The significance of bone marrow cell morphology and its correlation with cytogenetic features in the diagnosis of MDS-RA patients.

Besides cytopenia, dysplasia is crucial characteristic of MDS-RA. To summarize the morphological features that contribute to the diagnosis of MDS-RA, 48 RA patients with abnormal karyotype were analyzed for the features of morphological and cytogenetical abnormalities and the relationships between them. 46 MDS-RA patients with normal karyotype and 207 patients with non-MDS anemia were enrolled into control groups. More conspicuous and diverse dysplasia can be found in abnormal karyotype MDS-RA than those in control groups (P<0.05). Apparent dysplasia in granulocyte and megakaryocytoid lineages may provide valuable evidence for the diagnosis. Dysplasia occurred more frequently in patients with severe chromosome abnormalities.

[The morphological, immunophenotypical and cytogenetic characteristics study of blast crisis in chronic myeloid leukemia].

OBJECTIVE: To explore the morphological, immunophenotypical and cytogenetic (MIC) characteristics of chronic myeloid leukemia in blast crisis (CML-BC). METHODS: Marrow cells from 31 CML-BC patients were studied by bone marrow smears with morphological and cytochemical stain analysis. Immunophenotypic analysis was performed by micro blood method and flow cytometry. Cytogenetic analysis was performed using bone marrow cells prepared directly and or after 24 hours culture. RHG banding was used for karyotypic analysis. RESULTS: 23 patients (74.2%) were found to have myeloid blast crisis. 5 cases (16.1%) had a B-lymphoid immunophenotype and 4 of them expressed myeloid associated antigens at the same time. Another 3 cases were classified as acute undifferentiated leukemia (AUL 1 patient) and acute myeloid -lymphoid leukemia (AMLL 2 patients) with B and myeloid markers expression. A significant proportion (67.7%) of blast crisis cases were CD(34) positive. CD(34) was expressed in 80.0% of lymphoid and 65.2% of myeloid BC. Analysis of the relationship between CD(7) and CD(34) expression in blasts of myeloid crisis showed their dual expression in 8/23 (34.8%) of the cases. Cytogenetic analysis indicated that 14/27 (51.9%) of the patients had developed secondary cytogenetic abnormalities in addition to the Ph chromosome. These newly emerging abnormalities included a trisomy 8 in 3 patients, a double Ph chromosome in 3 patients, an iso chromosome 17q in 2 patients, loss of Y chromosome in 1 patent and other complicated translocation in 5 patients. CONCLUSION: Blast crisis of CML is a kind of disease of stem cell. The differentiation of blast cell is blocked in the early stage. The prognosis of such patients is poor. MIC showed its important values in diagnosis, judgement of prognosis and determination of therapeutic protocol of CML-BC.

[Clinical and cytogenetic studies of hematological malignancies with isolated trisomy 11].

OBJECTIVE: To evaluate the association between isolated trisomy 11 and the clinical, hematological, immunological, prognostic aspects in hematological malignancies. METHODS: Bone marrow cell cytogenetic analysis was performed by direct method and/or 24 h culture method. RHG banding was used for karyotype analysis. Immunophenotype analysis was carried out by flow cytometry. Ten patients with acute myeloid leukemia (AML) were treated with HA regimen chemotherapy and followed up. RESULTS: The isolated trisomy 11 was found in 11 of 1 * ! 763 hematological malignancies cases (0.6%). The diagnoses included 10 AML (6 M(2), 2 M(5), 1 M(1), 1 M(4)), and 1 myelodysplastic syndromes. Ten of them have no hepatosplenomegaly. The immunophenotypical analysis of leukemia cells showed positive for CD(13), CD(33) and CD(34) in 5 cases. Follow-up data were available in 10 cases. The complete remission rate was 40% with a median survival of 10 months. CONCLUSION: The isolated trisomy 11 was mainly seen in AML, especially in M(2) subtype. Their prognosis was poor.

A preliminary study on the early diagnosis of myelodysplastic syndromes.

OBJECTIVE: To evaluate the four techniques for clonal analysis in the early diagnosis of myelodysplastic syndromes (MDS). METHODS: Four techniques for clonal analysis were performed in bone marrow samples from fifty patients with suspected MDS: (1) Conventional cytogenetics (CC) for clonal chromosomal abnormalities; (2) BrdU-sister chromatid differentiation (BrdU-SCD) for cell cycle analysis; (3) Fluorescence in situ hybridization (FISH) for trisomy 8; (4) PCR-SSCP for N-ras mutation. RESULTS: The diagnosis of forty-five patients was compatible with FAB criteria of MDS, the other five patients didn't fully meet the FAB criteria. They had either only one lineage dyspoiesis or no any obvious dysplastic features and two of them were diagnosed as suspicious refractory anemia (RA), one as anemia with hypercellular bone marrow and two as chronic aplastic anemia. The results of the four techniques performed in them showed that four patients had clonal karyotype abnormalities, two had prolonged cell cycle, three had trisomy 8 of different proportions, and one had N-ras mutation. Thus, they were all diagnosed as RA. CONCLUSION: The untypical MDS patients can be diagnosed early by examination with combining several clonal analysis techniques.

Clinical and experimental study of two cases of myelodysplastic syndrome with t(3; 5) (q25; q34) translocation.

