1.Research advances in value of flow cytometric immunophenotyping in diagnosis and prognostic evaluation of myelodysplastic syndrome.
Journal of Experimental Hematology 2013;21(3):785-790
Myelodysplastic syndromes (MDS) are myeloid neoplasms characterized by dysplasia in one or more linages of cells and increased risk of development of acute myeloid leukemia (AML). Along with the deeply understanding of myelodysplastic syndrome, the diagnosis standards of this disease experienced a leap in essence: from a single standard of morphological test in FAB to multiple detecting means in WHO standard of 2008, flow cytometry has been proposed as an adjunctive diagnostic test in the 2007 Vienna standards and the 2008 WHO standards. Recently, A heterogeneous spectrum of immunophenotypic abnormalities have been reported in MDS, and some of which are of great significance to the diagnosis, classification, prognosis assessment, and treatment of the disease. In the year of 2003, a flow cytometric scoring system (FCSS) was built to evaluate the prognosis of MDS patients, which was able to qualify the phenotypic aberrancies in the myelomonocytic, erythroid, and megakaryocytic lineage. It filled the gap of the international prognostic scoring system (IPSS) and the WHO classification-based prognostic scoring system (WPSS), and was of great value to the clinical diagnosis and treatment of MDS. In this article, the value of MDS immunophenotyping in diagnosis and prognosis evaluation of MDS is reviewed in term of MDS immunophenotypic abnormalities and flow cytometric scoring system.
Flow Cytometry
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Humans
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Immunophenotyping
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Myelodysplastic Syndromes
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classification
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diagnosis
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immunology
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Prognosis
2.Role of flow cytometric immunophenotyping in diagnosis of myelodysplastic syndromes--review.
Journal of Experimental Hematology 2008;16(6):1477-1481
Bone marrow morphology and karyotyping are effective ways to diagnose myelodysplastic syndromes. However, there are still many patients without these two abnormalities. Recently, there are considerable amount of evidence showing that flow cytometric immunophenotyping is a good approach to detect the abnormal differentiation and maturation of different cell lines and maturation stages of bone marrow cells in MDS patients. The highly reproducible results allow to distinguish MDS from other non-clonal hematocytopenia disorders and have a diagnostic advantage especially in case of low-risk MDS. Meanwhile, it showed a good correlation with the morphology and karyotyping, and with the International Prognostic Scoring System (IPSS) playing a prognostic role. This article reviewed recent advances of research in this field such as the application of FCM in MDS detection, the correlation of FCM parameters with morphologic and cytogenetic diagnosis of MDS as well as the relationship of parameters with typing and prognosis of MDS and so on.
Flow Cytometry
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methods
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Humans
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Immunophenotyping
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Myelodysplastic Syndromes
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diagnosis
;
immunology
3.Significance and application value of multiparameter flow cytometry for differentiation of immunophenotype in chronic myelomonocytic leukemia, myelodysplastic syndrome and acute monocytic leukemia.
