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Objective: To evaluate the prognostic significance of minimal residual disease (MRD) monitoring by 10-color flow cytometry in multiple myeloma (MM) patients after treatment. Methods: 150 patients with MM who were admitted to the First Affiliated Hospital of Soochow University from July 2015 to July 2017 were retrospectively analyzed. Clinical data, MRD data monitoring by 10-color flow cytometry and prognosis were analyzed. Results: 39.1% (34/87) patients were MRD negative after induction chemotherapy, and 49.3% (34/69) patients were MRD negative within 1 year after autologous hematopoietic stem cell transplantation (ASCT) . MRD-negative patients after induction chemotherapy or after transplantation have better progress-free survival (PFS) than MRD-positive patients (P=0.022 and P<0.001) . According to the changes of MRD pre-ASCT and after ASCT, the patients were divided into 4 groups: patients with MRD continued negativity,improved from MRD positive to MRD negative, MRD continued positivity, transformed from MRD negative to MRD positive. The two-year PFS of the four groups were 83%, 82%, 44%, 0, respectively, (P=0.002) . Multivariate analysis showed that the level of MRD after induction chemotherapy was an independent factor for PFS (P=0.002) , HR=4.808 (95%CI 1.818-12.718) . Conclusion: Patients with MRD negative after treatment is a better prognosis marker than complete remission or even the best marker, which can evaluate prognosis by combining R-ISS and cytogenetic changes.
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Humanos , Citometría de Flujo , Mieloma Múltiple/diagnóstico , Neoplasia Residual , Pronóstico , Estudios Retrospectivos , Trasplante AutólogoRESUMEN
Objective: Early T-cell precursor acute lymphoblastic leukemia (ETP-ALL) is a recently recognized high-risk T lymphoblastic leukemia subgroup. The optimal therapeutic approaches to adult patients with ETP-ALL are poorly characterized. In this study, we explore the efficacy and outcome of allogeneic hematopoietic stem cell transplantation (allo-HSCT) for ETP-ALL. Methods: The clinical data of 23 patients with ETP-ALL receiving allo-HSCT from 2010 to 2018 were retrospectively analyzed. Patients with ETP-ALL were diagnosed based on the characteristic immunophenotypes. Second-generation sequencing was done in all patients. As to the donors, 12 patients had haploidentical donors (Haplo-HSCT) , 7 HLA-matched sibling donors (Sib-HSCT) and 4 HLA-matched unrelated donors (URD-HSCT) . Before transplantation, 19 patients achieved complete remission (CR) and 4 patients without. Results: The main clinical features of ETP-ALL included high white blood cell counts in 5 patients, splenomegaly in 14, lymphadenopathy in 19, and thymus masses in 5. According to cytogenetic and molecular characteristics, 11 patients had gene mutations related to myeloid tumors, and 7 with high risk Karyotype. After first induction regimen, 14/23 patients achieved CR. 5 patients reached CR after more than 2 cycles of chemotherapy, while another 4 patients did not reach CR. After allo-HSCT, 22 patients were successfully implanted. The median time of granulocyte and platelet reconstitution was +12 and +19 days. One patient died of transplant-related infection at +14 days. The estimated 18-month overall survival (OS) and relapse-free survival (RFS) rates were (55.0±14.4) % and (48.1±14.7) % respectively. Transplant-related mortality was 4.3%. The median OS in patients achieving CR before transplantation was 20 months, however, that in patients without CR was only 13 months. OS and RFS between haplo-HSCT and sib-HSCT were comparable (P=0.460 and 0.420 respectively) . Conclusions: Allo-HSCT is an effective therapy in some patients with ETP-ALL. Salvage HSCT cannot overcome the poor outcome. Haplo-HSCT and sib-HSCT in ETP-ALL patients have the similar clinical outcome.
