Objective To explore the influence of prozone effect on anti-nuclear antibodies (ANA) testing by indirect immunofluorescence assay (IIFA).Methods The samples with high titer of ANA (≥1∶1 000) were selected from 880fresh serum samples, and were subsequently diluted in 1∶100, 1∶1 000and 1∶10 000ratio.Prozone effect was defined as fluorescence intensity from 1∶1 000dilution was stronger than that from1∶100dilution.The samples with prozone effect were determined manually or by Sprinter XL and EUROPattern.The samples with prozone effect were further characterized by combinations of fluorescence patterns, fluorescence intensities and autoantibody specificities.Results A total of 880samples were tested.Importantly, 34samples displayed prozone effect (3.86％in total and 29.57％in samples with ANA≥1∶1 000).Interestingly, prozone effect was identified by manual detection as well as by Sprinter XL with similar fluorescence patterns and fluorescence intensities.Notably, EUROPattern can only select the central area for identification.Among all samples with prozone effect, 74.42％samples exhibited fluorescence intensities of≥1∶10 000.Speckled pattern was the most prevalent fluorescence patterns in samples with prozone effect (46.51％).In addition, anti-RNP antibodies (62.79％) were the most popular autoantibodies in samples with prozone effect, followed by anti-dsDNA antibodies (51.16％) and anti-SSA antibodies (51.16％).Conclusion Prozone effect was present in ANA testing, especially in samples with high titers, resulting in underestimating the titers.The study highlighted that special attention should be paid to the prozone effect in clinical practice.

Objective To investigate the clinical characteristics and the pathogens of recurrent urinary tract infection (RUTI) after renal transplantation.Methods The data of adult recipients with UTI from November 2011 to December 2016 were retrospectively analyzed.The recipients were divided into single UTI (SUTI) group and RUTI group.The clinical characteristics and pathogens were analyzed,and the independent risk factors of RUTI were analyzed using logistic regressionmodel.Results Fifty-three cases were selected,including 29 cases of SUTI and 24 cases of RUTI.The positive rate of blood culture (55％ vs.25％,P =0.042) and the concentration of FK506 in the peri-infection period (11.0 + 3.4 ng/mL vs.8.6 + 3.2 ng/mL,P =0.024) in the RUTI group were significantly higher than that those in the SUTI group at the first UTI.The increased concentration of FK506 in the peri-infection period at the first UTI was an independent risk factor for RUTI (β:0.282,95％ CI:1.026-1.713,P＜0.05).There were 86 infection events in 53 patients,and pathogenic microorganisms were cultured in blood culture and urine culture for 86 times.The positive frequency of culture in the RUTI group was higher than that in the SUTI group,but not significantly.The most common pathogenic microorganisms included Escherichia coli (17 times),pseudomonas aeruginosa (16 times),and Enterococcus (16 times).Conclusion Reduction of the FK506 concentration during the peri-infection period at the first UTI is the key to prevent RUTI after renal transplantation.The empirical antibiotics for RUTI should be sensitive for Escherichia coli (ESBL +)and pseudomonas aeruginosa.

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.

Objectives To explore the clinical significance of autoantibodiesin individuals who accept a routine physical examination.Methods From April to June 2012, the serum of 932 individualsincluding 649 males and 283 females, from department of routine physical examination center of Peking Union Medical College Hospitalwerecollected , it uesd IIF for ANA, line immunoassay ( LIA) for specific ANAs antibodies and ELISA for the other antibodies , includinganti-CCP antibodies , AMA-M2, ACL antibodies and anti-anti-β2GPⅠantibodies.Chi-square test was used for data measurement of positive rate of autoantibodies in men and women;Fisher′s exact test was used when the data not meet the conditions of Chi-square test.Individualswith high-risk of autoimmune disease according to the results ofautoantibodies ( Titersof autoantibodies≥2-fold cut-off and accompanied with other autoimmune diseases related laboratory abnormalities) were recalled to visit doctor.Results Of the 932 cases, the overall positive rate of ANA was 11.27%.The positive rate of ANA was19.79%inwomen, which was significantly higher than thatin men (7.09%)(χ2 =32.6, P<0.01); the overall positive rate of ANAswas 8.69%, and the positive rate of ANA was13.43%inwomen, whichwas significantly higher than that in men ( 6.63%) (χ2 =11.49, P <0.01);the overall positive rates of AMA-M2, anti-CCP antibodies , anti ACL antibodies and anti β2GPⅠantibodies were 3.22%, 0.54%, 2.90%and 0.21% respectively , which were 2.83%, 0.71%, 3.18%and 0.71 % in women , and 3.39%, 0.46%, 2.77% and 0.00% in men respectively , there was no statistically significant of positive rate between female and male 58 patients accounting for 6.22% in high-risk of autoimmune disease were recalled , of which 15 cases, accounting for 1.61% were diagnosed or highly suspected of autoimmune diseases (AID) of the 15 patients, 11 patients accounting for 1.18% were diagnosed AID, including 6 CTD, 3 pSS, 1 RA and 1 pSS/PBC;4 patients were highly suspected as AID , including 3 suspected CTD and 1 suspected pSS.The titers concentration of the positive antibodies in patients with confirmed or suspected AID ≥ 3 times cut-off.Conclusions The positive rate of autoantibodies in individuals of physical examination is high , but there is clinical significance when the titers concentration of positive autoantibodies ≥ 3 times of the cut-off.Positive-autoantibodies patients with high-risk of autoimmune disease need professional clinician to provide follow-up, consulting and health education for early discovery, timely diagnosis, and proper treatment of AID.

