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Objective:To investigate the expression and clinical significance of decoy receptor 3 (DcR3) and its signal pathway-related molecules in PBMCs of patients with ankylosing spondylitis (AS).Methods:Peripheral blood samples, clinical data and laboratory test results were collected from 100 patients with ankylosing spondylitis [50 patients with AS activity (ASA), 50 patients with AS stability (ASS)], 30 patients with osteoarthritis and 30 patients with gouty arthritis (as disease control group), and 60 healthy controls (HC). The mRNA expression levels of DcR3 and its signal pathway related genes (DR3, TL1A, Fas, FasL, LIGHT, LIGHTR, LTβR) were measured by real-time fluorescence quantitative polymerase chain reaction. Measurement data among the three groups in normal distribution were analyzed by t test or one-way analysis of variance, pairwise comparisons using LSD- t test, non-normal distribution data were analyzed by Mann-Whitney test or Kruskal-Wallis H test, χ2 test was used for correlation analysis of categorical variables. Correlation analysis between variables were analyzed using Spearman correlation analysis. Results:① By comparing the AS group, disease control group and HC group, the expression levels of DcR3 mRNA and DR3 mRNA in the AS group were lower than those in disease control group and HC group, and DcR3 mRNA and DR3 mRNA in disease control group were lower than those in the HC group {DcR3mRNA: [6.21 (3.89, 10.70)]×10 -4vs [9.51 (5.89, 16.65)]×10 -4vs [17.81 (11.27, 24.20)]×10 -4, H=55.28, P<0.001; DR3 mRNA: [41.05 (24.09, 66.95)]×10 -4vs [58.28 (28.41, 94.38)]×10 -4vs [94.79 (54.07, 144.51)]×10 -4, H=37.10, P<0.001}. The expression level of TL1A mRNA in the AS group was higher than that in disease control group {[14.71(4.91, 42.22)]×10 -4vs [4.00(1.07, 16.60)]×10 -4vs [7.70 (3.52, 27.83)]×10 -4, H=17.71, P<0.001}; The expression level of Fas mRNA in AS group and disease control group was lower than that in HC group {[20.99(4.63, 62.89)]×10 -4vs [23.97(15.82, 38.99)]×10 -4vs [78.45 (27.32, 146.46)]×10 -4, H=31.17, P<0.001}. The expression level of FasL mRNA in AS group was higher than that in disease control group and HC group {[42.87(6.57, 91.21)]×10 -4vs [5.45(2.83, 10.32)]×10 -4vs [6.88 (4.57, 23.79)]×10 -4, H=46.42, P<0.001}. The expression level of LIGHTR mRNA in AS group was lower than that in disease control group {[52.66 (7.20, 143.21)]×10 -4vs [98.80 (53.11, 166.24)]×10 -4vs [63.47(40.85, 138.07)]×10 -4, H=11.96, P<0.001}. There were no significant differences in LIGHT mRNA and LTβR mRNA among all groups ( H=0.86, P>0.05; H=3.18, P>0.05). ②The expression levels of DcR3 mRNA, DR3 mRNA and Fas mRNA in ASA group and ASS group were lower than those in HC group. DcR3 mRNA in ASA group was higher than that in ASS group, and DR3 mRNA in ASA group was lower than that in ASS group {DcR3 mRNA: [7.28 (4.92, 16.56)]×10 -4vs [4.59 (2.49, 7.03)]×10 -4vs [17.81 (11.27, 24.20)]×10 -4, H=62.63, P<0.001; DR3 mRNA: [30.93(16.18, 66.66)]×10 -4vs [47.17(29.91, 67.40)]×10 -4vs [94.79(54.07, 144.51)]×10 -4, H=41.48, P<0.001; Fas mRNA: [20.04(3.29, 62.30)]×10 -4vs [22.49(5.63, 64.79)]×10 -4vs [78.45(27.32, 146.46)]×10 -4, H=23.54, P<0.001}. The expression levels of TL1A mRNA and LTβR mRNA in the ASA group were higher than those in the ASS group and the HC group {TL1A mRNA: [32.36(10.09, 97.84)]×10 -4vs [9.98(1.29, 21.63)]×10 -4vs [7.70(3.52,27.83)]×10 -4, H=21.14, P<0.001; LTβR mRNA: [6.13(2.16,20.06)×10 -4vs [2.13(0.53,8.04)]×10 -4vs [2.72 (1.24,5.73)]×10 -4, H=12.86, P<0.001}. The expression level of FasL mRNA in the ASA group and the ASS group was higher than that in the HC group {[60.70 (8.16, 106.16)]×10 -4vs [30.14 (5.37, 78.40)]×10 -4vs [6.88 (4.57, 23.79)]×10 -4, H=18.99, P<0.001}. The expression level of LIGHTR mRNA in ASS group was lower than that in HC group {[49.79(10.75, 168.48)]×10 -4vs [15.92(3.27, 