The detection of autoimmune liver disease (AILD) relevant autoantibodies is of important value in the diagnosis and treatment of AILD and especially in autoimmune hepatitis and primary biliary cirrhosis.With the increasing of patients clinically diagnosed AILD,the detection of AILD relevant autoantibodies is gradually clinically concerned and appreciated.As the detection of AILD relevant autoantibodies affected by various factors,there are still many problems in the detection and clinical applications of AILD relevant autoantibodies.We should promote the universal clinical application of AILD relevant autoantibodies,emphasis on the quality management and improve the quality of detection constantly,attend to the standard detection and rational application of AILD relevant autoantibodies.

The detection of autoantibodies is of great value in the diagnosis and treatment of autoimmune diseases.The popular autoantibody screening promotes the rapid development of the clinical awareness,diagnosis and treatment of autoimmune diseases,which in turn results in the increasing demand of autoantibody detection and continuous improvement the detection quality.In order to make better use of autoantibodies results during the diagnosis and treatment of autoimmune diseases,the workers of autoantibodies detection should understand various affecting factors of the autoantibodies detection completely,emphasis on the autoantibodies quality management and improve the quality of detection constantly ; Aware of the complexity and specificity of autoantibodies fully and participate in the selection of autoantibodies detection and correct interpretation of the clinical significance of autoantibodies actively;Emphasis on the clinical application of autoantibodies and promote the universal clinical application of autoantibodies.

Objective Matrix-assisted laser desorption ionization-time of might-mass spectrometry (MALDI-TOF-MS) was utilized to analyze the protein fingerprint in brain-gut interaction of irritable bowel syndrome (IBS) model rats' colon, so as to find the clues for IBS. Methods Fourteen healthy male adult Wistar rats were selected and divided into a control and a chronic and acute stress ( CAS) group. Colon motility, visceral sensation and behavior changes of rats were detected to evaluate the model. MALDI-TOF-MS was used to observe the overall view of protein in colon so as to study whether there are abnormalities of protein levels in IBS. Results As compared with those in the control group, the number of fecal pellets [ (6. 00 ± 1. 69 ) pellets/1 h vs ( 1. 14 ± 0. 69 ) pellets/1 h, P < 0. 01 ] and frequency of abdominal contraction induced by colorectal distention (CRD) increased, while the amount of weight gain [ (298. 88 ± 18.61)gvs (348. 00±12. 44)g, P<0.01] and consumption of sucrose solutions [ (13. 63 ± 1. 69) ml/1 h vs (19.00±3.06) ml/1 h, P<0.05] decreased in the CAS group (P <0. 05). As far as protein/peptide quality different peak was concerned, CAS rats had 12 different peaks compared with the control rats. The different proteins could be divided into 4 types, which were related to iron secretion, protein synthesis, G protein system and immunity. The protein levels of the model group were higher than those in the control group (P < 0. 05). Conclusions The CAS rats integrate the major characteristics of IBS such as altered colon motility, higher visceral hypersensitivity and psychiatric disorder and can mimic the brain-gut interaction of IBS partly. The detection of differential proteins provides reference for the pathogenesis and treatment of IBS.

Objectives To investigate the ratios of peripheral blood CD4+CD8+ and CD4+CD25+ regulative T cells, and explore the association with hepatic damnification and anti-AMA-M2 antibodies.Methods The percentage of CD4+CD8+T cells and CD4+CD25+T cells in peripheral blood from patients with primary biliary cirrhosis(PBC) (n=27)、26 patients with other hepatic desease、30 normal individuals were measured by flowcytometry.Results Patients with PBC had statistically higher levels of CD4+CD25+T cells than the patients with other hepatic disease (P

Objective: To investigate the ratios of peripheral blood CD4+CD25+,CD4+CD8+ regulative T cells of systemic lupus erythematosus(SLE) patients, and explore the association with disease active stage,nephropathy,serum anti-ds-DNA antibody,and both IgG and C3 levels. Methods: The percentage of CD4+CD25+T cells and CD4+CD8+T cells of peripheral blood from patients with systemic lupus erythematosus(SLE)(30 females and 7 males),30 rheumatism controls and 30 normal individuals were measured by flowcytometry. Results: Patients with active disease had statisitically lower levels of CD4+CD25+T cells than did normal controls(P

