In the present study, we report two novel causative mutations of the F10 gene in a Chinese proband with severe FX deficiency EGFR inhibitor and mild clinical symptoms.

Furthermore, the molecular mechanisms of the two mutations were analysed. The proband, a 36-year-old Chinese male patient born from non-consanguineous parents, was diagnosed with FX deficiency in routine preoperative coagulation assay. He has exhibited numerous bleeding episodes since early childhood with recurrent epistaxis and gums bleeding after brushing his teeth and dental extraction. However, he had not experienced severe bleeding diathesis. One of his brothers had similar bleeding symptoms, but other family members had no history of bleeding. After informed consent, blood from the proband and family members was collected in 0.109 m trisodium citrate. FX:C assay was performed based on both prothrombin time (PT) and activated partial thromboplastin time (APTT) using

a one-stage clotting method on ACL-TOP automatic coagulometer (HemosILTM, IL, USA). FX amidolytic activity based on RVV was performed using a chromogenic assay kit (Hyphen Biomed, Neucille suroise, France) according to the manufacturer’s instructions. FX:Ag level was measured with a sandwich enzyme-linked immunoadsorbent assay (ELISA) using rabbit anti-human polyclonal FX antibody (Dako, Glostrup, Denmark) as a capture antibody, and horseradish peroxidase (HRP)

conjugated antibody (Dako) as a 上海皓元 detection antibody. Both FX:C and FX:Ag values are expressed Epacadostat supplier as the percentage of pooled plasma levels obtained from 30 healthy, unrelated individuals. Screening for inhibitors was performed by APTT and PT mixing assays. Genomic DNA of the proband and family members was extracted from peripheral blood leucocytes using a standard protocol. Genetic defect analysis of the F10 gene was performed as previously described [3]. TA-cloning with the pMD19-T Simple vector (TaKaRa, Shiga, Japan) and DNA sequencing were used to detect for the heterozygous deletion. All variants were confirmed by reverse sequencing using a second amplicon. The variant was reported in accordance with standard international nomenclature guidelines as recommended by the Human Genome Variation Society (HGVS, http://www.hgvs.org/mutnomen/recs.html) with nucleotide +1 as the A of the ATG translation initiation codon. The genomic DNA (GenBank:12738260) and cDNA (GenBank: M57285) sequences of the F10 gene were used as reference sequences. Ectopic transcription was used to analyse the splice pattern of the IVS5+1G>A mutation in the F10 gene. Briefly, total RNA of the proband and one healthy control was isolated from peripheral leucocytes using the TRIzol reagent (Invitrogen, Carlsbad, CA, USA).