Yun-Xiu WANG ; Ji-Hong ZHANG ; Yan-Ping HU ; Fang-Fang CAO ; Nan ZHANG ; Fang CHEN ; Xuan LIU ; Min-Yu ZHANG
Journal of Experimental Hematology 2012;20(4):857-862
This study was purposed to analyse the immunophenotypic characteristics of chronic myelomonocytic leukemia (CMML), myelodysplastic syndromes (MDS) and acute monocytic leukemia (AML-M5b) by using multiparameter flow cytometry, and to explore its significance in diagnosis and differential diagnosis. The immunophenotypic characteristics of bone marrow samples from 14 CMML patients, 48 MDS patients, 46 AML-M5b patients and 18 normal persons were analyzed and compared by multiparametric flow cytometry. The results showed that the ratio of monocytes in CMML patients was obviously higher than that in MDS, AML-M5b patients and normal persons (P < 0.05), but there was no statistically significant difference between bone marrow samples of MDS and AML-M5b patients as well as normal persons. The ratio of blast cells in MDS patients was obviously higher than that in normal persons (P < 0.05), but did not show significant difference as compared with CMML patients. The ratio of mature granulocytes in AML-M5b patients was obviously lower than that in CMML and MDS patients as well as normal person bone marrow (P < 0.05). Certain differences of CD45/SSC characteristics in MDS, AML-M5b and CMML patients were found in comparison with normal persons. The abnormal expression of CD2, CD56, and CD14 tailing phenomenon were observed in CMML patients in comparison with bone marrow samples of MDS, AML-M5b and normal persons (P < 0.05). Lack and decrease of CD15 expression in MDS and CMML patients was significant different from AML-M5b and normal persons marrow, abnormal expression rate of CD15 in CMML patients was higher than that in MDS patients (P < 0.05), the CD13/CD11b/CD16 abnormal expression of granulocytes was seen in both CMML and MDS patients, but there was no statistically significant difference between them. Other antigens showed abnormality of varying degrees, but did not have any statistical significance. It is concluded that MDS, CMML and AML-M5b displayed a certain degree of similarity, and also possess their own immunophenotype characteristics. Comprehensive analysis of immunophenotype by multiparameter flow cytometry may be important for differential diagnosis among CMML, MDS and AML-M5b. High percentage of monocytes, abnormal coexpression of CD2, CD56 and CD14 tailing phenomenon, lack or decrease of CD15 as well as abnormal expression of CD13/CD11b/CD16 in granulocytes may play important roles in diagnosis of CMML.
Case-Control Studies
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Flow Cytometry
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methods
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Humans
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Immunophenotyping
;
methods
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Leukemia, Monocytic, Acute
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diagnosis
;
immunology
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Leukemia, Myelomonocytic, Chronic
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diagnosis
;
immunology
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Myelodysplastic Syndromes
;
diagnosis
;
immunology
4.Immunologic characteristics and prognosis of myelodysplastic syndrome new subtype: refractory anemia with excess blasts-II.
Jia WEI ; Xiao-Fen ZHOU ; Fei ZHAO ; Jian-Feng ZHOU ; Yan CHEN
Journal of Experimental Hematology 2009;17(1):111-116
This study was aimed to investigate the immunologic characteristics of refractory anemia with excess blasts-II (RAEB-II) which belongs to a new subtype of World Health Organization (WHO) classification of myelodysplastic syndrome (MDS) and to screen out the independent immunologic prognostic factors of MDS. 35 cases of adult patients with de novo MDS were investigated. The immunofluorescent analysis by multiparameter flow cytometry was performed at the double gating of CD45/SSC to determine the immunophenotype of MDS cells in all cases. All patients were followed up. 47 cases of acute myeloid leukemia (AML) M1, 51 cases of AML-M(2) and 38 cases of acute lymphocytic leukemia (ALL) were selected as control. Software SPSS 13.0 was applied to analyze all the related data. The results showed that the positive expression rate of HLA-DR in RAEB-II was 100%, which was high in sensitivity and specificity. CD13 (94.74%), CD33 (84.21%) and CD117 (78.95%) were also highly expressed in RAEB-II. CD13 in RAEB-II was significantly higher than that in refractory cytopenia with or without multilineage dysplasia (RA/RCMD) (p < 0.01) and REAB-I (p < 0.05); CD33, CD117 (p < 0.05) and stem cell antigen CD34 (p < 0.01) in RAEB-II were significantly higher than that in RCMD (p < 0.01), but no statistically significant difference was found as compared with RAEB-I (p > 0.05). Compared with AML-M(1) and AML-M(2), no significant difference of CD13 and CD117 in RAEB-II was found (p > 0.05). CD33 (p < 0.01) and CD34 (p < 0.05) were significantly lower than that in AML-M(1), but no significant difference was found as compared with AML-M(2) (p > 0.05); CD15 (p < 0.01) and CD11b (p < 0.05) was significantly lower than that in M(2), but no significant difference was found as compared with AML-M(1) (p > 0.05); MPO was significantly lower than that in AML-M(1) and M(2) (p < 0.05); HLA-DR was significantly higher than that on AML-M(2) (p < 0.05), but no significant difference was found as compared with AML-M(1) (p > 0.05). RAEB-II did not express CD2, CD3, CD5 and CD8 (positive rate 0%, p < 0.01) when compared with T-ALL; CD4 (p < 0.05) and CD7 (p < 0.01) were significantly lower than that in T-ALL. RAEB-II did not express CD19 and CD20 (positive rate 0%, p < 0.01) as compared with B-ALL; CD10, CD22 and cCD79a were significantly lower than that in B-ALL (p < 0.05). CD117 (p = 0.0197) and MPO (p = 0.0085) were the two prognostic immunological antigens as regards the overall survival (OS) of MDS; CD117 (p = 0.003) was the single parameter in Cox regression. It is concluded that RAEB-II expresses mainly myeloid antigen without or with little expression of lymphoid antigen. Unique individual immunophenotypic features can be detected in patients with RAEB-II. HLA-DR can be a specific parameter to distinguish the other subtypes of MDS. CD117 may be an independent prognostic immunological antigen as regards OS of MDS.