105.91)]×10 -4vs [63.47(40.85, 138.07)]×10 -4, H=11.80, P<0.001]. There was no significant difference in LIGHT mRNA among all groups ( H=4.15, P>0.05). ③Spearman correlation analysis showed that DcR3 level was positively correlated with BASDAI score and hsCRP in AS patients ( r=0.52, P<0.001; r=0.35, P<0.01), and DR3 level was negatively correlated with BASDAI score, ESR and hsCRP level ( r=-0.28, P<0.001; r=-0.25, P<0.001; r=-0.31, P<0.001). TL1A was positively correlated with BASDAI score, ESR and hsCRP level ( r=0.23, P=0.046; r=0.26, P=0.015; r=0.25, P=0.017). Conclusion:DcR3 and its signal pathway-related molecules are differentially expressed in PBMCs of patients with AS, suggesting that they may participate in the occurrence and development of AS.
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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.
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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.
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Objective:To explore the three long non-coding RNA (long non-coding ribonucleic acid, the expression of lncRNA NR_002578, NR_038264 and NR_046252) in peripheral blood mononuclear cells (PBMCs) of patients with primary gout arthritis (GA) and their clinical value.Methods:Peripheral venous blood, clinical data and laboratory data were collected from 60 gout patients (including 30 AG patients in acute stage and 30 IG patients in intermittent stage) and 50 healthy subjects (HC group). Quantitative reverse transcription PCR (RT-qPCR) was used to detect the expression levels of PBMCs of 3 lncRNAs in GA and HC groups, and the differences of 3 lncRNAs expression levels in different groups were compared and the correlation analysis was conducted with clinical indicators. Receiver operating characteristic curve (ROC) was constructed to evaluate the possible efficacy of lncRNAs in gout diagnosis. The measurement data conforming to normal distribution were tested by t test or variance analysis, and non-normal distribution were tested by Mann-Whitney U test or Kruskal-Wallis H test. The comparison among the three groups was conducted by SNK. Results:① The expression of NR_002578 in GA was significantly lower than that in HC [60.2(16.8, 100.1)×10 -3vs 149.5 (92.6, 221.8)×10 -3, Z=-5.75, P<0.001], subgroup analysis showed that the expression of NR_002578 in AG was significantly lower than that in IG and HC [34.3(8.6, 72.8)×10 -3vs 88.3(47.7, 109.6)×10 -3vs 149.5(92.6, 221.8)×10 -3, H=40.12, P<0.001], and lower in IG than that in HC ( P<0.001). The expression of NR_046252 in GA was significantly higher than that in HC [6.5(2.1, 21.5)×10 -3vs 2.1(1.2, 3.5)×10 -3, Z=-4.21, P<0.001]. The expression of NR_046252 in AG and IG were higher than that of the HC group [6.3(2.0, 12.0)×10 -3vs 7.2(2.4, 30.6)×10 -3vs 2.1(1.2, 3.5)×10 -3, H=21.33, P<0.001], but there was no significant difference between the AG and IG group ( P>0.05). ② Spearman correlation analysis showed that NR_002578 expression was negatively correlated with erythrocyte sedimentation rate (ESR) ( r=-0.29, P=0.024)and hypersensitive C-reactive protein (hs-CRP) ( r=-0.35, P=0.006) in gout patients. ③ The areas under ROC curve of NR_002578 and NR_046252 for diagnosing gout were 0.819 and 0.750, respectively. Conclusion:The abnormal expression of NR_002578 and NR_046252 in gout patients suggests that NR_002578 may be involved in the pathogenesis of gout.
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Autoinflammatory diseases (AIDs) are a group of disorders characterized by dysfunction of innate immunity which caused by gene mutations leading to coded proteins changes, finally causing uncontrolled systemic inflammation. AIDs are a group of rare rheumatic and inflammatory diseases. Here, Chinese Rheumatology Association summarized manifestations of the main AIDs, and to standardize the methods for diagnosis of AIDs.