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Adulto , Humanos , Trasplante de Células Madre Hematopoyéticas , Leucemia-Linfoma Linfoblástico de Células Precursoras , Células Precursoras de Linfocitos T , Inducción de Remisión , Estudios RetrospectivosRESUMEN
OBJECTIVE@#To summarize the clinical characteristics of patients with acute myeloid leukemia-type M (AML-M) and analyze the factors affecting the prognosis.@*METHODS@#One hundred eighty-eight AML-M patients were retrospectively analyzed for the following parameters including peripheral blood, immune phenotypes, fusion genes and cytogenetics to explore their significance for the overall survival (OS) and progression-free survival (PFS). The prognostic factors were also analyzed.@*RESULTS@#Among 188 patients with AML-M, the chromosomal abnormality with t (8;21), normal chromosome and other abnormalities accounted for 37% (70/188), 41% (77/188) and 22% (41/188), respectively. For the immunopheno typing of M patients, the hematopoietic progenitor cell differentiation antigen CD117 (96.1%) were mainly expressed, CD34 (81.6%) and HLA-DR (55.9%), and myeloid-associated antigen of CD13 (90.5%) and CD33 (89.4%) were also highly expressed. There were lymphoid-associated antigens expressed in some patients, among which the positive expression rate of CD19 was highest (29.6%), and the next was CD7 (28.5%). The most common accompanied mutations was FLT3 mutation (30.2%). The univariate analysis showed that the patients at age<50 years old, without extramedullary infiltration, with positive expression of CD19, NPM-1 (-), CEBPA double mutation(+), and HSCT were significant superior in OS and PFS (P<0.05); the multivariate analysis showed that the patient at age<50 years old, without extramedullary infiltration, with positive expression of CD19 and CEBPA double mutation (+) were significant superior in OS and PFS (P<0.05). The analysis indicated that the Karytypes affected only OS (P<0.05), while the NPM-1 gene mutation positive affected only PFS (P<0.05). The univarate analysis of factors affecting the survival in 70 AML-M patients with t (8;21) abnormatity showed that the C-KIT gene mutation was a poor factor for OS and PFS.@*CONCLUSION@#The clinical characteristics are different between M patients with different karyotype, and prognostic analysis shows that the karytypes have an impact on overall survival; age, extramedullary infiltration, CD19 expression and CEBPA double mutation are also the main factors impacting the prognosis of patients.
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Humanos , Persona de Mediana Edad , Antígenos HLA-DR , Inmunofenotipificación , Leucemia Mieloide Aguda , Mutación , Pronóstico , Estudios RetrospectivosRESUMEN
<p><b>OBJECTIVE</b>To explore the differentiation-inducing potentiality of Pulsatilla saponin A on K562 cells.</p><p><b>METHODS</b>Pulsatilla saponin A of different concentrations was used to treat K562 cells; the benzidine staining and the hemoglobinometry were applied to measure the change of hemoglobin content; the flow cytometry (FCM) was used to detect the expression of CD71 and GPA on K562 cells.</p><p><b>RESULTS</b>K562 cells treated with 4 µg/ml pulsatilla saponin A differentiated into the erythroid lineage. With the treatment of pulsatilla saponin A, the hemoglobin content in K562 cells increased significantly; CD71 and GPA expression on the K562 cell surface were up-regulated.</p><p><b>CONCLUSION</b>Pulsatilla saponin A can induce K562 cells to differentiate into erythroid lineage.</p>
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Humanos , Antineoplásicos , Diferenciación Celular , Linaje de la Célula , Células Eritroides , Células K562 , SaponinasRESUMEN
<p><b>OBJECTIVE</b>To explore the autophagy activity of CD34+ cells in bone marrow of MDS patients and its clinical significance.</p><p><b>METHODS</b>The activity of autophagy in bone marrow CD34+ cells from 20 MDS patients, 20 non-malignant anemia patients and 5 AML patients admitted in our hospital from October 2012 to March 2014 was detected by flow cytometry (FCM).</p><p><b>RESULTS</b>The autophagy activity in low risk MDS patients and non-malignant anemia patients were both significantly higher than that in both high risk MDS and AML patients (P<0.05), and more interestingly, the autophagy activity in MDS negatively correlated with World Health Organization classification-based prognostic system (WPSS) score (r=-0.877) .</p><p><b>CONCLUSION</b>The autophagy activity CD34+ cells in the patients with MDS is higher than that in AML patients, and negatively correlated with WPSS scores, indicating that the decrease of autophagy activity maybe accelerate the genesis and development of MDS and relate with the prognosis of MDS patients.</p>
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Humanos , Antígenos CD34 , Metabolismo , Autofagia , Células de la Médula Ósea , Biología Celular , Patología , Citometría de Flujo , Leucemia Mieloide Aguda , Patología , Síndromes Mielodisplásicos , Patología , PronósticoRESUMEN
