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The production and oxidation mechanism of reactive oxygen species (ROS) are out of balance in rheumatoid arthritis (RA). However, the correlation between ROS and T cell subsets in RA remains unclear. Peripheral blood mononuclear cells (PBMCs) from patients with RA (n = 40) and healthy controls (n = 10) were isolated from whole blood samples. Synovial tissues (n = 3) and synovial fluid (n = 10) were obtained from patients with RA. The repartition of T cell subsets and expression of ROS and cytokines were examined according to RA severity. Fibroblast-like synoviocytes (FLSs) from patients with RA were stimulated with PBMCs and the expression of inflammation-related molecules were measured by RT-PCR and cytokine array. Regulatory T cells from patients with moderate (5.1 &gt, DAS28 ≥ 3.2) RA showed the highest expression of mitochondrial ROS among the groups based on disease severity. Although ROS levels steadily increased with RA severity, there was a slight decline in severe RA (DAS28 ≥ 5.1) compared with moderate RA. The expression of inflammatory cytokines in RA FLSs were significantly inhibited when FLSs were co-cultured with PBMCs treated with ROS inhibitor. These findings provide a novel approach to suppress inflammatory response of FLSs through ROS regulation in PBMCs.

There is growing evidence that apurinic/apyrimidinic endonuclease 1/redox factor-1 (APE1/Ref-1) regulates inflammatory responses. Rheumatoid arthritis (RA) is an autoimmune disease, which is characterized with synovitis and joint destruction. Therefore, this study was planned to investigate the relationship between APE1/Ref-1 and RA. Serum and synovial fluid (SF) were collected from 46 patients with RA, 45 patients with osteoarthritis (OA), and 30 healthy control (HC) patients. The concentration of APE1/Ref-1 in serum or SF was measured using the sandwich enzyme-linked immunosorbent assay (ELISA). The disease activity in RA patients was measured using the 28-joint disease activity score (DAS28). The serum APE1/Ref-1 levels in RA patients were significantly increased compared to HC and OA patients (0.44 ± 0.39 ng/mL for RA group vs. 0.19 ± 0.14 ng/mL for HC group, p &lt, 0.05 and vs. 0.19 ± 0.11 ng/mL for OA group, p &lt, 0.05). Likewise, the APE1/Ref-1 levels of SF in RA patients were also significantly increased compared to OA patients (0.68 ± 0.30 ng/mL for RA group vs. 0.31 ± 0.12 ng/mL for OA group, p &lt, 0.001). The APE1/Ref-1 concentration in SF of RA patients was positively correlated with DAS28. Thus, APE1/Ref-1 may reflect the joint inflammation and be associated with disease activity in RA.

To evaluate the efficacy of intravitreal aflibercept monotherapy for submacular hemorrhage secondary to neovascular age-related macular degeneration (AMD) and polypoidal choroidal vasculopathy (PCV). This prospective, phase 4 clinical trial included 29 patients diagnosed with fovea-involving submacular hemorrhage secondary to neovascular AMD (7 patients) or PCV (22 patients). Patients were initially administered 3 monthly aflibercept injections, followed by 1 injection every 2 months. The primary outcome measure was changes in Early Treatment Diabetic Retinopathy Study (ETDRS) best-corrected visual acuity (BCVA) during the 56-week study period. Other key outcome measures were the proportion of patients who exhibited changes in BCVA of ≥ 15 ETDRS letters from baseline and changes in central retinal thickness (CRT). The mean size of hemorrhage was 6.2 ± 4.8-disc-diameter area. The mean BCVA significantly improved from 52.9 ± 17.8 ETDRS letters at week 0 (baseline) to 71.8 ± 16.1 letters at week 56 (P

Objectives This study aims to examine the possible associations of mitochondrial single nucleotide polymorphisms (SNPs) and Behcet's disease (BD) in a larger patient group. Patients and methods Whole blood or buffy coat was collected from 98 BD patients (31 males, 67 females; mean age 48±2.8 years; range 20 to 60 years) from four university hospitals located in the Chung-Cheong district of the Republic of Korea, and 196 age- and sex-matched healthy controls (HCs) (62 males, 134 females; mean age 46.91±12.90 years; range 20 to 68 years) from Konyang University Hospital. Twenty targeted mitochondrial deoxyribonucleic acids (DNAs) were genotyped and compared using the revised Cambridge Reference Sequence. Chi square and Fisher's exact tests were used to analyze association of mitochondrial DNA SNPs with BD susceptibility and its clinical characteristics. Results There were no differences for m.248A>G, m.304C>A, m.709G>A, m.3010G>A, m.3970C>T, m.4883C>T, m.5178C>A, m.6392T>C, m.6962G>A, m.10310G>A, m.10609T>C, m.12406G>A, m.12882C>T, m.13928G>C, m.14668C>T, m.16129G>A, and m16304T> between patient and HC groups. However, m.16182A>C and m.16183A>C were more frequently observed in the patient group than the HC group (22 [22.4%] vs. 24 [12.2%], p=0.061 and 32 [32.7%] vs. 42 [21.4%], p=0.092) but without statistical significance. m.4883C>T and m.5178C>A were associated with posterior location of oral ulcers (p=0.025 for each) and m.16183A>C was associated with deep oral ulcers (p=0.001), while m.16189T>C was associated with deep oral ulcers and thrombosis (p=0.042, 0.048, respectively). Conclusion m.16182A>C and m.16183A>C may be associated with BD in the Korean population.

Rheumatoid arthritis (RA) is a common autoimmune disease characterized by immune cell infiltration of the synovium, leading to the loss of cartilage, bone, and joint function. Although regulatory T (Treg) cells are thought to modulate the initiation and progression of RA, a consensus has yet to be reached regarding the function and composition of Treg cells in RA patients. To address these discrepancies, we analyzed not only the total Treg frequency but also that of Treg subpopulations in the peripheral blood of RA patients and healthy controls by flow cytometry. We found that the total Treg population was not significantly different between RA and control subjects. However, the effector Treg cell subgroup, defined as CD45RA−CD25hi, showed markedly decreased frequency in RA patients. In addition, the total Treg population from RA patients showed a significant decline in the expression of CD25. Both the naïve and effector Treg subgroups also showed marked reduction of CD25 expression in RA patients compared to controls. These data suggest that the decreased frequency of effector Treg cells and overall reduction of CD25 expression in Treg cells in the peripheral blood may be evidence of altered Treg homeostasis associated with RA pathogenesis.

Monocytes are important cellular effectors of innate immune defense. Human monocytes are heterogeneous and can be classified into three distinct subsets based on CD14 and CD16 expression. The expansion of intermediate CD14+CD16+ monocytes has been reported in chronic inflammatory diseases including rheumatoid arthritis (RA). However, the mechanism underlying induction of CD16 and its role in monocytes remains poorly understood. Here, we demonstrate that activated platelets are important for induction of CD16 on classical CD14+CD16- monocytes by soluble factors such as cytokines. Cytokine neutralization and signaling inhibition assays reveal that sequential involvement of platelet-derived TGF-β and monocyte-derived IL-6 contribute to CD16 induction on CD14+CD16- monocytes. Activated platelet-induced CD16 on monocytes participates in antibody-dependent cellular phagocytosis (ADCP) and its level is positively correlated with phagocytic activity. CD14+CD16- monocytes treated with activated platelets preferentially differentiate into M2 macrophages, likely the M2c subset expressing CD163 and MerTK. Lastly, the amount of sCD62P, a marker of activated platelets, is significantly elevated in plasma of RA patients and positively correlates with clinical parameters of RA. Our findings suggest an important role of activated platelets in modulating phenotypical and functional features of human monocytes. This knowledge increases understanding of the immunological role of CD14+CD16+ cells in chronic inflammatory diseases.