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Objective:To screen for circle RNA (circRNA) differentially expressed in peripheral blood mononuclear cells (PBMCs) of patients with ankylosing spondylitis (AS), and to analyze its expression profile to explore the role of circRNAs in the pathogenesis of AS.Methods:CircRNA microarray chip technology was used to detect the expression of circRNAs in PBMCs of 3 patients with active AS, 3 patients with stable AS and 3 healthy controls (HC), and then screening for differentially expressed circRNAs by fold change (FC) and P value. Then differentially expressed circRNAs among the circRNAs with the highest differential expression were selected randomly to verify the chip results by real-time fluorescence quantitative polymerase chain reaction (qPCR); Differentially expressed circRNAs were subjected to Gene Ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, and microRNA (miRNA) target prediction software was used to predict the circRNA/miRNA interaction relationship. Finally, the data were statistically analyzed by t test and Mann-Whitney U test. Results:① Chip text results showed that there were 800 circRNAs with significantly different expression (FC>1.5, P<0.05) in active AS than HC, of which 466 were up-regulated and 334 were down-regulated; the stable AS had a total of 1 149 significantly differentially expressed when compared with the HC (FC>1.5, P<0.05) circRNAs, of which 589 were up-regulated and 560 were down-regulated. 233 circRNAs were significantly differentially expressed (FC>1.5, P<0.05) between active AS and stable AS, of which 145 were up-regulated and 88 were down-regulated. ②The RT-qPCR verification results suggested that the expression trends of the four differentially expressed circRNAs were consistent with the results of the chip. ③ GO analysis results suggested that these differentially expressed circRNAs were mainly involved in nonsense-mediated mRNA decay, Rho GTPase binding and other processes. The analysis showed that the KEGG pathway were enriched in Th17 cell differentiation and chemokine signaling pathways. The results of miRNA target prediction software analysis suggested that differentially expressed circRNAs might play a role by targeting miR-650, let-7b-5p and other miRNAs. Conclusion:Compared with HC group, there were differentially expressed circRNAs in Peripheral Blood Mononuclear Cells (PBMCs) of AS patients, The results of this study suggest that these circRNAs may be involved in the pathogenesis of AS.
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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.
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Objective:To analyze the expression of circular RNA (circRNA) in peripheral blood mononuclear cells (PBMCs) of patients with gout and to explore the possible mechanism of circRNA in the pathogenesis of gout.Methods:Peripheral blood samples of 24 patients with acute gout (AG), 24 patients with intermittent gout (IG) and 24 healthy control subjects (HC) were collected. Three cases of AG, IG, and HC were randomly selected, and the differentially expressed circRNA in PBMCs was screened by human circNA microarrays. The 6 circRNAs with large differences between the two comparison groups were selected, and the relative expression levels of 6 circRNAs in all the collected 72 PBMCs of the study subjects were detected by real-time fluorescent quantitative polymerase chain reaction (RT-qPCR). The significantly differentially ex-pressed circRNA (fold change>1.5, P<0.05) was analyzed by GO analysis, Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis, and its interaction with microRNA (miRNA) was predicted. The median (interquartile range) was used to describe the data, and the Mann-Whitney U test was used for comparison between groups. Results:(1) The microarray analysis results showed that compared with the HC group, the AG group and the IG group had 116 and 41 significantly differently expressed circRNAs, respectively; com-pared with the IG group, the AG group had 105 significantly differently expressed circRNAs. (2) Among the 6 circRNAs verified by PT-qPCR, the expression trends of 5 were consistent with the microarray results. The expression of hsa_circRNA_105034 in the AG group [5.17(4.60)] was statistically significantly different com-pared to the IG [1.68(2.39)] and HC [0.90(0.73)] groups (AG vs IG: Z=-4.413, P<0.01; AG vs HC Z=-5.052, P<0.01). (3) Bioinformatics analysis: ① GO analysis found that differential circRNA swere mainly involved in DNA transcriptional regulation, positive cell regulation and protein modification, etc. ② KEGG pathway analysis revealed that differential circRNA might be involved in the immune response mediated by the mitogen-activated protein kinase signaling pathway. ③ CircRNA might affect its inflammatory response by targeting molecules such as miRNA-146a, miRNA-302b and miRNA-23a. Conclusion:There are differentially expressed circRNAs in PBMCs of patients with gout, which may be closely related to the occurrence and development of gout.