<p><b>OBJECTIVE</b>To summarize the clinical characteristics as well as diagnosis and treatment in 1 case of acute myeloid leukemia(AML) with coexpression of Ph and inv(16).</p><p><b>METHODS</b>A series of clinical tests, the cellular morphological, immunological, cytogenetic and molecular biological examinations of leukemia cells were performed.</p><p><b>RESULTS</b>The clinical characteristics of this patient were very common. The cellular morphology is similar to the AML with inv(16). The leukemia cells were stained positively for CD13, CD33, CD34, CD117 and HLA-DR. Karyotypic analysis showed a complex chromosome abnormality including inv(16) and Ph, and the FISH analysis showed that the percentage of rearrangement of CBFβ allele was over that of the BCR-ABL fusion signals. The obvious adverse events did not occur in this patient within 3 years.</p><p><b>CONCLUSION</b>Ph as secondary aberration of inv(16) rarely occures in primary AML cases, and so far there have not been the clear criteria of diagnosis and treatment. The cytogenetic and molecular biology could provide the basis for diagnosis. Moreover, autologous hematopoietic stem cell transplantation combined with imatinib probably is one of the effective treatment methods.</p>
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Humanos , Aberraciones Cromosómicas , Trastornos de los Cromosomas , Inversión Cromosómica , Proteínas de Fusión bcr-abl , Antígenos HLA-DR , Leucemia Mieloide Aguda , Cromosoma FiladelfiaRESUMEN
<p><b>OBJECTIVE</b>To investigate the alteration of microparticles (MP) in the recipients following hematopoietic stem cell transplantation (HSCT) and its significance, and to search the early diagnostic indicators of thrombotic complications after transplantation.</p><p><b>METHODS</b>According to the occurrence of transplantation-associated complications, 94 allo-HSCT patients were divided into 4 groups: thrombotic group (VOD n = 7, TMA n = 2), acute graft-versus-host disease (aGVHD) group (n = 27), infection group (n = 41) and non-complication group (n = 17). Alterations of serum concentration of tissue factor positive microparticles (TF(+) MP) and endothelial microparticles (EMP) were analyzed by flow cytometry during the process of conditioning treatment and the early stage after transplantation. The relation of these 2 kinds of MP with complications was analysed.</p><p><b>RESULTS</b>(1) The levels of TF(+) MP and EMP of patients undogoing allo-HSCT before conditioning treatment were obviously higher than those in normal controls, and showed some elevation during different times, but there was no significant statistical difference. Although the levels of TF(+) MP and EMP at the end of conditioning treatment were some higher than those before conditioning treatment, but there was no statistical difference between them. (2)The levels of TF(+) MP and EMP in thrombotic group were obviously higher than those in aGVHD group and infection group (P < 0.05). (3)The levels of TF(+) MP and EMP in thrombotic group at different times were significant differences from those in other groups (P < 0.05), and the levels of TF(+) MP and EMP were no significant difference from those in non-complication group.</p><p><b>CONCLUSION</b>The increase of the TF(+) MP and EMP levels may be associated with occurrence of thrombosis after transplantation, indicating occurrence of the thrombotic complications, like hepatic vein occulusive disease (HVOD). The dynamically monitoring levels of TF(+) MP and EMP contributes to early discovery of thrombotic complications.</p>
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Humanos , Micropartículas Derivadas de Células , Citometría de Flujo , Enfermedad Injerto contra Huésped , Trasplante de Células Madre Hematopoyéticas , TrombosisRESUMEN
The purpose of this study was to summary the clinical and laboratorial features in 15 adult cases of mixed phenotypic acute leukemia with Ph chromosome and/or BCR-ABL fusion gene positive (Ph(+)MPAL), 15 adult patients with Ph(+)MPAL were defined by WHO-2008 classification. The clinical characteristics, results of morphology, immunology, cytogenetics and molecular genetic detections and results of follow-up in 15 adult patients with Ph(+)MPAL were analyzed retrospectively. The results showed that 15 patients among 87 cases of MPAL demonstrated Ph(+)MPAL (17.2%; 15/87) (7 males and 8 females), their median age was 51 (range 16-81) year old and median WBC count at diagnosis was 69 (12.7-921)×10(9)/L. Based on FAB criteria, these patients showed different morphologic types, including AML (13.3%; 2/15), ALL (40.0%; 6/15), HAL (46.7%; 7/15). Immunologic analysis indicated that 15 cases of Ph(-)MPAL were all classified as B-lymphoid +myeloid mixed immunophenotype. Except one patient, all expressed CD34 antigen on the surface of leukemia cells with 64.3% strong positive, only Ph (53.3%; 8/15), Ph with additional chromosomal abnormalities (33.3%; 5/15) and normal karyotype (13.3%; 2/15) were cytogenetically identified. BCR-ABL fusion gene transcript positive were detected by multiplex reverse transcription PCR in all cases, with e1a2 subtype (p190) (40.0%; 6/15) and b2a2 or b3a2 (p210) subtype (60.0%; 9/15). Four out of 7 (57.1%) patients were found to have IKZF1 gene deletion, without other common gene mutations. Seven out of 10 cases (70.0%) achieved complete remission (CR) after one cycle of induction chemotherapy. In the induction stage, CR rate seemed higher when tyrosine kinase inhibitors (TKI) were added to chemotherapy (83.3%:50.0%; P = 0.206). Overall survival (OS) in 4 patients received allogeneic hematopoietic stem cell transplantation (allo-HSCT) was longer than that in 4 patients received chemotherapy alone (P = 0.004). It is concluded that Ph(+)MPAL mainly is expressed as B+My phenotype. The majority of patients is older and has CD34 overexpression. In the aspect of molecular genetics, the Ph(+)MPAL is similar to other acute leukemia with Ph chromosome. Ph(+)MPAL is a subtype of acute leukemia with poor prognosis. WBC count at diagnosis is an independent prognostic factor. The combination of TKI and allo-HSCT can improve their long-term survival, which needs to be confirmed through carrying out a prospective and multicenter clinical trial for newly diagnosed Ph(+)MPAL.
