Thrombophilia Caused by Beta2-Glycoprotein Ⅰ Deficiency: In Vitro Study of a Rare Mutation in APOH Gene / 华中科技大学学报（医学）（英德文版）

This study aimed to explore the mechanism of a novel mutation (p.Lys38Glu) in apolipoprotein H (APOH) gene causing hereditary beta2-glycoprotein Ⅰ (β2GPI) deficiency and thrombosis in a proband with thrombophilia.The plasma level of β2GPI was measured by ELISA and Western blotting,and anti-β2GPI antibody by ELISA.Lupus anticoagulant (LA) was assayed using the dilute Russell viper venom time.Deficiency of the major natural anticoagulants including protein C (PC),protein S (PS),antithrombin (AT) and thrombomodulin (TM) was excluded from the proband.A mutation analysis was performed by amplification and sequencing of the APOH gene.Wild type and mutant (c.112A＞G) APOH expression plasmids were constructed and transfected into HEK293T cells.The results showed that the thrombin generation capacity of the proband was higher than that of the other family members.Missense mutation p.Lys38Glu in APOH gene and LA coexisted in the proband.The mutation led to β2GPI deficiency and thrombosis by impairing the protein production and inhibiting the platelet aggregation.It was concluded that the recurrent thrombosis of the proband is associated with the coexistence ofp.Lys38Glu mutation in APOH gene and LA in plasma.

Thrombophilia Caused by Beta2-Glycoprotein Ⅰ Deficiency: In Vitro Study of a Rare Mutation in APOH Gene / 华中科技大学学报（医学）（英德文版）

This study aimed to explore the mechanism of a novel mutation (p.Lys38Glu) in apolipoprotein H (APOH) gene causing hereditary beta2-glycoprotein Ⅰ (β2GPI) deficiency and thrombosis in a proband with thrombophilia.The plasma level of β2GPI was measured by ELISA and Western blotting,and anti-β2GPI antibody by ELISA.Lupus anticoagulant (LA) was assayed using the dilute Russell viper venom time.Deficiency of the major natural anticoagulants including protein C (PC),protein S (PS),antithrombin (AT) and thrombomodulin (TM) was excluded from the proband.A mutation analysis was performed by amplification and sequencing of the APOH gene.Wild type and mutant (c.112A＞G) APOH expression plasmids were constructed and transfected into HEK293T cells.The results showed that the thrombin generation capacity of the proband was higher than that of the other family members.Missense mutation p.Lys38Glu in APOH gene and LA coexisted in the proband.The mutation led to β2GPI deficiency and thrombosis by impairing the protein production and inhibiting the platelet aggregation.It was concluded that the recurrent thrombosis of the proband is associated with the coexistence ofp.Lys38Glu mutation in APOH gene and LA in plasma.

Val247Leu and Trp316Ser poly morphisms of β2-glycoprotein Ⅰ in systemic lupus erythematosus patients and their associations with antiphospholipid antibodies and thrombotic complications / 中华风湿病学杂志

Objective To study the Val247Leu and Trp316Ser polymorphisms of β2-glycoprotein Ⅰ(β2GPⅠ) in systemic lupus erythematosus (SLE) patients and their associations with antiphospholipid antibodies and thrombotic complications.Methods We used DNA sequencing to detect the polymorphisms of Val247Leu and Trp316Ser in 378 SLE patients and 240 normal controls.Anti-β2GP Ⅰ antibodies and anticardiolipin (ACA) were tested by enzyme linked immunosorbent assay (ELISA).Lupus-type anticoagulants(LAC) was performed by diluted Russell's Viper Venom Test.Then the patient group was further analyzed according to APLs (Anti-β2GP Ⅰ antibody,LAC and ACA),thrombosis and obstetrical complications using Logistic regression analysis to confirm whether there are associations between β2GPⅠpolymorphism and those factors.Results For Va1247Leu,the predominant genotype was LL in healthy controls which accounted for 57.08％,while it was VL in SLE patients which accounted for 59.5％ (x2=45.01,P=0.000).Frequency of VV genotype was significantly higher in patients with thrombosis,anti-β2GP Ⅰ,ACA and obstetrical complications (OR=6.79,3.75,2.12 and 3.85,respectively;P=0.000,0.001,0.044 and 0.017,respectively).Those patients with VL genotype tended to have positive anti-β2GPI,LAC,ACA,thrombosis and also obstetrical complications (OR=2.95,1.88,2.47,2.97 and 2.74,respectively;P=0.000,0.007,0.000,0.001 and 0.016,respectively) than those negative ones.The predominant genotype of Trp316Ser was TT,then TS.No correlations could be found between Trp316Ser polymorphism and APLs,neither relation to thrombosis complications.Conclusion The polymorphism of Val247Leu is significantly associated with the presence of APLs,thrombosis and obstetrical complications.Both VV and VL genotype are risk factors for the generation of APLs,occurrence of thrombosis and obstetrical complications.The VV genotype is a high risk factor for thrombosis.Trp316Ser polymorphism does not contribute to the APLs production and also have no correlations with thrombotic complication.

