Yuqiong Wu

Adalimumab significantly reduces inflammation and serum DKK‐1 level but increases fatty deposition in lumbar spine in active ankylosing spondylitis

To investigate whether adalimumab is effective for active ankylosing spondylitis (AS) patients and whether it has an impact on the formation of fatty deposition lesions (FDL) and serum Dickkopf homolog 1 (Dkk‐1) level in AS patients.

The Ankylosing Spondylitis Disease Activity Score is a highly discriminatory measure of disease activity and efficacy following tumour necrosis factor-α inhibitor therapies in ankylosing spondylitis and undifferentiated spondyloarthropathies in China

OBJECTIVE To validate the clinical value of the new Ankylosing Spondylitis Disease Activity Scores (ASDASs) in assessing the disease activity and efficacy of TNF-α inhibitor in AS and uSpA patients in China. METHODS Two hundred and thirty patients were included in our study. They consisted of patients with active AS (n = 87) and uSpA (n = 30) participating in a double-blind placebo-controlled randomized clinical trial of etanercept and patients with active AS (n = 58) and uSpA (n = 55) treated with infliximab. The disease activity and treatment effects were assessed by ASDAS, BASDAI, patient global and the acute inflammation score of lumbar and SI joints by MRI. Discriminatory ability of all the measures was analysed by standardized mean difference and t-score. RESULTS In both the AS and uSpA groups, ASDAS correlated well with patient global score (AS group: r = 0.65-0.72; uSpA group: r = 0.52-0.62), ESR (AS group: r = 0.57-0.81; uSpA group: r = 0.63-0.85) and CRP (AS group: r = 0.51-0.70; uSpA group: r = 0.61-0.76) both at baseline and in changes from baseline to 6 weeks after TNF-α inhibitor treatment. The ASDAS scores outperformed BASDAI, patient global score, ESR, CRP and the acute inflammation score by MRI in differentiating patients with different levels of disease activity and patients with different levels of change in both AS and uSpA groups. There was little difference in performance between the two versions of the ASDAS. CONCLUSION The new ASDAS is a highly effective measure in assessing disease activity and a great discriminatory measurement to assess the efficacy of TNF-α inhibitor in Chinese AS patients and uSpA patients.

Fractalkine stimulates cell growth and increases its expression via NF-κB pathway in RA-FLS

After the onset of rheumatoid arthritis (RA), fibroblast‐like synoviocytes (RA‐FLS) which are specialized types of fibroblasts, become tumor‐like, keeping their ability to increase proliferation and invasion. The mechanism of their tumor‐like growth is unclear. Fractalkine (FKN), also called CX3CL1, plays an important role in the proliferation of cells. FKN may stimulate the proliferation of RA‐FLS and the by nuclear factor κB (NF‐κB) pathway may be one of the steps in this process.

Higher risk of uveitis and dactylitis and older age of onset among ankylosing spondylitis patients with HLA-B*2705 than patients with HLA-B*2704 in the Chinese population.

Human leukocyte antigen (HLA)-B27 is closely associated with ankylosing spondylitis (AS). However, the exact correlation between HLA-B27 subtypes and AS manifestations remains unknown. This study aimed to investigate the correlation between HLA-B27 polymorphism and the clinical features of AS. This study included 846 patients with AS and 959 healthy controls. Direct sequencing was used to identify the HLA-B27 genotype. Clinical parameters, including age, age of onset, family history, low back pain, peripheral arthritis, hip joint involvement, dactylitis, uveitis, and sex ratio, were compared among patients with various HLA-B27 subtypes. In total, 741 AS patients (87.6%) and 39 healthy controls (4%) were HLA-B27-positive. The most prevalent subtypes were HLA-B*2704 (88%) and HLA-B*2705 (10.1%) in patients with AS. Compared with HLA-B*2704-positive patients, HLA-B*2705-positive patients demonstrated a significant increase in the incidence of uveitis (16% vs 6.13%, P = 0.002) and dactylitis (9.3% vs 3.8%, P = 0.028) and they had an older age of onset (22.9 ± 8.0 vs 20.7 ± 6.7 years, P = 0.028). Binary logistic regression analysis revealed that presence of uveitis was significantly associated with HLA-B*2705 (P = 0.008; odds ratio, 2.63; 95% confidence interval, 1.283-5.393). There were no significant differences in family history, low back pain, peripheral arthritis, or hip joint involvement among HLA-B27 subtypes. Specific HLA-B27 subtypes were positively associated with particular clinical features of AS. AS patients with HLA-B*2705 demonstrated an older age of onset and had a higher risk of uveitis and dactylitis than did AS patients with HLA-B*2704.

