Molecular Diagnosis Of One Pedigree With Protein S And Antithrombin Deficiency Risk Factors Analysis In617Hospitalized Patients With Deep Venous Thromboembolism

1.Objective To identify gene mutations for one patient and his family members with protein S and antithrombin deficiency.2.Methods ELISA were used to detect protein S （PS）, protein C （PC） and antithrombin （AT） activities for the proband and family members, respectively. The genomic DNA was extracted from the peripheral blood of proband and family members. All exons and their fianks of protein S gene and antithrombin gene were amplified by polymerase chain reaction （PCR）. The PCR products were sequenced directly. The mutation-related exons of his famliy members were amplified by PCR and sequenced directly.3.Results The proband was a49-year-old male. He presented with sudden left lower extremity swelling and pain without casues. Regular examination revealed that his APTT, PT, and TT were all in normal levels, but D-dimmer was5.62mg/L, Color doppler ultrasonography showed thrombosis in his left femoral vein. The activity of PS for his family members was Ⅱ10%,Ⅱ20%,Ⅱ30%, Ⅱ4130.8%,Ⅱ58.4%,Ⅲ10%,Ⅲ20%, and that of AT was Ⅱ1129.1%,Ⅲ251.9%,Ⅱ373.2%,Ⅱ4119.1%,Ⅱ5136.2%, Ⅲ165.5%and Ⅲ260.1%, respectively. The sequencing analysis showed that a heterozygous missense mutation G68395T （NG₀09813.1） was detected in Exon4of PS gene leading to the substitution of Arg90by Leu （NP₀00304.2） for the propositus. The heterozygous mutation （Arg90Leu） was also found in other family members. A heterozygous （nonsense） mutation G12444A （NG₀12462.1） was detected in Exon4of AT gene leading to Trp257Ter （NP₀00479.1） for the propositus. The mutation （Trp257Ter） was found in other family members with reduced activity of AT. These two mutations （G68395T in PS gene and G12444A in AT gene） were not reported before and were thus novel ones.4. Conclusion The novel mutation G68395T in PS gene and G12444A in AT gene might be the causes of deficiency of PS and AT for the family. 1.Objective To explore the prevalence, clinical features and risk factors of deep venous thromboembolism （DVT） in hospitalized patients.2.Methods The demographic features, acquired and inherited risk factors of in-patient cases diagnosed with DVT in Nanjing Gulou Hospital from2002to2011were analyzed retrospectively. Chi-square test was used for comparative analysis, the difference was statistically significant if P<0.05. ELISA were used to detect protein C （PC）, protein S （PS） and antithrombin activities for some patients of them, respectively.3.Results617cases of DVT patients were enrolled. Among them, male to female ratio was1:1.225and the median age was57year old （12-92）.550（89.14%） patients were at their first diagnosis with the peak ages between50and70for men and40and60for women. The age distribution of the constituent ratio of man and women was no significant difference （0.25<P<0.5）. More common acquired risk factors were surgery （46.2%）, age （>70）（20.7%） and malignancies （7.3%）.29.6%of the patients had multiple acquired risk factors. The activities of protein C （PC） and protein S （PS） were measured in104patients, and the activities of antithrombin were measured in34patients of them. Among the anticoagulant deficiencies, PC deficiency accounted for13.5%, combined deficiency of PC and AT accounted for14.7%, combined deficiency of PC and PS and AT accounted for14.7%.26%of the104patients had inherited plus acquired risk factors. Relevant laboratory tests found that mean platelet volume （MPV）, hematocrit, hemoglobin content and red blood cell count, which affect the hemodynamic parameters, were reduced, but the incidence of DVT was increasing with the increased value of these four parameters. It was found that most DVT patients was with elevated triglycerides and reduced H-cholesterol.4. Conclusions Age for the first event of DVT is between40and70, and the age for women is about10years ahead of that for men. The major acquired risk factors are surgery, age （>70） and malignancies, and inherited risk factors were PC deficiency, combined PC and AT deficiencies and combined PC, PS and AT deficiencies. Most DVT patients have elevated triglycerides and reduced H-cholesterol. It seems that the coexistence of multiple risk factors plays an important role in triggering DVT.