Objective: To analyze the clinical manifestations and molecular pathogenesis of 18 patients with inherited protein S (PS) deficiency. Methods: Eighteen patients with inherited PS deficiency who were admitted to the Institute of Hematology & Blood Diseases Hospital from June 2016 to February 2019 were analyzed: activity of protein C (PC) and antithrombin (AT) , PS activity were measured for phenotype diagnosis; high throughput sequencing (HTS) was used for screening of coagulation disease-related genes; Sanger sequencing was used to confirm candidate variants; Swiss-model was used for three-dimensional structure analysis. Results: The PS:C of 18 patients ranged from 12.5 to 48.2 U/dL. Among them, 16 cases developed deep vein thrombosis, including 2 cases each with mesenteric vein thrombosis and cerebral infarction, and 1 case each with pulmonary embolism and deep vein thrombosis during pregnancy. A total of 16 PROS1 gene mutations were detected, and 5 nonsense mutations (c.134_162del/p.Leu45*, c.847G>T/p.Glu283*, c.995_996delAT/p.Tyr332*, c.1359G> A/p.Trp453*, c.1474C>T/p.Gln492*) , 2 frameshift mutations (c.1460delG/p.Gla487Valfs*9 and c.1747_1750delAATC/p.Asn583Wfs*9) and 1 large fragment deletion (exon9 deletion) were reported for the first time. In addition, the PS:C of the deep vein thrombosis during pregnancy case was 55.2 U/dL carrying PROC gene c.565C>T/p.Arg189Trp mutation. Conclusion: The newly discovered gene mutations enriched the PROS1 gene mutation spectrum which associated with inherited PS deficiency.
Antithrombin III/genetics* ; Female ; Genetic Testing ; Humans ; Mutation ; Pregnancy ; Protein C/genetics* ; Protein S/genetics* ; Protein S Deficiency/genetics*

OBJECTIVE: To analyze the phenotype and genotype of a patient affected with inherited antithrombin deficiency. METHODS: All exons and exon-intron boundaries of the AT genes were subjected to PCR amplification and Sanger sequencing. The influence of variants on the disease was predicted using bioinformatic software (MutationTaster). RESULTS: The results of all coagulation tests were normal, though the antithrombin activity and antigen content of the proband and his father have decreased significantly (34%, 48% and 12.97 mg/dL, 15.60 mg/dL, respectively). His mother was normal. Genetic analysis revealed that the proband and his father both carried a heterozygous g.2736dupT variant of the AT gene. Bioinformatic analysis suggested that the variant may be pathogenic. CONCLUSION The proband and his father both had type I hereditary antithrombin deficiency caused by a g.2736dupT variant of the AT gene. The variant was unreported previously.
Antithrombin III/genetics* ; Antithrombin III Deficiency/genetics* ; DNA Mutational Analysis ; Genetic Testing ; Heterozygote ; Humans ; Male ; Mutation ; Pedigree

Antithrombin and protein C are two crucial members in the anticoagulant system and play important roles in hemostasis. Mutations in SERPINC1 and PROC lead to deficiency or dysfunction of the two proteins, which could result in venous thromboembolism (VTE). Here, we report a Chinese 22-year-old young man who developed recurrent and serious VTE in cerebral veins, visceral veins, and deep veins of the lower extremity. Laboratory tests and direct sequencing of PROC and SERPINC1 were conducted for the patient and his family members. Coagulation tests revealed that the patient presented type I antithrombin deficiency combined with decreased protein C activity resulting from a small insertion mutation c.848_849insGATGT in SERPINC1 and a short deletion variant c.572_574delAGA in PROC. This combination of the two mutations was absent in 400 healthy subjects each from southern and northern China. Then, we summarized all the mutations of the SERPINC1 and PROC gene reported in the Chinese Han population. This study demonstrates that the combination of antithrombin deficiency and decreased protein C activity can result in severe VTE and that the coexistence of different genetic factors may increase the risk of VTE.
Antithrombin III ; genetics ; Antithrombin III Deficiency ; etiology ; genetics ; China ; Female ; Humans ; Male ; Middle Aged ; Mutation ; Pedigree ; Protein C ; genetics ; metabolism ; Venous Thromboembolism ; complications ; genetics ; Young Adult