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Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Femenino , Humanos , Masculino , Persona de Mediana Edad , Adulto Joven , Antígenos CD34 , Metabolismo , Proteínas de Fusión bcr-abl , Genética , Metabolismo , Trasplante de Células Madre Hematopoyéticas , Cariotipificación , Fenotipo , Leucemia-Linfoma Linfoblástico de Células Precursoras , Diagnóstico , Genética , Terapéutica , Pronóstico , Inhibidores de Proteínas Quinasas , Usos Terapéuticos , Estudios Retrospectivos , Tasa de SupervivenciaRESUMEN
Objective of this study was to detect the level of tissue factor-positive microparticles (TF(+)MP) by flow cytometry (FCM) and to analyze its clinical significance in the haemostatic disorder. TF(+) MP was detected by FCM using antibody CD142-PE in 25 cases of acute promyelocytic leukemia (APL), 20 cases of hemostatic diseases and 20 healthy adults as controls. The differences of TF(+) MP between various groups were determined. The results showed that the level of TF(+) MP in the patients with thrombotic complications was significantly higher than that in the healthy adults (P < 0.05). The TF(+) MP level was higher in the patient with APL than that in the healthy adults, especially in course before therapy (P < 0.01), but the difference was not statistically significant in the patient with APL after therapy and the healthy adults. Among these patient with APL, the level of TF(+) MP in the 18 patients who complicated with disseminated intravascular coagulation (DIC) was also higher than that in the healthy adults (P < 0.05), but the level of TF(+) MP in the other 7 patients who did not complicate with DIC was similar before and after treatment. It is concluded that the method of TF(+) MP detection by FCM is feasible and simple, it is useful for the diagnosis of thrombotic disorder, and helps evaluation for the prognosis of APL patient.
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Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Femenino , Humanos , Masculino , Persona de Mediana Edad , Adulto Joven , Trastornos de la Coagulación Sanguínea , Sangre , Estudios de Casos y Controles , Citometría de Flujo , Leucemia Promielocítica Aguda , Sangre , Pronóstico , Tromboplastina , MetabolismoRESUMEN
Objective To observes the change of early effective biomarkers of endothelial injury with lowarsenic exposure in drinking water. MethodsNinety rurad residents, who had lived in Yanhe village, Xuyi county and Jiangsu province for at least 10 years, were recruited by simple random sampling in this study. The level of arsenic in their household shallow well were divided into three groups, which were < 10 (32 people), 10 - 50(28 people) and > 50 μg/L(30 people). Blood samples from individuals were collected. Malondialdehyde(MDA) in human plasma, which is considered as the most important marker for monitoring lipid peroxidation, was determined as conjugate with tetrabutylammonium hydrogen sulphate(TBA). The level of anti-superoxide anion radical(O-·2),C-reactive protein(CRP) and NO in human plasma was measured with colorimetry, turbidimetry and nitric acid reductase, respectively. The number of circulating endothelial progenitor cells(CEPCs) in peripheral blood was analyzed by CD133+/KDR+ antibodies and flow cytometry. Results Ninety cases underwent questionnaires. Between the groups, the difference of the levels of MDA (61.1, 65.5, 67.5 μmol/kg), O-·2 (4774.6, 5143.3, 4736.0 U/kg) ,CRP[(5.92 ± 2.44), (5.11 ± 2.40), (5.55 ± 2.96)mg/L], and NO[(659.8 ± 387.5), (667.4 ± 486.6), (762.1 ±763.2)μmol/kg], was not statistically significant (F =0.00, 0.46, 0.80, 0.47, all P > 0.05). The difference of the number of CEPCs in different groups of arsenic in drinking water was statistically significant(0.96 x 10-5, 0.77 x 10-5,1.59 x 10-5, F=5.08, P< 0.05), where < 10, 10 - 50 μg/L groups were significantly lower than > 50 μg/L group (q =4.58, 6.65, all P < 0.05). ConclusionsThe number of CEPCs in peripheral blood changes significantly with lower-arsenic exposure, whereas there are no obvious changes with the markers of oxidized damage and inflammation. This is the first human demonstration showing that lower-arsenic exposure may cause endothelial injury.