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 To identify biomarkers in cerebrospinal fluid (CSF) by proteomic technology and develop a diagnostic model for neuropsychiatric lupus (NPSLE).Methods CSF proteomic spectra of 27 patients with NPSLE before and after treatment,and 27 controls including 17 patients with scoliosis,and 10 SLE patients without neuropsychiatric manifestation (non-NPSLE) were generated by matrix assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) combined with weak cationic exchange (WCX) magnetic beads.Data were analyzed with t test,non-parametric Kruskal-Wallis H test or Wilcoxon sign-rank test.A decision tree model for NPSLE classification was built based on the discriminating peaks.In addition,CSF samples of 12 patients with NPSLE,12 patients with lumbar disc herniation and 9 patients with other neurological conditions were employed as blind test group to verify the accuracy of the model.Results Twelve discriminating mass-to-charge (m/z) peaks were identified between NPSLE and controls.The diagnostic decision tree model,built with a panel of m/z peaks 8595,7170,7661,7740 and 5806,recognized NPSLE with the sensitivity and specificity of 92.6％ and 92.6％ based on training group samples,91.7％ and 85.7％ based on blind test group,respectively.Conclusion Potential CSF NPSLE biomarkers are identified by proteomic technology,the novel diagnostic model is sensitive and relatively specific for the diagnosis of NPSLE.

Objective To investigate genetic polymorphisms of IRF7/KIAA1542 (rs4963128, rs2246614) and STAT4 (rs7574865) and their relationships with lupus nephritis and various autoantibodies present in Chinese Han population of SLE patients. Methods A total of 748 SLE patients and 750 healthy controls belonging to the Chinese population were enrolled into this study. They were genotyped using MALDI-TOF-MS method. Autoantibodies including anti-SSA, anti-SSB, anti-Sm, anti-RNP and anti-dsDNA were determined either by indirect immunofluorescence or double immunodiffusion methods. Results In the healthy group, rs7574865 (STAT4) T/T, T/G, G/G genotype frequency and T, G allele frequencies were as follows: 9.4% , 45. 6% , 45. 0% , 32. 2% , 67. 8% , the corresponding case group as follows: 17.0% , 48.1%, 34.9%, 41.0%, 59.0%, genotype and allele frequencies were significantly different (x2 = 26.30, P<0.01). Compared with the control group, in the case group, T/T genotype frequency and T allele frequency were significantly increased, and in three genetic models ( additive model, dominant model, recessive model), the genotype frequencies were significant difference (P <0. 01). Two polymorphic loci of rs4963128 and rs2246614 (IRF7/KIAA1542) were not statistically significant (x2 =4.49,5.32,P>0.05) in case group and control group, but the rs2246614 genotype frequencies had a statistically significant in recessive model (P <0. 05) , whereas rs4963128 genotype frequencies was no significant difference in the three genetic model (P=0.068, 0.958, 0.067, respectively). In the clinical subphenotype analysis, IRF7/KIAA1542 (rs4963128) in lupus nephritis group (OR = 2. 69, 95% CI = 1. 89-3. 82, P < 0.01) ,anti-SSA antibody group ( OR = 0. 61, 95% CI = 0. 43-0. 87, P < 0. 05 ) and anti-SSB antibody group ( OR =0. 36, 95% CI = 0. 23-0. 56, P < 0.01) of the analysis were statistically significant. At the same time, IRF7/KIAA1542 (rs2246614) in the joint comparison of positive and negative symptoms were also statistically significant (OR=1.34, 95% CI = 1. 06-1. 69, P < 0. 05). Conclusions This findings provide strong evidence suggesting that STAT4 ( rs7574865 ) is the susceptible factor of SLE in Chinese Han population. However, there is not a significant relationships between IRF7/KIAA1542 (rs4963128, rs2246614) polymorphisms and the risk of SLE, but the associations of IRF7/KIAA1542 (rs4963128, rs2246614) with the a variety of clinical subphenotypes, such as lupus nephritis, joint symptoms and production of anti-SSA antibody and anti-SSB antibody implicates IRF7/KIAA1542 as a putative candidate gene of SLE.

