Current state of technologies and recognition of anti-SSA/Ro antibodies in China: A multi-center study.

BACKGROUND: Previous studies have demonstrated that Ro60 and Ro52 have different clinical implications, and anti-Ro52 antibodies are an independent serum marker of systemic autoimmune diseases, including Sjögren's syndrome. Many different assays have been adopted to detect anti-Sjögren's syndrome antigen A (SSA)/Ro antibodies, while to date no specific approach has been recommended as optimal for anti-SSA/Ro antibody testing. Herein, we performed a multi-center study to explore the current clinical utility of different strategies for anti-SSA/Ro antibody testing in China. METHODS: Twenty-one tertiary care centers were included in this questionnaire-based study. The self-administered questionnaire mainly includes testing methods for anti-SSA/Ro antibodies, reporting system of results, and interpretation of results by clinicians. RESULTS: Six different methods were applied to detect anti-SSA/Ro antibodies in the 21 centers. Line immunoassay (eight different commercial kits) was the most frequently adopted method (21/21, 100%), with different cutoff values and strategies for intensity stratification. There were two reporting systems: One was reported as "anti-SSA antibodies" and "anti-Ro52 antibodies" (12/21, 57%), while the other was "anti-SSA/Ro60 antibodies" and "anti-SSA/Ro52 antibodies" (9/21, 43%). Notably, six centers (29%) considered either positive anti-Ro60 or anti-Ro52 antibodies as positive anti-SSA antibodies, all of which adopted the latter reporting system. CONCLUSION: Significant variabilities existed among anti-SSA/Ro assays. Nearly 30% of centers misinterpreted the definition of positive anti-SSA antibodies, which may be attributed to the confusing reporting systems of line immunoassay. Therefore, we advocate standardization of the nomenclature of anti-SSA/Ro antibodies, changing the "anti-SSA/Ro52" label in favor of the "anti-Ro52" antibodies for a clear designation.

Identification of Novel Biomarkers for Behcet Disease Diagnosis Using Human Proteome Microarray Approach.

Behcet disease (BD) is a chronic systemic vasculitis and considered as an autoimmune disease. Although rare, BD can be fatal due to ruptured vascular aneurysms or severe neurological complications. To date, no known biomarker has been reported for this disease, making it difficult to diagnosis in the clinics. To undertake this challenge, we employed the HuProt arrays, each comprised of ∼20,000 unique human proteins, to identify BD-specific autoantibodies using a Two-Phase strategy established previously. In Phase I, we profiled the autoimmunity on the HuProt arrays with 75 serum samples collected from 40 BD patients, 15 diagnosed autoimmune patients who suffer from Takayasu arteritis (TA; n = 5)), ANCA associated vasculitis (AAV; n = 5), and Sjogren's syndrome (SS; n = 5), and 20 healthy subjects, and identified 20 candidate autoantigens that were significantly associated with BD. To validate these candidates, in Phase II we constructed a focused array with these 20 candidate BD-associated antigens, and use it to profile a much larger cohort, comprised of serum samples collected from 130 BD patients, 103 autoimmune patients (i.e. 40TA, 40 AAV and 23 SS), and 110 healthy controls. This allowed us to validate CTDP1 (RNA polymerase II subunit A C-terminal domain phosphatase)as a BD-specific autoantigen. The association of anti-CTDP1 with BD patients was further validated using the traditional Western blotting analysis. In conclusion, anti-CTDP1 antibody serves a novel autoantibody for Behcet disease and is expected to help more accurate clinical diagnosis.

Telangiectasia as a potential clinical marker of microvascular lesions in systemic sclerosis patients from EUSTAR data in China.

