OBJECTIVETo investigate the gene mutations in a pedigree with inherited prothrombin (FII) deficiency.

METHODSThe activated partial thromboplastin time (APTT), prothrombin time (PT), FII activity (FII:C) and FII antigen (FII:Ag) test were used for phenotype diagnosis. The genomic DNA was extracted from the peripheral blood of the propositus. All the 14 exons, intron/exon boundaries and the 5' and 3' untranslated regions (UTR) of the prothrombin gene were amplified by polymerase chain reaction (PCR). The PCR products were screened by direct sequencing and the mutations detected were further confirmed by restricted enzyme digestion. One hundred and three healthy blood donors were used as controls.

RESULTSThe phenotype of the propositus was prothrombin deficiency (type I). With reference to the prothrombin nucleotide sequence published by Degen & Dacie, three variations were found in the FII gene of the propositus. Among them, the novel mutation was a homozygous A601G subtitution in exon 2.

CONCLUSIONThe prothrombin deficiency of the propositus is caused by a homozygous Glu29 to Gly mutation in the prothrombin gene.

Blood Coagulation ; Child ; Female ; Humans ; Hypoprothrombinemias ; blood ; genetics ; Point Mutation ; Prothrombin ; genetics

Vitamin K is the cofactor for the hepatic carboxylation of glutamic acid residues in a number of proteins including the procoagulants factors ll, Vll, lX, and X. The role of vitamin K in normal bone function is not fully understood. Inherited deficiency of vitamin K dependent coagulation factors is a rare bleeding disorder reported only in a few patients. Here we present an 18-month old child who presented with osteopeni due to inherited vitamin K deficiency. While the patient had high bone specific alkaline phosphatase and parathyroid hormone levels and low osteocalcin and bone mineral density values, with the regular supplementation of vitamin K all the mentioned parameters returned to normal values.
Bone Density ; Bone Diseases, Metabolic/etiology ; Human ; Infant ; Male ; Osteocalcin/blood ; Prothrombin Time ; Vitamin K Deficiency/blood/complications/*genetics

OBJECTIVETo investigate the correlation between prothrombin G20210A polymorphism and the risk for idiopathic sudden sensorineural hearing loss (ISSNHL) using Meta-analysis methodology.

METHODSDatabases, including PUBMED, EMBASE, Cochrane Library and CBM, were searched to collect the case control studies on the correlation between prothrombin G20210A polymorphism and idiopathic sudden sensorineural hearing loss. Only high quality studies were included. All analysis were conducted with Review Manager Version 4.2 software.

RESULTSA total of 9 studies were included, involving 735 cases and 1230 controls. The quality assessment involved 3 parts, 8 scores (totally 8 stars). The results showed the included studies were high-quality. Two studies were 8 stars of quality, three studies were 7 stars, one study was 6 stars, one study was 5 stars, and two studies were 4 stars. Meta-analysis showed that the prothrombin G20210A mutation frequencies of the genotypes and alleles showed significant statistically difference between cases and controls [P = 0.03, OR = 1.79, 95% CI = (1.06, 3.01); P = 0.03, OR = 1.77, 95% CI = (1.06, 2.97), respectively].

CONCLUSIONSThe prothrombin G20210A polymorphism might be a genetic risk factor for sudden hearing loss. However, this conclusion remains to be confirmed by high-quality, large-scale studies.

Case-Control Studies ; Genotype ; Hearing Loss, Sensorineural ; epidemiology ; genetics ; Hearing Loss, Sudden ; epidemiology ; genetics ; Humans ; Polymorphism, Genetic ; Prothrombin ; genetics ; metabolism ; Risk Factors ; Thrombophilia

PURPOSE: The precise molecular mechanisms culminating in coronary artery disease (CAD) are not well understood, despite a wealth of knowledge on predisposing risk factors and pathomechanisms. CAD and myocardial infarction (MI) are complex genetic diseases; neither the environment alone, nor a single gene, cause disease, rather, a mix of environmental and genetic factors lead to atherosclerosis of the coronary arteries. MATERIALS AND METHODS: In the present study, our aim was to investigate the roles of prothrombin G20210A mutation and Factor VLeiden mutation in atherosclerotic coronary artery disease. 287 subjects (106 control subjects, who were angiographically normal, and 181 angiographically documented coronary atherosclerotic patients who exhibited coronary artery narrowing to a degree of > or = 50%) were included in this study. The mutations were assessed with LightCycler Real-Time PCR mutation detection kits (Roche Diagnostics, GmbH, Germany). RESULTS: 6.6% of control subjects, and 6.1% of patients with (50% coronary artery narrowing were determined to have the Factor VLeiden heterozygote mutation. 6.6% of control subjects had the Prothrombin G20210A heterozygote mutation, while 7.7% of patients with (50% coronary artery narrowing had this mutation. The OR for Factor VLeiden was 1.52 (CI: 0.240-9.602) and for Prothrombin G20210A mutation, the OR was 1.415 (CI: 0.287-6.962). CONCLUSION: Although both the heterozygote Factor VLeiden and Prothrombin gene mutations were more frequent in patients with CAD than in control subjects, there was no statistical relationship found to exist between coronary artery disease and the Factor VLeiden and Prothrombin G20210A mutations.
Aged ; Coronary Artery Disease/*genetics ; Factor V/*genetics ; Female ; Gene Frequency ; Genotype ; Humans ; Male ; Middle Aged ; *Polymorphism, Single Nucleotide ; Prothrombin/*genetics