Unprovoked deep vein thrombosis (DVT) is uncommon in pediatric patients and, among those, combined hereditary thrombophilia is particularly rare. We present a 9-year-old Korean boy who developed lower extremity pain with swelling, and was diagnosed with unprovoked DVT due to hereditary (combined hereditary thrombophilia). Coagulation test revealed antithrombin III and protein S deficiency. The genetic work up confirmed the first case of combined antithrombin III deficiency and protein S deficiency by SERPINC1 heterozygous termination mutation [c.685C>T (p.Arg229*)] and PROS1 heterozygous missense mutation [c.1597G>A (p.Val533Met)]. He was treated with continuous heparin and catheter intervention but those were ineffective or transiently effective. His DVT gradually improved only after prolonged anticoagulation.
Antithrombin III Deficiency ; Antithrombin III* ; Catheters ; Child ; Heparin ; Humans ; Lower Extremity ; Male ; Mutation, Missense ; Protein S Deficiency* ; Protein S* ; Thrombophilia* ; Venous Thrombosis*

OBJECTIVETo explore the phenotype, genotype and molecular mechanism for two pedigrees affected with hereditary antithrombin (AT) deficiency.

METHODSClinical diagnosis was validated by assaying of coagulation parameters including prothrombin time, activated partial thromboplastin time, thrombin time, fibrinogen, antithrombin activity (AT:A) and specific antigen (AT:Ag), protein C activity, as well as protein S activity. To detect potential mutations in the probands, all exons, exon-intron boundaries and the 3', 5' untranslated regions were amplified by PCR and subjected to direct sequencing. Suspected mutation was confirmed by reverse sequencing and silver staining. The effect of mutations on the AT protein was analyzed with bioinformatics software.

RESULTSThe AT:Ag of pedigree 1 was normal, but its AT:A has reduced to 30%. A heterozygous c.235C>T mutation in exon 2 causing p.Arg47Cys, in addition with two single nucleotide polymorphisms (c.981G>A, c.1011G>A) in exon 5 were identified in the patient. His four children, except for the elder daughter, were heterozygous for the mutations. The plasma levels of AT:A and AT:Ag in proband 2 have decreased to 39% and 103 mg/L, respectively. A heterozygous deletion (g.5890-5892delCTT) leading to loss of p.Phe121 was also detected in his father. Bioinformatic analysis suggested that the missense mutation Arg47Cys can affect the functions of AT protein. Meanwhile, lacking of Phe121 will result in loss of hydrogen bonds with Ala124, Lys125 and the cation π interactions with Lys125, Arg47, which may jepordize the stability of the protein.

CONCLUSIONThe proband 1 had type II AT deficiency, while proband 2 had type I AT deficiency. The p.Arg47Cys and g.5890-5892delCTT mutations of the AT gene are significantly correlated with the levels of AT in the two probands, respectively.

Adult ; Aged, 80 and over ; Antithrombin III ; genetics ; metabolism ; Antithrombin III Deficiency ; enzymology ; genetics ; physiopathology ; Exons ; Female ; Genetic Testing ; Genotype ; Humans ; Male ; Mutation ; Partial Thromboplastin Time ; Pedigree ; Phenotype ; Protein C ; genetics ; metabolism ; Protein S ; genetics ; metabolism

Antithrombin (AT) is the main inhibitor of blood coagulation proteases. Hereditary AT deficiency is an autosomal-dominant thrombophilic disorder caused by a SERPINC1 abnormality, it represents a risk factor for thromboembolic disease. A 25-day-old male infant was referred to Severance Children's Hospital for hemorrhagic cerebral infarction with hydrocephalus. The initial laboratory study showed 11% AT activity. An approximate 4 mm-x-3 mm-size thrombosis was also found in the right ventricle by echocardiography. We found two deletion in the coding and flanking sequences of SERPINC1 c.235C>T (p.Arg79Cys) and c.442T>C (p.Ser148Pro) at 10 months of age. The p.Ser148Pro mutation was found in his mother but the other was not. This case is a rare thrombotic event that occured early year in due to AT deficiency. Our patient had side effects after heparin treatment, so aspirin therapy was employed. No thrombotic events occurred until 1 year of age.
Antithrombin III Deficiency ; Aspirin ; Blood Coagulation ; Cerebral Infarction ; Clinical Coding ; Echocardiography ; Heart Ventricles ; Heparin ; Humans ; Hydrocephalus ; Infant ; Male ; Mothers ; Peptide Hydrolases ; Risk Factors ; Thrombosis

OBJECTIVETo study the molecular mechanisms of inherited antithrombin (AT) deficiency caused by AT L99 mutation.

