β2GP/anti-β2GP complex inhibits oxLDL-mediated lipid accumulation and FAK activation in THP-1 macrophages / 临床检验杂志

Objective@#To investigate the effects of β2 glycoprotein Ⅰ/anti-β2 glycoprotein Ⅰ complex (β2/aβ2) on oxidized low density lipoprotein (oxLDL)-mediated lipid accumulation and focal adhesion kinase (FAK) activation in THP-1 macrophage, as well as the role of Toll-like receptor 4 (TLR4) during the process. @*Methods@#THP-1 cells were differentiated into THP-1 macrophage by PMA (100 ng/mL). THP-1 macrophages were treated with RPMI 1640 medium, oxLDL, oxLDL+β2/aβ2 or oxLDL+lipopolysaccharide (LPS). The mRNA expressions of lipid transportation molecules, ACAT1, ABCA1 and ABCG1 were detected by RT-qPCR. Intracellular total cholesterol (TC) and free cholesterol (FC) in THP-1 macrophages were evaluated by Trinder assay, then the content and proportion of intracellular cholesteryl ester (CE) were calculated. The expression and phosphorylation of FAK were detected by immune fluorescence, RT-qPCR and western blot. To evaluate the role of TLR4, THP-1 macrophages were pre-treated with or without TLR4 inhibitor TAK-242 (1 μg/mL). @*Results@#β2/aβ2 treatment significantly inhibited oxLDL-mediated lipid accumulation and FAK expression and phosphorylation in THP-1 macrophages, which could be reversed by TLR4 blockage. @*Conclusion@#β2/aβ2 inhibits the oxLDL-mediated lipid accumulation and FAK activation of THP-1 macrophage, which is related to the function of TLR4.

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.

Anti-β2GPⅠantibodies accelerate the formation of atherosclerosis in high fat diet fed ApoE deficient mice / 中华心血管病杂志

Objective@#To investigate the effects of anti-β2 glycoprotein Ⅰ(β2GPⅠ) antibody on atherosclerosis in ApoE deficient mice.@*Methods@#A total of 24 male ApoE deficient mice of specific pathogen free level(six to eight-week old)were divided into normal control group, high fat diet group, high fat diet with anti-β2GPⅠ group, high fat diet with homologous control antibody group (n=6 each group). During the feeding period, mice were weighed every 2 weeks and were intraperitoneally injected with anti-β2GPⅠIgG (100 μg/per) and homologous control IgG (100 μg/per) according to grouping once a week. At the 16th week, the carotid arterial lipid deposition was observed by small animal magnetic resonance imaging, and blood samples were collected from internal vein of eyeball and the concentrations of TC, TG, HDL-C and LDL-C in plasma were measured after EDTA anticoagulant treatment. AI was calculated. The mice were then sacrificed and carotid arteries were removed, hematoxylin-eosin staining was used to observe the atherosclerotic lesions near the bifurcation of carotid artery and to calculate lesion size.@*Results@#(1) The body weight of mice was significantly higher in the high fat diet group compared to other 3 groups(all P<0.05), which was similar among high fat diet+ anti-β2GPⅠantibody group, high fat diet+ homologous control IgG group and normal diet control group (P>0.05). (2) After 16 weeks, plasma concentrations of TC and LDL-C in high fat diet group, high fat diet+ anti-β2GPⅠantibody group and high fat diet+ homologous control IgG group were significantly higher than in normal diet group (all P<0.05), there was no significant difference among high fat diet groups. The level of HDL-C was significantly higher in high fat diet control group than in normal diet control group. The concentration of TG was similar among groups. However, the value of AI in high fat+ anti-β2GPⅠ antibody group was significantly higher than in other groups (all P<0.05). (3) After 16 weeks, magnetic resonance imaging revealed that mice in high fat diet+ anti-β2GPⅠ antibody group had more obviously lipid deposition in the carotid arteries, it was significantly higher than that in the other groups, and the cross sections of carotid arteries stained with HE also demonstrated obviously carotid lumen stenosis and the percentage of carotid plaque area to carotid artery was (37.545±1.351)% in the high fat diet+ anti-β2GPⅠ antibody group, it was significantly higher than normal diet group ((1.235±0.460)%), high fat diet control group((11.635±2.751)%) and high fat diet+ homologous control IgG group ((11.815±2.623)%), all P<0.01. In high fat diet+ anti-β2GPⅠ antibody group, the area of carotid plaque was (3.121±0.124)×104 μm2, it was also significantly higher than normal diet group ((0.094±0.015)×104 μm2), high fat diet control group ((1.309±0.147)×104 μm2) and high fat diet+ homologous control IgG group ((1.027±0.228)×104μm2), all P<0.01.@*Conclusion@#Anti-β2GPⅠ antibody can promote atherosclerotic plaque formation in high fat diet fed ApoE deficient mice.