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<p><b>OBJECTIVE</b>To investigate the clinical and biological characteristics and prognosis of mixed phenotype acute leukemia (MPAL).</p><p><b>METHODS</b>Thirty two patients were diagnosed as MPAL by bone marrow examination, immunophenotyping, cytogenetic and molecular assay and were treated with combined chemotherapy regimens for both acute lymphoblastic and acute myeloid leukemia. Two cases were received allogeneic hematopoietic stem cell transplantation (allo-HSCT).</p><p><b>RESULTS</b>(1) The incidence of MPAL in acute leukemias was 2.6%. There were 16 cases (50.0%) of mixed myeloid and B-lymphoid (M/B), 14(43.8%) myeloid and T-lymphoid (M/T), one each (3.1%) of trilineage (M/B/T) and B- and T-lymphoid (B/T) phenotype. (2) The positive rates of CD34 and HLA-DR were 87.5% and 62.5%, respectively. (3) Abnormal karyotypes were detected in 70.0% of 30 MPAL patients, which were structural and numerical abnormalities including t(9;22), 11q23 and complex karyotypes. (4) The total complete remission (CR) rate was 75.0% and the overall survival (OS) and disease-free survival (DFS) at 2 years were 14.8% and 14.2% respectively. The CR rates for M/B and M/T cases were 75.0% and 71.4% respectively. No statistical difference was observed in OS and DFS between M/B and M/T cases.</p><p><b>CONCLUSIONS</b>MPAL is a rare type of acute leukemia with a high heterogeneity. The unfavorable indicators of MPAL may be factors such as abnormal karyotypes, high expression of CD34 and extramedullary infiltration. Combined regimens and more intensive therapy including allo-HSCT might contribute to improving survival.</p>
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Adolescente , Adulto , Niño , Preescolar , Femenino , Humanos , Masculino , Persona de Mediana Edad , Adulto Joven , Inmunofenotipificación , Cariotipo , Leucemia Bifenotípica Aguda , Clasificación , Genética , Alergia e Inmunología , Leucemia Mieloide Aguda , Genética , Alergia e Inmunología , Leucemia-Linfoma Linfoblástico de Células Precursoras , Genética , Alergia e Inmunología , PronósticoRESUMEN
This study was aimed to investigate the immunophenotypic characteristics of 109 cases of B-cell chronic lymphoid leukemia (B-CLL) so as to provide evidences for the diagnosis and therapy of B-CLL, and for the detection of the minimal residual disease and its prognosis. Immunophenotyping was performed in 109 patients of B-CLL by two/three color multiparameter flow cytometry analysis using a panel of monoclonal antibodies. The results showed that in 109 cases of B-CLL, all cases expressed CD19, the positive ratios of other B lineage antigen such as CD20, CD22 and CD23 were 95.40%, 94.50%, 86.20% respectively. None of the B-CLL cases expressed CD10. The expression ratio of FMC-7 and CD38 in 105 cases of B-CLL were 28.60% and 36.20%. Among the B-CLL cases the CD5(+) cells amounted to 86.23%, CD5(-) cells amounted to 13.76%, ZAP-70 protein was expressed in 12 out of 50 patients. It is concluded that immunophenotypic data are very useful for the diagnosis and detection of minimal residual disease of B-CLL, and the relationship between the immunophenotypic characteristics and the prognosis of B-CLL needs further study.
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Adulto , Anciano , Anciano de 80 o más Años , Femenino , Humanos , Masculino , Persona de Mediana Edad , Citometría de Flujo , Inmunofenotipificación , Leucemia Linfocítica Crónica de Células B , Diagnóstico , Alergia e Inmunología , Neoplasia Residual , Diagnóstico , Alergia e Inmunología , PronósticoRESUMEN
<p><b>OBJECTIVE</b>To explore the expression of CD34 in patients with acute promyelocytic leukemia (APL) and investigate the clinical and laboratory features of CD34(+) APL patients.</p><p><b>METHODS</b>262 APL patients diagnosed by chromosome analysis and/or fusion gene examination in the last five years were retrospectively analyzed in this study. To survey the expression of CD34 in those patients, all the cases were divided into two groups (CD34(+) APL vs. CD34(-) APL). The clinical features including age, gender, abnormal values of the peripheral hemogram before treatment, the complete remission (CR) rate and the incidence of DIC and laboratory data such as the results of morphology, immunology, cytogenetics and molecular biology (MICM) between those two groups were compared.