OBJECTIVES: To investigate the prevalence and clinical relevance of telangiectasia in Chinese patients with systemic sclerosis (SSc). METHODS: Data from 230 SSc EUSTAR patients from Peking Union Medical College Hospital (2009-2011) that fulfilled the 1980 American College of Rheumatology SSc classification criteria were prospectively collected. Demographic, clinical, and laboratory data were calculated between groups with and without telangiectasia, and a six-minute walk test, pulmonary function test (PFT), transthoracic echocardiography (TTE), right heart catheterisation (RHC) and modified Rodnan skin score (mRSS) were performed. RESULTS: 96 patients (41.7%) were diagnosed with telangiectasia. There were no significant differences between patients with and without telangiectasia based on gender, age at onset, Raynaud's phenomenon (RP) duration, or SSc classification. Disease duration both from RP onset of patients and from first non-RP manifestation of patients with telangiectasia was significantly longer than patients without (p<0.05). RP (97.9% vs. 90.3%), finger/toe sclerosis (96.9% vs. 88.1%), facial sclerosis (68.8% vs. 53.7%), digital ulcers (DUs; 40.6% vs. 23.1%), digital pitting (49.0% vs. 33.8%), joint contracture (20.8% vs. 10.4%) and erythrocyte sedimentation rate elevation (26.7% vs. 14.8%) were significantly greater in telangiectasia patients (p<0.05). There were no differences in autoantibody development between patients with and without telangiectasia (p>0.05). PFT showed that forced vital capacity (77.0±17.26 vs. 83.05±16.53, p=0.005) and diffusion capacity for CO of the lung (58.9±19.4 vs. 65.7±19.7, p=0.030) were lower, while forced expiratory volume ratio (87.02±7.8 vs. 84.33±7.1, p=0.029) was higher in SSc with telangiectasia. Pulmonary artery hypertension (PAH) prevalence (25.0% vs. 14.2%) was significantly greater in patients with telangiectasia. CONCLUSIONS: Telangiectasia are common in Chinese SSc patients and usually associated with DUs, RP, and PAH. Telangiectasia could be a clinical marker of microvascular disease in SSc.

Identification of new autoantigens for primary biliary cirrhosis using human proteome microarrays.

Primary biliary cirrhosis (PBC) is a chronic cholestatic liver disease of unknown etiology and is considered to be an autoimmune disease. Autoantibodies are important tools for accurate diagnosis of PBC. Here, we employed serum profiling analysis using a human proteome microarray composed of about 17,000 full-length unique proteins and identified 23 proteins that correlated with PBC. To validate these results, we fabricated a PBC-focused microarray with 21 of these newly identified candidates and nine additional known PBC antigens. By screening the PBC microarrays with additional cohorts of 191 PBC patients and 321 controls (43 autoimmune hepatitis, 55 hepatitis B virus, 31 hepatitis C virus, 48 rheumatoid arthritis, 45 systematic lupus erythematosus, 49 systemic sclerosis, and 50 healthy), six proteins were confirmed as novel PBC autoantigens with high sensitivities and specificities, including hexokinase-1 (isoforms I and II), Kelch-like protein 7, Kelch-like protein 12, zinc finger and BTB domain-containing protein 2, and eukaryotic translation initiation factor 2C, subunit 1. To facilitate clinical diagnosis, we developed ELISA for Kelch-like protein 12 and zinc finger and BTB domain-containing protein 2 and tested large cohorts (297 PBC and 637 control sera) to confirm the sensitivities and specificities observed in the microarray-based assays. In conclusion, our research showed that a strategy using high content protein microarray combined with a smaller but more focused protein microarray can effectively identify and validate novel PBC-specific autoantigens and has the capacity to be translated to clinical diagnosis by means of an ELISA-based method.

Clinical characteristics of systemic sclerosis patients with digital ulcers in China.

OBJECTIVES: To investigate the clinical characteristics of SSc patients with DUs in China. METHODS: The data of 267 consecutive SSc patients based on the EUSTAR DATABASE from Peking Union Medical College Hospital from February 2009 to March 2012 were prospectively collected. The patients with DUs were compared to those without DUs. RESULTS: Seventy-nine patients (29.6%) had DUs out of 267 SSc patients analysed. There were significant differences between patients with and without DU based on sex (female/male: 65/14 vs. 174/14), age of onset of SSc (32.3±11.7 vs. 40.4±12.6 y), age of onset of Raynaud's phenomenon (31.8±12.3 vs. 38.7±12.2) (p<0.05). In addition, there was a higher rate of diffuse SSc, gastrointestinal involvement, (especially esophageal involvement), and pericardial effusion, higher mRodnan score, and more anti-scl70 antibody positivity in patients with DU (p<0.05). A multivariate analysis identified anti-Scl70 antibody positivity, gastrointestinal involvement and a younger age at disease onset as three risk factors for developing DUs in SSc patients. CONCLUSIONS: The occurrence of DUs in Chinese SSc patients is frequent. It is possible that SSc patients with DUs were influenced by the disease earlier in life, which should be detected early for effective intervention.

