Background The purpose of this study was to identify biomarkers for the diagnosis of gout in Chinese Han males using methylation microarray profiling. Methods We screened for differentially methylated genes (DMGs) in gout using a methylation microarray and analyzed the functions of the DMGs using gene ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. We verified gene methylation levels by pyrosequencing and protein levels by enzyme‐linked immunosorbent assays (ELISAs). Statistical analyses were performed using SPSS. Two‐sided p values <0.05 were deemed to be statistically significant for all analyses. Results We identified 20,426 significant differential methylation sites (5719 high‐methylation sites and 14,707 low‐methylation sites). Bioinformatics analysis showed that the DMGs were mainly involved in 43 biological functions, 13 cellular components, 18 molecular functions, and 35 KEGG pathways. We selected opioid receptor delta 1 ( OPRD1 ) for verification of methylation levels between 50 gout patients and 50 controls. The methylation levels of OPRD1 (Chr1:29,139,121) were significantly lower in the gout group ( p < 0.05), while OPRD1 protein levels were significantly higher in the gout group ( p < 0.05). In addition, the AUC of the combination of OPRD1 (Chr1:29,139,121) methylation and OPRD1 protein levels was 0.796 (0.710, 0.883) with a high sensitivity of 82% and a specificity of 68% ( p < 0.001). Conclusions The combination of OPRD1 (Chr1:29,139,121) hypomethylation and high levels of OPRD1 protein is a potential biomarker for gout diagnosis.

Background Ankylosing spondylitis (AS) is a chronic inflammatory arthritic disease, and sacroiliitis, enthesitis, and propensity for sacroiliac and spinal fusion are characteristic pathological features. Interleukin-34 (IL-34) plays a role in the induction and differentiation of osteoclasts. Other inflammatory factors are not directly involved in the induction and differentiation, but play an indirect role by modulating the level of receptor activator of nuclear factor-κB (RANKL) and other molecules during the process of inflammatory bone destruction in AS. However, to our knowledge, the relationship between enthesitis and bone erosion, and IL-34 and RANKL in AS has not yet been elucidated. Objective To determine the correlation between serum IL-34, RANKL, and disease severity including enthesitis and bone erosion in patients with AS and develop multivariable predictive model. Methods We conducted a cross-sectional study of 40 patients with AS, compared with 40 patients with osteoarthritis, and 40 healthy volunteers. Their serum levels of IL-34 and RANKL were measured using enzyme-linked immunosorbent assays (ELISAs). Enthesitis and bone erosion were assessed with real-time ultrasonography. Spearman rank correlation coefficients were determined to analyze the relationship between the variables. Multiple logistic regression was used to determine associations and receiver operating characteristic (ROC) curve analyses were conducted to determine the diagnostic performance of cytokine levels. Results In patients with AS, serum levels of IL-34 (878.9 ± 116.4 pg/mL) and RANKL (155.6 ± 13.8 pg/mL) were significantly (P < 0.01) higher than those in patients with osteoarthritis (626.6 ± 79.0 and 138.1 ± 15.3 pg/mL, respectively) or a healthy group (612.9 ± 61.1 and 104.9 ± 15.4 pg/mL, respectively). Serum levels of IL-34 were not significantly correlated with the levels of RANKL. In patients with AS, serum levels of IL-34 and RANKL adjusted for age and weight were significantly correlated with enthesitis (0.798, P < 0.01; 0.347, P < 0.05, respectively) and bone erosion (0.822, P < 0.01; 0.368, P < 0.05, respectively). The area under the ROC curve (AUC) for the serum levels of IL-34 was 0.995 between patients with AS and healthy individuals. When serum level of IL-34 was >697.1 pg/mL, the sensitivity (SE) was >99% and specificity (SP) was 95.0%. The AUC for IL-34 was 0.982 between patients with AS and patients with osteoarthritis. When serum IL-34 was >688.4 pg/mL, the SE was >99% and SP 85.0%. IL-34 correlation with the number of bone erosions of enthesis was rs = 0.795, P < 0.01. The AUC for serum RANKL was 0.993 between patients with AS and healthy individuals. When serum RANKL was >126.2 pg/mL, the SE was 97.5% and SP 97.5%. The AUC for serum RANKL was 0.798 between patients with AS and patients with osteoarthritis. When serum RANKL was >149.3 pg/mL, the SE was 70% and SP was 80.0%. Conclusions In patients with AS, serum levels of IL-34 and RANKL may be useful indicators of enthesitis, especially for bone erosions. IL-34 is associated with AS-associated enthesis damage and is a potential biomarker for predicting subsequent progression in patients with AS.