Objective To explore the prevalence of autoimmune liver disease-related antibodies in patients with PBC, and study the clinic significance of autoimmune liver disease related antibody profiles in patients with PBC. Methods The anti-AMA in 247 specimens from patients with liver disease, including 173 PBC, 37 AIH and 37 LDC were detected by IIF. Anti-AMA-M2, anti-GP210, anti-SP100, anti-SLA, anti-LCI and anti-LKM-1 antibodies were measured by ELISA. Results The positive rates of anti-AMA, anti-AMA-M2, anti-GP210, anti-SPl00, anti-LC1, anti-SLA and anti-LKM-1 antibodies were92. 5% (160/ 173), 86.7% (150/173), 35. 8% (62/173), 24. 3% (42/173), 0.6% (1/173), 0% (0/173) and 0.6%(l/173) in PBC group, 18.9% (7/37), 5.4% (2/37),8.1% (3/37),13. 5% (5/37),0% (0/ 37 ) ,5.4% (2/37 ) and 2. 7% (1/37 ) in AIH group and 5.4% (2/37), 2.7% (1/37 ) , 5.4% (2/37 ) , 10. 8% (4/37) , 0% (0/37) , 0% (0/37) and 0% (0/37) respectively in LDC group. Anti-AMA, anti-AMA-M2 and anti-GP210 was detected more frequently in patients with PBC group than AIH group (x~2 =101.3,100.8 and 11.0,P<0.01) while anti-SLA was detected more frequently in patients with AIH group than PBC group (x~2 = 9. 4, P < 0.01). The levels of ALT, TBIL, DBIL, GGT and ALP were higher in patients known to have positive anti-GP210 ( U = 1212.0,1199.0,1218.0,1074.0,1030. 0,P < 0. 01) and the levels of IgM were higher in patients known to have positive AMA ( U = 94.0, P <0.05). Conclusions Anti-LCI, anti-SLA and anti-LKM-1 antibodies in PBC and AIH are detected at a very low frequency in the corhort. Anti-GP210 antibody is found to be associated with the severity of liver damage while AMA is found to be associated with immunologic function in patients with PBC. There is little significance for screening anti-LCI, anti-SLA, anti-LKM-1 antibodies in patients with autoimmune liver diseases. It is of importance to detect anti-AMA and anti-GP210 antibodies for diagnosis of PBC.

Objective To detect the value of autoantibedy profile in primary biliary cirrhosis (PBC). Methods 96 serum samples of patients with primiary biliary cirrhosis, 100 serum samples of other antoimmune disease and 49 serum samples of healthy were tested for anti- M2, anti-3E(BPO), anti-Sp100,anti-PML,anti-gp210,anti-LKM-1 ,anti-LC-1 ,anti-SLA/LP by EUROLine. Results The positive of the anti-M2,anti-BPO, anti-Sp100, anti-PML and anti-gp210 for PBC was 76. 0%, 84.4%, 32. 3%, 28. 1% and 35.4% ,respectively. The positive of the anti-M2, anti-BPO, anti-Sp100, anti-PML and anti-gp210 for other autoimmune disease was 13.0% ,9. 0% ,3.0% ,2.0% and 1. 0%, respectively. The sensitivity of the anti-M2 for PBC was 76. 0% ,with specificity of 87. 0%. The sensitivity of the anti-BPO for PBC was 84. 4%, with specificity of 91.0% ;the sensitivity of the anti-Sp100 for PBC was 32. 3%, with specificity of 97.0%. The sensitivity of the anti-PML for PBC was 28. 1% ,with specificity of 98.0%. The sensitivity of the anti-gp210 for PBC was 35.4%, with specificity of 99. 0% . Anti-LKM-1, anti-LC-1, anti-SLA/LP positive patients with PBC were not detected;the incidence rate of liver function failure in anti-gp210 positive serum higher than anti-gp210 negative serum (χ2 = 11.17, P < 0. 01). Conclusions Multiple autoantibedies can be detected in the sera of PBC patients. The detection of autoantibody profile is useful for the diagnosis and differential diagnosis of PBC, and may he helpful for therapy and prognosis of PBC.

