ABSTRACT
Purpose: Diagnosis of myelodysplastic syndrome (MDS) primarily relies on the detection of morphological dysplasia in bone marrow. It is subjective and many studies have reported lack of interobserver agreement in reporting. Biopsy is preferred specimen for megakaryocyte assessment. We studied 43 bone marrow biopsies from 40 suspected MDS patient having persistent undiagnosed cytopenia. Utility of immunohistochemistry (IHC) with CD61 and p53 in detecting low-grade MDS was analyzed over routine morphology. Method and Results: Total number of megakaryocytes and number of dysplastic megakaryocytes seen on CD61 IHC was significantly higher than that on H and E stain (P value < 0.05) Out of total 43 biopsies, 13 [30.2%] cases showed dysplastic megakaryocytes that were confirmed by interobserver agreement after IHC. From 30 cases with no significant dysplasia on morphology, 21/43 [48.8%] cases showed >10% dysplastic megakaryocytes on CD61 (P value 0.0001). Nine cases showed no significant dysmegakaryopoiesis with either H and E or CD61 IHC. Fourteen cases could meet higher cut off (30%) of dysmegakaryopoiesis with CD 61 IHC. Out of total 34 cases showing significant dysplasia 7 cases (20.6%) showed positivity for p53 on IHC, which is little less than that reported in low-grade MDS. Conclusion: CD61 IHC is helpful in making correct diagnosis of MDS in cases with minimal dysplasia and should be performed before excluding possibility of MDS on morphology in a patient with undiagnosed cytopenia. IHC is cost effective tool for MDS diagnosis in developing world where access to extensive flow cytometery and molecular testing is limited.
ABSTRACT
OBJECTIVES@#To observe the changes of the formation time of venous thrombus in rats, and to provide new ideas and methods for the estimation on thrombus formation time of the forensic cases died from thrombosis.@*METHODS@#Totally 80 rats were randomly divided into 10 groups （0 h, 3 h, 6 h, 12 h, 1 d, 3 d, 1 week, 2 weeks, 3 weeks and 4 weeks after operation）. A vein thrombosis model was established by the "narrow" method. The processes of thrombosis, organization, recanalization and the features of change on hemosiderin and calcium salt were observed by HE stain, Perls stain and Von Kossa stain. The expression changes of CD61, α-SMA and CD34 were observed by immunohistochemical staining technique.@*RESULTS@#Platelets adhered to the exposed blood vessel intima 3 h after operation, and platelet trabeculae were formed by the repeated accumulation of platelets 1 d after operation. The thrombus organization formed through the fibroblasts from vessel wall that grew into the interior of the thrombus 3 d after operation. Endothelial cells covered the surface of thrombus and then the new blood vessels were reformed, and the vessels were reconstructed. The expression of CD61 upregulated at the stages of the thrombus formation （3 h） and thrombus reformation （4 weeks）, and reached the peak 1 d after thrombus formation. The release of hemosiderin and the initial expression of α-SMA were detected 3 d later. Calcium deposit and expression of CD34 were observed 1 week later.@*CONCLUSIONS@#The hemosiderin, calcium salt, CD61, α-SMA and CD34 show time-dependent changing characteristics, which is expected to provide a reference for the estimation on thrombus formation time of the forensic cases died from thrombosis.
Subject(s)
Animals , Rats , Antigens, CD34/analysis , Hemosiderin/metabolism , Venous Thrombosis/pathologyABSTRACT
BACKGROUND: The automated hematologic analyzer has increased the precision and accuracy for platelet counting. However, spurious elevations of automated platelet counts occur occasionally in patients receiving chemotherapy or radiotherapy for solid organ tumors, leukemia, and other lymphomas. The CELL-DYN Sapphire analyzer (Abbott, USA) detects platelets with a CD61 monoclonal antibody and uses both impedance and optical technologies; thus, it is expected to present more accurate platelet counts. METHODS: We evaluated platelet counts obtained with the CELL-DYN Sapphire impedance, optical, and CD61 methods and compared them with the results obtained with the XE-2100 analyzer (Sysmex, Japan). We analyzed 111 samples from hospitalized patients with various hematologic diseases, who were receiving chemotherapy or radiotherapy. RESULTS: The results from the impedance, optical, and CD61 methods of CELL-DYN Sapphire and those from XE-2100 showed significant linearity, with correlation coefficients greater than 0.99. Three cases had significantly different platelet counts among the different methods used. Microscopic examination of these three cases showed very low platelet counts that corresponded with the low counts from the CD61 methods. It should be noted that because the automated blood counter assesses cell populations by their dimensions, many cellular fragments that were of the same size or smaller than platelets were thus counted as platelets. CONCLUSIONS: The CELL-DYN Sapphire analyzer has good precision, linearity and performance, comparable with the XE-2100 analyzer. As the CD61 methods of CELL-DYN Sapphire is specific for platelet, this method may reduce the interference from other blood components and count the exact platelet numbers.