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Objective:To investigate the clinical features, laboratory characteristics and risk factors of systemic sclerosis (SSc) patients with hematologic damages.Methods:The clinical data of 180 SSc patients were collected from January 2010 to April 2020, at the Affiliated Hospital of North Sichuan Medical College. The demographic information, laboratory tests, and clinical symptoms were analyzed retrospectively. Statistical Product and Service Solutions (SPSS) 19.0 was used for t-test, non-parametric Mann-Whitney U test, Chi-squared test, Logistics regression analysis. Results:① Among 180 SSc patients, 70(38.9%) cases were complicated with hematologic damages. Fifty-one (72.9%) cases had anemia, 24 cases (34.3%) had leukopenia, 24 cases (34.3%) had thrombocytopenia, and 22 cases had hematologic damages associated with more than two cell line involvement. ② Clinical symptoms: arthritis was significantly higher in the hematologic damage group than patients without( χ2=4.815, P=0.028), however, there was no significant difference in gender, age, disease course, respiratory symptoms, gastrointestinal symptoms, Raynaud's phenomenon, interstitial lung disease and pulmonary hypertension ( P>0.05). ③ Laboratory tests: erythrocyte sedimentation rate (ESR) and C reactive protein (CRP) were increased in the hematologic damage group, while the albumin decreased ( Z=-2.448, P=0.014; Z=-2.450, P=0.014; Z=-4.773, P<0.01). The positive rates of anti-dsDNA antibody and anti-ribosomal P protein anti-body was higher in the hematologic damage group ( χ2=5.428, P=0.020; χ2=8.169, P=0.004). ④ Prognosis: during follow-up, leukopenia was more likely to recover, while the thrombocytopenia was more difficult to recover. ⑤ Logistics regression analysis showed that positive of anti-ribosomal P protein antibody might bea risk factor for SSc complicated with hematologic damages [ OR=3.930, 95% CI(1.130, 13.666, P=0.031)]. Conclusion:SSc complicated with hematologic damages is common, and patients with hematological damage have more serious clinical symptoms, some of whom have difficulty in recovery. Anti-ribosomal P protein anti-body may be a risk factor for SSc hematologic damages.
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Objective:To investigate the possible role of miR-146a in the patho-genesis of inflammation in primary gout arthritis.Methods:① The RAW264.7 mouse macrophage was stimulated with 200 μg/ml monosodium urate (MSU) crystal for 0 h, 3 h, 6 h, and 12 h. Then cells and super-natants were collected. The miR-146a was detected by TaqMan probe method. The expression of interleukin-1 receptor-associated kinase 1 (IRAK 1), tumor necrosis factor receptor-associated factor 6 (TRAF6), nuclear factor-kappa B (NF-κB), interleukin (IL)-1β, tumor necrosis factor-α (TNF-α) mRNA were detected by real-time (RT)-quantitative polymerase chain reaction (qPCR). The concentration of IL-1β was measured in the culture supernatant by enzyme-linked immunosorbent assay (ELISA). The protein expression levels of TRAF6, NF-κB and IL-1β were detected by Western-blotting. ② The RAW264.7 mouse macrophage was transfected with miR-146a mimics, miR-146a mimic control, miR-146a inhibitor, and miR-146a inhibitor control. After stimulating each group of cells with 200 μg/ml MSU crystals for 6 h, the expression of miR-146a, IRAK1, TRAF6, NF-κB, IL-1β mRNA and TRAF6, NF-κB, IL-1β protein were measured. The measurement data were compared by Independent sample t test. and repeated measures analysis of variance (ANOVA). Results:① After MSU crystals stimulated RAW264.7 cells, we found that the expression level of miR-146a in the stimulation group at 3 h, 6 h, and 12 h was lower than that in the control group ( t=-10.234, -17.059, -26.204, P<0.01), and then, IL-1β protein concentration at 6 h, 12 h was higher ( t=7.552, 9.007, P<0.01). Meanwhile, IRAK1, TRAF6, NF-κB and IL-1β mRNA in the stimulation group at 3 h and 6 h were higher than those in the control group ( t=9.847, 6.147, P<0.01; t=3.49, 3.32, P<0.05; t=3.643, 8.471, P<0.05; t=8.726, 49.68, P<0.01). TNF-α mRNA at three time points in the stimulation group was high ( t=4.691, 11.115, 