The details can be found in the right-most column of Table ?Table22) Four of 22 mutations identified in the gene are located in the PH domain, 2 in the TH domain, 1 in the SH3 domain, 4 in the SH2 domain and 8 in the kinase domain

The details can be found in the right-most column of Table ?Table22) Four of 22 mutations identified in the gene are located in the PH domain, 2 in the TH domain, 1 in the SH3 domain, 4 in the SH2 domain and 8 in the kinase domain. PCR . Results We evaluated the clinical symptoms of 22 XLA patients and investigated genetic mutations present, identifying six novel mutations in the gene: 2 missense mutations (c.23G? ?T and c.112?T? ?C), 2 frameshift mutations (c.522_523insC and c.1060delA), 1 large deletion (deletion Ehk1-L of exon 2 to PROTAC Mcl1 degrader-1 5), and 1 splice-site mutation (c.1631?+?2?T? ?C). Prenatal diagnoses were performed in six families (F10, F11, F15, F18, F20 and F21), with the following results: the male fetus in Family 10 (F10) did not carry the c.922_923delGA mutation; PROTAC Mcl1 degrader-1 the male fetus in Family 15 (F15) did not carry the c.1631?+?1G? ?T splicing mutation; the female fetus in Family 20 (F20) did not carry the c.1931?T? ?C mutation; the female fetus in Family 21 (F21) did not carry the large deletion mutation. Hence, these four fetuses are not likely to develop XLA. Male fetuses with c.1060delA and c.1684C? ?T mutations were identified in Family 11 and Family 18, respectively. The pregnant woman in F18 chose to terminate the pregnancy, whereas the pregnant woman in F11 chose to continue the pregnancy. Conclusion We confirmed the diagnosis of 22 XLA patients from 22 unrelated families and detected six new pathogenic mutations. Prenatal diagnosis was performed in six families. Early genetic diagnosis and routine lifelong immunoglobulin replacement therapy can prevent and treat infections in XLA children, saving their lives. gene is located at Xq21.3-Xq22; the gene is 37.5?kb and comprises 19 exons. The protein encoded by the gene is a cytoplasmic tyrosine kinase that contains five different functional domains: pleckstrin homology (PH), Tec homology (TH), Src homology 3 (SH3), SH2, and kinase (TK) domains [5]. The N-terminal PH domain binds to membrane phosphatidylinositol (3,4,5)-trisphosphate (PIP3), and the TH, SH3, and SH2 domains are involved in protein-protein interactions. Y223 and Y551 are two tyrosine phosphorylation sites in the SH3 and TK domains, respectively [6]. BTK activates many major signaling pathways, including the phosphoinositol-3 kinase (PI3K)-AKT pathway, phospholipase-C (PLC), protein kinase C, and nuclear factor kappa B (NF-kB) [7]. BTK also participates in B cell receptor (BCR) engagement by antigens and induces a range of protein interactions as well as recruitment of signaling molecules, resulting in B cell survival, proliferation and differentiation and the production of antibodies [8]. Methods Patients and study design From 2016 to 2019, 22 male XLA patients from 22 unrelated families in Henan Province of China were enrolled in this study. XLA was diagnosed according to the diagnostic criteria for XLA developed by the Joint European Society for Immunodeficiencies Committee [9]. After determining gene mutations in the proband, the fetal villi or amniotic fluid of high-risk pregnant women were used for prenatal diagnosis. Mutation analysis of the fetal genome was carried out by DNA sequencing. The study was approved by the Ethics Committee of the First Affiliated Hospital of Zhengzhou University. The patients 16?years of age and over signed informed consent forms. A written informed consent was obtained from the parents or legal guardians of any participant under the age of 16. Routine immunological analysis Serum was separated from 3?mL of peripheral PROTAC Mcl1 degrader-1 venous blood without anticoagulant treatment. Immunoglobulins were examined by rate scatter immunoturbidimetry using a Siemens BN II automatic protein analyzer. CD19+ was detected with a FACSCanto II flow cytometer using 3?mL of EDTA-treated blood. Genetic testing Genomic DNA was extracted from 2?mL of EDTA-treated peripheral venous blood from each proband and mother using Blood DNA Midi Kit D3494 (Omega Biotek, USA) with nucleic acid automatic extraction equipment (Eppendorf epMotion 5075?m, Germany). Amniotic fluid cell DNA was extracted and cleaned using QIAamp Blood DNA Midi Kit (250, Germany) and Genomic DNA Clean & Concentrator (Zymo Research, USA). The DNA sequence of the gene obtained from the NCBI database was used as a reference. PCR amplification was carried out using relevant primers (Table S1) under conventional PCR conditions. The PCR product was confirmed by 2% agarose gel and was purified for two-way sequencing. The sequencing product was separated.