collected from 745 healthy subjects and 132 CML patients with complete remission. Genotypes were determined with gene scan technology and multiplex PCR with fluorescence-labeled primers. Allelic polymorphisms of 15 STR loci were compared between the two groups. Potential genes related with CML were predicted with statistical analysis of differences in allelic frequencies.

RESULTSAllelic frequencies of 3 loci, including CSF1PO, vWA and TPOX, showed a significant difference (P<0.05) between the two groups.

CONCLUSIONCSF1PO, vWA and TPOX loci may be related with CML, albeit that the exact biologic mechanisms is unclear.

Gene Frequency ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; Microsatellite Repeats ; Polymorphism, Genetic

No abstract available.
Gene Rearrangement* ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive* ; Polymerase Chain Reaction*

P210(bcr/abl) transgene mouse is a good model to research the chronic myelogenous leukemia (CML), but the P210(bcr/abl) gene has a lethal effect on embryogenesis if driven by the constitutive promoter. So, the use of promoter which induces the special expression in hematopoietic tissue is the key to construct CML transgenic mice. This study was purposed to investigate the TEC promoter mediated P210(bcr/abl) gene expression in BaF3 cells. The CMVie promotes of IRES2-eGFP vector was replaced with the -364-+22 domain of TEC promoter cloned from mouse genome, and the P210(bcr/abl) gene was inserted into the EcoR I site of TEC-IRES2-eGFP vector. Then, the constructed vector was transfected into the BaF3 cells and 293 cells respectively. The expression levels of eGFP gene and P210(bcr/abl) gene in BaF3 and 293 cells were detected. The results showed that with fluorescent microscopy and flow cytometry, the eGFP gene was found to be expressed in the BaF3 cells, the expression rate was 7.10%, 23.35%, 64.61% at 6, 24, 72 h respectively after transfection, but the fluorescence was not seen in 293 cells. A 372 bp fragment of BCR/ABL mRNA was amplified by RT-PCR in BaF3 cells, but not in 293 cells. It is concluded that the -364-+22 domain of TEC promoter can mediate high-effective and specific expression of related genes in hematopoietic tissue, which can be used to construct P210(bcr/abl) transgene mice model.
Animals ; Fusion Proteins, bcr-abl ; genetics ; Gene Expression ; Genetic Vectors ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; Mice ; Mice, Transgenic ; Promoter Regions, Genetic ; Protein-Tyrosine Kinases ; genetics

Four cases of atypical chronic myeloid leukemia (aCML), which were compatible with the FAB guideline for the classification of chronic myeloid leukemia (CML), are presented. All 4 patients showed the onset in old age, leukocytosis with an increase in the number of immature granulo-cytes, monocytosis, a low basophil count, and a dysgranulopoiesis in the peripheral blood, a nega-tivity of the bcr-abl gene rearrangement, and a hypercellular marrow with marked granulocytic hyperplasia and dyshemopoietic features. Two patients died within 3 months and the other 2 are currently under observation after a partial response to hydroxyurea. aCML is known to have a poor therapeutic response and outcome without a blastic crisis. A greater deal of concern regarding aCML is required for an accurate diagnosis and classification.
Basophils ; Bone Marrow ; Classification ; Diagnosis ; Gene Rearrangement ; Humans ; Hydroxyurea ; Hyperplasia ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; Leukemia, Myeloid, Chronic, Atypical, BCR-ABL Negative* ; Leukocytosis

Adolescent ; Adult ; Female ; Fusion Proteins, bcr-abl ; genetics ; Gene Expression ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; Leukemia, Myeloid ; genetics ; Male ; Oncogene Proteins, Fusion ; genetics ; Retrospective Studies

The aim of this study was to investigate the apoptosis-inducing effect of cinnamic aldehyde (CA) on chronic myeloid leukemic (CML) cells and its mechanism. K562 cells and primary bone marrow mononuclear cells (MNC) from patients with CML were treated by various concentrations of CA. Flow cytometry was employed to measure the apoptosis of K562 cells and primary CML bone marrow MNC. Western blot was used to determine the expression of C-MYC and the phosphorylation of CrkL in K562 cells, and real-time polymerase chain reaction (real-time PCR) was used to quantify the expression of BCR-ABL mRNA in K562 cells. The results indicated that CA induced the apoptosis of K562 cells in a time- and dose-dependent manner. CA induced apoptosis of CML MNC dose-dependently. CA inhibited the expression of BCR-ABL mRNA and C-MYC, reduced CrkL phosphorylation levels in K562 cells. It is concluded that CA induces apoptosis of CML cells in vitro. Down-regulation of the expression and function of BCR-ABL may be one of its most important anti-leukemia mechanisms.
Acrolein ; analogs & derivatives ; pharmacology ; Apoptosis ; drug effects ; Fusion Proteins, bcr-abl ; metabolism ; Gene Expression Regulation, Leukemic ; Humans ; K562 Cells ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; metabolism ; pathology

OBJECTIVETo construct a vector containing protein transduction domain (PTD) and bcr/abl fusion gene of chronic myelogenous leukemia and express PTD-bcr/abl fusion protein in E. Coli.

METHODSDNA fragment encoding PTD was synthesized and fused to PCR-amplified bcr/abl gene fragment, then inserted into plasmid pET-16b to get the expression vector pEPb containing PTD-bcr/abl fusion gene, which was transfected and expressed in E. Coli LB21. PTD-bcr/abl fusion protein was purified by affinity chromatography.

RESULTS523 bp bcr/abl fusion gene was effectively amplified. The PTD-bcr/abl gene sequencing showed the same sequence as scheduled. The fusion peptide was successfully expressed in E. Coli and purified.

CONCLUSIONThe results may provide a new PTD-bcr/abl fusion peptide for the immunotherapy of CML.

Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; genetics ; Fusion Proteins, bcr-abl ; genetics ; metabolism ; Gene Expression ; Gene Products, tat ; genetics ; metabolism ; Genetic Vectors ; genetics ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; Recombinant Fusion Proteins ; genetics ; metabolism