Castleman Disease ; virology ; Epstein-Barr Virus Infections ; complications ; Herpesvirus 4, Human ; Humans

Objective: To analyze the mutation type of FⅧ gene in children with hemophilia A and to explore the relationship among hemophilia gene mutation spectrum, gene mutation and clinical phenotype. Method: Sixty-two children with hemophilia A from Department of Pediatric Hematology, Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology between January 2015 and March 2017 were enrolled. All patients were male, aged from 4 months to 7 years and F Ⅷ activity ranged 0.2%-11.0%. Fifty cases had severe, 10 cases had moderate and 2 cases had mild hemophilia A. DNA was isolated from peripheral blood in hemophilia A children and the target gene fragment was amplified by PCR, in combination with the second generation sequencing, 22 and 1 introns were detected. Negative cases were detected by the second generation sequencing and results were compared with those of the international FⅧ gene mutation database. Result: There were 20 cases (32%) of intron 22 inversion, 2 cases (3%) of intron 1 inversion, 18 cases (29%) of missense mutation, 5 cases (8%) of nonsense mutation, 7 cases (11%) of deletion mutation, 1 case(2%)of splice site mutation, 2 cases (3%) of large fragment deletion and 1 case of insertion mutation (2%). No mutation was detected in 2 cases (3%), and 4 cases (7%) failed to amplify. The correlation between phenotype and genotype showed that the most common gene mutation in severe hemophilia A was intron 22 inversion (20 cases), accounting for 40% of severe patients, followed by 11 cases of missense mutation (22%). The most common mutation in moderate hemophilia A was missense mutation (6 cases), accounting for 60% of moderate patients. Conclusion The most frequent mutation type in hemophilia A was intron 22 inversion, followed by missense mutation, again for missing mutation. The relationship between phenotype and genotype: the most frequent gene mutation in severe hemophilia A is intron 22 inversion, followed by missense mutation; the most frequent gene mutation in medium hemophilia A is missense mutation.

Objective:To investigate the clinical features of acute megakaryocytic leukemia (AMKL) in children.Methods:The clinical data of 14 children with AMKL in Tongji Hospital of Tongji Medical College of Huazhong University of Science and Technology from January 2012 to July 2021 were retrospectively analyzed, and the related literature was reviewed.Results:Among 14 children with AMKL, there were 5 males and 9 females, and the median age of onset was 19 months (0.1-109 months); 1 case was Down syndrome-related AMKL, and 13 cases were non-Down syndrome-related AMKL. Most of the children presented with fever, anemia or bleeding symptoms, and a few patients presented with joint pain as the primary symptom. Some children were accompanied by extramedullary infiltration such as hepatomegaly, splenomegaly or lymphadenovarix. Initial investigations of 14 children showed that the median white blood count, hemoglobin concentration and platelet count were 10.67×10 9/L [(6.56-83.62)×10 9/L], 84 g/L (55-121 g/L), 37×10 9/L [(8-1443) ×10 9/L], respectively, and the median proportion of naive cells in peripheral blood was 0.09 (0.00-0.79). Bone marrow smear showed that the primitive megakaryocytes were characterized by various size and irregular form, a few of which had cytoplasmic vacuoles, and the median proportion of bone marrow primitive megakaryocytes was 0.636 (0.332-0.976); the nuclei were round or irregular, with multiple nucleoli or hidden nucleoli. RAS staining was partially positive, and immunohistochemical assay showed that POX, AS-DNCE and α-NBE were negative. Detection of megakaryocyte-associated antigens by flow cytometry showed 12 children expressed CD41a or CD61, and 10 children expressed CD42b. Among 3 children who completed chemotherapy, 1 case of Down syndrome-related AMKL and 1 case of non-Down syndrome-related AMKL were event-free survival, and 1 case of non-Down syndrome-related AMKL died after bone marrow relapse. Conclusions:The clinical manifestations and biological characteristics of children with AMKL are complicated and the prognosis is poor. Some children can achieve disease-free survival through chemotherapy alone.

Objective To study the clinical features and prognostic analysis of 36 children with relapsed acute lymphoblastic leukemia(ALL)treated with the ALL 2006 protocol.Methods The data of 308 children who were new-ly diagnosed as ALL at the Department of Pediatric Hematology,Tongji Hospital,Tongji Medical College,Huazhong Uni-versity of Science Technology treated with the ALL 2006 protocol between January 2007 and December 2012 were col-lected,and the clinical features and prognosis of 36 children with relapsed ALL were retrospectively analyzed.The date included initial treatment time,age,gender,immunophenotyping,the white blood cell count,the risk classification,the chromosome,the fusion gene(29 kinds of fusion genes including MLL rearrangement,BCR/ABL,E2A/PBX1,TEL/AML1)in relapsed patients with ALL on the initial diagnosis and recurrence time,recurrence site,as well as whether to get second complete remission(CR2),follow-up time,follow-up deadline condition.Results After treatment with ALL 2006 protocol,the recurrence rate was 11.7%(36/308 cases);the 3 years overall survival rate was(38.0 ± 9.0)%.The recurrence happened almost in the very early stage,about 75.0%(27/36 cases),and the recurrence rate of the early stage and the late stage patients was 16.7%(16/36 cases)and 8.3%(3/36 cases),respectively.The re-lapsed sites were mainly in the bone marrow alone(66.7%,24/36 cases),and the extramedullary recurrence and the combination of bone marrow with extramedullary recurrence was 16.7%(6/36 cases).The 3-year OS was(16.4 ± 8.0)% and(80.0 ± 18.0)% for those relapsed in the very early stage and early stage,respectively(P=0.002).The 3-year OS of the high-risk,medium-risk relapsed patients were(21.0 ± 11.0)%,(51.9 ± 16.0)% and(64.3 ± 21.0)%(P=0.022).Conclusions After the treatment with ALL 2006 protocol,the patients with recurrence were almost at the very early stage.The relapsed sites were mainly in bone marrow alone.The factor associated with survival time of children with relapsed ALL is the relapse at the very early stage.