The MIC Study Of Experimental Diagnosis In Myelodysplastic Syndroms

Objective: Myelodysplastic syndrome(MDS) is a group of clonal bone marrow disorders characterized by ineffective hematopoiesis and dysplasis.It is easier to progress for acute leukemia and/or bone marrow failure.The incidence of MDS is about 0.004 percentage of all the population in our country,growing year after year.It has become a serious malignant of hematopoietic system tumors,which is a destroyer of old people.Because of tremendous heterogeneity of MDS, the gold standard for diagnosing MDS has not been established.So far the diagnosis of MDS is important and difficulty in blood diseases. In this study,we producted MIC as a primary or adjunctive means to diagnosed MDS,which can provide the basis for clinical diagnosis.Method: Using retrospective study to analysis 41 cases of MDS patients and 18 cases of suspected patients,who were defined according to 2006 Vienna minimum criteria for the diagnosis of MDS and the 2008 World Health Organization(WHO) classification subcategorizes MDS.The results were retrospectively correlated with morphologiy bone marrow biopsy, cytogenety and cells flow classification. An additional 30 patients with aplastic anemia(AA) in comparison to MDS patients as a control cohort.SPSS17.0 statistical software used for statistical analysis of dataResults: 1 The clinical data Among 59 patients, 35 had dizziness, fatigue, pale, 24 of fever, 6 had bleeding, 6 hepatosplenomegaly, 1 lymphadenopathy. 2 Cell morphology 2.1 Peripheral Blood59 patients, the erythrocyte mean was 2.13 × 1012 / L;the leukocyte mean was 2.74 × 109 / L;the hemoglobin average number was 82 g / L, the platelet mean was 91 × 109 / L; MCV mean was 100.62fl; RET mean was 0.5%. 2 patients had unilineage cytopenia,14 patients had biolineage cytopenia,43 cases were multilineage cytopenia. A 200 cell differential count was performed by oil microscope.Among 59 patients, mean percentage of blasts was 2.7%,35 were detected immature granulocytes, which are myeloblasts, promyelocytic, myelocytes and metamyelocyte. The number of them were 23(65.7%),1(2.9%), 10(28.6%), 1(2.9%). The immature granulocytes of respectively stages values range were(0~ 15),(0 ~ 1),(0 ~ 18),(1 ~ 24). 23 cases were hypogranularity cytoplasm,one was false pseudoPelger-Huet. 13(48.1%) patients were detected circulating nucleated red blood cells,values range(0-18). 27 cases apparent shaped red blood cells, visibility, target-shaped, oval, teardrop-shaped red blood cells and giant.No megakaryocytes, 17 cases seen large platelets. 2.2 Bone marrow aspirate 59 patients, bone marrow of 55 cases were hypercellular or apparently hypercellular,4 cases were hyporcellular.The proportion of erythroid dysplasia was 66.1%.12 cases of immature red blood cells appeared cytoplasmic vacuoles. 39 cases appeared dual-core.7 cases were triple-core. 3 cases were nuclear petals, irregular nucleus. The proportion of myeloid dysplasia was 81.4%, 17 patients had cytoplasm hypogranularity, 6 cases of cytoplasmic granules increased significantly.24 were pseudo- Pelger-Huet. 2 cases had excessive leaf.3 cases appeared Auer bodies.Megakaryocytic dysplasia ratio was 20.3%,10 patients were characterized by micromegakaryocytes with hypolobated nuclei and megakaryocytes of all sizes with monolobated nuclei.2 patients were characterized by megakaryocytes with multiple, widely separated nuclei.18 patients periodic acid-Schiff reaction was positive.27 patients extracellular ferric stain were positive(++ ~ +++),23 patients Intracellular iron were positive,and 8 patients appeared RS in each subtype.3 Bone marrow biopsy In MDS patients, bone marrow biopsy showed hypercellular or apparently hypercellular of 46 cases. 2 cases reticulin increased; small blood vessels of 17 cases marked hyperplasia; 8 cases appeared abnormal localization of immature precursor cells(ALIP); 15 cases CD34+ cells were increased, 32 cases were detected the micro-megakaryocytes. 4 Conventional karyotype analysis 25 had abnormal chromosomal findings of 59 patients.The karyotype abnormalities accounted for 36.0%.Subtype distribution 1 RCUD, 4 MDS-RCMD, 3 MDS-RAEB-I, 1 MDS-RAEB-Ⅱ. Simple karyotype abnormalities was almostly appeared in MDS-RCMD.2 cases were trisomy 8.-5,5q- were each two cases.-7,7q- were each two cases, 20q- was 1 case. 5 Flow Cytometric Immunophenotyping 5.1 Myeloid precursor cells abnormal phenotype We defined CD34、CD117、HLA-DR、CD11b、CD15 Monoclonal antibody combinations as parameters that should be analyzed cells in MDS.This experiment detected CD34 + blasts abnormal phenotype in MDS group listed below: 23 patients with the proportion of immature cells was increased;11 cases reduced CD45 expression; 7 expressed mature antigen CD11 b, CD15; 12 cases expressed lymphocyte-associated antigen. 5.2 Myeloid phenotype abnormalities CDl3 / CDl6 and CD11 b / CDl6 as monoclonal antibody combination detected and analied in myeloid phenotype abnormalities during differentiation and maturation phase.MDS patients, 39 cases SSC reduced;34 patients with CD13 / CD16, CD11 b / CD16 relationship mode changed; 28 cases expressed abnormaly CD56. 5.3 Mononuclear cell phenotype abnormalities 59 MDS patients, 12 patients had abnormal expression of CD56; 5 cases were abnormal expression of CD34; 11 cases reduced CD33 expression; 19 cases reduced CD14 expression; 1 case expressed lymphoid-associated antigens.5.4 Erythrocyte phenotype abnormalities Used CD71, Gly A monoclonal antibody combination to detect red blood cell group. MDS group CD45 expression was decreased in 28 cases; 10 cases of CD71 expression increased; 9 cases Gly A enhanced expression; 1 case of abnormal expression of CD117.Conclusion: Cell morphology, cytogenety, pathology and flow cytometric immunophenotype for myeloproliferative syndrome are of great significance, but their had different characteristics, different emphases and complemented each other. Cell morphology is the key and foundation in MDS diagnosis and typing.Conventional karyotype examination plays an important role in MDS patients, especially in suspected MDS patients with newly diagnosed and follow-up. Bone marrow biopsy is important for assessing cellularity,fibrosis,stromal alteration, and megakaryocytic dysplasia. Flow cytometry immunophenotyping for assessing abnormal hematopoietic cells with high sensitivity and good repeatability, and less affected by the specimen quality and subjective factors.In particular, there is persistent cytopenia which had no reason,and no typical morphological and cytogenetic abnormalities suspected MDS patients.FCI had significant value for them. Dysplasia morphological, chromosomal abnormalities and flow cytometry is not unique in MDS, and MDS patients do not have all of the above abnormal changes, so MIC is helpful to support and except diagnosis of MDS.