Objective:To explore the expression and clinical significance of late autophagy in per-ipheral blood mononuclear cells (PBMCs) of patients with primary gouty arthritis (GA).Methods:Peripheral blood, clinical data, and laboratory tests were collected from 30 patients with acute gout (AG), 30 patients with intermittent gout (IG), and 50 healthy controls (HC). Quantitative polymerase chain reaction (RT-qPCR) was used to detect mRNA expression levels of autophagy-related genes (ATG5, ATG12, ATG16, ATG3, ATG7, ATG10, ATG4B, LC3-2/LC3B). Measurement data conformed to normal distribution were tested using t test or analysis of variance (ANOVA), and non-normal distribution data were tested using Mann-Whitney test or Kruskal-Wallis H test. SNK was used for pairwise comparison among the three groups. Correlation between variables was tested by Spearman correlation analysis. Results:① The expression level of ATG5 mRNA,ATG12 mRNA, ATG16 mRNA, ATG10 mRNA and LC3-2 mRNA in the AG group was lower than that of the IG group and the HC group, and the expression level of the IG group was lower than that of the HC group[9.16×10 -3(6.04×10 -3, 15.00×10 -3) vs 14.48×10 -3(9.95×10 -3, 21.38×10 -3) vs 0.08×10 -3(12.21×10 -3, 42.79×10 -3), H=19.377, P<0.001; 18.89×10 -3(13.85×10 -3, 24.92×10 -3) vs 21.13×10 -3(12.11×10 -3, 28.06×10 -3) vs 33.57×10 -3(13.11×10 -3, 49.89×10 -3), H=7.545, P=0.023; 8.72×10 -3(4.96×10 -3, 13.74×10 -3) vs 10.62×10 -3(7.48×10 -3, 24.71×10 -3) vs 20.07×10 -3(11.99×10 -3, 39.56×10 -3), H=20.962, P<0.001; 1.05×10 -3(0.73×10 -3, 1.84×10 -3) vs 1.60×10 -3(0.93×10 -3, 2.58×10 -3) vs 1.69×10 -3(1.05×10 -3, 3.54×10 -3), H=8.193, P=0.017; 2.31×10 -3(1.22×10 -3, 3.53×10 -3) vs 2.78×10 -3(1.68×10 -3, 5.96×10 -3) vs 3.68×10 -3(2.00×10 -3, 5.67×10 -3) , H=7.135, P=0.028]. The expression level of ATG4B mRNA in the AG and IG group was higher than that in HC group, and there was significant difference between IG group and AG group, IG group and HC group[9.95×10 -3(6.32×10 -3, 12.23×10 -3) vs 10.86×10 -3 (8.80×10 -3, 17.03×10 -3) vs 8.07×10 -3(5.52×10 -3, 11.63×10 -3), H=8.531, P=0.014]. There was no significant difference between the ATG3 mRNA and ATG7 mRNA groups ( H=0.539, 3.739, bothall P values >0.05). ② The results of Spearman correlation analysis suggested that in patients with acute gout, ATG3 was negatively correlated with PDW and MPV ( r=-0.499, P=0.006; r=-0.463, P=0.011); ATG4B was positively correlated with HDL-C ( r=0.408, P=0.048); ATG7 was negatively correlated with GLOB ( r=-0.554, P=0.001); ATG10 was positively correlated with ALB ( r=0.412, P=0.024) and negatively correlated with Crea and hsCRP ( r=-0.459, P=0.011; r=-0.375, P=0.045); ATG12 was negatively correlated with MO ( r=-0.434, P=0.017); ATG16 was negatively correlated with ALT and AST ( r=-0.389, P=0.034; r=-0.366, P=0.047); LC3-2 was positively correlated with UA ( r=0.381, P=0.041) and negatively correlated with MPV and PDW ( r=-0.413, P=0.026; r=-0.449, P=0.015). In patients with intermittent gout, ATG3 and ATG4B were negatively correlated with apoB100 ( r=-0.555, P=0.011; r=-0.462, P=0.040); ATG5 was negatively correlated with Crea ( r=-0.456, P=0.011); ATG10 was negatively correlated with TC, LDL-C, and apoB100 ( r=-0.526, P=0.017; r=-0.556, P=0.011; r=-0.515, P=0.020). Conclusion:Autophagy is involved in the development of gout, and is correlated with ibflammatory and metabolic indicators, suggesting that autophagy is an important feature in the pathogenesis of GA.