Adolescent
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Adult
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Aged
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Anemia, Refractory
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complications
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diagnosis
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immunology
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Blast Crisis
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complications
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diagnosis
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immunology
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Female
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Humans
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Male
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Middle Aged
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Myelodysplastic Syndromes
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complications
;
diagnosis
;
immunology
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Prognosis
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Young Adult
5.Diagnostic significance of immunophenotyping of bone marrow cells in myelodysplastic syndrome without an increase of marrow blasts.
Juan XU ; Wei ZHANG ; Yan LIU ; Sui-Gui WAN ; Xue-Jing SUN ; Hong ZHAO
Journal of Experimental Hematology 2009;17(6):1477-1481
This study was aimed to investigate the characteristics of immunophenotypes in the patients with myelodysplastic syndrome (MDS) without an increase of marrow blasts, and to confirm their diagnostic significance. Marrow cells from 222 patients with pancytopenia, dysplastic changes in one or more hematopoietic lineages and blast cells less than 5% were analyzed by multiparametric flow cytometry(FCM). The abnormal immunophenotypes were evaluated in asynchronous antigen expression (CD34 or CD117 in mature granulocytes or mature monocytes, HLA-DR in mature granulocytes), in cross-lineage antigen expression (CD7 or CD56 in granulocytes or monocytes), in aberrant light-scatter (CD45/SSC in mature granulocyte or monocyte) and in abnormal expression of differentiation antigen (CD13/CD16 pattern in granulocytes and HLA-DR under-expression in monocytes). The sensitivity and specificity of abnormal immunophenotypes were determined on diagnosis. Among 222 cases, 127 cases were diagnosed as MDS by traditional diagnostic method and 95 cases were non-MDS (drug-related neutropenia, autoimmune cytopenia and idiopathic thrombocytopenia). In mature granulocyte gate, the sensitivity of asynchronous, cross-lineage antigen expression, aberrant light-scatter of CD45/SSC and abnormal expression of differentiation antigen were 31.5%, 30.7%, 49.6% and 60.6% respectively, and the specificity were 100%, 100%, 88.4% and 52.6% respectively. In monocyte gate, the sensitivity of asynchronous, cross-lineage antigen expression, aberrant light-scatter of CD45/SSC and abnormal expression of differentiation antigen were 2.3%, 11%, 37% and 12.6% respectively. The specificity was 100% in all of them. Among 8 above mentioned items, sensitivity of more than 2 abnormalities was 77.9%, and specificity was 95.8%. The positive predictive value was 96.1%. It is concluded that the abnormal expression of asynchronous, cross-lineage antigen expression, aberrant light-scatter of CD45/SSC have a high specificity and a low sensitivity for diagnosis of MDS. The abnormal expressions of differentiation antigens have a high sensitivity and a low specificity; however, the detection of multiple expression abnormalities possesses the high sensitivity and specificity for diagnosis of MDS.