</p><p><b>RESULTS</b>Of the 262 APL patients, 38 (14.5%) cases were positive for CD34 expression. There were no statistically significant differences between CD34(+) APL and CD34(-) APL groups in gender and age (P > 0.05). Before treatment, the median level of WBC in CD34(+) APL was 25.92 x 10(9)/L, which was significantly higher than that of CD34(-) APL (5.3 x 10(9)/L, P < 0.05). CD34(+) APL by morphology classification were mostly of the subtypes M3b and M3v (65.8%), while these subtypes in CD34(-) APL (40.3%) were significantly less (P < 0.01). There were no statistically significant differences between the two groups compared in respect of complete remission (CR) rate and the incidence of DIC (P > 0.05). The expression level of CD34 in APL had correlation to the expression level of CD2, CD7 and CD117; the latter three phenotypes in CD34(+) APL were significantly higher than those in CD34(-) APL (P < 0.01). No significant difference was found between those two groups by chromosome analysis, but there was more PML-RAR-alpha transcript short form in CD34(+) APL than that in CD34(-) APL (P < 0.05).</p><p><b>CONCLUSION</b>CD34(+) acute promyelocytic leukemia is a unique subtype of APL with different biological characteristics.</p>
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Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Niño , Femenino , Humanos , Masculino , Persona de Mediana Edad , Adulto Joven , Antígenos CD34 , Sangre , Antígenos CD7 , Sangre , Antineoplásicos , Usos Terapéuticos , Antígenos CD2 , Sangre , Coagulación Intravascular Diseminada , Inmunofenotipificación , Leucemia Promielocítica Aguda , Quimioterapia , Genética , Alergia e Inmunología , Proteínas Nucleares , Metabolismo , Fenotipo , Proteína de la Leucemia Promielocítica , Proteínas Proto-Oncogénicas c-kit , Sangre , Receptores de Ácido Retinoico , Metabolismo , Inducción de Remisión , Receptor alfa de Ácido Retinoico , Estudios Retrospectivos , Factores de Transcripción , Metabolismo , Translocación Genética , Tretinoina , Usos Terapéuticos , Proteínas Supresoras de Tumor , MetabolismoRESUMEN
<p><b>OBJECTIVE</b>To investigate the effect of thalidomide on Annexin II (AnxA2) gene regulation in multiple myeloma cell line RPMI8226 and human microvascular endothelial cell line HMEC-1 cells in vitro, and explore the potential mechanism of thrombosis induced by thalidomide.</p><p><b>METHODS</b>RPMI8226 and HMEC-1 cells were cultivated in vitro. Real time quantitative PCR (RQ-PCR) was used to detect the influence of thalidomide at different concentration on the expression of AnxA2 mRNA, flow cytometry (FCM) and confocal microscopy were used to detect the cell surface protein level after the samples were stimulated with different concentrations of thalidomide.</p><p><b>RESULTS</b>AnxA2 mRNA level in RPMI8226 cells treated with thalidomide at 12.5 microg/ml, 25.0 microg/ml and 50.0 microg/ml was decreased compared with the control group (0.60+/-0.15, 0.33+/-0.14, 0.42+/-0.16, vs 1.07+/-0.16, respectively, P<0.05) and did so in HMEC-1 cells (0.21+/-0.20, 0.08+/-0.08, 0.17+/-0.16 vs 1.16+/-0.24, respectively, P<0.05). The AnxA2 protein level in RPMI8226 cells treated with above mentioned concentrations of thalidomide was also decreased compared with the control (3.39+/-0.32, 2.82+/-0.28, 3.21+/-0.23 vs 5.53+/-0.32, respectively, P<0.05) and that did so in HMEC-1 cells (0.72+/-0.11, 0.64+/-0.08, 0.67+/-0.08 vs 1.40+/-0.15, respectively, P<0.05).</p><p><b>CONCLUSIONS</b>Thalidomide can inhibit the expression of AnxA2 mRNA and protein in RPMI8226 and HMEC-1 cells, which may be one of the mechanisms for the development of thrombosis induced by thalidomide in multiple myeloma patients.</p>
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Humanos , Anexina A2 , Genética , Metabolismo , Línea Celular , Células Endoteliales , Metabolismo , Endotelio Vascular , Biología Celular , Mieloma Múltiple , Metabolismo , Patología , ARN Mensajero , Genética , Talidomida , FarmacologíaRESUMEN
<p><b>OBJECTIVE</b>To investigate the effects of Annexin II (AnxA2) gene silencing by siRNA on proliferation and invasive potential of lymphoma cell line Jurkat cells.</p><p><b>METHODS</b>A synthesized siRNA duplex targeting to AnxA2 was transfected into Jurkat cells. Transfection efficiency was analyzed by real-time PCR and flow cytometry. MTT assay for cell proliferation and transwell plates for invasive potential were performed.</p><p><b>RESULTS</b>Compared with the negative controls, the cell proliferation inhibitory rate of the AnxA2 siRNA transfected Jurkat cells was significantly increased at 24 h, 48 h and 72 h [(17.4 +/- 2.3)%, (22.4 +/- 3.8)%, (37.6 +/- 1.5)% vs (-1.3 +/- 5.1)%, (-5.5 +/- 4.4)%, (-10.8 +/- 5.5)%, respectively, P<0.05]. The cell invasive potential of the transfected Jurkat cells was inhibited remarkably at 48 h (11.3 +/- 4.2 vs 54.3 +/- 8.7, P<0.01).</p><p><b>CONCLUSION</b>AnxA2 gene silenced by siRNA can inhibit the proliferation and the invasive potential of Jurkat cells remarkably.</p>