Disorders of MicroRNAs in Peripheral Blood Mononuclear Cells: As Novel Biomarkers of Ankylosing Spondylitis and Provocative Therapeutic Targets

Background. MicroRNAs can potentially regulate every aspect of cellular activity. In this study, we investigated whether AS pathogenesis involves microRNAs disorders. Result. The expression of 2 microRNAs, hsa-miR-126-3p and hsa-miR-29a, was significantly lower in active AS group before etanercept therapy than in control group. Marched fold changes of them were 3.76 and 16.22. Moreover, expressions of hsa-miR-126-3p and hsa-miR-29a were dramatically upregulated after 12-weeks etanercept treatment. Fold changes were 2.20 and 3.18. All regulations of microRNAs expression mentioned before were statistically significant (fold change >2 and P < 0.05). The expression disorders of the 2 microRNAs did not statistically significantly correlated with BASDAI, CRP, and ESR. Conclusion. AS pathogenesis involved dysregulation of microRNAs. Hsa-miR-126-3p and hsa-miR-29a will probably become the potential biomarkers and provocative therapeutic targets of AS.

A Genome-Wide SNP Linkage Analysis Suggests a Susceptibility Locus on 6p21 for Ankylosing Spondylitis and Inflammatory Back Pain Trait

Objectives To screen susceptibility loci for ankylosing spondylitis (AS) using an affected-only linkage analysis based on high-density single nucleotide polymorphisms (SNPs) in a genome-wide manner. Patients and Methods AS patients from ten families with Cantonese origin of China were enrolled in the study. Blood samples were genotyped using genomic DNA derived from peripheral blood leukocytes by Illumina HumanHap 610-Quad SNP Chip. Genotype data were generated using the Illumina BeadStudio 3.2 software. PLINK package was used to remove non-autosomal SNPs and to further eliminate markers of typing errors. An affected-only linkage analysis was carried out using both non-parametric and parametric linkage analyses, as implemented in MERLIN. Result Seventy-eight AS patients (48 males and 30 females, mean age: 39±16 years) were enrolled in the study. The mean age of onset was 23±10 years and mean duration of disease was 16.7±12.2 years. Iritis (2/76, 2.86%), dactylitis (5/78, 6.41%), hip joint involvement (9/78, 11.54%), peripheral arthritis (22/78, 28.21%), inflammatory back pain (IBP) (69/78, 88.46%) and HLA-B27 positivity (70/78, 89.74%) were observed in these patients. Using non-parameter linkage analysis, we found one susceptibility locus for AS, IBP and HLA-B27 in 6p21 respectively, spanning about 13.5Mb, 20.9Mb and 21.2Mb, respectively No significant results were found in the other clinical trait groups including dactylitis, hip involved and arthritis. The identical susceptibility locus region spanning above 9.44Mb was detected in AS IBP and HLA-B27 by the parametric linkage analysis. Conclusion Our genome-wide SNP linkage analysis in ten families with ankylosing spondylitis suggests a susceptibility locus on 6p21 in AS, which is a risk locus for IBP in AS patients.

Serum from patients with ankylosing spondylitis can increase PPARD, fra-1, MMP7, OPG and RANKL expression in MG63 cells

OBJECTIVES: To explore the effects of serum from patients with ankylosing spondylitis on the canonical Wnt/β-catenin pathway and to assess whether the serum has an osteogenic effect in MG63 cells. METHODS: MG63 cells were cultured with serum from 45 ankylosing spondylitis patients, 30 healthy controls, or 45 rheumatoid arthritis patients. The relative PPARD, fra-1, MMP7, OPG and RANKL mRNA levels were measured using quantitative real-time polymerase chain reaction. Associations between gene expression and patient demographics and clinical assessments were then analyzed. RESULTS: MG63 cells treated with serum from ankylosing spondylitis patients had higher PPARD, fra-1, MMP7 and OPG gene expression than did cells treated with serum from controls or rheumatoid arthritis patients (all p<0.05). RANKL expression was higher in MG63 cells treated with serum from patients with ankylosing spondylitis or rheumatoid arthritis than in those treated with serum from controls (both p<0.05). The OPG/RANKL ratio was also higher in MG63 cells treated with serum from ankylosing spondylitis patients than in those treated with serum from controls (p<0.05). No associations were found between the expression of the five genes and the patient demographics and clinical assessments (all p>0.05). CONCLUSIONS : Serum from ankylosing spondylitis patients increases PPARD, fra-1, MMP7, OPG and RANKL expression and the OPG/RANKL ratio in MG63 cells; these effects may be due to the stimulatory effect of the serum on the Wnt pathway.