Up-regulation of TGF-ß1 mRNA expression in peripheral blood mononuclear cells of patients with chronic fatigue syndrome.

BACKGROUND/PURPOSE: It has been shown that the abnormality in immune cells in chronic fatigue syndrome (CFS) patients is closely associated with the participation of TGF-ß. In order to study the relationship between TGF-ß1 and CFS, we investigated the mRNA levels of TGF-ß1 in peripheral blood mononuclear cells (PBMCs) in patients with CFS. METHODS: Fluorescent quantitative real time reverse-transcription polymerase chain reaction (FQ-RT-PCR) was performed to test TGF-ß1 mRNA expression in PBMCs in 63 cases of CFS, 50 cases of disease controls, and 50 cases of healthy controls. RESULTS: The mean value of TGF-ß1 mRNA expression in CFS patients was ΔΔCt=1.17±0.58, which was significantly higher than the disease controls (ΔΔCt=0.07±1.08, df=111, p < 0.01) and the healthy controls (ΔΔCt=0.00±1.63, df=111, p < 0.01). No significant difference was detected between disease and healthy controls (p > 0.05). CONCLUSION: The expression of TGF-ß1 in PBMCs is significantly elevated in patients with CFS. It might be correlated to the pathogenesis of the disease.

Effectiveness of iguratimod as monotherapy or combined therapy in patients with rheumatoid arthritis: a systematic review and meta-analysis of RCTs.

BACKGROUND: This study aims to evaluate the efficacy and safety of the iguratimod (IGU) as monotherapy or combined therapy in patients with rheumatoid arthritis (RA) by using meta-analysis. METHODS: We searched Medline, EMBASE, Cochrane library, CNKI, Wanfang medical network from initial to 30 June, 2020, for randomized clinical trials (RCTs). Two authors independently screened the studies via reading the title, abstract, and full text. The risk of bias in individual studies was assessed using the Cochrane Risk of Bias tool. STATA 12.0 was used for pooled analysis of all included studies. RESULTS: A total of 23 RCTs were included in this analysis. Meta-analysis showed that patients in the IGU monotherapy or combined therapy group had significantly higher ACR20 (OR = 1.97, 95% CI 1.29 to 3.00, P = 0.002), lower DAS28-CRP (SMD = -3.49, 95% CI -5.40 to -1.58, P < 0.001) and DAS28-ESR (SMD = -2.61, 95% CI -3.64 to -1.57, P < 0.001), as well as shorter duration of morning stiffness (SMD = -2.06, 95% CI -2.86 to -1.25, P < 0.001) and lower HAQ score (SMD = -0.91, 95% CI -1.61 to -0.21, P = 0.011), than those received other disease-modifying antirheumatic drugs (DMARDs) monotherapy (primarily comprising methotrexate). For the safety profile, IGU monotherapy had similar risks for gastrointestinal reactions (P = 0.070), leucopenia (P = 0.309), increment in transaminase (P = 0.321), increase of ALT (P = 0.051), and liver damage (P = 0.182) to methotrexate monotherapy, and IGU combined with other DMARDs therapy did not increase the risks of these AEs (P > 0.05). CONCLUSIONS: Our evidence suggests that IGU is effective and tolerant as monotherapy or combined therapy especially with methotrexate in patients with active RA. IGU may be regarded as a potential alternative to methotrexate, and a preferable choice when combined with other DMARDs for the treatment of RA.

Higher levels of CCL20 expression on peripheral blood mononuclear cells of chinese patients with inflammatory bowel disease.

This study aimed at characterizing the levels of CCL20 mRNA transcripts in peripheral mononuclear blood cells (PMBC) of 56 Chinese patients with inflammatory bowel disease (IBD), 30 other intestinal diseases and 30 healthy controls by quantitative real time polymerase chain reaction. The levels of CCL20 mRNA transcripts in PBMC of patients with IBD were significantly higher than that of patients with non-IBD intestinal diseases and healthy controls (p < 0.01) and the CCL20 expression in active IBD patients was significantly higher than that in remission patients (p < 0.01). Importantly, the levels of CCL20 expression in PBMC were significantly correlated with the degrees of disease severity, the levels of erythrocyte sedimentation rate and C-reaction protein, but not hemoglobin, in patients with IBD (p < 0.01). Furthermore, the levels of CCL20 expression in active IBD patients after treatment with salazosulphapyridine or prednisone were significantly reduced, as compared with before treatment (p < 0.01). Therefore, analysis of CCL20 expression in PBMC may be used as a surrogate measure for evaluation of IBD activity, disease progression and therapeutic efficacy in Chinese IBD patients.