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Humanos , Anexina A2 , Genética , Metabolismo , Proliferación Celular , Quimiotaxis , Genética , Silenciador del Gen , Células Jurkat , ARN Interferente Pequeño , Genética , TransfecciónRESUMEN
<p><b>OBJECTIVE</b>To report a case of pure erythroid leukemia.</p><p><b>METHODS</b>The clinical features, treatment and prognosis of a rare case of pure erythroid leukemia were reported, and the related literature was reviewed.</p><p><b>RESULTS</b>The pure erythroid leukemia patient was diagnosed by 90.4% pronormoblasts in bone marrow, 99.5% for erythroid antigen CD71, 67.4% for glycophorin A were detected, while no differentiation antigen of myeloid, lymphoid and megakaryocyte lineages were observed. HAG (homoharringtonine + Cytarabine and G-CSF) regimen were administered with no effect. The patient developed multiple organ failure and died soon.</p><p><b>CONCLUSION</b>Pure erythroid leukemia has a fulminant clinical course with poor response to chemotherapy and worse prognosis.</p>
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Humanos , Masculino , Persona de Mediana Edad , Leucemia Eritroblástica Aguda , Terapéutica , PronósticoRESUMEN
<p><b>OBJECTIVE</b>To explore the characteristics of morphology, immunophenotype and cytogenetics (MIC) of myeloid surface antigen-expressing acute lymphoblastic leukemia (My+ ALL).</p><p><b>METHODS</b>One hundred and twenty untreated acute lymphoblastic leukemia (ALL) patients were diagnosed by standard bone marrow smear morphologic analysis and peroxidase staining. Flow cytometry and myeloid monoclonal antibodies (McAb) were used to analyze immunophenotype. Chromosome karyotypes were analyzed by R-band technique.</p><p><b>RESULTS</b>Of 120 cases, 66 (55%) were My+ ALL, including 50 cases of My+ B-ALL (56.8% of B-ALL ), 14 cases of My T-ALL (50% of T-ALL) and 2 cases of My+ T and B-ALL (50% of T and BALL). Of 66 My+ ALL, 10 cases (15.1%) were misdiagnosed as acute non-lymphoblastic leukemia (ANLL), the other 54 My- ALL cases were correctly diagnosed. The inconsistent rate between morphological and immunophenotype classifications was higher in My+ ALL than in My- ALL , and there were more atypical morphology cases in My+ ALL than in My- ALL (P < 0.01). In My+ ALL cases 95.5% expressed CD13, 81.8% CD33, 77.3% CD13 and CD33 simultaneously, and 1.5% CD117, but none CD14, CD15 and MPO. CD34 expression rate in My+ ALL cases was significantly higher than that in My- ALL (P < 0.01 ). Cytogenetic abnormalities rates in My+ ALL and My- ALL were 72.3% and 66.7% (P > 0.05) respectively. t(9;22) and t(9;22) plus other cytogenetic abnormalities were detected more frequently in My+ LL cases than in My- B-ALL (P < 0. 01), and not in My+ T-ALL and My- T-ALL cases. The complete remission (CR) rates was 83.9% in My+ ALL and 79% in My- ALL(P > 0.05).</p><p><b>CONCLUSION</b>My+ ALL had a specific characteristics in morphology, immunophenotype and cytogenetics. Some cases have a myeloid morphologic appearance and might be misdiagnosed as acute myeloid leukemia (AML). My+ ALL have a higher CD34 expression rate than My- ALL. t(9;22) abnormality was more frequently observed in My B-ALL than in My- B-ALL. There was no significant difference in CR rate between My+ ALL and My- ALL.</p>
Asunto(s)
Adolescente , Adulto , Anciano , Niño , Femenino , Humanos , Masculino , Persona de Mediana Edad , Inmunofenotipificación , Cariotipificación , Leucemia-Linfoma Linfoblástico de Células Precursoras , Clasificación , Genética , Alergia e InmunologíaRESUMEN
The study was aimed to investigate the immunophenotypic and cytogenetic features of chronic lymphocytic leukemia (CLL) in order to provide an evidence for diagnosis and therapy. Immunophenotypic analysis was performed by using a panel of monoclonal antibodies and three-color immunofluorescence staining methods of flow cytometry in 51 patients with CLL, and the cytogenetic features were analyzed by R-banding technique. The results indicated that among 51 CLL cases, the positive rate of CD19 and CD23 was 96.1%, followed by CD15 (94.1%), CD20 (82.4%) and CD22 (78.4%). The positive rate of CD38 was 23.5%. Forty-six patients expressed both CD5 and CD19. Seven main clonal chromosomal abnormalities were detected by conventional cytogenetics (CC) in eighteen cases (35.3%), with three cases of +12, two cases of 13q(-), other chromosomal abnormalities included +14, 6q(-), t (11; 14), t (14; 18) and t (2; 7). Expression of the antigens had no relationship with chromosomal abnormalities. It is concluded that typical CLL express CD5, CD19 and CD23, and the positive rate detected by CC in CLL is low. Immunophenotyping in combination with cytogenetic technique plays an important role in the diagnosis and prognosis of CLL.