We report a case of intolerance to warfarin dosing due to impaired drug metabolism in a patient with CYP2C9*3/*4. A 73-yr-old woman with atrial fibrilation was taking warfarin. She attained a high prothrombin time international normalized ratio (INR) at the standard doses during the induction of anticoagulation and extremely low dose of warfarin (6.5 mg/week) was finally chosen to reach the target INR. Genotyping for CYP2C9 revealed that this patient had a genotype CYP2C9*3/*4. This is the first Korean compound heterozygote for CYP2C9*3 and *4. This case suggests the clinical usefulness of pharmacogenetic testing for individualized dosage adjustments of warfarin.
Aged ; Anticoagulants/pharmacology ; Aryl Hydrocarbon Hydroxylases/*genetics ; Atrial Fibrillation/*drug therapy/*genetics ; Female ; *Genotype ; Heterozygote ; Humans ; International Normalized Ratio ; Pharmacogenetics ; Polymorphism, Genetic ; Prothrombin Time ; Warfarin/*pharmacology

OBJECTIVETo screen potential mutation and explore the underlying mechanism for a consanguineous pedigree featuring hereditary coagulation factor Ⅴ (FⅤ) deficiency.

METHODSClinical diagnosis was validated by coagulant parameter assays of prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (FIB), FⅤ procoagulant activity (FⅤ:C) and FⅤ antigen (FⅤ:Ag). Potential mutations of the F5 gene in the proband and his family members were analyzed by direct DNA sequencing of PCR products of all exons, exon-intron boundaries and 3', 5' untranslated regions. Suspected mutation was confirmed by reverse sequencing.

RESULTSThe PT and APTT in the proband were significantly prolonged, which measured 23.5 s (reference range 11.8-14.8 s) and 50.5 s (reference range 27.0-41.0 s), respectively. FⅤ activity and FⅤ antigen of the proband were significantly reduced to 8% and <1%, respectively. PT and APTT in the younger sister of the proband were also significantly prolonged (24.1 s and 62.4 s, respectively). Her FⅤ activity and FⅤ antigen were also significantly decreased (7% and <1%, respectively). PT and APTT of other family members were within the normal range. The homozygous missence mutation causing T→C transition at position 29170 in exon 5 of F5 gene has resulted in a Phe190Ser substitution in the proband. His younger sister was also homozygous for Phe190Ser. Heterozygosity for Phe190Ser was confirmed in his elder brother, elder sister, two daughters and niece, and their FⅤ activity were slightly decreased (57%, 73%, 72%, 66% and 75%, respectively). A normal wild type was observed in two younger brothers of the proband, and their FⅤ activity and FⅤ antigen were in the normal range.

CONCLUSIONHomozygous missence mutation of Phe190Ser has been found in above family featuring hereditary FⅤ deficiency. The homozygous missence mutation was inherited from the parents by consanguineous marriage. Phe190Ser probably underlies may underlie the pathogenesis of hereditary FⅤ deficiency in this pedigree.

Adult ; Consanguinity ; Factor V ; genetics ; Factor V Deficiency ; blood ; genetics ; Female ; Humans ; Male ; Middle Aged ; Mutation, Missense ; Partial Thromboplastin Time ; Pedigree ; Prothrombin Time ; Sequence Analysis, DNA

OBJECTIVETo identify potential mutation underlying coagulation factor X (FX) deficiency in a consanguineous Chinese pedigree.

METHODSProthrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen, FX activity (FX:C) and other coagulant parameters were determined with a one-stage clotting assay. The FX antigen (FX:Ag) was determined with an ELISA assay. All coding exons and exon-intron boundaries of the F10 gene were amplified with PCR and subjected to direct sequencing. Suspected mutation was confirmed by reverse sequencing and analyzed with CLC Genomics Workbench 7.5 software.

RESULTSThe PT and APTT in the proband were prolonged to 67.2 s and 102.9 s, respectively. Further study showed that her FX:C and FX:Ag were reduced by 1% and 8%, respectively. The PT of her father, mother, and little brother were slightly prolonged to 14.5 s, 14.4 s and 14.4 s, respectively. The FX:C and FX:Ag in her father, mother and little brother were all slightly reduced. Genetic analysis of the proband has revealed a homozygous G>A change at nucleotide 27881 in exon 8 of the F10 gene, which predicted a p.Val298Met substitution. The proband's father, mother, and little brother were all heterozygous for the p.Val298Met mutation. The proband has inherited the homozygous mutation from her parents by consanguineous marriage. Other family members were all normal. Bioinformatics analysis has indicated that this mutation may result in changes in the secondary structure of the FX protein.