12.816, P<0.01). Moreover, the results showed that the relative expression of TRAF6 and NF-κB protein in the stimulation group at 6 h and 12 h was higher than that in the control group ( t=8.052, 8.119, P<0.01, t=22.454, 5.845, P<0.01), IL-1β protein in the stimulation group increased at all three time points compared with the control group ( t=18.561, 4.74, 8.432, P<0.01). ② After trans fection, the miR-146a mRNA expression of the mimics group was significantly higher than the mimics control group ( t=31.769, P<0.01); the inhibitor group was significantly lower than the inhibitor control group ( t=-4.22, P<0.05). ③miR -146a overexpression group was stimulated with 200 μg/ml MSU crystals for6 h, the expression levels of IRAK 1, TRAF 6, NF-κB and IL-1β mRNA in the mimic group were lower than those in the mimic control group ( t=-14.754, -21.201, -19.381, -17.323, P<0.01), the expression levels of TRAF 6, NF-κB and IL-1β protein were also lower than those in the mimic control group ( t=-3.137, -32.974, -18.789, P<0.05), while the inhibitor group had good results. Conclusion:① Overexpression of miR-146a can reduce the expression of IRAK1, TRAF6, NF-κB, IL-1β and inhibit MSU crystal-mediated inflammation, while inhibition of miR-146a expression can aggravate inflammation, suggesting that miR-146a participates in the negative feedback regulation of gout inflammation. ② miR-146a may target the NF-κB signaling pathway and participate in spontaneous remission of gouty arthritis.
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Hyperuricemia/gout is a common metabolic disease in China, which is a serious threat to people′s health. In clinical practice, the standardization of prevention and diagnosis and the rate of treat-to-target need to be improved. There is still a lack of education for the patients about the understanding of clinical guidelines, the disease knowledge and the importance of cooperating with doctors to carry out diagnosis and treatment. From the most concerned issues of the patients, we established the hyperuricemia/gout patient practice guideline working group with multidisciplinary physicians and patients. Seventeen opinions, as the hyperuricemia/gout patient practice guidelines, are proposed in accordance with the relevant principles of the "WHO guidelines development manual" , and with the international normative process, aiming to improve the patients compliance, improve the level of health management of the disease.
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Objective To investigate the role of long noncoding RNA-AK001903 in the pathogenesis of primary gout arthritis (GA).Methods The subjects were divided into four groups:30 acute gout patients (AGA),24 non-acute gout patients (NAGA),24 healthy controls and 24 hyperuricemia (HUA).Real-time quantitative polymerase chain reaction (RT-qPCR) was employed to examine the expression of AK001903 in peripheral blood mononuclear cells (PBMCs) from four groups.100 μg/ml monosodium (MSU) was used to stimulate the peripheral blood of NAGA and healthy control patients.Then the expression of AK001903 was detected by RT-qPCR.Kruskal-Wallis test,Mann-Whitney test,Spearman correlations were used for statistical analysis.Results The expression level of AK001903 in the AGA group (0.079±0.022) and the NAGA group (0.071±0.021) were higher than the healthy control group (0.014±0.004).There was no significant difference between the NAGA group and the NAGA group (Z=-0.655,P＞0.05).Those of the GA group (0.078±0.018) and the HUA group(0.047±0.016) was higher than the healthy control group (0.014±0.004) (Z=-2.887,Z=-4.157;P＜0.05).Compared with the control group,the expression of AK001903 in NAGA and the healthy control group which were stimulated by MSU was significantly increased.The Spearman correlation analysis found that the AK001903 expression levels in the GA groups were correlated with TG (r=0.938,P＜0.05),VLDL (r=0.873,P＜0.05),GLU9 (r=0.671,P＜0.05) and were negatively correlated with apoA1 (r=-0.661,P＜0.05).Conclusion Altered expression of AK001903 may be involved in the process of imbalance between lipid metabolism and hyperuricemia,and takes part in the pathogenesis of GA.