Background Reactive oxygen species (ROS) regulate the migration and invasion of fibroblast-like synoviocytes (FLS), which are key effector cells in rheumatoid arthritis (RA) pathogenesis. Nicotinamide adenine dinucleotide phosphate oxidase 4 (NOX4) induces ROS generation and, consequently, enhances cell migration. Despite the important interrelationship between RA, FLS, and ROS, the effect of NOX4 on RA pathogenesis remains unclear. Methods FLS isolated from RA (n = 5) and osteoarthritis (OA, n = 5) patients were stimulated with recombinant interleukin 17 (IL-17; 10 ng/ml) and tumor necrosis factor alpha (TNF-α; 10 ng/ml) for 1 h. Cell migration, invasion, adhesion molecule expression, vascular endothelial growth factor (VEGF) secretion, and ROS expression were examined. The mRNA and protein levels of NOX4 were analyzed by RT-qPCR and western blotting, respectively. The NOX4 inhibitor GLX351322 and NOX4 siRNA were used to inhibit NOX4 to probe the effect of NOX4 on these cellular processes. Results Migration of RA FLS was increased 2.48-fold after stimulation with IL-17 and TNF-α, while no difference was observed for OA FLS. ROS expression increased in parallel with invasiveness of FLS following cytokine stimulation. When the expression of NOX was examined, NOX4 was significantly increased by 9.73-fold in RA FLS compared to unstimulated FLS. Following NOX4 inhibition, cytokine-induced vascular cell adhesion molecule 1 (VCAM1), VEGF, and migration and invasion capacity of RA FLS were markedly decreased to unstimulated levels. Conclusion NOX4 is a key contributor to cytokine-enhanced migration and invasion via modulation of ROS, VCAM1, and VEGF in RA FLS.

OBJECTIVES To investigate the impact of European Scleroderma Trials and Research (EUSTAR) standardized training on the accuracy of modified Rodnan skin score (mRSS) in patients with systemic sclerosis (SSc). METHODS Eight SSc patients (four diffuse, four limited) and 10 physicians (4 fellows, 6 professors) were included. Gold-standard mRSS was performed by a senior instructor. Training comprised a video presentation and a live demonstration. Each physician performed mRSS with no clinical information in all patients: (a) before training; (b) after video session; and (c) after live demonstration. Primary outcome was the change in scoring accuracy, which was defined as the difference from the gold-standard skin score, as analyzed using a linear mixed model. RESULTS Mean (standard deviation) difference from the gold-standard score in all measurements by participants before the training was 7.7 (9.5). Completion of training significantly enhanced mRSS accuracy (adjusted β = -7.61; 95% CI: -11.91 to -3.32). This was largely attributable to the video presentation (adjusted β = -5.47; -9.16 to -1.78), although the live demonstration was associated with numerical reduction in the difference from the gold-standard score (adjusted β = -2.15; -5.84 to 1.55). Effect of training was prominent in fellows whereas professors showed an increase in the difference from gold-standard score after training (P value for interaction

Objective.We aimed to investigate demographic and clinical features and predictors of mortality in Korean patients with systemic sclerosis (SSc).Methods.We performed a retrospective multicenter medical chart review in Korean patients diagnosed with SSc from 1986 to 2016 at 11 university hospitals representing each geographic area of Korea. SSc patients were defined according to the American College of Rheumatology preliminary classification criteria and subtyped as limited cutaneous (lcSSc) or diffuse cutaneous (dcSSc) SSc.Results.We enrolled 751 patients (female, 86.7%; mean age at diagnosis, 48.9 yrs). The most common organ involvement was interstitial lung disease (52.7%), followed by gastroesophageal reflux disease (32.9%) and pulmonary arterial hypertension (13.6%). Patients with lcSSc were more common than those with dcSSc (64.8 vs 35.2%), whereas anti-Scl-70 and anticentromere antibody positivity were identified in 302 (42.5%) and 175 (25.5%) patients, respectively. In the 46 (6.1%) patients who developed a malignancy, lung cancer (23.9%) was the most common diagnosis, followed by gastric (13%) and breast cancer (13%). During the study period, 57 (7.6%) patients died, and the 5- and 10-year survival rates were 94% and 87%, respectively. Increased age at diagnosis, cardiovascular involvement, and anti-Scl-70 antibody positivity were significant predictors of death.Conclusion.Clinical manifestations and survival rates in Korean SSc patients are similar to those of other populations. However, the prevalence of anti-Scl-70 antibody is higher in Korean SSc patients compared with whites, while the prevalence of anticentromere antibody is lower.

PURPOSE Although cardiac manifestation of Behcet disease (BD) has been described in sporadic reports, its timely diagnosis remains difficult. The objective of this study was to describe early cardiac manifestations of BD. We also performed a comprehensive classification of systemic BD activity and compared their cardiac manifestations. METHODS A prospective screening using speckle tracking echocardiography was performed in 85 patients with BD who had no history of heart disease. After excluding subjects with left ventricular (LV) ejection fraction (LVEF)

Background Rheumatoid arthritis (RA) is characterized by inflammation and cellular proliferation in the synovium. Activated lymphocytes and proinflammatory molecules are important in the pathogenesis of RA. CD70 belongs to the tumor necrosis factor (TNF) ligand superfamily and is typically present on activated B and T lymphocytes, natural killer cells and mature dendritic cells. CD70 expressing CD4+ T cells are enriched in the peripheral blood and synovial fluid of patients with RA and promote autoimmunity via co-stimulatory CD70-CD27 interaction. CD70 expression is associated with aggressive phenotype of cancer cells and it is mediated by hypoxia inducible factor 2α (HIF-2α). Objectives In this study, we examined the presence of CD70 on the surface of fibroblast-like synoviocyte (FLS) of patients with RA (RA-FLS) and investigate the role of CD70 in the pathogenesis of RA associated with HIF-2α. Methods RA FLS were obtained from 7 patients with RA who were undergone operation like total knee replacement or synovectomy. All patients were fulfilled the 2010 ACR-EULAR classification criteria for RA. CD70 and HIF-2α messenger ribonucleic acid (mRNA) were analyzed in RA-FLS by quantitative polymerase chain reaction (qPCR). CD70 and CD27 on the surface of RA-FLS were stained by PE-Anti CD70 antibody and PerCP-Cy5.5-CD27 antibody respectively and evaluated by flow cytometry. Same experiments were performed after treatment with interleukin (IL)-17, TNF-α and HIF-2α blocking antibody (Anti HIF-2α antibody). Results CD70 and HIF-2α mRNA in the RA-FLS were elevated after treatment with IL-17 and TNF-α (Figure 1, 2). The level of CD70 expression on the surface of RA-FLS was elevated after stimulation with IL-17 and TNF-α (Figure 3). And it was lowered after treatment with HIF-2α blocking antibody as dose dependent pattern (Figure 3). CD27 wasn’t present on the surface of RA-FLS (Figure 4). Conclusion We identified the expression of CD70 on the surface of RA-FLS. And in inflammatory conditions like stimuation with IL-17 and TNF-α, both CD70 and HIF-2α mRNA were increased. The level of CD70 on the surface of RA-FLS also elevated by treatment with IL-17 and TNF-α. The result of decreased level of CD70 after treatment with anti HIF-2α antibody suggest that CD70 expression on the surface of RA-FLS is associated with HIF-2α. From these results, we expect that CD70-targeted therapy associated with HIF-2α may be effective for treatment with RA. References [1] Han, B. K., Olsen, N. J., & Bottaro, A. (2016). The CD27-CD70 pathway and pathogenesis of autoimmune disease. Semin Arthritis Rheum, 45(4), 496-501. [2] Kitajima, S., Lee, K. L., Fujioka, M., Sun, W., You, J., Chia, G. S. Poellinger, L.(2018). Hypoxia-inducible factor-2 alpha up-regulates CD70 under hypoxia and enhances anchorage-independent growth and aggressiveness in cancer cells. Oncotarget, 9(27), 19123-19135. [3] Lee, W. W., Yang, Z. Z., Li, G., Weyand, C. M., & Goronzy, J. J. (2007). Unchecked CD70 expression on T cells lowers threshold for T cell activation in rheumatoid arthritis. J Immunol, 179(4), 2609-2615. [4] Oflazoglu, E., Boursalian, T. E., Zeng, W., Edwards, A. C., Duniho, S., McEarchern, J. A. Grewal, I. S. (2009). Blocking of CD27-CD70 pathway by anti-CD70 antibody ameliorates joint disease in murine collagen-induced arthritis. J Immunol, 183(6), 3770-3777. [5] Park, J. K., Han, B. K., Park, J. A., Woo, Y. J., Kim, S. Y., Lee, E. Y., Song, Y. W. (2014). CD70-expressing CD4 T cells produce IFN-gamma and IL-17 in rheumatoid arthritis. Rheumatology (Oxford), 53(10), 1896-1900. [6] Ruf, M., Moch, H., & Schraml, P. (2015). Interaction of tumor cells with infiltrating lymphocytes via CD70 and CD27 in clear cell renal cell carcinoma. OncoImmunology, 4(12), e1049805. Disclosure of Interests None declared

