Anti-mutant citrullinated vimentin antibody in the diagnosis of rheumatoid arthritis / 中华检验医学杂志

Objective This study aimed to assess the diagnostic value of anti-mutated citrullinated vimentin (MCV) antibodies in rheumatoid arthritis (RA) and its correlation with disease progression, extra-articular manifestations and overlap syndrome. Methods Retrospective Studies. Clinical data of 837 patients in PekingUnionMedicalCollegeHospitalfrom June to August 2017 were collected, including the result of anti-MCV, anti-cyclic citrullinated peptide (anti-CCP) antibodies, rheumatoid factor (RF), erythrocyte sedimentation rate (ESR) and High-sensitivity-C-reactive protein (CRP). According to the 1987 American College of Rheumatology classification criteria for rheumatoid arthritis, there were 323 patients diagnosed with RA, including 59 males and 264 females with the average age of 51 years. According to whether the RA patients have overlap syndrome with other autoimmune disease (AID) or have extra-articular manifestations, 258 cases were categorized into RA group, including 47 males and 211 females with the average age of 50 years; 14 cases were categorized into the group of overlap syndrome, including 1 male and 13 females with the average age of 36 years;51 cases were categorized into the group of extra-articular manifestations, including 11 males and 40 females with the average age of 59 years.According to 2010 rheumatoid arthritis classification criteria for destruction in joints, the radiographic changes were divided into 4 stages. There were 203 casesenrolled in our study, 88 caseswere fitted into early stage group (stage I)including 21 males and 67 females with the average age of 48 years; 115 caseswere fitted into progressive stage group, which compromisedstageⅡ (interim stage), stage Ⅲ (severe stage) and stage Ⅳ(final stage) cases, including 19 males and 96 females with the average age of 53 years. Mann-Whitney U test, x2 test, Receiver operating characteristic (ROC) curves and Spearmancorrelation coefficientwere used in Statistical analysis. Results Ⅰ Amongdiagnosed RA patients, 199 (61.6％) cases were positive for anti-MCV, anti-CCP and RFsimultaneously, 42 (13％) cases were positive for anti-MCV, which was higher than anti-CCP positive (1 cases, 0.3％) or RF positive (7cases, 2.2％). The difference was statistically significant(P<0.001, P<0.001). ⅡROC was calculated and MCV=35.95 U/ml was used as best-fit cut-off value. The AUC for anti-MCV was 0.867, while the sensitivity was 80.5％and specificity was 80.9％.ⅢThe detection levels of anti-MCV (682.8 (106.4-1000.0)), anti-CCP (407 (4.0-1536.0)) and RF (82.8 (21.1-244.9)) in the group of progressive stage were higher than those in the group of early stage (114.5 (28.5-1000.0), 62.5 (5.0-1020.7), 50.1 (6.7-127.1)), which showed a significant difference(P<0.05, P<0.05, P<0.05). The anti-MCV, anti-CCP and RF were positively related to the degree of joint destruction (r=0.229, P<0.05;r=0.187, P<0.05;r=0.167, P<0.05);anti-MCV and anti-CCP were positively related to extra-articular manifestation (r=0.152, P<0.05;r=0.136, P<0.05). Conclusion Anti-MCV antibodies are more sensitive in patients with RA, and have complementary diagnostic value for anti-CCP and RF-negative patients; high levels of anti-MCV and anti-CCP in RA patients are associated with RA progression and extra-articular involvement.

Research of SSA epitopes in patients of primary biliary cirrhosis and primary Sj(o)gren's syndrome / 中华风湿病学杂志

Objective To analyze the difference between primary biliary cirrhosis (PBC) and primary Sj(o)gren's syndrome (pSS) and primary biliary cirrhosis complicating with Sj(o)gren's syndrome (SS) in clinical features,anti-60 000 SSA multiple antigenic peptides (MAPs) detection rate,and explore the potential mechanisms of PBC with SS.Methods MAPs were artificially synthesized.Enzyme-linked immunosorbent assay (ELISA) was done to detect anti-MAPs antibodies in the sera of the three groups of patients.The detection rates of anti-MAPs antibodies were compared among groups and the relations of anti-MAPs antibodies with clinical features were analyzed.Chi-square test,Fisher's exact test,t test or Wilcoxon signed rank test were conducted in this study.Results There was no significant difference in clinical features,liver function tests and antibody profiles between PBC and PBC with SS.Significant difference of anti-MAP20 antibody detection rote was detected between pSS and PBC with SS groups [25％(7/28) vs 0(0/25),x2=7.201 1,P=0.007 3],and anti-MAP3,7 and 17 antibody detection rates in PBC with SS patients [4％(1/25),4％(1/25),8％(2/25)] were similar to pSS [4％(1/28),7％(2/28),7％(2/28)].The anti-MAP7 antibody and anti-MAP12 antibody detection rates were significantly higher in patients with splenomegaly than patients without splenomegaly [38％(3/8) vs 2％(2/84),38％(3/8) vs 4％(3/84); P=0.039 4,P=0.039 4],while the antiMAP17 antibody detection rate was significantly lower in patients with salivary gland injury than patients without salivary gland injury [5％(3/64) vs 39％(5/13); x2=4.431 8,P=0.035 3].Conclusion There is significant difference in the anti-MAPs detection rates among the three patients groups,and the detection rate may be higher in patients with certain clinical manifestations.