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Adulto , Anciano , Anciano de 80 o más Años , Femenino , Humanos , Masculino , Persona de Mediana Edad , Anticuerpos Monoclonales , Antígenos CD , Metabolismo , Antígenos CD19 , Metabolismo , Antígenos CD20 , Metabolismo , Aberraciones Cromosómicas , Cromosomas Humanos Par 12 , Cromosomas Humanos Par 13 , Citometría de Flujo , Métodos , Inmunofenotipificación , Leucemia Linfocítica Crónica de Células B , Genética , Alergia e Inmunología , Antígeno Lewis X , Metabolismo , Translocación Genética , GenéticaRESUMEN
<p><b>OBJECTIVE</b>To explore the diagnostic value of intracellular antibody combination in acute leukemia (AL) expressing cross-lineage cell-surface antigens.</p><p><b>METHODS</b>Flow cytometric immunophenotyping using intracellular antibody combination (cMPO/cCD79alpha/cCD3/CD45) was performed additionally in 60 patients who expressed cross-lineage antigens from 269 previously untreated adult AL.</p><p><b>RESULTS</b>Fifty-four of 269 previously untreated adult AL patients who expressed only one kind of intracellular antigen were diagnosed as cross-lineage AL, the percentage of cross-lineage AL in T cell acute lymphoblastic leukemia (T-ALL), B-ALL and acute myeloid leukemia (AML) was 28.6%, 43.6% and 13.4%, respectively. The positive rate of CD7, CD19, CD5 and CD20 in cross-lineage AML was 65.4%, 15.4%, 11.5%, and 7.7%, respectively. The positive rate of CD13, CD33 and CD15 in cross-lineage ALL was 89.3%, 21.4% and 3.6%, respectively. Six (2.3%) patients expressed two-lineage intracellular antigens were diagnosed as biphenotypic AL: 2 of T/B type and 4 B/M (B/myeloid) type.</p><p><b>CONCLUSION</b>Intracellular antibodies possess lineage specificity and four-color combination flow cytometric immunophenotyping can provide fast and multi-parameter data. To ensure accuracy of the results, CD45/SSC gating and normal cells as internal reference should be used in the immunophenotyping of abnormal cells.</p>
Asunto(s)
Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Femenino , Humanos , Masculino , Persona de Mediana Edad , Anticuerpos Monoclonales , Antígenos de Superficie , Alergia e Inmunología , Complejo CD3 , Alergia e Inmunología , Antígenos CD79 , Alergia e Inmunología , Reacciones Cruzadas , Citometría de Flujo , Inmunofenotipificación , Antígenos Comunes de Leucocito , Alergia e Inmunología , Leucemia-Linfoma Linfoblástico de Células Precursoras , Diagnóstico , Alergia e Inmunología , Sensibilidad y EspecificidadRESUMEN
<p><b>OBJECTIVE</b>To report abnormal expression of cCD79a/cCD22 in four cases of acute myeloid leukemia (AML) with t (8;21).</p><p><b>METHODS</b>The characteristics of morphology, immunophenotype, chromosome karyotype (MIC) and clinical manifestations of 4 AML patients with t (8;21) expressing cCD79a/cCD22 were analyzed.</p><p><b>RESULTS</b>The features of the 4 patients were: (1) no difference in gender; (2) young age; (3) exmedullary infiltration may be present; (4) normal number of white blood cells in peripheral blood; (5) morphology showed acute myeloid leukemia with high percentage of blast cells; (6) B-lymphoid and myeloid immunophenotype, and high expression of CD34; (7) frequent depletion of Y chromosome and complex changes of chromosomes; (8) positive for AML1/ETO fusion gene; (9) response well to chemotherapy regimen which simultaneously treated myeloid and lymphocytic leukemia.</p><p><b>CONCLUSION</b>Abnormal expression of cCD79a/cCD22 in AML with t (8;21) (q22;q22) suggested that this kind of leukemia might be related with abnormal expression gene of B cell.</p>