Background: There was some reports of seasonal association with myopsitis onset. With the discovery of new myositis-specific autoantibodies (MSA), detailed grouping of idiopathic inflammatory myopathies was possible. Therefore, we evaluated the seasonal patterns in the onset of idiopathic inflammatory myositis (IIM) with MSA in Korea. Objectives: To evaluate the correlation between MSA and seasonal patterns of IIM in Korea. Methods: A total of 90 patients who met the criteria for probable or definite PM or DM and for whom data were collected from 7 referal centers in korea. 16 myositis-specific autoantibodies were detected by immunoblot with patient’s serum. Statistical analyses were performed using a Poisson model that assessed associations of sex, MSA, and month of onset of symptoms or month of diagnosis. Results: There were no significant seasonal patterns of disease onset in total IIM patients. Among MSAs, anti-synthetase (n=18), anti-transcriptional intermediary factor 1 γ (TIF1γ) (n=13), anti-melanoma differentiation-aasocianted protein 5 (anti-MDA5) (n=12), and anti–signal recognition particle (SRP) (n=12) were analyzed. Among 55 patients with dermatomyositis, patients with anti-MDA5 showed a significant peak in winter (n=12, P=0.05). This seasonal association was significant in women (n =10; P=0.045). Patients with anti–synthetase, anti-TIF1 γ or anti-SRP antibodies did not have a significant seasonal onset patterns. There were no significant seasonal patterns in patients without myositis-specific autoantibodies. Conclusion: Patients with anti-MDA5 showed a seasonality of myositis onset, in winter. Disclosure of interests: None declared

Background SinceBohan and Peter first described their diagnostic criteria foridiopathic inflammatorymyopathies (IIM) in 1975, new discoveries suchas myositis-specific and myositis-associated autoantibodies (Abs) have been made. Objectives To investigate correlations between specificmyositis Abs and their frequenciesand clinical associationsacross different IIM groups, collectively demonstrating theutility of the new clinicoserologic classification in Koreanadult patients with IIM. Methods We conducted a multicenter cohort study including 67adult patients (age≥18 years) who have been diagnosed as IIM by ENMC criteria.Immunoblot assay with Euroline strip(EUROIMMUN, Germany)was performed using the sera of definite deramatomyositis (DM, n=36), definite polymyositis (PM, n=25), amyopathic DM (n=4), DM sine dermatitis (n=1), and immune mediated necrotizing myopathy (IMNM, n=1). Patients were classifiedbased on three classifications: 1) novel clinicoserologic classification suggested by Troyanov et al. in 2017. 2) 2017 EULAR/ACR classification criteria. 3) 2004 European neuromuscular center (ENMC) criteria.Associations ofmyositis Absand clinical subsets of IIM were investigated. Results The distribution of the various IIM differed strikingly from those using the 3 classifications (Fig1). According to the 2004 ENMC classification and 2017 EULAR/ACR classification criteria, DM and PM was the most and the second frequent entities (DM: 55.2%, 56.7%; PM: 35.8%, 37.3%). But, using the new clinicoserologic classification,overlapmyositis(OM) is the major type of IIM and the frequency of PM is significantly decreased.Anti-ARS Abs specificity included anti-Jo-1(16.4%), -OJ(4.6%), -EJ(6.2%) -PL-7(3.1%), and -PL-12(4.6%). Interstitial lung disease was closely associated with anti-MDA5,and anti-ARS Abs, while DM-specific skin lesion was frequently observed in patients with anti-TIF1γ and anti-ARS Abs. Sevenpatients with cancer-associated DM were identified. They were positive for anti-TIFγ (5/7) and anti- SRP(3/7) (table 1). Conclusion Novel classification based on distinctive features and new myositis Absreflects the clinical phenotype of IIM better. Establishment of a system routinely available to screen myositis Abs is needed. Thiswill be beneficial to providemore precise diagnosis and proper management for patients with IIM. Reference: [1] Jean-Luc Senecal, Jean-Pierre Raynauld, Yves Troyanov. A New Classification of Adult Autoimmune Myositis. Arthritis Rheum2017;69:878-884. Disclosure of Interests None declared

Background T cells play a regulatory role in rheumatoid arthritis (RA) through inducing the homeostasis maintenance and self-tolerance [1]. Specially, the production and the oxidation mechanism of reactive oxygen species (ROS) were out of balance. Objectives The aim of the study was to compare ROS productions in T cell subset, which are helper T (TH) cell, cytotoxic T (TC) cell, T helper 17 (TH17) cell and regulatory T (Treg) cell in peripheral blood mononuclear cells (PBMC) of RA patients with RA activity. Methods Blood samples were collected from 30 RA patients and 10 healthy adult volunteers under IRB approval. RA activity was divided according to clinical parameter DAS28 [2]. PBMC cells were obtained from the whole blood using lymphocyte separation medium density gradient centrifugation. For separation between the live and dead cell populations, PBMC was stained with Live/Dead stain dye. After PBS washing, cells were incubated with antibodies for CD3, CD4, CD8, and CD25. Following fixation and permeabilization, and further stained with antibodies for FoxP3 and IL-17A. For ROS staining, CellRox and MitoSox were used. Results The frequency of TH cell was increased and that of TC cell was decreased in the peripheral blood of RA patients. TH17 and Treg cell population were significantly increased more than about 2-3 folds in active and inactive RA than healthy control. When the whole of cellular ROS production was measured, only Treg cell population was significantly increased in RA than control. Although ROS level was steadily increased with RA activity, there was a slight decline in severe RA compared to moderate and low RA. This difference is lager in mitochondrial specific ROS than total cellular ROS. The mitochondrial complex inhibitor reduced Treg cell frequency in PBMC from RA patients. Conclusion Treg is the most sensitive to ROS production among T cell subsets in RA. These findings provide a novel approach to regulate Treg function in RA through mitochondrial metabolism related ROS production. References [1] Szekanecz, Z., et al., New insights in synovial angiogenesis. Joint Bone Spine, 2010. 77(1): p.13-9. [2] Prevoo, M.L., et al., Modified disease activity scores that include twenty-eight-joint counts. Development and validation in a prospective longitudinal study of patients with rheumatoid arthritis. Arthritis Rheum, 1995. 38(1): p.44-8. Disclosure of Interests None declared