THU0421 Female primary gout had its unique ultrasound features

Background Primary gout is a metabolic disease occurred in male and post-menopause female in most cases. Though the ultrasound features of gout had been discovered for several years, no reports illuminated whether there would be difference presentations between different genders in the joints. Objectives We employed ultrasound instead of dual-energy CT to explore more refined pathological manifestations of primary gout in different genders. Methods All cases were confirmed as gout fulfilling 1997 ACR classification criteria. All cases excluded secondary gout induced by drug, tumor, hypertension, diabetes mellitus, renal failure. Ultrasound was performed during chronic stage of gout but not at acute attack. The process was done by 2 observers blinded to each other, blood tests. Final diagnosis was determined by the third US expert if the 2 observers got the different conclusion. Bilateral toes, dorsal feet, ankle, knee, wrist, fingers, elbow, and shoulder were detected to find 6 features of gout suggested by OMERCT: tophus, “snow storm”-like effusion, cloudy synovium in grey scale, double-contour sign, bone erosion and Power Doppler (PD) signal. Each above positive presentations in each above range would get 1 point and the sum scores would be ranged from 0 to 84. Serum uric acid (UA) was recorded too. Results 1) 23 female and 139 male were recruited in the program. The female-male ratio was 1:6. Mean age and disease duration of the female subjects were elder than male ones (female:male=57.2±14.1: 44.7±14.7 years old) with longer disease duration to confim the diagnosis (female:male = 10.9:1.2 months). The average serum UA level in female was lower than male group (female:male = 413.8±162.1umol/L, 515.5±156.9umol/L, sig<0.05). 2) The intra-observer reliability from 20% samples random seclection showed an overall agreement of 80%, 92%, 96%, 87%, 80%, 73% for tophus, “snow storm”-like effusion, cloudy synovium in grey scale, positive double-contour sign, bone erosion and PD signal with kappa value of 0.78, 0. 92, 0.95, 0.86, 0.79, 0.72, respectively. 3) The difference showed female gout had higher frequency of tophus, bone erosion and lower frequency of effusion while the other indexes were equal:topus scores (female:male=87%:,74.1%), cloudy synovium grey scale scores (female:male=65.2%:62.6%), effusion scores (female:male=17.4%,31.7%), bone erosion scores (female:male=30.4%,16.5%), power dopplar scores (female:male=34.8%,41%), positive double-contour signs (60.9%:59.9%. The top 2 affected ranges were ankle (female:male=69.6%:55.3%), knee (female:mlae=60.9%:54.0%). Furthermore, female gout had more frequently occurred not in the less typical ranges such as fingers (female:male=34.7%:19.4%), elbow (8.7%:2.7%), which might be the cause for delayed diagnosis. Conclusions Though the level of serum UA was lower, Female gout had its unique ultrasound features with more tophus, bone erosion and less effusion compared to male gout. The less typical ranges were recommended for US examinations. Disclosure of Interest None declared

AB0100 IL-22-Mediated BCL-2 Expression in Rheumatoid Arthritis Fibroblast-like Synoviocytes through STAT3 Activation

Background Interleukin-22 (IL-22) is a novel cytokine that is mostly produced by T cells and innate lymphoid cells but selectively stimulates non- immune cells, which immediately caught the attention of many fields such as tumor, chronic inflammatory disease and infectious disease. Considering the persistent biological abnormalities in rheumatoid arthritis fibroblast-like synoviocytes (RA-FLSs) and transient effects of current best treatment such as anti-TNF biologics, other cytokines might be involved and we focus here on the role of IL-22. Objectives To test the effect of IL-22 on the survival of RA-FLSs and to investigate whether signal transducer and activator of transcription 3 (STAT3) is implicated in this process. Methods Bcl-2 mRNA and Bcl-xL mRNA expression in the RA-FLSs, osteoarthritis-FLSs and trauma-FLSs were determined using the Real-time quantitative polymerase chain reaction (RT-qPCR), Bcl-2 protein expression in FLSs were detected by the Western Blotting analysis. The effects of recombinant human interleukin-22 (rhIL-22) at concentrations of 0, 1, 10, 100 ng/mL and STA-21 (a STAT3 inhibitor at concentrations of 0, 25μM, 50μM) on the protein levels of Bcl-2 and p-STAT3 in the RA-FLSs were determined by Western blotting analysis. RA-FLSs were co-cultured with rhIL-22 in the presence or absence of STA21 for 24 h, and the protein levels of Bcl-2 and p-STST3 were evaluated by Western blotting analysis. Results The anti-apoptotic Bcl-2 (P<0.05) and Bcl-xL (P<0.01) are increased in RA-FLSs compared with osteoarthritis-FLSs and trauma-FLSs, and so is Bcl-2 protein (P<0.01). IL-22 increased, and STA-21 decreased the protein levels of Bcl-2 and p-STAT3 (IL-22: FBcl-2=48.63, PBcl-2<0.01; FP-STAT3=25.7, PP-STAT3<0.01 STA-21: FBcl-2=49.74, PBcl-2<0.01l; FP-STAT3=109.97, PP-STAT3<0.01) in the RA-FLSs obviously. Additionally, treatment with STAT3 inhibitor STA-21 reversed the effect of IL-22-induced Bcl-2 upregulation in the RA-FLSs (FBcl-2=84.67, PBcl-2<0.01; FP-STAT3=82.46, PP-STAT3<0.01). Conclusions STAT3 is critical in the process which IL-22-induced Bcl-2 upregulation in the RA-FLSs. The effects of IL-22/STAT3 confer on them the potential to be used for therapeutic applications in RA. Disclosure of Interest None declared