Adolescent
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Adult
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Aged
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Aged, 80 and over
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Bone Marrow Cells
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immunology
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Female
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Flow Cytometry
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Humans
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Immunophenotyping
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Male
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Middle Aged
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Myelodysplastic Syndromes
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diagnosis
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immunology
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Sensitivity and Specificity
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Young Adult
6.Analysis of immunophenotypic features of blasts in patients with myelodysplastic syndrome by flow cytometry and its diagnostic significance.
Jia-Feng SUN ; Bo YANG ; Li ZHANG ; Yan HUANG ; Jia YANG
Journal of Experimental Hematology 2012;20(3):632-635
This study was purposed to analyse the characteristics of blast immunophenotype in patients with refractory anemia with excess blasts, type 1 (RAEB-1) and refractory anemia with excess blasts, type 2 (RAEB-2) by flow cytometry and investigate its diagnostic significance, as well as to compare the blast rate detected by FCM and bone marrow smear (BMS). FCM was used to count the blast rate and detect the expression of its antigens in 29 cases of MDS. The result indicated that: (1) The rate of the blasts detected by FCM and BMS was not statistically significant between patients with RAEB-1 and with RAEB-2 (P > 0.05); (2) Out of 13 patients with RAEB-1, the blasts of 10 cases, 12 cases, 8 cases, 11 cases, 11 cases, 3 cases, 7 cases, 0 case, 0 case, 3 cases and 2 cases patients had positive expressions of CD34, HLA-DR, CD117, CD33, CD13, CD15, CD11b, CD3, CD19, CD7 and CD56, separately. The blasts of 12 cases, 13 cases, 3 cases, 12 cases, 15 cases, 7 cases, 5 cases, 0 case, 1 case, 4 cases and 2 cases among 16 patients with RAEB-2 positively expressed CD34, HLA-DR, CD117, CD33, CD13, CD15, CD11b, CD3, CD19, CD7 and CD56, respectively. However, there was no significant difference in the expression of antigens in blasts of the bone marrow between the patients with RAEB-1 and with RAEB-2 (P > 0.05); (3) The positive expression for CD15 and CD11b in blasts was found in 10 cases and 12 cases, respectively; (4) The positive expression for CD19, CD7 and CD56 was observed in 1, 7 and 4 cases, respectively. None of the 29 cases of MDS was positive for CD3. It is concluded that FCM can reveal the characteristics of immunophenotypic abnormalities of the blasts in patients with MDS and provide the important information for diagnosis and differential diagnosis of MDS.
Adolescent
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Adult
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Aged
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Aged, 80 and over
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Bone Marrow
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pathology
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Female
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Flow Cytometry
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Humans
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Immunophenotyping
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Male
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Middle Aged
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Myelodysplastic Syndromes
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diagnosis
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immunology
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pathology
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Prognosis
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Young Adult
7.CD4+CD25highFoxP3+ Regulatory T-cells in Hematologic Diseases.
Hee Won MOON ; Bo Hyun KIM ; Chul Min PARK ; Mina HUR ; Yeo Min YUN ; Sung Yong KIM ; Mark Hong LEE
The Korean Journal of Laboratory Medicine 2011;31(4):231-237
BACKGROUND: CD4+CD25+ regulatory T-cells (Tregs) play a critical role in immune responses. We explored the status of Tregs in neoplastic and autoimmune hematologic diseases. We also evaluated the technical aspects of Treg measurement in terms of sample type and detection markers. METHODS: A total of 68 subjects were enrolled: 11 with AML, 8 with MDS, 10 with autoimmune diseases, and 39 controls. Tregs were analyzed in peripheral blood (PB) and bone marrow (BM) samples from each subject. Flow cytometry and the Human Regulatory T cell Staining Kit (eBioscience, USA) for CD4, CD25, and FoxP3 (forkhead box P3) were used. RESULTS: The CD4+CD25high/CD4 and CD4+CD25highFoxP3+/CD4 populations were significantly correlated (P<0.0001). The AML and high-risk MDS groups had significantly larger CD4+CD25high/CD4 and CD4+CD25highFoxP3+/CD4 populations in PB than the autoimmune (P=0.007 and 0.012, respectively) and control groups (P=0.004 and 0.006, respectively). Comparable findings were observed in BM. The CD4+CD25highFoxP3+/CD4 population was significantly larger in PB than in BM (P=0.0003). CONCLUSIONS: This study provides comparison data for Tregs in AML, MDS, and autoimmune hematologic diseases, and would be helpful for understanding the different immunologic bases of various hematologic diseases. Treg measurement using CD4, CD25, and/or FoxP3 in PB rather than in BM seems to be practical for routine hematologic purposes. Large-scale analysis of the diagnostic role of Treg measurement is needed.