Background:Liver kinase B1 (LKB1) is known as a tumor suppressor gene and also inhibits reactive oxygen species (ROS) levels. Intracellular ROS are catalyzed by the enzyme complex nicotinamide adenine dinucleotide phosphate (NADPH) oxidase (NOX). We previously reported that NOX4 induced the migration and invasion of fibroblast-like synoviocytes (FLS) in rheumatoid arthritis (RA). Although LKB1 is expected to alleviate synovial inflammation through ROS regulation, the role of LKB1 in RA has not been examined.Objectives:To explore whether LKB1 affects RA inflammation, we transfected LKB1 siRNA and analyzed related gene expressions in RA FLS.Methods:Synovial tissues were obtained from RA patients who were undergoing synovectomy or joint replacement. The isolated cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum, 100 U/ml penicillin and 100 mg/ml streptomycin and maintained in a 5% CO2 incubator at 37 °C. FLS were used for experiments after four to six passages. Cells were transfected with lipofectamine transfection reagent and LKB1 siRNA duplex targeting constructs. After incubation for 24 h, downregulation of target expression was evaluated by real-time PCR and western blot analysis.Results:RA FLS was transfected with LKB1 siRNA and 90% of LKB1 mRNA expression was decreased. LKB1 knock-down also caused the decreased expression of mechanistic target of rapamycin (mTOR; 0.38 fold) and serine/threonine kinase (AKT) 2 (0.40 fold), which are downstream targets of LKB1. NOX4 was significantly increased (4.94 fold) by LKB1 inhibition. On the other side, the down regulated NOX4 induced significantly elevated LKB1 mRNA expression in RA FLS. When the expressions of proinflammatory cytokines were examined, IL-1β, IL-6, TNF-α were highly increased by LKB1 deficiency. FLS migration-related chemokines, IL-8 and MMP-3 were also enhanced compared to control.Conclusion:There was a negative correlation between NOX4 and LKB1 in RA FLS. As LKB1 deficiency induced the expression of proinflammatory cytokines and migration related chemokines, LKB1 could play a critical role in RA pathogenesis.References:[1]Bartok B, Firestein GS. Fibroblast-like synoviocytes: key effector cells in rheumatoid arthritis. Immunol Rev. 2010;233(1):233–55.[2]Mateen S, Moin S, Khan AQ, Zafar A, Fatima N. Increased reactive oxygen species formation and oxidative stress in rheumatoid arthritis. PLoS One. 2016;11(4):e0152925.Disclosure of Interests:None declared.

BACKGROUND Pulmonary arterial hypertension (PAH) is a major cause of morbidity and mortality among patients with systemic sclerosis (SSc). Early detection and prompt treatment of PAH associated with SSc (SSc-PAH) result in better prognosis. We conducted echocardiographic study to presume the prevalence of PAH in Korean adult SSc patients and to diagnose SSc-PAH in their early stages with right heart catheterization (RHC). METHODS We performed free of charge echocardiographic study including 37 adult SSc patients at the Chungnam National University Hospital. The possibility of PAH is determined by the estimation of pulmonary arterial pressure by peak tricuspid regurgitation velocity of > 3.0 m/s. Patients with possible PAH were recommended to undergo RHC to confirm the diagnosis. RESULTS In 37 patients, 8 patients were suspected with PAH. Among them, 6 patients agreed to be examined with RHC, and 4 were confirmed with PAH. The prevalence of possible PAH was 21.6% (8 of 37 patients), and that of confirmed PAH was 10.8% (4 of 37 patients). Four patients who were confirmed with SSc-PAH through RHC have been treated with specific pulmonary vasodilators and maintained stable. CONCLUSION Eight patients (21.6%) were possible PAH and 4 (10.8%) were diagnosed as SSc-PAH by RHC after the echocardiographic screening study of 37 adult SSc patients.

Aim There is growing evidence that cold-inducible RNA-binding protein (CIRP) promotes inflammatory responses. This study investigated the relationship between CIRP and rheumatoid arthritis (RA). Methods Peripheral blood and synovial fluid were collected from 15 patients with RA and from 16 patients with osteoarthritis (OA). The concentration of CIRP was measured with the sandwich enzyme-linked immunosorbent assay (ELISA). Results The concentration of serum CIRP was significantly elevated in the RA patient group (RA patients = 26.39 ± 10.48 pg/mL, OA patients = 17.14 ± 7.24 pg/mL, P = 0.009). Furthermore, the RA patient group had a significantly higher CIRP concentration than that of the OA patient group in synovial fluid (153.56 ± 108.93 pg/mL vs. 23.63 ± 16.18 pg/mL, P < 0.001). The mean synovial fluid concentration of CIRP was significantly higher than that of the serum concentration in the RA patient group (serum concentration = 26.39 ± 10.48 pg/mL, synovial fluid = 153.56 ± 108.93 pg/mL, P < 0.001). Disease Activity Score of 28 joints (DAS28)-ESR (erythrocyte sedimentation rate) and DAS28-CRP (C-reactive protein) were positively correlated with the synovial fluid concentration of CIRP (DAS28-ESR: r = 0.582, P = 0.023; DAS28-CRP: r = 0.541, P = 0.037). Conclusion The serum and synovial concentrations of CIRP in the RA patients were increased compared to the OA patients. Additionally, the synovial concentration of CIRP in RA patients correlated well with disease activity, that is, the DAS28-ESR/CRP. Based on these results, CIRP mediates inflammation and is a potential marker for synovial inflammation.

Background:Reactive oxygen species (ROS) and T helper 17 (TH17) cells have been known to play an important role in the pathogenesis of rheumatoid arthritis (RA). However, the interrelationship between ROS and TH17 remains unclear in RAObjectives:To explore whether ROS affect TH17 cells in peripheral blood mononuclear cells (PBMC) of RA patients, we analyzed ROS expressions among T cell subsets following treatment with mitochondrial electron transport chain complex inhibitors.Methods:Blood samples were collected from 40 RA patients and 10 healthy adult volunteers. RA activity was divided according to clinical parameter DAS28. PBMC cells were obtained from the whole blood using lymphocyte separation medium density gradient centrifugation. Following PBMC was stained with Live/Dead stain dye, cells were incubated with antibodies for CD3, CD4, CD8, and CD25. After fixation and permeabilization, samples were stained with antibodies for FoxP3 and IL-17A. MitoSox were used for mitochondrial specific staining.Results:The frequency of TH17 cells was increased by 4.83 folds in moderate disease activity group (5.1>DAS28≥3.2) of RA patients compared to healthy control. Moderate RA activity patients also showed higher ratio of TH17/Treg than healthy control (3.57 folds). All RA patients had elevated expression of mitochondrial specific ROS than healthy control. When PBMC cells were treated with 2.5uM of antimycin A (mitochondrial electron transport chain complex III inhibitor) for 16 h, the frequency of TH17 cells was significantly decreased.Conclusion:The mitochondrial electron transport chain complex III inhibitor markedly downregulated the frequency of TH17 cells in moderate disease activity patients with RA. These findings provide a novel approach to regulate TH17 function in RA through mitochondrial metabolism related ROS production.References:[1]Szekanecz, Z., et al., New insights in synovial angiogenesis. Joint Bone Spine, 2010. 77(1): p. 13-9.[2]Prevoo, M.L., et al., Modified disease activity scores that include twenty-eight-joint counts. Development and validation in a prospective longitudinal study of patients with rheumatoid arthritis. Arthritis Rheum, 1995. 38(1): p. 44-8.Disclosure of Interests:None declared

