| Aims: Chronic myeloid leukemia (CML) is a myeloproliferative disease, the mortality rate increased year by year, China has moved up to No.3. CML diagnosis, classification and prognosis depend on the morphology and cytogenetic testing which has proposed by WHO. But for chronic myeloid leukemia patients with early primitive myeloid cells, immature granulocytes with normal morphology difference is not obviously.Therefore, the accuracy of the morphologic diagnosis of certain limitations. With the progress of technology, single-cell analysis confirmed the content of a single cell types or a component of chemical components will be produced with the different stages of the cell changes. Studies have shown that the occurrence of myeloperoxidase associated with many diseases. However, the existence of chronic myeloid leukemia between MPO and a relationship is not yet clear. Early this study is to design a single neutrophil capable of analyzing the microfluidic chip to explore the relationship between MPO levels and corresponding changes in the activity of neutrophils and chronic myeloid leukemia among guide diagnosis.Methods:(1)95cases chronic myeloid leukemia patients were collected from the peripheral blood and bone marrow aspirates. As the Health control group of50patients, peripheral blood were collected.(2) Use SYSMEX XE-5100hematology analyzer to measure the number of blood RBC, WBC, Hb and the number of PLT.(3) Preparation of the blood and bone marrow smears using Wright’s dye staining of hematopoietic cells, according to cell morphology School for all types of blood cells count, calculate the system cells and immature and mature myeloma cells Quantities.(4) The various stages of neutrophil myeloperoxidase staining be done morphology.(5) Able to design a single neutrophil cell analysis microfluidic chips.(6) Use to design a chip, neutrophil chronic myeloid leukemia patients and the control group were ingle-cell analysis, determination of myeloperoxidase activity.(7) To investigate the role of several important factors affecting. Results:The first part of the application of the general method to check the situation of chronic myeloid leukemia and myeloperoxidase(1) Blood test results for95patients with blood test results:WBC:10×109/L the following two cases,(20-50)×109/L11, average42.1×109/L;(31-60) g/L (severe anemia)15cases;(115-150) g/L (normal)11cases; platelets:<120×109/L16例;>350×109/L20例;(120-350)×109/L (normal)59cases; eosinophils:>8%in62cases; addicted basophils:>1%of the59cases; neutrophils:>70%of the59cases.(2) The results of blood smears of white blood cells were significantly increased, the various stages of myeloid cells can be seen to neutral myelocytes.(3) The results of bone marrow smears of bone marrow cell proliferation was active in36cases,59cases of hyperactivity, grain red ratio (5-60):1, easy to see the original granulocytes, myeloblasts<4%,15cases seen monocyte-macrophage cells.(4) Myeloperoxidase staining results in healthy controls and patients with CML in promyelocytic late promyelocytic, rod-shaped nucleus and leaf neutrophils were seen MPO positive, and varying degrees of staining various stages of MPO, granular uniform distribution, darker colored patients, the relative activity was significantly higher than the control group.The second part of the application microfluidic chip in a single cell measuring myeloperoxidase(1) Chip has three parts cover sheet, the channel layer and the substrate. Microchannel layer using the traditional "十" shaped channel design, width100μm, deep30μm, the channel length of49mm.(2) The use of microfluidic chip to analyze the expression of a single neutrophil MPO analysis CML Group neutrophil MPO expression, the use of the microscope, CCD image capture technology, found in CML patients promyelocytic cells.(3) The case of using the microfluidic chip select neutrophil CCD image capture device to record MPO positive cells at a constant speed in the forward flow in the microchannel, uniform electric field across.(4) Compared to the situation mean neutrophil MPO expression analysis of each patient mature granulocytes and immature granulocytes each100cells, was measured using the micro-fluidic chip NE cells per unit of MPO activity relative degree.Conclusion:(1) Hematology analyzer are classified according to the size of the volume of blood cells, although it can distinguish eosinophils, basophils and monocytes, but can not identify immature cells, different leaching concentration of nucleated red blood cells and granulocytes becomes toxic.(2) Hematology analyzer of red blood cells, platelets internal structure can not be subtle observation and identification.Microscopic examination should be as complete blood cell analysis (CBC) a very important quality control, but also the last one off.(3) Design a microfluidic chip can accurately identify the various stages of granulocytes.(4)suggesting that the average MPO activity of myeloid leukemia may be used as diagnostic markers and individual indicators of chronic myeloid check. |
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