Objective To evaluate the practicability of using flow cytometry analysis for diagnosis of non-hodgkin′s lymphoma ( NHL) among children with serous effusion.Methods Twelve children who were diagnosed with malignant lymphoma from February 2011 to November 2013 at Shanghai Children′s hos-pital were recruited in this study.Pleural effusion and ascites samples were collected from those children who showed serous effusion as initial symptoms and analyzed by using flow cytometry based immunophenotyping. The antibodies used for immunophenotyping included CD45, CD10, CD33, CD7, CD1a, MPO, cCD3, CD79a, CD22, CD19, CD20, CD5, CD3,κ,λ,αβ,γδ,CD56 and other common markers for T, B and NK cells.Anti-CD30 antibody was used when necessary.Results All of the twelve cases with serous effusion were diagnosed with aggressive NHL.Six out of the twelve children including five cases with ascites and one case with pleural effusion showed high expression of CD20 and were classified as NHL-B type by flow cytom-etry.Three children with pleural effusion and one child with both pleural effusion and ascites were typed as NHL-T as characterized by monoclonal expression of αβorγδ.The other two children with pleural effusion were diagnosed with anaplastic large cell lymphoma with positive expression of CD30 and morphological het-erogeneity.Conclusion Flow cytometry analysis based immunophenotyping could be used as an auxiliary method for rapid and accurate diagnosis of lymphoma in children with serous effusions.

Objective To evaluate the diagnostic and prognostic significance of 43 fusion gene screening in combination with immunophenotyping and cytogenetics in childhood hematologic neoplasms. Methods A total of 176 children from Shanghai Children′s hospital with acute leukemia newly or recurrently diagnosed from May.2015 to Apr.2017 were enrolled into this study.There were 99 males and 77 females,aged between 4 months to 15 years old.Application of real-time fluorescent probe PCR assay for detection of 43 fusion genes including the common type of BCR-ABL,AML1-ETO, CBFβ-MYH11, MLL and RaRα related fusion genes in ALL(Acute lymphoblastic leukemia)and AML(Acute myeloid leukemia).Results The fusion genes were detected at positive rate of 28.4 %(50/176).The final diagnosis included 110 cases of ALL-B,11 cases of ALL-T and 46 cases of AML,the other 9 cases were as follows:1case of neuroblastoma,2 cases of NHL -B,1case of NHL -T,1case of AML transformed from CML,2 cases of M DS,1 case of CML and 1 case of anemia.Positive fusion gene of ALL -B(27/110, the positive rate of 24.5%),included:TEL-AML1 14 cases,E2A-PBX1 6,MLL-AF10 2 and MLL -AF4 2,MLL-AF9 1,MLL -ENL 1 and BCR-ABL 1 cases.ALL -T positive rate 27.3%(3/11):1 case with MLL-ENL and 2 cases of SIL -TAL1.In AML patients the positive rate was 37%(17/46), included AML-ETO 7,MLL -AF10 1, MLL -AF10 1, PML-RaRa 4, DCK -CAN 3and CBFB -MYH11 1 case respectively.The other included BCR-ABL 2 cases, E2A-PBX1 1 case.Conclusion The 43 fusion gene screening contribute to leukemia diagnosis and differential diagnosis, which helps to evaluate the risk stratification and prognosis effectively.

Objective:To analyze the relationship between molecular cytogenetic abnormalities and clinical characteristics of acute lymphoblastic leukemia (ALL) in childhood .Methods:A total of 403 patients newly diagnosed with ALL in the Department of Hematology, Shanghai Children′s Hospital from January 2009 to December 2018 were enrolled in this study.All the patients had completed the test of bone marrow smear cytology, immunotyping, karyotype analysis, and fluorescence in situ hybridization (FISH).Results:(1)There were 240 males (59.6%) and 163 females (40.4%) aged (5.31±3.46)years.There were 374 patients(92.8%) with B cell acute lymphoblastic leukemia (B-ALL)and 29 patients(7.2%) with T cell acute lymphoblastic leukemia (T-ALL). (2)Cytogenetics: A total of 311 cases (77.2%) showed mitosis in the chromosomal karyotype analysis, of which 126 cases were abnormal (abnormality detection rate was 40.5%), including 15.4% (48/311cases) hyperdiploid.(3)Fusion gene: Positive fusion genes were found in 110 cases (27.3%), including TEL/AML1 gene in 70 cases (17.4%), BCR/ ABL in 13 cases (3.2%), MLL in 19 cases (4.7%). From 2015-2018, 8 cases (4.0%) of PBX1/TCF3 fusion gene, 1 case of EBF1-PDGFRB fusion gene, 6 cases of SIL/TAL1 fusion gene were detected, SIL/TAL1 positive patients which were accounting for 33.3% of T-ALL improved the detection rate of T-ALL molecular abnormalities.Patients with positive BCR/ ABL were older than those with positive TEL/AML1 and positive MLL[(8.01±3.11) years vs.(3.89±1.84) years, (1.56±1.25) years, P<0.001]; patients with positive PBX1/TCF3 [6.58±4.83) years]were older than those with positive TEL/AML1 and positive MLL (all P<0.05); patients with positive MLL were younger than those with positive TEL/AML1 [(1.56±1.25) years vs.(3.89±1.84) years, P=0.001]; the white blood cell (WBC) count of positive MLL patients was higher than that of positive TEL/AML1 and positive BCR/ ABL patients [(76.97±19.87)×10 9/L vs.(16.94±2.28)×10 9/L, P=0.002; (76.97±19.87)×10 9/L vs.(20.53±6.49)×10 9/L, P<0.05]; the WBC count of PBX1/TCF3 positive children was higher than that of positive TEL/AML1 patients [(85.75±30.32)×10 9/L vs.(16.94±2.28)×10 9/L, P=0.002]. The immunotyping of positive MLL patients was dominated by early precursor B-ALL (14/19 cases), while the immunotyping of TEL/AML1 and BCR/ABL positive patients were dominated by common-B-ALL(57/70 cases and 11/15 cases). (4)The detection rates of chromosome karyotype analysis, FISH, and polymerase chain reaction (PCR) were used to detect molecular genetic abnormalities in primary ALL patients, the detection rate was 40.5% (126/403 cases), 69.2% (279/403 cases), and 29.7% (60/202 cases), respectively.The difference was statistically significant ( P<0.001). There was no significant difference in the abnormality detection rate between chromosome karyotype analysis and PCR ( P=0.71). (5)There was no significant difference in the detection rate of molecular cytogenetic abnormalities between different genders and age groups ( P=0.651, 0.721). There was a significant difference between the WBC count ≥50 × 10 9/L group and <50 × 10 9/L group(37/51 cases vs.107/352 cases, P<0.001). The detection rate of B-ALL genetic abnormalities was higher than that of T-ALL genetic abnormalities(275/374 cases vs.14/29 cases, P=0.005). Conclusions:There are a higher proportion of hyperdiploidy chromosomes in children ALL.The distribution of fusion genes is related to age, primary white blood cell count, and immunotyping.The three detection methods complement each other and greatly improve the detection rate of genetic abnormalities.The detection rate of T-ALL genetic abnormality is low, and new detection methods may be needed.