Background:Rheumatoid arthritis (RA) is a progressive, chronic inflammatory autoimmune disease. Pro-inflammatory molecules, activated lymphocytes, and the migration of inflammatory cells are important in the development of RA. There are many unknown causes of RA. And there are many patients who are refractory to treatment with known disease-modifying anti-rheumatic drugs. So, unknown cause of RA needs to be elucidated.CD70 is a member of the tumor necrosis factor (TNF) superfamily and a ligand for CD27. The interaction of CD70 with its receptor CD27 promotes expansion and differentiation of memory and effector T cells as well as B-cell expansion and plasma cell differentiation. Hypoxia is an important micro-environmental factor in RA synovium. Hypoxia induces activation of hypoxia inducible factor (HIF). The expression of HIF-2α is up-regulated in human RA synovium. Reactive oxygen species (ROS) has been implicated in the pathophysiology of RA.Objectives:In this study, we tried to examine the presence of CD70 in RA synovium and investigate the role of CD70 in the development of RA associated with HIF-2α and ROS.Methods:Fibroblast-like synoviocyte (FLS), peripheral blood (PB) and synovial fluid (SF) were used for experiments. FLS was stimulated with recombinant human (rh)-IL-17 and rh-TNF-α. N-acetyl-L-cysteine (NAC) was used as a ROS scavenger. HIF-2α inhibitor (PT-2385) was used for examine the effect of HIF-2α in RA-FLS. RT-PCR, qPCR, western blotting, flow-cytometry, ELISA, cell migration assay, and scratch wound assay were performed.Results:CD70 mRNA is present and elevated by stimulation with IL-17 and TNF-α in both RA-FLS and osteoarthritis (OA)-FLS (Fig 1). CD70 also expresses on the surface of RA-FLS and OA FLS (Fig 2). CD70 expression on the surface of FLS is elevated by stimulation with IL-17 and TNF-α in both RA and OA. Soluble CD27 is present higher in the supernatant of RA-SF than OA-SF (Fig 3). HIF-2α mRNA, HIF-2α protein, and the amount of ROS were all elevated after treatment with IL-17 and TNF-α in RA-FLS (Fig 4, Fig 5). CD70 expression and the amount of ROS were lowered by treatment with HIF-2α inhibitor in RA-FLS (Fig 6). Decreased amount of ROS results in decreased CD70 expression on the RA-FLS (Fig 7). CD70 influenced on cell migration directly or by HIF-2α (Fig 8).Conclusion:In this study, we found the function of CD70 in RA-FLS associated with HIF-2α and ROS. First, CD70 on RA-FLS interacts with CD27 in the RA-SF and this interaction produces sCD27 (Fig. 9) and CD70 has an influence on the migration of RA-FLS. Second, IL-17 and TNF-α are critical factors to trigger the expression of CD70, HIF-2α and ROS in RA synovium. Third, CD70 is regulated by HIF-2α associated with ROS. From these results, we suggest that CD70 may be a new therapeutic target of RA. And sCD27 also may be an important diagnostic maker of RA.References:[1]Lundy SK, Sarkar S, Tesmer LA, Fox DA. Cells of the synovium in rheumatoid arthritis. T lymphocytes. Arthritis Res Ther. 2007;9(1):202.[2]Nevius E, Gomes AC, Pereira JP. Inflammatory Cell Migration in Rheumatoid Arthritis: A Comprehensive Review. Clin Rev Allergy Immunol. 2016;51(1):59-78.[3]Bowman MR, Crimmins MA, Yetz-Aldape J, Kriz R, Kelleher K, Herrmann S. The cloning of CD70 and its identification as the ligand for CD27. J Immunol. 1994;152(4):1756-61.[4]Kitajima S, Lee KL, Fujioka M, Sun W, You J, Chia GS, et al. Hypoxia-inducible factor-2 alpha up-regulates CD70 under hypoxia and enhances anchorage-independent growth and aggressiveness in cancer cells. Oncotarget. 2018;9(27):19123-35.[5]Gaber T, Dziurla R, Tripmacher R, Burmester GR, Buttgereit F. Hypoxia inducible factor (HIF) in rheumatology: low O2! See what HIF can do! Ann Rheum Dis. 2005;64(7):971-80.Disclosure of Interests:None declared

Background Behcet’s disease(BD) is an inflammatory multi-genetic disorder with unknown etiology. In the previous study, we sequenced whole mitochondrial nucleotides from blood of 20 BD patients and 10 sex-, age-matched healthy controls, m.248A>G, m.709G>A, m.3970C>T, m.6392T>C, m.6962G>A, m.10310G>A, m.10609T>C, m.12406G>A, m.12882C>T, m.13928G>C, m.16129G>A, and m16304T>C were more frequently observed in patients group than healthy control without statistical significance. While, m.304C>A, m.3010G>A, m.4883C>T, m.5178C>A, and m.14668C>T were more frequent in control group (p=0.008, 0.026, 0.007, 0.007, and 0.026, respectively). m.16182A>C, m.16183A>C, m.16189T>C were associated with uveitis (p=0.041, 0.022, and 0.014, respectively). Objectives We performed a follow-up study to validate these possible associations in larger groups. Methods Whole blood or buffy coat were collected from 98 BD patients from four university hospitals located in Chung-Cheong district of Republic of Korea, 196 age-, sex-matched healthy controls from Konyang University Hospital. Above mentioned 20 targeted mitochondrial DNA(mtDNA) genomes were analysed using MassARRAY® system(Agena Bioscience, Inc., San Diego, CA, USA) for m.709, m.3010, m.4883, m.6392, m.6962, m.10310, m.10609, m.12406, m.12882, m.13928, and m.14668; Sanger sequencing for m.248, m.304, m.5178, m.16129, m.16182, m.16183, m.16189, and m.16304; and TaqMan-based genotyping assay(Applied Biosystems, Foster City, CA, USA) for m.3970. Results were compared with the revised Cambridge Reference Sequence (rCRS). Chi square or Fisher’s exact test were used to analyse association of mtDNA alterations between groups and between mtDNA alterations and clinical/laboratory characteristics. Results Presence of m.248A>G, m.304C>A, m.709G>A, m.3010G>A, m.3970C>T, m.4883C>T, m.5178C>A, m.6392T>C, m.6962G>A, m.10310G>A, m.10609T>C, m.12406G>A, m.12882C>T, m.13928G>C, m.14668C>T, m.16129G>A, and m16304T>C were not differentiated by the groups. However, m.16812A>C was more frequently observed in the patient group than control [22 (22.4%) vs. 24 (12.2%), p=0.061], and it was significantly associated with HLA-B51 positivity (p=0.011), arthralgia (p=0.043) and methotrexate use (p=0.02), and was not associated with uveitis in the follow-up study. Among clinical and laboratory characteristics in BD patients, thrombosis was more frequently observed in male patients than female patients [7 (22.6%) vs. 0 (0%), p Conclusions We performed a follow-up study to validate possible associations between BD and 20 mtDNA alterations. m.16812A>C could be associated with BD and its several clinical or laboratory characteristics a Korean population. References [1] Xavier JM, Shafiee NM, Ghaderi F, Rosa A, Abdollahi BS, Nadji A, et al. Association of mitochondrial polymorphism m.709G>A with Behcet’s disease. Ann Rheum Dis2011;70:1514–6. [2] Kwon MH, Joung CI. Genetic Associations of Mitochondrial DNA Polymorphisms with Behcet’s Disease in a Korean Population: A Pilot Study. J Rheum Dis 2016;23(1):23–9. Acknowledgements The current study was supported by the National Research Foundation of Korea funded by the Korean Government (grant no. NRF-2017R1C1B2008199). Disclosure of Interest None declared

Background In osteoarthritis (OA), the activation of inflammation response involving the interaction of cartilage and synovial hyperplasia may contribute to disease progression. However, inflammatory cells in OA synovial fluid (OASF) have been rarely studied. Objectives To investigate the phenotype of CD14 +cells and the secretion of proinflammatory cytokines by these cells Methods Immunohistochemistry staining in OA synovium was performed using anti-CD14, anti-CD16 and anti-CD56 antibody. The OASF was obtained through arthrocentesis. Mononuclear cells from OASFs were stained with anti-CD3, anti-CD4, anti-CD14, anti-CD16, anti-TLR4 or anti-TLR2 and analysed using flow cytometry. CD14 +CD16+and CD14+CD16 mononuclar cells in OASF were selected with magnetic microbeads. In the supernatant of these cells culture, the concentration of IL-1β, IL-6, IL-8, TNFα, MMP-1,–3 were measured by Luminex. Results In OA synovium, CD14 or CD16 was stained, but CD56 was not expressed. In OASF, there was a substantial number of CD14 +cells (36.6%±25.2%), CD3 +cells (37.4%±12.9%), with a rarity of CD90 +cells (1.7%±1.3%). The proportion of CD14 +cells was increased significantly in recurred synovial fluid, compared with the proportion in initial synovial fluid. Among CD14 +cells in OASF, CD14 +CD16+monocyte subpopulation (21.2%±21.8%) was more abundant than CD14 +CD16− monocyte subpopulation (10.9%±10.0%). TLR4 and TLR2 expressions were higher in CD14 +CD16+cells than in CD14 +CD16− cells. The concentration of IL-8 and MMP-3 was more increased in the supernatant of CD14 +CD16+cells than in that of CD14 +CD16− cells. Conclusions In OASF, the proportion of CD14 +cells was increased in recurred synovial effusion. Compared to CD14 +CD16− monocyte subpopulation, CD14 +CD16+monocyte subpopulation released more cytokine such as IL-8 and MMP-3, and had higher expressions of TLR4 and TLR2. Disclosure of Interest None declared