METHODSWild type (WT), L99V, L99A, L99I and L99S AT were purified from drosophila expression system. The binding capacity of AT and the low molecular weight heparin sodium was analyzed by the heparin binding assay. Surface plasmon resonance (SPR) was used to detect the binding ability of AT to thrombin (FIIa) or AT to coagulation factor Xa (FXa). The activity of AT(AT∶A)was detected by chromogenic assay.

RESULTSThe purified WT and mutant AT were at the same size. No additional band was observed by coomassie blue staining and western blot assay. Compared to the WT AT, the binding abilities of the low molecular weight heparin sodium to the AT L99V, L99A, L99I and L99S were (44.8±3.6)%, (118.9±14.0)%, (15.2±8.8)%, and(23.0±8.2)%, respectively. The binding abilities of FIIa to AT L99V, L99A, L99I and L99S were 13%, 57%, 3%, and 29%, while the binding of FXa to AT L99V, L99A, L99I and L99S were 7%, 51%, 1%, and 25%. The AT∶A of WT, L99V, L99A, L99I and L99S AT were 146.5%, 21.4%, 120.9%, 10.8%, and 39.0%, respectively.

CONCLUSIONThe binding abilities of AT to heparin, FIIa and FXa were damaged by the L99 mutation, which resulted in decreased AT∶A and inherited AT deficiency.

Amino Acids ; genetics ; Animals ; Antithrombin III ; genetics ; Antithrombin III Deficiency ; genetics ; Antithrombins ; Drosophila ; Factor Xa ; genetics ; Genetic Vectors ; Humans ; Mutation

Adolescent ; Adult ; Antithrombin III Deficiency ; genetics ; Antithrombins ; Female ; Humans ; Male ; Mutation ; Pedigree ; Phenotype

PURPOSE: Neonatal seizures are one of the most common neurologic manifestations in neonates and could be the important clinical sign of underlying brain disorders. The aim of this study is to review the clinical characteristics and to find the prognostic factors related to the outcomes of neonatal seizures. METHODS: We reviewed medical records retrospectively in 23 patients with neonatal seizures who admitted to Dankook University Hospital from July 2007 to June 2009. RESULTS: During the study period, neonatal seizures were diagnosed in 23/1,474 (1.56%) neonates. Nineteen of them (82.6%) were term and 4 were preterm. The main cause of neonatal seizures was hypoxic ischemic encephalopathy (n=8, 35%). Other various causes included metabolic disorders (n=4, 17%, carnitine palmitoyl transferase 1 deficiency, severe hypernatremic dehydration, prolonged severe hypoglycemia, and pyridoxine dependent seizure), intracranial hemorrhages (n=2, 9%), congenital brain anomaly (callosal dysgenesis, hemimegalencephaly) (n=2, 9%), and infection (congenital syphilis, early neonatal sepsis, n=2, 9%). Among nineteen neonates (82.6%) who were treated with anticonvulsants, we could not stop the anticonvulsants in six of them (32%). They had severe HIE, prolonged severe hypoglycemia with residual encephalomalatic changes, sinovenous thrombotic hemorrhages due to antithrombin III deficiency, congenital brain anomaly, and septic shock, respectively. CONCLUSION: Approximately one third of neonatal seizures were caused by HIE, and moderate to severe HIE had more serious outcomes. Neonatal seizure could be a significant clinical sign indicating specific underlying etiologies such as stroke, metabolic disturbances or congenital brain anomalies. Therefore, intensive workup and prompt management for neonatal seizures should be considered for better outcomes.
Anticonvulsants ; Antithrombin III Deficiency ; Brain ; Brain Diseases ; Carnitine ; Dehydration ; Hemorrhage ; Humans ; Hypoglycemia ; Hypoxia-Ischemia, Brain ; Infant, Newborn ; Intracranial Hemorrhages ; Medical Records ; Neurologic Manifestations ; Prognosis ; Pyridoxine ; Retrospective Studies ; Seizures* ; Sepsis ; Shock, Septic ; Stroke ; Syphilis ; Transferases

Hypercoagulability state is the result of the interplay of genetic predisposition and risk factors. Many key enzymes and reactions in coagulation and anti-coagulation system are involved. Hereditary antithrombin deficiency is one of the major risk factors of venous thromboembolic disease (VTE), whereas hyperhomocysteinemia may also play a role. This article reviews the recent researches on the contributions of these two factors to VTE.
Antithrombin III Deficiency ; complications ; Humans ; Hyperhomocysteinemia ; complications ; Venous Thrombosis ; etiology