Adolescent
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Adult
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Aged
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Aged, 80 and over
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Autoimmune Diseases/diagnosis/immunology
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Bone Marrow Cells/cytology
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Female
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Flow Cytometry
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Forkhead Transcription Factors/*metabolism
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Hematologic Diseases/*diagnosis/immunology
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Humans
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Interleukin-2 Receptor alpha Subunit/*metabolism
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Leukemia, Myeloid, Acute/diagnosis/immunology
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Leukocytes, Mononuclear/cytology
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Male
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Middle Aged
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Myelodysplastic Syndromes/diagnosis/immunology
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T-Lymphocytes, Regulatory/immunology/*metabolism
8.Myelodysplastic Syndrome Mimicking Idiopathic Thrombocytopenic Purpura.
Yusun HWANG ; Jung Won HUH ; Yeung Chul MUN ; Chu Myong SEONG ; Wha Soon CHUNG
The Korean Journal of Laboratory Medicine 2010;30(2):105-110
BACKGROUND: In patients with isolated thrombocytopenia, but without significant dysplasia, diagnosis of idiopathic thrombocytopenic purpura (ITP) rather than myelodysplastic syndrome (MDS) may be taken into account. It is important to make an accurate diagnosis because different treatments are used for ITP and MDS. The purpose of this study was to investigate the clinical and hematologic features of patients who were initially diagnosed as ITP but had cytogenetic abnormalities. METHODS: We retrospectively reviewed cytogenetic studies of 100 patients who were diagnosed as ITP from 2004 to 2009 at Mokdong Hospital of Ewha Womans University based on clinical features and hematologic studies. Bone marrow pathology was re-evaluated based on 2008 WHO classification. Cytogenetic analysis was performed by 24-48 hr culture of bone marrow aspirates without using mitogens and 20 metaphases were analyzed. RESULTS: Of the 100 patients diagnosed as ITP initially, three patients (3%) had cytogenetic abnormalities. They had no thrombocytopenia-related symptoms and thrombocytopenia was found accidentally. The numbers of megakaryocytes in bone marrow were increased and dysplasia was not found in megakaryocyte, erythroid, and myeloid cell lineages. The proportion of blasts was within normal limits. Clonal chromosomal abnormalities found were der(1;7)(q10;p10), add(9)(q12), or t(7;11)(p22;q12). Presumptive diagnosis of MDS or diagnosis of idiopathic cytopenia of undetermined significance (ICUS) was made according to 2008 WHO classification. During the follow up, disease progression was not found. CONCLUSIONS: In patients with suspected ITP, cytogenetic analysis should be done. If specific clonal chromosomal abnormality is found, presumptive diagnosis of MDS has to be considered and close follow up is needed.
Adult
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Bone Marrow Cells/cytology
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Cell Lineage
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Chromosome Aberrations
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Diagnosis, Differential
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Female
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Humans
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Male
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Megakaryocytes/immunology/pathology
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Middle Aged
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Myelodysplastic Syndromes/*diagnosis/genetics/pathology
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Purpura, Thrombocytopenic, Idiopathic/*diagnosis/genetics/pathology
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Retrospective Studies
9.Disseminated Mycobacterium kansasii Infection Associated with Skin Lesions: A Case Report and Comprehensive Review of the Literature.