Background The recent studies about gout demonstrated the correlation between gout and cardiovascular disease (CVD). The urate lowering therapy (ULT) ameliorates the outcomes of CVD. The poor adherence with ULT in patients with gout is the main obstacle. It is induced mainly due to a lack of adequate information about the efficacy of ULT. Objectives This study was performed to analyse the effects of the education for patients with gout. Methods 116 patients with gout were enrolled and categorised by two groups, education and non-education. The face to face education was conducted by the specially educated nurse. And all participants in two groups received an leaflet about general information of gout including lifestyle advice, nutrition and drugs of ULT. The patients in non-education group were also educated by nurse on their second visit (after two or three month from first visit). The score of patients’ satisfaction using the visual analogue scale (from 0 to 100 mm) and questionnaire about satisfaction and questionnaire of patients’ knowledge about disease (gout) were assessed. And we analysed patients’ serum uric acid level and drug compliance. Results A total of 116 patients were randomised to education or non-education group equally. Patients’ satisfaction in visual analogue scale was significantly higher in education group (education group: 82.7±21.0 mm, vs. non-education group: 72.8±20.7 mm, p Patients’ satisfaction questionnaire was significantly higher in education group (education group: 3.89±0.5, vs. non-education group: 3.69±0.4, p=0.017). The level of knowledge about gout was higher in education group (education group: 7.4±2.0, vs. non-education group: 6.2±2.3, p=0.004). The serum uric acid level on second visit is decreased after education (baseline: 5.97±1.93 mg/dL, vs. second visit: 5.32±1.35 mg/dL, p=0.001). Besides drug compliance on second visit is improved after education (baseline: 89.6%±16.4%, vs. second visit: 94.3%±9.9%, p=0.018). Conclusions The face to face education for gout improved the patients’ satisfaction, drug adherence and serum uric acid level. The education is very important part in treatment of gout. References [1] Singh JA, Strand V. Gout is associated with more comorbidities, poorer health-related quality of life and higher healthcare utilisation in US veterans. Ann Rheum Dis2008;67:1310–6. [2] Reesi F, Jenkins W, Doherty MPatients with gout adhere to curative treatment if informed appropriately: proof-of-concept observational study. Ann Rheum Dis201372: 826–830. [3] Cho SK, Kim D, Choi J, et al. Impact of Patient Education on the Satisfaction of Rheumatoid Arthritis Patients: A Randomized Trial of Nurse-led Versus Medical Docter-led Education. J Rheum DisVol. 23, No. 2, April, 2016. Disclosure of Interest None declared

OBJECTIVE Anti-transcriptional intermediary factor 1 (TIF1) antibody is associated with idiopathic inflammatory myopathies (IIMs). The aim of this study was to investigate the expression of TIF1s in IIMs. METHOD TIF1α, β or γ expression in the skin and muscle of patients and controls was studied by immunohistochemistry. Serum myositis-specific autoantibodies were detected by immunoblot. RESULTS TIF1α was expressed in the skin of most dermatomyositis (DM) patients but not in the controls (80% vs 0%, P

B7-H3, a newly identified B7 family member, has functional duality as a co-stimulator and co-inhibitor that fine-tunes T cell-mediated immune responses. Given that B7-H3 expression on human monocytes and dendritic cells is enhanced by inflammatory cytokines, its potential inmmunoregulatory role at sites of inflammation has been suggested. Further, monocytes play crucial roles in the pathophysiology of various inflammatory disorders including autoimmune diseases; however, the immunological role of B7-H3 in rheumatoid arthritis (RA) has not been defined. Thus, we aimed to investigate the possible roles of monocyte B7-H3 in the pathogenesis of RA. Synovial monocytes, but not peripheral monocytes, in RA patients predominantly express surface B7-H3. The 4Ig isoform of B7-H3 is exclusively induced on the cell surface, whereas the 2Ig B7-H3 isoform is constitutively expressed in the intracytoplasmic region of both peripheral and synovial monocytes. B7-H3 knockdown experiments reveal that surface B7-H3 has an inhibitory effect on IFN-γ production in CD4 memory cells. Moreover, surface B7-H3 expression on synovial monocytes inversely correlates with RA clinical parameters. Our findings demonstrate that activation-induced B7-H3 expression on synovial monocytes has the potential to inhibit Th1-mediated immune responses and immunomodulatory roles affecting RA pathogenesis.

Purpose To evaluate the incidence of fellow-eye neovascularization in retinal angiomatous proliferation (RAP) in a Korean population and associated risk factors. Method This retrospective, observational study included 81 eyes (81 patients) diagnosed with unilateral RAP who were followed up for ≥12 months. The RAP diagnosis was based on an indocyanine green angiography reviewed by two retinal specialists. In fellow eyes experiencing neovascularization, the period between RAP diagnosis and neovascularization was compared between eyes with and without reticular pseudodrusen. Results The mean age (±standard deviation) of the 81 patients was 74.7 ± 6.1 years. The mean follow-up period was 27.8 ± 12.4 months. Fellow-eye neovascularization was noted in 31 patients (38.3%), and 24 of these (77.4%) was a RAP subtype. Fellow-eye involvement was noted within 12 months in 13 eyes (16.0%). The period between diagnosis and fellow-eye neovascularization was significantly shorter in eyes with reticular pseudodrusen (mean 13.8 ± 8.5 months) than in eyes without reticular pseudodrusen (mean 21.2 ± 9.1 months; p = 0.031). Conclusion In our cohort of unilateral RAP patients, fellow-eye neovascularization was noted in 38.3% in 27.8 months. The presence of reticular pseudodrusen in the fellow eye was closely associated with relatively early onset.

Rheumatoid arthritis (RA) is a chronic inflammatory autoimmune disease that primarily affect the joints and inflammatory cell migration into inflamed articular sites contribute to this disease. Among the inflammatory cells, human mucosal-associated invariant T (MAIT) cells were recently recognized as critical cellular component with a pathological role in RA. However, their migratory characteristics are poorly understood. The aim of this study was to determine whether human MAIT cells preferentially traffick to inflamed synovial sites in rheumatoid arthritis patients and to elucidate the underlying mechanism. First, we found that TNFα and IL-1β were elevated in synovial fluid (SF) of RA patients, which resulted in increased expression of E-selectin, ICAM-1 and V-CAM-1 on blood vessel endothelial cells. To understand whether TNFα and IL-1β in the SF facilitated MAIT cell migration, we analyzed CD161+ TCRα7.2+ MAIT and other CD3+ T cells for differences in migratory capacity. Collectively, our results demonstrate that TNFα and IL-1β in the SF facilitated MAIT cell migration dependent on expression of selectin ligand, sialyl LewisX (sLeX) and CCR6 on MAIT cells. We also showed that MAIT cells in the SF from RA patients equipped upregulated sLeX compared to the peripheral blood of RA patients and healthy persons, which suggest that TNFα and IL-1β mediated expression of E-selectin preferentially attract sLeX mediated MAIT cell migration into the SF of RA patients.