OBJECTIVE: To report two myelodysplatic syndromes (MDS) patients with t(3; 5) (q25; q34). METHODS: Chromosome specimens were prepared by short-term culture of bone marrow cells. Karyotype analysis was performed by R banding technique, chromosome painting (fluorescence in situ hybridization, FISH) by using whole chromosome 3 and 5 probes in case 1. RESULTS: The clinical and hematological findings were compatible with diagnosis of MDS. Karyotype analysis showed that both patients had identical t(3; 5) (q25; q34) translocation. A reciprocal translocation between chromosomes 3q and 5q was proved by FISH in one patient. CONCLUSIONS: t(3; 5) translocation is a rare chromosome abnormality specifically associated with MDS and frequently displays trilineage dysplasia. Chromosome painting technique is a reliable tool for detecting this translocation.

Acute biphenotypic leukemia in the adults.

OBJECTIVE: To study the clinical, biological features and prognosis of acute biphenotypic leukemia (BAL) in the adults. METHODS: Bone marrow specimens of 63 BAL patients were evaluated to prove the diagnosis and the classification by morphologic, cytochemical, immunologic and cytogenetic (MIC) examinations. These patients were treated with protocols suitable for acute myeloid leukemia (AML), or acute lymphoblastic leukemia (ALL), or both. RESULTS: No significant difference in clinical features was observed between BAL, AML or ALL. Morphologically, the subtypes of M(5), M(1) and M(2) were predominant in AML, as L(2) and L(1) were in ALL. Immunologically, coexpression of myeloid and B lineage associated antigens was predominant and CD(34) was hyperexpressed in BAL, which suggested that BAL might originate from malignant transformation of earlier hematopoietic cells. Cytogenetically, Ph chromosome was observed in 25.5% (13/51) of BAL patients. Prognostically, both the treatment response and the overall survival of BAL patients were poor. CONCLUSION: Patients with BAL have unique clinical, biological and prognostic features.

[Dual-color FISH study on myelodysplastic syndrome with 1;7 translocation].

OBJECTIVE: To study the myelodysplastic syndrome(MDS) with 1;7 translocation in five cases and to determine further the constitution and origin of centromere of the derivative chromosome resulting from 1;7 translocation. METHODS: Bone marrow chromosome preparation of five cases was made using direct method or short- term culture. Karyotypic analysis was carried out by R-banding technique. Dual-color fluorescence in situ hybridization(FISH) using Spectrum Red and Spectrum Green directly labeled chromosome 1-specific a-satellite DNA probe(red) and chromosome 7-specific a-satellite DNA probe(green) was performed in three patients of them. RESULTS: All of the five cases had 1;7 translocation. The centromere of the derivative chromosome 7p/1q was constituted with red and green signals in three of them. CONCLUSION: The result of dual-color FISH confirms that the centromere of the derivative chromosome resulting from 1;7 translocation originated from both centromeres of chromosome 1 and chromosome 7.

[Report of a patient with spontaneous aggregation of his giant and morphologically abnormal platelets].

OBJECTIVE: To study the pathological and clinical characteristics of a patient with spontaneous platelet aggregation of his giant and morphologically abnormal platelets. METHODS: Platelet size and structure were observed under light microscope and electron microscope. Platelet aggregation was measured turbidometrically. Platelet glycoproteins (GP) were analyzed using flow cytometry. PCR and DNA sequencing were performed to identify the gene abnormality. RESULTS: The patient had spontaneous platelet aggregation of giant platelets with thickened plasma membrane and increased number of granules in various shapes. Aspirin and ticlopidine did not affect the spontaneous aggregation. The expression of GP I b, GP II b, GP III a and P-selectin in the platelet membrane were in normal range. Results of gene analyses for GP I balpha, GP I bbeta and GPIX were also normal. CONCLUSION: Both morphological and functional abnormalities of the platelets from the patient were clearly distinguishable from that of other hereditary giant platelet disorders. It would probably represent a novel platelet disorder which had not been reported to date.

[Application of Metaphase-Fluorescence in Situ Hybridization to the Diagnosis of Acute Promyelocytic Leukemia and the Detection of Minimal Residual Disease]

To investigate the value of metaphase-fluorescence in situ hybridization (M-FISH) in the diagnosis of acute promyelocytic leukemia (APL) and the detection of its minimal residual disease (MRD), 10 cases of untreated acute myeloid leukemia (AML) (de novo APL 5 cases, relapsed APL 3 cases, AML-M(1) and AML-M(5) one case each) diagnosis by cell morphology at presentation and 10 cases of APL after complete remission (CR) were studied by M-FISH using a whole chromosome 17 painting probe labeled by digoxigenin and the results were compared with that of conventional cytogenetic examination and reverse transcription-polymerase chain reaction (RT-PCR). Among 10 untreated AML cases, 7 had positive M-FISH results, of whom 4 had t (15;17) translocation, 3 had normal karyotype. Six of them had PML/RARalpha fusion transcript except one, in whom RT-PCR did not be performed; 3 had negative M-FISH results, of whom one had del (2q) x 2 abnormalities, who was RT-PCR-positive for PML/RARalpha fusion transcript; one had complex karyotype abnormalities, whose RT-PCR was negative for PML/RARalpha fusion transcript; one had t (9;22) translocation, whose RT-PCR was negative for PML/RARalpha fusion transcript, but positive for BCR/ABL fusion transcript. Thus the diagnosis of AML-M(3) was revised as AML-M(2) for the latter two cases. 10 APL cases after CR had normal karyotype, but 12/15 M-FISH assays detected 1 - 5 t (15;17) positive cells in 9 of them. This finding is compatible with the results of RT-PCR assays. Leukemia relapse was seen in one case, and two positive M-FISH results were appeared in the 2 assays at a 13 months' interval. This study suggests that M-FISH had important practical value in the diagnosis of APL and the detection of MRD, and that it is less sensitive than RT-PCR, however, it seems to be more potential for prediction of the relapse of leukemia due to its capacity of detecting quantitatively the chromosome translocation in proliferative cells.