[A pilot study of protein fingerprinting in brain-gut interaction model of irritable bowel syndrome.].

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) g vs (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.

CD74 auto-antibodies display little clinical value in Chinese Han population with axial spondyloarthritis.

The European cohort study has indicated about CD74 IgG-autoantibodies as potential marker for axial spondyloarthritis (axSpA) diagnosis. However, multiple studies have questioned the diagnostic value of various disease-specific autoantibodies in different ethnic groups. Here, we have tried to assess the diagnostic value of anti-CD74 IgG and IgA autoantibodies in axSpA patients from Chinese Han population.The anti-CD74 IgG and IgA autoantibodies were analyzed using ELISA assay in a cohort of 97 axSpA patients, including 47 treatment-naïve axSpA patients never treated with steroids or immunosuppressants and 50 treated axSpA patients. The rheumatic disease control (RDC) group consisted of 40 rheumatoid arthritis, 25 systemic lupus erythematosus, 18 psoriatic arthritis patients, and 60 healthy controls (HC).Our data demonstrated the presence of anti-CD74 IgA auto-antibodies in 25.8% of the axSpA patients, 30.1% of the RDC group patients and none in HC. Similarly, anti-CD74 IgG autoantibodies were observed in 23.7% of the axSpA patients, 18.1% of the RDC patients and 18.3% of the HC. The sensitivity, specificity, and accuracy of IgA autoantibodies were 21.3%, 82.5%, & 67.4%, respectively, while for IgG, it was 27.7%, 81.8%, and 68.4%, in treatment-naïve axSpA patients. Furthermore, weak positive relationship between anti-CD74 IgA autoantibodies and bath ankylosing spondylitis disease activity index ( r = 0.253, P = .012) and functional index (bath ankylosing spondylitis functional index; r = 0.257, P = .011) was observed.Overall, our study demonstrated little clinical and predictive value of CD74 autoantibodies in the diagnosis of axSpA and its related manifestations, among Chinese Han population.

[Promising diagnostic model for systemic lupus erythematosus using proteomic fingerprint technology].

OBJECTIVE: To establish a diagnostic model for systemic lupus erythematosus (SLE) using proteiomic fingerprint techology. METHODS: Blood samples were collected from 64 cases of SLE, 30 cases of rheumatoid arthritis (RA), 30 cases of Sjogren's syndrome (SS), 25 cases of systemic sclerosis (SSc), as well as 83 healthy controls. Proteomic spectra of these 232 serum samples were generated by proteomic fingerprint technology. Diagnostic model was established by a machine learning algorithm called decision boosting. The sensitivity and specificity of the diagnostic model was validated with a blinded testing set. RESULTS: Sixty differential protein peaks (P<0.05) between SLE and control subjects were indicated, 28 of them were up regulated and 32 were down regulated in SLE patients. The algorithm identified a cluster pattern segregating SLE from non-SLE with sensitivity of 91% and specificity of 92%. The discriminatory diagnostic pattern correctly identified SLE. A sensitivity of 78% and specificity of 96% for the blinded test were obtained when comparing SLE vs non-SLE. CONCLUSION: This diagnostic model using proteiomic fingerprint techology appears to be a promising tools with high sensitivity and specificity in diagnosis of SLE.

[Study on proteomics of familial systemic lupus erythematosus patients in one family from Sichuan, China].