AB0114 PTEN Expression in Fibroblast-like Synoviocytes of Rheumatoid Arthritis

Background Fibroblast-like synoviocytes (FLS), with some tumor-like characters, play a key role in the pathogenesis of rheumatoid arthritis (RA) [1]. Phosphatase and tensin homolog (PTEN) is a tumor suppressor gene, and its mRNA expression has been reported to be absent in the lining layer of RA synovium, which is supposed to contribute to the invasive behavior of RA-FLSs [2,3]. It has been demonstrated in various kinds of tumor cells that the PTEN protein is a negative regulator of the PI3K/Akt signal pathway [4]. However, data of the PTEN protein level in RA-FLSs and its effect on the phosphorylation status of Akt (Thr308) is limited. Objectives To investigate the expression of PTEN gene in cultured human RA-FLSs and its effect on Akt (Thr308) phosphorylation status. Methods FLSs were isolated from synovial tissues obtained from patients with RA, osteoarthritis (OA) or joint trauma during joint replacement surgery or arthroscopy, and each group contains 3 patients. The mRNA expression of PTEN was detected by RT-qPCR. The protein levels of PTEN, pAkt (Thr308) and total Akt, were measured by western blotting. The phosphorylation status of Akt was determined by protein expression ratio of pAkt (Thr308)/total Akt. Results The expression of PTEN mRNA was significantly lower in RA-FLSs compared with OA-FLSs and joint trauma-FLSs (P<0.01), while no statistically significant difference was observed between those of OA-FLSs and joint trauma-FLSs (P>0.05). Similarly, the PTEN protein level in RA-FLS was much lower than OA-FLSs and joint trauma-FLSs (P<0.05), and there was no difference between the latter two (P>0.05). Moreover, the phosphorylation level of Akt (Thr308) in RA-FLSs was significantly higher than those in the other two control groups (P<0.01), and that in OA-FLSs was much lower than joint trauma-FLSs (P<0.01). Conclusions The mRNA and protein expression of PTEN are both decreased in cultured RA-FLSs, suggesting that low expression level of PTEN is conserved in RA-FLSs out of the inflammatory environment in vitro throughout passages. And it may contribute to the increased phosphorylation level of Akt (Thr308). References Bottini N, Firestein GS. Duality of fibroblast-like synoviocytes in RA: passive responders and imprinted aggressors. Nature reviews Rheumatology. 2013 Jan; 9(1):24–33. Pap T, Franz JK, Hummel KM, Jeisy E, Gay R, Gay S. Activation of synovial fibroblasts in rheumatoid arthritis: lack of Expression of the tumour suppressor PTEN at sites of invasive growth and destruction. Arthritis research. 2000; 2(1):59–64. Wang CR, Shiau AL, Chen SY, Lin LL, Tai MH, Shieh GS, et al. Amelioration of collagen-induced arthritis in rats by adenovirus-mediated PTEN gene transfer. Arthritis and rheumatism. 2008 Jun; 58(6):1650–1656. Song MS, Salmena L, Pandolfi PP. The functions and regulation of the PTEN tumour suppressor. Nature reviews Molecular cell biology. 2012 May; 13(5):283–296. Acknowledgement The research was supported by Guangdong natural science foundation (No. 2014A030313080). Disclosure of Interest None declared