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Objective To investigate the role of high mobility group box l protein (HMGB1) in the pathogenesis of ankylosing spondylitis (AS). Methods Enzyme-linked immuno sorbent assay (ELISA) was used to test the levels of plasma HMGB1 levels in 58 patients with active AS [bath ankylosing spondylitis disease activity index (BASDAI)>6, or 6>BASDAI>4 and erythrocyte sedimentation rate (ESR)>22 mm/1 h, 6>BASDAI>4 and hypersensitive C reactive protein (hsCRP)>9 mg/L], 73 cases of stable AS (BASDAI<4) and 70 healthy control. Twelve patients who were treated with TNF-alpha antagonist for 6 month were followed-up. Their plasma levels of HMGB1 were detected before and after treatment. Quantitative data were described by, while qualitative data were described by case number. Variance analysis or rank sum test was adopted for the difference between measurement data groups, LSD method was adopted for further pair-wise comparison. The correlation between variables was analyzed by using Spearman correlation analysis. Results The levels of plasma HMGB1, ESR, hsCRP, White blood cell WBC, GR, Mo and GLOB were significantly higher in the AS patients than those in the healthy control group (P<0.001), and the level of plasma HMGB1 in the AS patients was significantly positively correlated with BASDAI, Bath ankylosing spondylitis functional index (BASFI), ESR, hsCRP, WBC, GR, Mo, and GLOB (r=0.288, 0.174, 0.308, 0.243, 0.261, 0.301, 0.279, 0.289; P=0.004, 0.047, 0.000, 0.005, 0.003, 0.000, 0.001 ,0.001). The level of plasma HMGB1, BASDAI, BASFI, ESR, hsCRP, WBC, GR, GLOB were significantly higher in the active AS group than in the stable group (Z=-3.598,-9.456,-5.907, -2.562, -3.178, 4.134, -2.574, -4.582; P=0.000, 0.000, 0.000, 0.012, 0.002, 0.000, 0.011, 0.000). The level of plasma HMGB1 was not found statistically poor in the patients with different expressions of HLA-B27, or hip involvement and history of vuvitis (P>0.05). The plasma HMGB1 level, BASDAI, BAIFI, ESR, hsCRP and GLOB in the 12 followed-up patients were significantly decreased (P=0.034, 0.002, 0.002, 0.005, 0.004, 0.004) after being treated with biological agents for 6 months. Conclusion HMGB1 might play a vital role in the pathogenesis of ankylosing spondylitis,and the HMGB1 might be used as a clinical indicator to evaluate the activity of AS and to assess the clinical efficacy.
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Objective This study is aimed to investigate the possible role of Th1/Th2 cell in the pathogenesis of primary gout arthritis. Methods The peripheral blood of 21 acute gout patients (AG), 20 intermittent gout patients (IG) and 20 healthy controls (HC) were collected. The clinical data and laboratory indicators of them were enrolled. The percentages of Th1 and Th2 cells were detected by flow cytometry (FCM). The expression of GATA-3, T-bet, IL-4 and interferon (IFN)-γ mRNA in Peripheral blood mononuclear cells (PBMCs) were measured using Real time quantitative polymerase chain reaction (PCR). The protein expression levels of IL-4 and IFN-γ in serum were detected by enzyme-linked immuno sorbent assay (ELISA). The measurement data were compared by one factor analysis of variance test. The correlation between variables was used by Spearman correlation analysis. Results The percentage of Th1 cells in peripheral blood of the AG group was [(23.2 ±8.3)％], and the IG group was [(20.5 ±9.3)％], which were significantly higher (F=6.520, P<0.05) than the percentage of HC group [(14.8±3.8)％]. There was no significant difference between AG group and IG group. The percentage of Th2 cells in peripheral blood of AG group were [(1.9 ±0.7)％], which was significantly lower (F=8.267, P<0.05) than and the percentage of HC group [(3.4±1.8)％] and IG group [(3.3± 1.2)％]. There was no significant difference between the IG group and the HC group. And the proportion of TH1/Th2 cells in the AG group was higher than that of the IG group and the HC group (F=10.406, P<0.01). The expression of T-bet mRNA and IFN-γ mRNA in the AG group and IG group were higher than that in the HC group (F=4.942, P=0.010)、(F=4.458, P=0.016). However, there was no significant difference between IG group and AG group. The expression of GATA-3 mRNA and IL-4 mRNA was significantly lower (F=3.564, P=0.035) (F=5.385, P=0.007) in the AG group and IG group when compared to the HC group. And there was no significant difference between the AG and the IG group. The IFN-γ level increased in the AG and IG group compared to the HC group (F=7.659, P=0.001). The IL-4 levels in the AG group was lower (F=7.099, P=0.002) than those of the IG and HC group. Conclusion Th1 cells in the peripheral blood of patients with GA are increased and accompanied with the decrease of Th2 cells. The results of this study suggest that the imbalance of Th1/Th2 cells in the inflammatory and immune response plays a critical role in the pathogenesis of GA.