Objective:To explore the expression and clinical significance of Janus protein tyrosine kinase/signal transducer and activator of transcription (JAK3/STAT5) signaling pathway in peripheral blood mononuclear cells (PBMCs) of patients with primary gouty arthritis (GA).Methods:Peripheral blood, clinical data and laboratory tests were collected from 50 patients with acute gout (AG), 50 patients with intermittent gout (IG) and 50 healthy controls (HC). Quantitative real-time-polymerase chain reaction (RT-qPCR) was used to detect mRNA expression level of JAK3/STAT5 related genes (JAK3, STAT5a, STAT5b). Enzyme linked immune sorbent assay (ELISA) was used to detect interleukin-2 (IL-2) concentration in subject′s plasma. Measurement data among the three groups that was in accordance with normal distribution was analyzed by one-way analysis of variance, pairwise comparisons using LSD, non-normal distribution data was analyzed by Mann-Whitney test or Kruskal-Wallis H test, and correlation analysis between variables was analyzed using Spearman correlation analysis. Results:① The mRNA expression levels of JAK3, STAT5a and STAT5b were significantly different among the three groups ( F=50.13, P<0.01; F=7.573, P=0.000 7; F=12.14, P<0.01), of which the JAK3 mRNA expression level in the HC group [(606±65)×10 -4] was significantly higher than that in the AG group [(103±13)×10 -4] and IG group [(114±24)×10 -4], and the difference was statistically significant (both P<0.01), while the STAT5a mRNA expression level in the AG group [(89±9)×10 -4] was significantly higher than that in the IG group [(59±4)×10 -4] and HC group [(61±4)×10 -4], and the difference was statistically significant ( P=0.002, P=0.003 9), and the expression level of STAT5b mRNA in HC group [(60±5)×10 -4] was significantly lower than that in AG group [(95±7)×10 -4] and IG group [(98±7)×10 -4], and the difference was statistically significant ( P=0.000 2, P<0.000 1). ② The difference of IL-2 concentration in plasma among the three groups was statistically significant ( F=22.87, P<0.01), and the serum IL-2 concentration in the AG group [(87.9±8.4) pg/ml] was significantly higher than that in the IG group [(32±4) pg/ml] and HC group [(44±4) pg/ml], and the difference was statistically significant (both P<0.01). ③ Spearman correlation analysis showed that the mRNA expression of STAT5a and STAT5b was positively correlated with the absolute value of neutrophils in patients with gout ( r=0.282, P<0.05; r=0.257, P<0.05). Conclusion:The IL-2/JAK3/STAT5 signaling pathway is involved in the occurrence and development of gout, suggesting that this pathway may play a key role in the pathogenesis of gout.