Sang Hoon HAN ; Kyoung Min KIM ; Bum Sik CHIN ; Suk Hoon CHOI ; Han Sung LEE ; Myung Soo KIM ; Su Jin JEONG ; Hee Kyoung CHOI ; Chang Oh KIM ; Jun Yong CHOI ; Young Goo SONG ; June Myung KIM
Journal of Korean Medical Science 2010;25(2):304-308
Mycobacteruim kansasii occasionally causes disseminated infection with poor outcome in immunocompromised patients. We report the first case of disseminated M. kansasii infection associated with multiple skin lesions in a 48-yr-old male with myelodysplastic syndrome. The patient continuously had taken glucocorticoid during 21 months and had multiple skin lesions developed before 9 months without complete resolution until admission. Skin and mediastinoscopic paratracheal lymph node (LN) biopsies showed necrotizing granuloma with many acid-fast bacilli. M. kansasii was cultured from skin, sputum, and paratracheal LNs. The patient had been treated successfully with isoniazid, rifampin, ethmabutol, and clarithromycin, but died due to small bowel obstruction. Our case emphasizes that chronic skin lesions can lead to severe, disseminated M. kansasii infection in an immunocompromised patient. All available cases of disseminated M. kansasii infection in non HIV-infected patients reported since 1953 are comprehensively reviewed.
Antitubercular Agents/therapeutic use
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Clarithromycin/therapeutic use
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Glucocorticoids/therapeutic use
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Humans
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Immunocompromised Host
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Isoniazid/therapeutic use
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Male
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Middle Aged
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Mycobacterium Infections, Nontuberculous/*diagnosis/drug therapy/immunology
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*Mycobacterium kansasii/isolation & purification
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Myelodysplastic Syndromes/drug therapy
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Rifampin/therapeutic use
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Skin Diseases, Bacterial/*diagnosis/immunology/pathology
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Sputum/microbiology
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Sweet Syndrome/diagnosis
10.A Case of Inguinal Sparganosis Mimicking Myeloid Sarcoma.
Jin Yeob YEO ; Jee Young HAN ; Jung Hwan LEE ; Young Hoon PARK ; Joo Han LIM ; Moon Hee LEE ; Chul Soo KIM ; Hyeon Gyu YI
The Korean Journal of Parasitology 2012;50(4):353-355
We report here a case of inguinal sparganosis, initially regarded as myeloid sarcoma, diagnosed in a patient undergone allogeneic hematopoietic transplantation (HSCT). A 56-year-old male patient having myelodysplastic syndrome was treated with allogeneic HSCT after myeloablative conditioning regimen. At day 5 post-HSCT, the patient complained of a painless palpable mass on the left scrotum and inguinal area. Pelvic magnetic resonance imaging and computed tomography revealed suspected myeloid sarcoma. Gun-biopsy was performed, and the result revealed eosinophilic infiltrations without malignancy. Subsequent serologic IgG antibody test was positive for sparganum. Excisional biopsy as a therapeutic diagnosis was done, and the diagnosis of sparganosis was confirmed eventually. This is the first report of sparganosis after allogeneic HSCT mimicking myeloid sarcoma, giving a lesson that the physicians have to consider the possibility of sparganosis in this clinical situation and perform adequate diagnostic and therapeutic approaches.
Animals
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Diagnosis, Differential
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Hematopoietic Stem Cell Transplantation
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Humans
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Larva
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Magnetic Resonance Imaging
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Male
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Middle Aged
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Myelodysplastic Syndromes/complications/surgery
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Republic of Korea
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Sarcoma, Myeloid/diagnosis
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Scrotum/parasitology/*pathology
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Sparganosis/parasitology/*pathology/radiography
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Sparganum/*immunology/isolation & purification
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Tomography, X-Ray Computed
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Transplantation, Homologous