Detection and clinical significance of anti-cardiolipin and anti-β2 glycoprotein Ⅰ antibodies in missed abortion women / 中国综合临床

Objective To investigate the correlation of the anti-cardiolipin(aCL) antibodies and anti-β2 glycoprotein Ⅰ(anti-β2GPI) antibodies in missed abortion women.Methods Fourteen patients diagnosed missed abortion were chose as trial group,while 20 normal females received induced abortion in the same time as control group.Chemiluminescence method was used to measure the serum levels of IgG/IgM aCL and IgG/IgM anti-β2GPI antibodies in the two groups.Results The median levels of serum IgM aCL antibodies and serum IgM anti-β2GPI antibodies in trial group were significantly higher than those of control group(21.3(2.2-39.4)KU/L vs.6.0(1.5-10.7) KU/L,11.9(1.2-25.4) KU/L vs.1.9(1.1-4.0) KU/L;u=2.031,2.912;P ＜0.05).There were no significant differences in terms of the levels of serum IgG aCL antibodies and serum IgG anti-β2GPI antibodies between trial group and control group (P＞0.05).Conclusion There is correlation between IgM aCL as well as IgM anti-β2GPI antibodies and missed abortion,so in reducing the incidence of missed abortion,early and dynamic detection of anti-cardiolipin and anti-β2 glycoprotein I antibodies in childbearing women has a certain clinical significance.

Detection and Clinic Application of Antiphospholipid Antibody and Anti-β2-GPⅠ in Female Infertility / 现代检验医学杂志

Objective To analyze the correlation of different antiphospholipid antibody isotypes in female infertility.Methods Selected 296 patients with female infertility and 31 normal female between May 2015 to August 2015 in Dalian Women’s and Children’s Medical Centre.All the antibodies were detected by chemilumininescence.The positiverate of antipholipid an-tibodies in female infertility patients and health controls were calculated.The concentrations of antiphospholipid antibodies in different groups were compared with Mann-Whitney U test.Results The positive rate of aCL IgG of female infertility and normal female were 0.34% and 0.00%.The positive rate of aCL IgM of female infertility and normal female were 1.35%and 0.00%.The positive rate ofβ2 GPⅠ IgG of female infertility and normal female were 0.00% and 0.00%.The positive rate ofβ2 GPⅠ IgM of female infertility and normal female were 0.34% and 0.00%.aCL IgG were differet beween female infertility and normal female (P=0.013),others had on significant difference.Conclusion Need to detect LA,combined with aCL and anti-β2 GPⅠ,exclusively diagnose APS,layed the foundation of sequential treatment.

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.

Research Progress ofβ2 GlycoproteinⅠ and Anti-β2GPⅠ Antibodies and Antiphospholipid Syndrome / 现代检验医学杂志

Antiphospholipid syndrome (APS)is an autoimmune multisystem disorder characterized clinically by recurrent thrombosis and pregnancy morbidity.β2-glycoproteinⅠ (β2GPⅠ)is the major antigen in the APS.Anti-β2GPⅠ antibody is the autoantibody which targets toβ2GPⅠ.There is overwhelming evidence thatβ2GPⅠand anti-β2GPⅠ antibody play an important role in the pathogenesis of APS.Now the diagnosis and treatment of anti-β2GPⅠ antibody are more and more get people's attention,detection of anti-β2GPⅠDⅠ antibody can specifically diagnose APS and research of the molecular level may provide a new therapeutic target for APS.In this paper,β2GPⅠ,anti-β2GPⅠ antibody and APS associated development were reviewed.

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.

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.

Relationship between recurrent spontaneous abortion and anti-?_2-glycoprotein Ⅰ antibody / 吉林大学学报(医学版)

0.05).Conclusion The(anti-?_2-GP Ⅰ) antibody is related to RSA,and it may be regarded as a immunological assistant diagnostic index for RSA.

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

Study of relationship between some autoantibodies and chronic liver disease / 中华消化杂志

Objective To investigate the relationship between some autoantibodies including anti-? 2-glycoprotein Ⅰ (anti-? 2GPⅠ) and chronic liver disease. Methods A cohort of patients with chronic hepatitis B (CHB) and post-hepatitis B cirrhosis (PHBC) was studied for the serum anti-? 2GPⅠ levels by ELISA with purified ? 2GPⅠ as antigen. The serum dsDNA, smooth muscle antibody (SMA) and ribonucleoprotein antibody (RNPA) were also detected. Results High positive rate was observed in patients with CHB or PHBC (20.9%, 9/43; 49.3%,35/75) comparing with that in control group (3.1%, 1/32) ( P