Objective:To detect the expression of interleukin 2 (IL-2)/Janus kinase 3/signal transduction and transcriptional activator 5 (JAK3/STAT5) signaling pathway in peripheral blood of patients with ankylosing spondylitis (AS) and explore its mechanism in the development and progression of AS.Methods:Clinical data, peripheral blood and laboratory tests of 30 patients with active AS (ASA), 30 patients with stable AS (ASS) and 50 healthy subjects (HC) were collected. The mRNA expression levels of JAK3, signal transduction and transcription activator 5a (STAT5a) and signal transduction and transcription activator 5b (STAT5b) were detected by quantitative real-time-polymerase chain reaction (RT-qPCR). The expression levels of JAK3, STAT5a and STAT5b proteins and phosphorylated proteins were detected by Western-blot. Plasma IL-2 concentration was determined by enzyme-linked immunosorbent assay (ELISA). Two independent samples t-test or one-way analysis of variance were used for measurement data consistent with normal distribution, LSD- t test was used for pairwise comparison between the three groups, Mann-Whitney U test or Kruskal-Wallis H test was used for non-normal distribution, χ2 test was used for correlation analysis of categorical variables. Spearman correlation analysis was used for correlation analysis between variables, and receiver operating characteristic (ROC) curve was used to evaluate the value of JAK3, STAT5a and STAT5b mRNA expression levels in monitoring AS activity. Results:① The mRNA expression levels of JAK3, STAT5a and STAT5b were significantly different among the three groups ( F=65.98, P<0.001; F=21.15, P<0.001; F=13.67, P<0.001). JAK3 mRNA expression in ASA group (2.5±0.9) was significantly higher than that in ASS group (1.1±0.4) and healthy subjects (1.0±0.5), the difference was statistically significant (both P<0.001). The mRNA expression level of STAT5a in ASA group (1.4±0.3) was significantly higher than that in ASS group (0.9±0.3) and healthy subjects group (1.0±0.3), the difference was statistically significant (both P<0.001). STAT5b mRNA expression level in ASA group (1.5±0.6) was significantly higher than that in ASS group (1.0±0.4) and healthy subjects (1.0±0.4), the difference was statistically significant (both P<0.001). The expression level of JAK3 mRNA in HLA-B27 positive group (1.9±1.0) was higher than that in HLA-B27 negative group (1.4±0.6), and the difference was statistically significant ( t=-2.22, P=0.032). The phosphorylation levels of JAK3, STAT5a and STAT5b showed statistically significant differences among the three groups ( F=91.56, P<0.001; F=25.15, P< 0.001; F=178.59, P<0.001). The phosphorylation level of JAK3 protein in ASA group (1.04±0.08) was significantly higher than that in ASS group (0.568±0.019) and healthy subjects (0.536±0.064), the difference was statistically significant (both P<0.001). The phosphorylation level of STAT5a protein in ASA group (1.166±0.096) was significantly higher than that in ASS group (0.923±0.018) and healthy subjects (0.911±0.017), the difference was statistically significant (both P<0.001). The phosphorylation level of STAT5b protein in ASA group (0.81±0.05) was significantly higher than that in ASS group (0.21±0.03) and healthy subjects (0.24± 0.07), the difference was statistically significant (both P<0.001). The difference of plasma IL-2 concentration among the three groups was statistically significant ( F=3.32, P=0.040). The IL-2 concentration in the ASA group [(110±40) pg/ml] was significantly higher than that in the ASS group [(89±40) pg/ml] and the healthy group [(88±39) pg/ml], the difference was statistically significant ( P=0.044, P=0.016). ② Spearman correlation analysis showed that STAT5a mRNA expression level was positively correlated with platelets in AS patients ( r=0.353, P=0.006). JAK3 mRNA expression level in ASA group was positively correlated with IL-2 concentration ( r=0.766, P<0.001), and negatively correlated with estimated glomerular filtration rate ( r=-0.485, P=0.007). STAT5a mRNA expression level was positively correlated with erythrocyte sedimentation rate ( r= 0.680, P<0.001), and STAT5b mRNA expression level was positively correlated with hypersensitive C-reactive protein (CRP) ( r=0.823, P<0.001). ③ The ROC curve showed that JAK3 mRNA expression level predicted the area under ROC curve (AUC) of ASA with a 95% CI of 0.920 (0.853, 0.987), sensitivity and specificity of 86.7% and 90.0%, respectively. STAT5a mRNA expression level predicted the AUC 95% CI of ASA was 0.874 (0.787, 0.961), and the sensitivity and specificity were 96.7% and 66.7%, respectively. STAT5b mRNA expression level predicted the AUC 95% CI of ASA was 0.749 (0.617, 0.881), and the sensitivity and specificity were 73.3% and 80.0%, respectively. Conclusion:This study suggests that IL-2/JAK3/STAT5 may be involved in the pathogenesis of AS, and JAK3 mRNA can be used as a biological indicator to monitor the activity of AS disease.