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Objective To investigate the role of long noncoding RNA-AJ227913 in the pathogenesis of primary gout arthritis (GA). Methods The subjects were divided into three groups:30 acute gout patients (AGA), 30 non-acute gout patients (NAGA), 30 healthy controlsand 30 hyperuricemia patients (HUA). Real-time quantitative polymerase chain reaction (RT-qPCR) was employed to examine the expression of AJ227913 in peripheral blood mononuclear cells(PBMCs) from four groups. 100 μg/ml monosodium urate (MSU) was used to stimulate the peripheral blood of NAGA and healthy controls patients. Then the expression ofAJ227913 was detected by RT-qPCR. Kruskal-Wallis test, Mann-Whitney test, Spearman correlations were used for statistical analysis. Results The expression level of AJ227913 in the AGA group (0.0557 ±0.0156) was higher than that in the NAGA group (0.0223±0.018) and healthy controls group (0.0038±0.0013). There was significant difference between the NAGA group and healthy controls group (P>0.05). Compared with the control group, the expression of AJ227913 in NAGA group which were stimulated by MSU was significantly increased. The Spearman correlation analysis found that the AJ227913 expression levels in GA groups were correlated with UREA (r=0.608, P<0.01), CREA (r=0.337, P<0.05), CYSC (r=0.422, P<0.01). Conclusion Altered expression of AJ227913 may be involved in the inflammatory process of GA and the balance of uricacid.
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Objective To investigate the role of miR-223 in the pathogenesis of acute gouty inflammation.Methods The subjects were divided into 3 groups:65 acute gout patients (AG),50 inter-critical gout patients (IG),and 45 healthy controls (HC).The peripheral blood mononuclear cells (PBMCs) and the clinical laboratory parameters were all collected.The expression of miR-223 in the PBMCs was detected using realtime fluorescent quantitative polymerase chain reaction (RT-qPCR) (TaqMan probe).The PBMCs of 5 healthy people were stimulated with monosodium urate (MSU) (100 μg/ml) for 12 h,and then,miR-223,NLRP3 mRNA and IL-1β production were all measured using RT-qPCR and ELISA respectively.All data were analyzed by SPSS 17.0 statistical software,Wilcoxon rank sum test,t test and Spearman's correlations analysis were used for statistical analysis.Results ① The expression of miR-223 in AG and IG groups was both significantly decreased than that in the HC group (8±17 vs 26±76,P＜0.05;9±17 vs 26±76,P＜0.05;respectively),AG group was significantly decreased than that in the IG group [8(17) vs 9(17),Z=11.387,P＜0.01].② After stimulated with MSU in healthy controls,IL-1β production and NLRP3 mRNA were both significantly increased [(86±5) pg/ml vs (13±6) pg/ml,t=21.042,P＜0.01;5.2±0.4 vs 1.2±0.4,t=14.640,P＜0.01;respectively],while the expression of miR-223 was significantly decreased (0.34±0.20 vs 1.05±0.24,t=-5.164,P＜0.01).Conclusion The data suggests that miR-223 might be involved in the patho-genesis of spontaneous regulation,but further study is needed to discover the exact mechanism.
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Objective To investigate the current situation in Chinese rheumatologic physicians' clinical diagnosis and evaluation of Takayasu's arteritis (TA).Methods Nineteen rheumatology experts and three vascular surgery specialists in China were invited to make the nationwide investigation for the first time about the diagnosis and disease activity evaluation of TA in China,through the questionnaire survey on the internet.Weighted average was used to calculate the average scores of corresponding problems.Results Chinese experts mainly adopted 1990 American College of Rheumatology (ACR) classification criteria for clinical diagnosis of TA.In details,symptoms of age,limb claudication and amaurosis,signs including pulselessness or pulse weakening,vascular bruits,increasing bilateral pulse pressure and hypertension and acute phase reactants (APR) were critical to the clinical diagnosis of TA.Besides,noninvasive imaging examinations,such as computed tomography angiography (CTA),magnetic resonance angiography (MRA),vascular ultrasonography,and positron emission tomography (PET) were also of great importance.In the aspect of disease activity assessment,Chinese experts mainly used Kerr scoring tool.APR and noninvasive radiological examinations were considered with vital value.Some TA patients with carotid artery involvement were recommended using vascular ultrasonography,while others with pulmonary artery and thoracic/abdominal aorta trunk involvement were preferred CTA other than MRA.Conclusions APR and noninvasive imaging examinations were thought with great help to make clinical diagnosis and evaluation of TA for Chinese physicians.