The Correlation betweenβ2-GP1,VEGF and TF with Rat DVT Formation / 天津医药

Objective To build rat DVT inferior vena cava partial stasis (narrow) model, to detected the expression ofβ2-GP1, VEGF and TF in rat blood, and to investigat the correlation betweenβ2-GP1, VEGF and TF with DVT. Meth?ods SD rats (n=70) are divided into control group (n=10), sham operation group (n=30) and the model group (n=30) ran?domly and DVT model was built by the inferior vena cava partial stasis (narrow) after 2 h, 8 h and 24 h respectively. In each time point, ten rats were taken in each group, inferior vena cava blood were collected whileβ2-GP1, VEGF and TF expres?sion were detected by ELISA. Results In rat experiment, compared with control group, there was no significant change in?expression of β2-GP1, VEGF and TF in sham operation group (P > 0.05). Levels of β2-GP1, VEGF and TF were in?creased at the 2nd hour and 8th hour then peak at the 24th hour which was higher than those in the 24th hour control group and in Sham group and it was also higher than those in the 2nd hour and the 8th hour in model group with statistical signifi?cant difference (P<0.01). Conclusion Based on the above experimental data, in rat DVT formation process, β2-GP1, VEGF and TF may play an important role in promote DVT formation.

Clinical significance of different antiphospholipid antibody isotypes in systemic lupus erythematosus / 中华检验医学杂志

Objective To investigate the prevalence and clinical significance of different subtypes (IgG,IgM and IgA) of anticardiolipin antibodies (aCL) and anti-β2-glycoprotein Ⅰ antibodies (aβ 2GP1),as well as lupus anticoagulant (LA) in systemic lupus erythematosus (SLE).Methods IgG/IgM/IgA,IgG,IgM,IgA aCL and anti-β2GP1 were tested by enzyme-linked immunosorbent assay (ELISA) in 100 patients with SLE (42 patients were diagnosed as secondary antiphospholipid syndrome),44 healthy controls and 32 patients with other connective tissue diseases excluding SLE and antiphospholipid syndrome (APS).Meanwhile,LA was tested by modified Dilute Russell's viper venom time (dRVVT).The correlation between antiphospholipid antibodies and clinical manifestation was analyzed by Spearman correlation analysis.The postiverate of antiphospholipid antibodies in SLE patients,health controls and patients with other connective tissue diseases were compared by chi square test.The concentrations of antiphospholipid antibodies in different groups were compared using independent sample Kruskal Wallis test.The diagnostic efficacy of antiphospholipid antibodies in SLE patients was analyzed by crosstable using clinical diagnosis of APS as gold standard.P ＜ 0.05 was considered statistically significant.Results The prevalence of IgG aCL (x2 =15.031,P ＜ 0.001),IgA/G/M (x2 =11.678,P =0.003) and IgA (x2 =6.17,P =0.036) antiβ2GP1 were significantly higher in patients with SLE than in the other two groups.IgA/G/M (r =0.207,P=0.039),IgG (r=0.230,P=0.021) and IgA (r=0.217,P=0.030) aCL,IgA/G/M (r=0.218,P=0.029) and IgA (r =0.255,P =0.01) anti-β2GP1,as well as LA (r =0.233,P =0.02) were associated with thrombotic events.IgA/G/M anti-β2GP1 (r =0.22,P =0.029) and LA (r =0.254,P =0.011) were associated with pathological pregnancy.23.1％ (6/26) aCL positive SLE patients were IgM and/or IgA aCL positive.53.6％ (15/28) anti-β2GP1 positive SLE patients were IgM and/or IgA antiβ2GP1 positive.In SLE patients,the specificity and sensitivity of IgA/G/M aCL for APS were 98.3％ and 26.2％,respectively.The specificity and sensitivity of IgA/G/M anti-β2GP1 were 84.5％ and 40.5％,respectively.The specificity of at least two isotypes positive for APS (both aCL and anti-β2GP1 were 98.3％),was higher than IgG aCL (94.8％) or anti-β2GP1 (93.1％).The sensitivity of at least one isotype of aCL (47.6％) or anti-β2GP1 (42.9％) positive for APS were higher than IgG aCL (40.5％)and anti-β2GP1 (21.4％).Conclusions IgG and IgM aCL together would be better than IgA/G/M aCL for APS screening.IgA/G/M anti-β2GP1 would be better for APS screen due to higher sensitivity and strong association with thromboembolic events and pathologic pregnance.IgA aCL or anti-β2GP1 was associated with thromboembolic events.IgA aCL or anti-β2GP1 would be useful for APS diagnosis in IgG and IgM aCL or anti-β2GPl negative patients.

Examination and clinical significance of combination rate between beta2-glycoproteinⅠand platelet in the ulcerative colitis patients / 中国实用内科杂志

Objective To explore the level and clinical significance of combination rate between beta2-glycoproteinⅠand platelet in the ulcerative colitis patients.Methods For 67 ulcerative colitis(UC) patients from the First and the Second Hospital of Jilin University from the September 2003 to December 2004,using flow cytometry(FCM),we detected combination rates of between beta2-glycoproteinⅠand platelet in the UC patients and in the normal control subjects,respectively.Results There was significant difference in the combination rates of beta2-glycoproteinⅠand platelet in the two UC groups and normal control group(P