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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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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 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.