Objective To study the protection of vanillin derivative VND3207 on the cytogenetic damage of mouse bone marrow cell induced by ionizing radiation.Methods BALB/c mice were randomly divided into five groups:normal control group,2 Gy dose irradiation group,and three groups of 2 Gy irradiaiton with VND3207 protection at doses of 10,50 and 100 mg/kg,respectively.VND3207 was given by intragastric administration once a day for five days.Two hours after the last drug administration,the mice were irradiated with 2 Gy γ-rays.The changes of polychromatophilic erythroblasts micronuclei (MN),chromosome aberration (CA) and mitosis index (MI) of mouse bone marrow cells were observed at 24 and 48 h after irradiation.Results Under the protection of VND3207 at the dosages 10,50,100 mg/kg,the yields of poly-chromatophilic erythroblasts MN and CA of bone marrow cells were significantly decreased(t = 2.36-4.26,P ＜ 0.05),and the marrow cells MI remained much higher level compared with the irradiated mice without drug protection (t = 2.58,2.01,P ＜ 0.05).The radiological protection effect was drug dose-dependent,and the administration of VND3207 at the dosage of 100 mg/kg resulted in reduction by 50% and 65% in the yields of MN and CA,respectively.Conclusions VND3207 had a good protection effect of on γ-ray induced cytogentic damage of mouse bone marrow cells.

Objective To explore the prevalence of the anti-saccharomyces cerevisiae antibody (ASCA) in patients with primary biliary cirrhosis and evaluate it's clinical significance. Methods The subtypes of ASCA including IgA and IgG in blood samples from 162 patients with PBC, 44 patients with AIH,4-1 patients with other non-autoimmune liver diseases controls (LDC), 144 patients with inflammatory bowel disease (IBD) and 35 healthy controls were measured by ELISA. Chi-square test and Mann Whitney U test were used for statistical analysis. Results The positive rate of ASCA-IgA in PBC was 24.1%, which was higher than that in ulcerative colitis (UC) group ( 11.6%,χ2=5.5, P＜0.05 ) and healthy controls (0, χ2=10.5,P＜0.01 ). Compared with the AIH group (20.5%) or LDC group ( 14.6% ) or Crohn's disease (CD) (34.5%),there was no statistically significant difference (P＞0.05). The prevalence of ASCA-IgG in PBC was 11.1%,lower than the CD group (27.6%, χ2=8.9, P＜0.01 ), but higher than that in the healthy controls (0, χ2=10.5,P＜0.01 ). There was no statistically significant difference (P＞0.05) between PBC and the AIH group (15.9%)or LDC group (7.3%) or UC group (8.1% ). The positive rate of both ASCA-IgA and ASCA-IgG in PBC was only 6.2%, statistically lower than that of the CD group ( 17.2%, χ2=6.3, P＜0.05). The prevalence of ASCA-IgA or ASCA-IgG in PBC was 29.0%, which was statistically lower than that of the CD group (44.8%, χ2=4.8,P＜0.05), but higher than that of the UC group (χ2=5.9, P＜0.05) or healthy controls (χ2=13.3, P＜0.01).ASCA was detected more frequently in PBC patients with positive anti-GP210 antibody than in anti-GP210 antibody negative PBC patients (38.6% vs 23.8%,χ2=3.9, P＜0.05). The positive rate of ASCA between AMA positive and negative patients with PBC or anti-SP100 antibodies positive and negative patients with PBC was not significantly different. PBC patients with positive ASCA-IgA had higher level of TBIL, DBIL, TBA, LD,IgA, IgM, ESR and lower level of ALB, A/G, CHE than patients with negative ASCA-IgA. There was no statistically significant difference in liver injury indicators and immune function parameters between patients with positive ASCA-IgG and negative ASCA-IgG. Conclusion ASCA is not an IBD-specific antibody. There is a high prevalence of ASCA in patients with PBC, especially the subtype of ASCA-IgA. ASCA-IgA is found to be associated with the severity of liver damage and immune activity whereas ASCA-IgG is not associated with them.

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.