Construction and Identification of Recombinant Expressing of Human Sjogren's Syndrome Antigen A / 医学研究杂志

Objective To clone human Sjogren's syndrome antigen A(SSA)for expressing of antigen SSA-52kD and establishing a new clinical detecting method.Methods According to the human SSA-52kD cDNA sequence reported in GenBank,primers of human SSA-52kD cDNA were designed and synthesized.Human SSA-52kD cDNA was amplified from RNA of cultured Hela cell by reverse transcriptase polymerase chain reaction(RT-PCR).The production of amplification was ligated to PET-30a vector and then transformed into the competent bacteria DH5?to construct the recombinant plasmid PET-30a-SSA-52kD.The recombinant plasmid was digested with Bgl Ⅱ and Hind Ⅲ,and positive clones were sequenced.Results The Human SSA-52kD cDNA fragment containing 1447bp was amplified by RT-PCR.Restriction endonuclease mapping using Bgl II and Hind III showed that the target gene was inserted into the recombinant plasmid.The complete coding sequence of Human SSA-52kD was consistent with that of GenBank through DNA sequencing.Conclusions The full length of human SSA-52kD cDNA was successfully cloned and the recombinant plasmid PET-30a-SSA-52kD was constructed.

The diagnostic value of anti-cyclic citrullinated peptide antibody in patients with rheumatoid arthritis / 中华风湿病学杂志

Objective To explore the diagnostic value of anti cyclic citrullinated peptide (CCP) antibody (anti CCP) detected by ELISA in patients with rheumatoid arthritis (RA).Methods The synthesized CCP deduced from the known cDNA sequence was used as substrate in ELISA.The anti CCP was detected by ELISA in 294 RA,132 rheumatic diseases other than RA and 135 non rheumatic diseases.To compare the correlation among the anti CCP and APF (antiperinuclear factor),AKA (anti keratin antibody),RF (rheumatoid factor) and HLA DR4.Results One hundred and thirty seven (46 6%) RA sera,7 (5 3%) other rheumatic disease sera and 2 (1 8%) non rheumatic disease were found to be anti CCP positive by ELISA.The sensitivity of anti CCP was 46 6% with high specificity (96 9%) in RA.Anti CCP was associated with APF,AKA and HLA DR4 except RF.Conclusions Anti CCP presents a reasonable sensitivity (46 6%) with high specificity (96 6%) to RA and it is valuable to RA diagnosis.Anti CCP is associated with APF,AKA and HLA DR4 but not completely overlaps them.Anti CCP could be regarded as a new diagnostic marker in RA.

Preparation of Mi-2 antigen and significance of detecting anti-Mi-2 antibody in autoimmune connective tissue diseases / 中华风湿病学杂志

Objective To prepare Mi 2 antigen,detect anti Mi 2 antibody and investigate its significance in autoimmune connective tissue diseases (CTD).Methods Mi 2 antigen was prepared from rabbit thymus acetone powder and purified by DE52 chromatography.Anti Mi 2 antibodies were tested in 40 normal controls and 315 patients with CTD by double immunodiffusion.The distribution of anti Mi 2 antibodies in CTD was analysed.The clinical characteristics of dermatomyositis between anti Mi 2 positive group and negative group were compared.Results The precipitation line developed between the antigen and the standard anti Mi 2 serum.In DE52 chromatography,Mi 2 antigen was found in elute with 0 1 mmol/L and 0 2 mmol/L NaCl.The positive rate of anti Mi 2 antibodies was 26 1%(12/46) in dermatomyositis and 4 5%(2/44) in polymyositis respectively.There were no positive cases in 50 patients with rheumatoid arthritis,30 patients with primary Sjgren syndrome (SS),60 patients with systemic lupus erythematosus,50 patients with systemic sclerosis,35 patients with other CTD and 40 normal controls.The specificity of anti Mi 2 antibody in dermatomyositis was 99 4%.In dermatomyositis,the patients with positive anti Mi 2 antibody usually presented skin lesions at beginning,and most patients had V sign and/or shawl sign during the course of disease.On the other hand,the antibody negative ones manifested muscle involvements initially and easily got fever during the course of disease.There were statistic differences between the two groups in above features ( P