OBJECTIVE: To explore the genetic pathogenesis of X-linked agammaglobulinemia in two patients for clinical diagnosis and family counseling. METHODS: Data was collected from the patients' family including clinical information, blood immunoglobulin level, as well as classification and subgrouping of B lymphocytes. Gene mutations were screened by whole exome sequencing (WES) through next-generation sequencing (NGS), the result was verified with Sanger sequencing. RESULTS: A BTK c.1627T>C (p.Ser543Pro) variant was found in the pedigree. The phenotype and variant have co-segregated in the pedigree. The variant was not found in population database. The variant has affected in the kinase domain which contained no benign variants and is harmful as predicted through bioinformatic analysis. CONCLUSION BTK c.1627T>C (p.Ser543Pro) is a pathogenic variant contributing to X-linked agammaglobulinemia in this pedigree. Above finding has provided reproduction guidance for this family.
Agammaglobulinaemia Tyrosine Kinase/genetics* ; Agammaglobulinemia/genetics* ; DNA Mutational Analysis ; Genetic Diseases, X-Linked ; Humans ; Mutation ; Pedigree

Objective:To summarize the morphological characteristics of diffuse large B-cell lymphoma (DLBCL) cells and investigate the prognosis value of the characteristics and the number of DLBCL cells in bone marrow.Methods:Retrospective study. We collected 79 cases newly diagnosed with DLBCL in the First Affiliated Hospital of Zhengzhou University from January 2020 to August 2022. 30 cases newly diagnosed without bone marrow involvement were selected as controls, whose mean age 58 years (30-82 years). The DLBCL cells were evaluated by the bone marrow smear, biopsy and flow cytometry separetely.The detection rate of DLBCL cells in the bone marrow was compared, to analyse the relationship between the morphological characteristics of DLBCL in the smear, clinical characteristics and flow cytometry parameters, and the prognostic value of DLBCL detected in the bone marrow smear and its quantity was analyzed. Logistic regression was used to analyze the correlation between the detection of DLBCL cells in bone marrow smears and the age, clinical stage, and the number of extraderules involved organs. Multivariate Cox regression was used to analyze the influence of DLBCL cells detection and its number on the prognosis of patients.Results:(1) The positive rates of DLBCL cells in bone marrow biopsy, bone marrow smear and flow cytometry were 4.86%, 5.14% and 9.27% respectively. (2) The morphological characteristics of 79 cases in bone marrow smears were described: more than 2 times the volume of the cell body of the lymphocyte, the shape was different, round or quasi-round or irregular shape, can be seen pseudopodia or protrusion; The volume of cytoplasm was moderate, vacuoles were visible, and a few perinuclear areas were visible. The nucleus were different in shape, round or quasi-round or irregularly shaped, with a majority of them having multiple nuclei and a few of them having delicate and loose chromatin. Most nucleoli were medium or large obviously, with a majority of them having 1-2 nucleoli and a few having more than 3.Sergiosomes and hemophagocytosis were observed in some DLBCL cases, tumor cell aggregation phenomenon was observed in a few DLBCL cases, occasionally pathological mitosis.(3) DLBCL cells in bone marrow smear was positively related to the age of patients, clinical stage and the number of extranodal organs involved(regression coefficient were 2.012, 2.754, 2.028, P<0.05);The volume of DLBCL cells in bone marrow smear was positively correlated with the ratio of CD4 and CD8(regression coefficient is 2.545, P<0.05);The vacuoles in cytoplasm and the pseudopod of tumor were both negative relationship with the quantity of CD38 expressed on DLBCL cells(regression coefficient was -2.465, -3.045, P<0.05); (4) DLBCL cells in bone marrow smear was an independent risk factor for PFS and OS( RR=7.059, P<0.05); RR=5.409, P<0.05). Conclusion:The appearace of DLBCL cells in bone marrow smear with prognosis, and could be used for clinical staging.