OBJECTIVE: To investigate the proteomic characteristics of systemic lupus erythematosus (SLE) in a SLE family from Sichuan, China which consisting of 7 members with 3 SLE cases, and to find the proteins correlated with the heredity of SLE. METHODS: A total of 153 serum samples were collected from 7 members including 3 SLE sisters in this SLE family, 63 individual SLE patients, as well as 83 healthy controls. The diagnosis of SLE is based on the American College of Rheumatology criteria (1997). All serum samples were analyzed using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) combined with magnetic beads technology. Serum protein profiles were obtained by MALDI-TOF-MS combined with magnetic beads in order to identify predictive biomarkers of risk of suffering SLE. The resulting spectra were analyzed with Biomarker Wizard software 3.1.0. RESULTS: Four discriminative mass/charge (m/z) proteins serving as pathogenic biomarkers were identified on arrays for family SLE cases versus individual SLE and healthy controls. The protein level of peak intensities at m/z of 9342.23 was significantly greater in SLE family group compared with that in individual SLE patients and healthy controls (P<0.05), those of individual SLE patients were significantly greater compared with healthy controls (P<0.05); the proteins level of peak intensities at m/z of 4094.03, 5905.35 and 7973.53 in SLE family group were significantly lower compared with that in individual SLE patients and healthy controls (P<0.05), those of individual SLE patients were significantly lower compared with healthy controls (P<0.05). CONCLUSION: The proteins of m/z of 9342.23, 4094.03, 5905.35 and 7973.53 maybe play a great role in assemble pathogenesis of SLE and predict the risk of suffering SLE. The higher protein level of m/z of 9342.23 and the lower protein level of m/z of 4094.03, 5905.35 and 7973.53, the higher risk of sufferring with SLE.

Serum proteomes of hypertension patients with abundant phlegm-dampness.

OBJECTIVE: To study the serum proteomes of essential hypertension (EH) patients with abundant phlegm-dampness, and try to find special proteins associated with abundant phlegm-dampness syndrome. METHODS: Fifty-nine hypertension patients were included, and the patients were divided into abundant phlegm-dampness syndrome group (39 cases) and non-phlegm-dampness syndrome group (20 cases). To find the special proteins associated with abundant phlegm-dampness, the EH patients with non-phlegm-dampness and another 30 healthy persons were regarded as control. Weak cation nano-magnetic beads were used to capture proteins in serum, and proteomic fingerprint was made by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS). All the proteomic fingerprints were analyzed by Biomarker Wizard 3.1 Software. Then Biomarker Patterns Software (BPS) 5.0 was used to identify the differentiated proteins, which could induce phlegm-dampness. RESULTS: There were 102 differentiated protein peaks between abundant phlegm-dampness and the control group. The best markers of abundant phlegm-dampness were protein peaks with the mass to charge ratio (m/z) of 9,334.958 m/z (the expression increased), 9,280.191 m/z (the expression decreased), 8,030.794 m/z (the expression increased), and 2,941.551 m/z (the expression increased). These four protein peaks found by BPS could induce abundant phlegm-dampness. They could be used to separate the abundant phlegm-dampness syndrome from the healthy persons and the hypertension patients with non-phlegm-dampness. The sensitivity of the model was 93.103% (27/29), specificity was 92% (23/25), false positive rate was 8% (2/25), false negative rate was 6.897% (2/29) and Youden's index was 85.103%. Blind test data indicated a sensitivity of 90% (9/10) and a specificity of 88% (22/25), and the false positive rate was 12% (3/25), false negative rate was 10% (1/10), and Youden's index was 78%. CONCLUSION: The differentiated proteins between the abundant phlegm-dampness group and the control group are the material foundation of abundant phlegm-dampness. The selected differentiated proteins can be used to distinguish the EH patients with abundant phlegm-dampness from the healthy persons and the EH patients with non-phlegm-dampness. The molecular biology diagnosis model can offer an objective and accurate way for TCM syndrome differentiation.

[Establishment of serum protein pattern model for screening pancreatic cancers by SELDI-TOF-MS technique].

OBJECTIVE: To detect the serum specific proteins in pancreatic cancer patients and establish diagnostic model by surface enhanced laser desorption/ionization time of flight mass spectrometry (SELDI-TOF-MS) technique. METHODS: Twenty-nine serum samples from patients of pancreatic cancer were collected before surgery and an additional 57 serum samples from age and sex matched individuals without cancer were used as controls, SELDI-TOF-MS technique and WCX magnetic beads were used to detect the protein fingerprint expression of all the serum samples and the resulting profiles between pancreatic cancer patients and controls were analyzed with biomarker wizard system, established the model using biomarker patterns system software. A double-blind test was used to determine the sensitivity and specificity of the classification model. RESULTS: A panel of four biomarkers (relative molecular weight are 5705, 4935, 5318 and 3243 Da) were selected to set up a decision trees as the classification model for screening pancreatic cancer effectively. The result yielded a sensitivity of 100%, specificity of 97.4%. The double-blind test challenged the model with a sensitivity of 88.9% and a specificity of 89.5%. CONCLUSIONS: SELDI-TOF-MS offers a unique platform for the proteomic detection of serum in pancreatic cancer patients. It also offers a noninvasive method to further study the proteomic changes in the development and progression of pancreatic cancer.