Objective To clone and express the human asialoglycoprotein receptor(ASGPR) H1 subunit, purify and identify the immunoreactivity of the recombinant protein, and establish the enzyme linked immunosorbent assay (ELISA) to detect anti-ASGPR antibodies in diagnosis of autoimmune hepatitis. Methods The CRDHI cDNA (435 bp) was subcloned into eukaryotic vector PEGH, and the recombinant protein expression was induced by D (+)-Galactose. The recombinant CRDH1 was purified with Glutathione Sepharose 4B, and its immunoreactivity was identified by SDS-PAGE and western blot as well as MALDI-TOF. ELISA was established to detect the anti-ASGPR antibodies in serum samples of 45 patients with AIH, 30 patients with SLE, 30 patients with RA, 10 patients with SS and 30 normal controls. Results The sequencing of recombinant plasmid showed the CRDH1 gene was successfully inserted to the eukaryotic expression vector with correct sequence and open reading frame. The fusion protein showed a molecular weight of 42 500 Da on SDS-PAGE gel and confirmed to be the human ASGPR by MALDI-MS through peptide mass fingerprint analysis with Mascot in human protein database. It shared 98. 34% homology with ASGPR H1 subunit. Western blot analysis showed that the fusion protein had the same immunoreactivity as human ASGPR. The results of ELISA indicated that the positive rate of anti-ASGPR was 35.6% ( 16/45 ), but the ELISA was negative in other control. There was significant difference of positivity of the autoantibodies between AIH and non-AIH controls (χ2 = 31.85,P < 0. 01 ). Conclusions The human plasmid containing ASGPR is successfully clone into Saccharomyces cerevisiae Y258. The recombinant autoantigen owns good antigenicity and specificity. ELISA established with the purified protein shows good specificity for diagnosis of AIH.

Objective To investigate the relationship between the PTPN22 gene polymorphism and rheumatoid arthritis(RA).Methods Real time fluorescent quantitation PCR was used to detect the 1123G＞C polymorphism of the PTPN22 gene from 200 RA patients,100 others rheumatic diseases and 200 the normal controls.The results were analyzed by SPSS 11.0 software.Results The CC genotype frequencies of RA patients.others rheumatic diseases and the normal controls were 0.120,0.020 and 0.015 respectively.There was a significant difiefence between BA patients and others rheumatic diseases (X=18.708.P＜0.01).1'here Wag a significant difference between RA patients and the normal controls(X2=24.337,P＜0.01).There was not statistically significant between others rheumatic diseases and the normal controls(X2=1.066,P＞0.05).The C allele frequency of RA patients,others rheumatic diseases and the normal controls were 0.360.0.190 and 0.215 respectively.The results were significant difference.Conclusion The PTPN22 gene could be one of predisposing genes and the therapeutic target genes with RA patients.

Objective To study the relationship between the expression of eostimulating factor B7-H4 in patients with primary biliary cirrhosis (PBC) and the pathogenesis of PBC. Methods The expression of B7-H4 mRNA on peripheral blood mononuclear cell (PBMC) of 65 patients with PBC was tested by real-time PCR. Serum levels of IL-2 were assayed by ELISA. CD4+, CD8+ T lymphocytes expression level and B7-H4 expression rate before and after activation were measured by three-color flow cytometry (FCM). Re-sults (1) Expression of B7-H4 mRNA and BT-H4 percentage in PBC group were significantly lower than that in none-PBC group and healthy controls(P<0.01);(2)After 72 h activation, the percentage of CD4+, CD8+, and CD4+ CD8+T lymphoeytes and serum levels of IL-2 decreased (P<0.05), and the percentage of CD4+, CD4+ CD8+T lymphocytes and serum levels of IL-2 were significantly higher than that of none-PBC group and healthy controls(P<0.01);(3)Levels of alanine aminotransferase(ALT), aspartate amin-otransferase(AST), alkaline phosphatase(ALP) and γ-glutamyl transpeptidase(GGT) of patients with posi-tive anti-mitochondrial antibody(AMA)-M2 rose. There was no significant difference of B7-H4 expressions on T cells between patients either with or without AMA-M2 antibody. Conclusion The costimulating factor B7-H4 can express on T lymphocytes which is activated by phytohemagglatinin(PHA) aad plays a negative role on T cells responses.