Purpose: To investigate 12-month treatment outcomes of anti-vascular endothelial growth factor therapy in eyes with typical exudative age-related macular degeneration with good baseline visual acuity. Methods: This retrospective observational case series included 18 eyes (18 patients) with typical exudative age-related macular degeneration with a baseline best-corrected visual acuity of 20 / 25 or better. Patients were treated with anti-vascular endothelial growth factor monotherapy during the 12-month follow-up period. Baseline visual acuity and central foveal thickness were compared to the values at 12 months. Results: Patients received an average of 4.4 ± 1.3 intravitreal anti-vascular endothelial growth factor injections. The mean logarithm of minimum angle of resolution visual acuity was 0.08 ± 0.04, 0.08 ± 0.07, 0.12 ± 0.09, and 0.16 ± 0.11 at baseline, three months, six months, and 12 months, respectively. Visual acuity at 12 months was significantly worse than the baseline value at diagnosis (p = 0.017), and the mean central foveal thickness at the defined time points was 270.2 ± 55.6, 204.4 ± 25.4, 230.1 ± 56.3, and 216.8 ± 48.7 µm, respectively. The central foveal thickness at 12 months was significantly less than the baseline value at diagnosis (p = 0.042). Conclusions: Deterioration in visual acuity was noted in eyes with typical exudative age-related macular degeneration with good baseline visual acuity, suggesting the need for close patient monitoring and prompt treatment even in patients with good baseline visual acuity.

The genome of the thermotolerant methylotrophic yeast Hansenula polymorpha reveals the presence of five PMT homologues (HpPMT1, HpPMT2, HpPMT4, HpPMT5, and HpPMT6) encoding protein O-mannosyltransferases. Here, we report on the systematic characterization of HpPMT5 and HpPMT6, encoding novel PMT1 and PMT2 subfamily members, respectively. Although no apparent growth defects were detected in the Hppmt5Δ and Hppmt6Δ single mutants, the single mutants showed dramatic sensitivity to the Pmt1p inhibitor, and the Hppmt1pmt5Δ and Hppmt1pmt6Δ double mutants displayed increased susceptibility to cell wall-disturbing reagents. Activation of the cell wall integrity signaling pathway in the double mutant strains was further indicated by the markedly induced phosphorylation of MAP kinases, such as HpMpk1p and HpHog1p. Noticeably, O-mannosylation of the surface glycoproteins HpWsc1p and HpMid2p became severely defective only in the double mutants, supporting the involvement of HpPmt5p and HpPmt6p in O-mannosylation of these sensor proteins. On the other hand, co-immunoprecipitation experiments revealed only marginal interaction between HpPmt5p and HpPmt2p, even in the absence of HpPmt1p. Taken together, our results suggest that the functions of HpPmt5p and HpPmt6p are minor but become crucial upon the loss of HpPmt1p for protein O-mannosylation, which is essential for cell growth, cell wall integrity, and stress resistance in H. polymorpha.

Lung diseases (LD) are common extra-articular manifestations in rheumatoid arthritis (RA). However, little is known about factors associated with susceptibility to rheumatoid arthritis-related lung diseases (RA-LD). The aim of the present study was to investigate whether the single nucleotide polymorphisms (SNPs) of PADI4 and HLA-DRB1 alleles were associated with RA-LD. Blood samples and clinical data were collected from 116 consecutive RA patients who satisfied the 1987 American College of Rheumatology classification criteria. RA-LD was diagnosed using high-resolution computed tomography of the chest. All patients were genotyped for SNPs of PADI4 and HLA-DRB1 alleles and analyzed for full amino acid sequence of the HLA protein corresponding to a 4-digit HLA typing. Data were analyzed by independent t test (or Mann–Whitney test) for continuous variables, Chi-square test (or Fisher’s exact test) and trend test for categorical variables, and logistic regression analysis. Ninety-four (81.0 %) RA patients had LD, of which eight (6.9 %) had interstitial lung disease (ILD) and 92 (79.3 %) had airway abnormalities in which 64 (55.2 %) showed bronchiectasis and 47 (40.5 %) revealed bronchial wall thickening. The recessive genotype of padi4_92 was susceptible to airway abnormalities (OR = 2.22, 95 % CI = 1.05–4.49, p = 0.034). Tryptophan at position 9 of HLA-DRB1 sequence was associated with the susceptibility to RA-ILD (OR = 22.89, 95 % CI = 1.20–432.56, p = 0.037). PADI4 polymorphisms and HLA-DRB1 alleles could attribute differently to the development of airway abnormalities and ILD, respectively, in RA.

Non-small cell lung cancer (NSCLC) is the leading cause of cancer related deaths. Most patients were presented with advanced disease at the time of diagnosis. In advanced NSCLC, it is almost impossible to anticipate complete remission by using only cytotoxic chemotherapy or molecularly targeted agents. In our case, two patients were diagnosed as advanced NSCLC and received chemotherapy. They achieved complete response (CR). After finishing treatment, disease recurred. They were retreated with the same regimens and achieved second CR. Until now, they have received each regimen, continuously, and the CR state has been maintained.

Dermatomyositis (DM) and polymyositis (PM) are representative idiopathic inflammatory myopathies characterized by symmetric and progressive proximal muscle weakness. Especially, DM is identified by characteristic skin lesions and has many extramuscular manifestations including various cardiac abnormalities, interstitial lung disease, and malignancy. However, involvement of peripheral nervous system in DM/PM is very rare and less known. The term “Neuromyositis” was introduced by Senator in 1893 to describe the concomitant involvement of the peripheral nervous system in DM/PM. Since then, a very few cases of neuromyositis have been reported mainly in the United States and Europe. Therefore, the pathogenetic mechanism and disease progression are unclear. In recent years, a few more cases were reported in Asia, specifically, China and Japan; however, none in Korea. Here, we describe a case of DM-associated neuromyositis in a 42-year-old man in Korea and review previous publications through literature research. (J Rheum Dis 2019;26:211-218)

Background: Glutamate is a well-known central nervous system (CNS) excitatory neurotransmitter that plays a role in memory and learning. However, in excess, it leads to a process called excitotoxicity resulting in cell death. To investigate the mechanism of glutamate cytotoxicity, the apoptosis signaling pathway of primary rat astrocytes was explored in vitro. With this study, we hope to improve the prevention and treatment of ischaemic strokes and various central nervous system disorders. Methods: To produce a model of cell injury, primary rat astrocytes were treated with glutamate. Results: Treatment with glutamate induced death by apoptosis in primary rat astrocytes. This was evidenced by an increase in sub-G0/G1 fraction of the cell cycle with a loss of cell viability. Glutamate also increased the intracellular accumulation of Ca ions, the expression levels of glucose-regulated protein 78 (Grp78) and C/EBP homologous protein (CHOP) protein, and the phosphorylation of protein kinase r-like endoplasmic reticulum kianse (PERK). However, the expression pattern of activating transcription factor (ATF)4 protein did not change and the 90 kDa ATF6 was cleaved to 50 kDa along with a reduced amount of Bcl-2 protein in a time-dependent manner. Interestingly, pre-treatment with glutathione markedly suppressed the reactive oxygen species (ROS) generation and the sub-G0/G1 fraction. However, the intracellular concentration of Ca did not change. Conclusion: Our findings suggest that glutamate injures primary rat astrocytes through the endoplasmic reticulum (ER) stressmediated apoptotic signaling pathway, as well as, ROS generation. More specifically, through Grp78, PERK, and CHOP, glutamate activates the ER stress-mediated signaling pathway in astrocytes and activates ATF6 to reduce the expression of the Bcl-2 proteins contributing to apoptosis. In addition, the ER stress-mediated signaling pathway is closely related to the transformation of intracellular ROS. This information should be applied to research for the prevention and treatment of strokes and other CNS conditions.

Free amine chitooligosaccharides (FACOS, Kitto Life, Seoul, Republic of Korea) with an average molecular mass of 3.5 kDa were efficiently produced using an ultrafiltration membrane reactor system. To evaluate the effect of chitooligosaccharides on serum cytokine levels in elderly adults after oral intake, 5.1 g/day of FACOS was given to elderly (age range, 74-86 years; mean, 80 +/- 3 years) volunteers during an 8-week experimental period. Interleukin (IL)-12 and interferonã levels were significantly higher in the FACOS group than in the control group (P.05). However, levels of the inflammatory cytokines IL-1beta and tumor necrosis factor-alpha decreased after FACOS intake during the experimental period. The results of this study suggest that the oral intake of chitooligosaccharides may have beneficial effects on specific cell-mediated immunity while also acting as an anti-inflammatory agent in aged subjects.