[The contribution for the diagnosis of the tumor-like polypoid lesions of the gallbladder by SELDI-TOF-MS].

OBJECTIVE: To detect the serum specific proteins in tumor-like polypoid lesions of the gallbladder patients and establish diagnostic model. METHODS: Surface enhanced laser desorption/ionization time of flight mass spectrometry (SELDI-TOF-MS) technique and WCX Magnetic Beads were used to detect the serum proteomic patterns of 23 patients with tumor-like PLG, 21 patients with non tumor-like PLG and 26 normal persons. Biomarker Wizard and Biomarker Patterns Software were used in combination to analyze the data. RESULTS: Preliminary screening out 22 representative specific proteins for the diagnosis of the tumor-like PLG. Analysis system under the conditions set selected 3 specific proteins to establish diagnostic model for the tumor-like PLG. The sensitivity and specificity of the model for the diagnosis of the tumor-like PLG were 100% and 89.4%, respectively. CONCLUSION: SELDI-TOF-MS technique can select specific protein of the tumor-like PLG, and establish diagnostic model of the tumor-like PLG.

Exploring pathogenesis of primary biliary cholangitis by proteomics: A pilot study.

AIM: To explore the pathogenesis of primary biliary cholangitis (PBC) by identifying candidate autoantibodies in serum samples by proteomics and bioinformatics. METHODS: Nine antimitochondrial antibody (AMA)-positive PBC patients and nine age- and sex-matched AMA-negative PBC patients were recruited. Antigen enrichment technology was applied to capture autoantigens of human intrahepatic biliary epithelial cells (HiBECs) that are recognized by autoantibodies from the sera of PBC patients. Candidate autoantigens were identified by label-free mass spectrometry. Bioinformatics analysis with MaxQuant software (version 1.5.2.8), DAVID platform, and Cytoscape v.3.0 allowed illustration of pathways potentially involved in the pathogenesis of PBC. RESULTS: In total, 1081 candidate autoantigen proteins were identified from the PBC patient pool. Among them, 371 were determined to be significantly differentially expressed between AMA-positive and -negative PBC patients (P < 0.05). Fisher's exact test was performed for enrichment analysis of Gene Ontology protein annotations (biological processes, cellular components, and molecular functions) and the Kyoto Encyclopedia of Genes and Genomes pathways. Significantly different protein categories were revealed between AMA-positive and -negative PBC patients. As expected, autoantigens related to mitochondria were highly enriched in AMA-positive PBC patients. However, lower levels of AMA were also detected in AMA-negative PBC patients. In addition, autoantigens of AMA-negative PBC patients were mainly involved in B-cell activation, recognition of phagocytosis, and complement activation. CONCLUSION: AMA-negative PBC individuals may not exist, but rather, those patients exhibit pathogenesis pathways different from those of AMA-positive PBC. Comprehensive research is needed to confirm these observations.

Primary biliary cirrhosis: what do autoantibodies tell us?

Primary biliary cirrhosis (PBC) is a chronic, progressive, cholestatic, organ-specific autoimmune disease of unknown etiology. It predominantly affects middle-aged women, and is characterized by autoimmune-mediated destruction of small- and medium-size intrahepatic bile ducts, portal inflammation and progressive scarring, which without proper treatment can ultimately lead to fibrosis and hepatic failure. Serum autoantibodies are crucial tools for differential diagnosis of PBC. While it is currently accepted that antimitochondrial antibodies are the most important serological markers of PBC, during the last five decades more than sixty autoantibodies have been explored in these patients, some of which had previously been thought to be specific for other autoimmune diseases.

[The comparison of the three anti-dsDNA antibody detecting test].