CONCLUSIONA homozygous mutation g.27881G>A(p.Val298Met) of the F10 gene has been identified, which probably accounts for the low FX concentrations in this pedigree.

Adult ; Amino Acid Sequence ; Consanguinity ; Factor X ; genetics ; Factor X Deficiency ; genetics ; Female ; Homozygote ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Mutation, Missense ; Pedigree ; Prothrombin Time

OBJECTIVETo analyze the gene mutation in two pedigrees of inherited coagulation factor VII (FVII) deficiency, and investigate the relationship between the genotype and phenotype.

METHODThe coagulation function and coagulation factors activity of probands were detected for phenotype diagnosis, all exons and junctions of FVII gene from the family members' genomic DNA were amplified using polymerase chain reaction (PCR), and detected the gene mutation by direct sequencing. Mutations were confirmed by reverse sequencing.

RESULTThe prothrombin time (PT) of proband 1 was 265.2 s, FVII:C was 22% and the PT of proband 2 was > 120 s, FVII:C was 1%. Homozygous 17844G→A mutation in No. 8 exon of FVII gene was identified in the proband 1 resulting in Gly343Ser, and heterozygosity for the same mutations were confirmed in his parents and a sister. The proband 2 was compound heterozygous, one mutation was the same as the proband 1 but was a heterozygosity that can also found in his mother and brother; the other heterozygosity mutation was located on No. 8 exon 18055G→A that resulted in Gln413Arg which was inherited from his father.

CONCLUSIONNo. 8 exon of FVII gene encodes catalytic domain. Mutation found in those domain could change the FVII catalytic domain spatial structure, affected FVII function and stability, and the sufferer of homozygote and compound heterozygous may have clinical bleeding tendency. Almost no clinical findings in simple heterozygotes, however, a few of heterozygotes could have a tendency of bleeding because of genetic polymorphism which would reduce the FVII:C.

Blood Coagulation Disorders ; blood ; genetics ; Child, Preschool ; DNA Mutational Analysis ; Factor VII ; genetics ; Factor VII Deficiency ; blood ; genetics ; Heterozygote ; Homozygote ; Humans ; Infant ; Male ; Molecular Sequence Data ; Mutation ; Pedigree ; Polymerase Chain Reaction ; Prothrombin Time

OBJECTIVETo identify gene mutations of a pedigree with inherited factor V (FV) deficiency.

METHODSThe activated partial thromboplastin time (APTT), prothrombin time (PT), FV activity (FV:C) and FV antigen (FV:Ag) tests were performed for phenotypic diagnosis. The genomic DNA was extracted from the peripheral blood of the proband and all the 25 exons and their flanks of FV gene were amplified by polymerase chain reaction (PCR). The PCR products were screened by direct sequencing and the mutations were further confirmed by restriction enzyme digestion.

RESULTSAPTT, PT, TT, FV:C, FV:Ag of the proband were 249.2 s, 46.6 s, 17.9 s, 0.1% and 1.5%, respectively. FII, FVII, FVIII, FIX, FX activities, vWF and Fg were within normal ranges. Taking the GenBank Z99572 sequence as the reference, four mutations were identified in FV gene of the proband. They were a heterozygous two bases deletion in exon 13 (2238 approximately 2239delAG) introducing a frameshift and a premature stop at codon 689, and a heterozygous missense mutation in exon 23 (G6410T) resulting in the substitution of Gly for Val at codon 2079, respectively. The proband's father and mother were heterozygous for G6410T and for 2238 approximately 2239delAG, respectively.

CONCLUSIONThe severe FV deficiency of the proband is caused by a frameshift mutation of 2238 approximately 2239delAG and a missense mutation of G6410T, which haven't been identified before.

Adult ; Base Sequence ; DNA Mutational Analysis ; Exons ; genetics ; Factor V ; genetics ; metabolism ; Factor V Deficiency ; genetics ; Female ; Frameshift Mutation ; Heterozygote ; Humans ; Infant ; Male ; Mutation, Missense ; Partial Thromboplastin Time ; Pedigree ; Phenotype ; Prothrombin Time ; Thrombin Time

Biomarkers ; blood ; Biomarkers, Tumor ; blood ; Carcinoma, Hepatocellular ; blood ; diagnosis ; genetics ; Early Diagnosis ; Humans ; Insulin-Like Growth Factor II ; analysis ; Liver Neoplasms ; blood ; diagnosis ; genetics ; Protein Precursors ; blood ; Prothrombin ; RNA, Messenger ; analysis ; alpha-Fetoproteins ; metabolism