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Objective To investigate the expression profile variation of long non-coding RNAs (lncRNAs) in ankylosing sporidylitis (AS) and explore the role of lncRNAs in the pathogenesis of AS.Methods The peripheral blood mononuclear cells of AS patients and health controls (HC) were used to detect for differently expressed lncRNAs by microarray.The roles of lncRNAs were predicted with GO and pathway analysis.The results were verified by real time-polymerase chain reaction (PCR).Results A total of 148 lncRNAs and 134 mRNAs were detected,which had more than 2-fold differentially expressed in AS patients.Bioinformatics analysis found that GO term enrichment included protein binding,regulation of transcription,metabolism,signal transduction,et al.and might involve in toll-like receptor pathway,protein kinase,complement pathway,notch signaling pathway and so on.The expressions of three lncRNAs were estimated by real time-PCR which found that consistent with that of microarrays.Among these,D90064 was the most aberrantly expressed lncRNAs.Conclusions Several lncRNAs expression was changed significantly in AS patients in comparison with HC,which implies that those different lncRNAs may have an important role in the development and progression of AS.
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Objective To investigate the mRNA and protein expression levels of telomeric-repeat binding factor-1 (TRF1) and TRF2 in the peripheral blood mononuclear cells (PBMCs) of patients with systemic lupus erythematosus (SLE),and the relations between these gene expression levels and clinical data of SLE patients were explored.Methods According to disease activity,these SLE patients were divided into the active group (40 cases) and the stable group (67 cases).These patients were also grouped as renal damage group (46 cases) and renal damage-free group (61 cases) based on their renal conditions.Healthy individuals (41 cases) were also included as control.Real-time quantitative polymerase chain reaction (RT-qPCR) was employed to study the mRNA expression of TRF1 and TRF2.The protein levels of TRF1 and TRF2 were measured by Western Blot (WB).Independent-Samples t test or one-way analysis of variance (ANOVA) in conjunction with the Least-Significant Difference method (LSD method) wasperformed if the data were in normal distributions;otherwise,the Kruskal-Wallis test was applied.Spearman's correlation analysis was also used for statistical analysis.Results The mRNA and protein expression levels of TRF1 and TRF2 in the PBMCs of the active group (TRF1:0.003 1±0.003 3;TRF2:0.010 5±0.064 8) and renal damage group (TRF1:0.002 3 ±0.002 6;TRF2:0.004 3 ±0.003 3) were significantly increased compared to the stable group (TRF1:0.001 2±0.001 1;TRF2:0.004 2±0.008 6),the renal damage-free group (TRF1:0.001 3±0.001 8;TRF2:0.003 4±0.007 2) and healthy (TRF1:0.001 2±0.003 0;TRF2:0.003 4±0.002 7) individuals respectively (P＜0.05).In SLE patients,the expression levels of TRF1 mRNA were correlated with erythrocyte sedimentation rate (r=0.365,P＜0.05);the expression levels of TRF2 mRNA were correlated with SLEDAI score (r=0.270,P＜0.05),erythrocyte sedimentation rate (r=0.304,P＜0.05),creatinine (r=0.258,P＜0.05) and 24-hour urinary protein (r=0.344,P＜0.05).Conclusion Altered expression of TRF1 and TRF2 might be involved in the pathogenesis of Systemic lupus erythematosus.The positive correlation between TRF2 and SLEDAI score,24-hour urinary protein suggest that TRF2 might be usedas a biomarker for disease activity or renal damage in
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The long noncoding RNAs refers to RNA transcripts more than 200 nucleotides in length,and do not encode proteins.In the end of the 20th century,the lncRNAs were found to play crucial role in many important biological processes,including embryonic development,cell differentiation,species evolution,metabolism,and disease occurrence by the scientific community.Currently,multiple studies have indicated that long noncoding RNAs have participated in rheumatic diseases,immune response,immune cell differentiation,and dynamic balance of the immune system.Therefore,summary of the roles of lncRNAs in rheumatic diseases could be beneficial to understand the pathogenesis of autoimmune diseases.This review article attempts to highlight the recent progresses regarding IncRNAs studies and the relationship between long noncoding RNA and rheumatic diseases by taking systemic lupus erythematosus,rheumatoid arthritis,arthritis and other typical diseases as examples.