OBJECTIVE: To compare the three Anti-dsDNA antibody detecting test (IIF, ELISA, Farr) with 200 serum samples to evaluate which one has higher sensitivity and specificity. METHODS: 200 serum samples including 120 serum samples of SLE, 20 serum samples of rheumatoid arthritis, 20 serum samples of MCTD, 20 serum samples of SS, 20 serum samples of PSS and 50 serum samples of healthy measured by IIF, Farr and ELISA. RESULTS: Detection the Anti-dsDNA antibody of the serum sample with the methods of IIF, ELISA and Farr. The positive percentage of Anti-dsDNA in SLE is 25%, 32% and 32%, while in RA is 0, 0 and 0; in PSS is 0, 0 and 5%; in SS is 0, 0 and 0; in healthy is 0, 0 and 0. CONCLUSION: Detection the Anti-dsDNA antibody with two method in the same time, especially with IIF and ELISA, will heighten the positive rate than with single method and will be helpful for the diagnosis of SLE.

Promising diagnostic biomarkers for primary biliary cirrhosis identified with magnetic beads and MALDI-TOF-MS.

(PBC) is not a rare disease worldwide. Most patients are diagnosed at the advanced stage, primarily because there are not yet any valid biomarkers available for early diagnosis. Useful biomarkers are absolutely necessary for early detection of PBC. Fortunately, the use of MALDI-TOF-MS and pattern recognition software has been successful in finding specific markers for the early detection of the disease. To screen for potential protein biomarkers in the serum for diagnosing PBC, MALDI-TOF-MS combined with magnetic beads and pattern recognition software was used to investigate 119 serum samples from 44 patients with PBC, 32 controls with other hepatic disease, and 43 healthy controls. A total of 69 discriminant m/z peaks were identified as being associated with PBC. Of them, the m/z peaks at 3445, 4260, 8133, and 16,290 were used to construct a model for the diagnosis of PBC. This diagnostic model can distinguish PBC from non-PBC controls with a sensitivity of 93.3% and a specificity of 95.1%. In our blind test, it demonstrated good sensitivity and specificity: 92.9% and 82.4%, respectively. These results indicate that useful serum biomarkers for PBC can be discovered by MALDI-TOF-MS combined with the use of magnetic beads and pattern recognition software. The pattern of multiple markers provides a powerful and reliable diagnostic method for PBC with high sensitivity and specificity.

Expression of matrix metalloproteinase-1 mRNA in peripheral blood mononuclear cells of systemic lupus erythematosus patients and its relationship with atherosclerosis.

BACKGROUND: Matrix metalloproteinase-1 (MMP-1) plays an important role in atherosclerosis. This study was to examine expression of MMP-1 mRNA in peripheral blood mononuclear cells (PBMCs) of patients with systemic lupus erythematosus (SLE), and to explore its relationship with atherosclerosis in SLE. METHODS: Fluorescent quantitative reverse transcription polymerase chain reaction (RT-PCR) was used to examine the expression of MMP-1 mRNA in PBMCs in 80 SLE patients, including 39 prone to atherosclerosis (Group A) and 41 unprone to atherosclerosis (Group B). Meanwhile, 30 patients who were free of cardiovascular diseases and 30 healthy individuals were selected as disease and normal control group (Groups C and D). The changes of MMP-1 gene expression were analyzed by differences of cycle threshold (DeltaCt), with the following formula: DeltaCt = Ct(target) gene - Ct(reference) gene. RESULTS: The expression level of MMP-1 mRNA in Group A was significantly higher than that of group B (DeltaCt = 8.64 +/- 2.43 vs DeltaCt = 12.09 +/- 2.26, t = 6.588, P < 0.01). The expression level of MMP-1 mRNA of SLE patients was significantly higher than that of Group C (DeltaCt = 10.41 +/- 2.90 vs DeltaCt = 12.29 +/- 2.51, t = 3.135, P < 0.01) and Group D (DeltaCt = 10.41 +/- 2.90 vs DeltaCt = 12.48 +/- 1.69, t = 3.675, P < 0.01). CONCLUSIONS: In comparison to disease and control group, expression of MMP-1 mRNA in PBMCs of SLE patients was significantly elevated, and significant difference of MMP-1 mRNA expression was also found between SLE patients prone and unprone to atherosclerosis, indicating that expression of MMP-1 mRNA may be correlated with the pathogenesis and activity of atherosclerosis in SLE.