Xanthii Fructus (XF) is an herb widely used in medicine for the treatment of a variety of inflammatory pathologies. In this study, using mouse peritoneal macrophages, we have examined whether XF affects nitric oxide (NO), tumor necrosis factor (TNF)-alpha, and interleukin (IL)-12p40 production induced by interferon (IFN)-gamma and lipopolysaccharide (LPS). XF inhibits IFN-gamma and LPS-induced NO production in a dose dependent manner. The decrease in NO synthesis was reflected as a decreased amount of inducible NO synthase protein. Furthermore, we also found that XF inhibits pro-inflammatory cytokine TNF-alpha production. However, treatment of XF in peritoneal macrophages had no effect on IL-12p40 production. These findings suggest that XF may be used in controlling macrophages-mediated inflammatory diseases.

We studied the inhibitory effect of Powerdental on the growth and acid production of Streptococcus mutans as well as secretion of pro-inflammatory cytokine tumor necrosis factor-alpha (TNF-alpha). The growth of Streptococcus mutans was reduced by the presence of the Powerdental (1 mg/ml) and NaCl (1 mg/ml) significantly, and the positive control group (1% NaF) also exhibited a significant antibacterial activity. The decrease of pH was significantly inhibited in the presence of Powerdental (1 mg/ml) compared to the control group. The decrease in pH was also inhibited in the presence of positive control (1% NaF), but the bamboo salt alone did not show inhibitory activity. We also found that Powerdental (0.01 mg/ml) inhibited significantly the secretion of TNF-alpha with 46.5+/-0.2% from human mast cells. Our results suggest that Powerdental contributes to the prevention or treatment of periodontitis and other oral diseases or inflammatory diseases.

Bucillamine is used for the treatment of rheumatoid arthritis. This study investigated the protective effects of bucillamine against cisplatin-induced damage in auditory cells, the organ of Corti from postnatal rats (P2) and adult Balb/C mice. Cisplatin increases the catalytic activity of caspase-3 and caspase-8 proteases and the production of free radicals, which were significantly suppressed by pretreatment with bucillamine. Bucillamine induces the intranuclear translocation of Nrf2 and thereby increases the expression of γ-glutamylcysteine synthetase (γ-GCS) and glutathione synthetase (GSS), which further induces intracellular antioxidant glutathione (GSH), heme oxygenase 1 (HO-1) and superoxide dismutase 2 (SOD2). However, knockdown studies of HO-1 and SOD2 suggest that the protective effect of bucillamine against cisplatin is independent of the enzymatic activity of HO-1 and SOD. Furthermore, pretreatment with bucillamine protects sensory hair cells on organ of Corti explants from cisplatin-induced cytotoxicity concomitantly with inhibition of caspase-3 activation. The auditory-brainstem-evoked response of cisplatin-injected mice shows marked increases in hearing threshold shifts, which was markedly suppressed by pretreatment with bucillamine in vivo. Taken together, bucillamine protects sensory hair cells from cisplatin through a scavenging effect on itself, as well as the induction of intracellular GSH.

A chronic inflammatory response associated with beta-amyloid (Abeta) and interleukin-1beta (IL-1beta) is responsible for the pathology of Alzheimer's disease (AD). Astrocytes are predominant neuroglial cells of the central nervous system and are actively involved in cytokine-mediated events in AD. To investigate the biological effect of water-soluble chitosan (WSC), we examined cytotoxicity, production of pro-inflammatory cytokines and inducible nitric-oxide synthase (iNOS) on human astrocytoma cell line CCF-STTG1 stimulated with IL-1beta and Abeta fragment 25-35 (Abeta[25-35]). In 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazoliumbromide colorimetric assay, WSC by itself had no effect on cell viability on human astrocytoma cells. The effects of WSC on tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) were evaluated with enzyme-linked immunosorbent assay and Western blotting. The production of TNF-alpha and IL-6 was induced by IL-1beta and Abeta[25-35] and synergistically amplified by the co-stimulation of IL-1beta and Abeta[25-35]. The secretion and expression of pro-inflammatory cytokines, TNF-alpha and IL-6, was significantly inhibited by pretreatment with WSC in human astrocytoma cells. The expression of iNOS was induced by IL-1beta and Abeta[25-35] and was partially inhibited by treatment with WSC. We demonstrate the regulatory effects of WSC in human astrocytes for the first time and suggest the anti-inflammatory effect of WSC may reduce and delay AD pathologic events.

Diallyl disulfide (DADS), a component of garlic (Allium sativum), has been known to exert potent chemopreventative activity against colon, lung, and skin cancers. However, its molecular mechanism of action is still obscure. The present study demonstrated that DADS induces apoptosis of human leukemia HL-60 cells in a concentration- and time-dependent manner with an IC50 for cell viability of less than 25 microM. DADS activated caspase-3 as evidenced by both the proteolytic cleavage of the proenzyme and increased protease activity. Activation of caspase-3 was maximal at 3 hr and led to the cleavage of 116 kDa poly(ADP-ribose) polymerase (PARP), resulting in the accumulation of an 85 kDa cleavage product. Both activation of caspase-3 and cleavage of PARP were blocked by pretreatment with either antioxidants or a caspase-3 inhibitor, but not a caspase-1 inhibitor. DADS increased the production of intracellular hydrogen peroxide, which was blocked by preincubation with catalase. These results indicate that DADS-induced apoptosis is triggered by the generation of hydrogen peroxide, activation of caspase-3, degradation of PARP, and fragmentation of DNA. The induction of apoptosis by DADS may be the pivotal mechanism by which its chemopreventative action against cancer is based.

Monocytes function as crucial innate effectors in the pathogenesis of chronic inflammatory diseases, including autoimmunity, as well as in the inflammatory response against infectious pathogens. Human monocytes are heterogeneous and can be classified into three distinct subsets based on CD14 and CD16 expression. Although accumulating evidence suggests distinct functions of monocyte subsets in inflammatory conditions, their pathogenic roles in autoimmune diseases remain unclear. Thus, we investigated the phenotypic and functional characteristics of monocytes derived from synovial fluid and peripheral blood in RA patients in order to explore the pathogenic roles of these cells. In RA patients, CD14+CD16+, but not CD14dimCD16+, monocytes are predominantly expanded in synovial fluid and, to a lesser degree, in peripheral blood. Expression of co-signaling molecules of the B7 family, specifically CD80 and CD276, was markedly elevated on synovial monocytes, while peripheral monocytes of RA and healthy controls did not express these molecules without stimulation. To explore how synovial monocytes might gain these unique properties in the inflammatory milieu of the synovial fluid, peripheral monocytes were exposed to various stimuli. CD16 expression on CD14+ monocytes was clearly induced by TGF-β, although co-treatment with IL-1β, TNF-α, or IL-6 did not result in any additive effects. In contrast, TLR stimulation with LPS or zymosan significantly downregulated CD16 expression such that the CD14+CD16+ monocyte subset could not be identified. Furthermore, treatment of monocytes with IFN-γ resulted in the induction of CD80 and HLA-DR expression even in the presence of TGF-β. An in vitro assay clearly showed that synovial monocytes possess the unique capability to promote Th1 as well as Th17 responses of autologous peripheral CD4 memory T cells. Our findings suggest that the cytokine milieu of the synovial fluid shapes the unique features of synovial monocytes as well as their cardinal role in shaping inflammatory T-cell responses in RA.

The production of recombinant therapeutic proteins is one of the fast-growing areas of molecular medicine and currently plays an important role in treatment of several diseases. Yeasts are unicellular eukaryotic microbial host cells that offer unique advantages in producing biopharmaceutical proteins. Yeasts are capable of robust growth on simple media, readily accommodate genetic modifications, and incorporate typical eukaryotic post-translational modifications. Saccharomyces cerevisiae is a traditional baker's yeast that has been used as a major host for the production of biopharmaceuticals; however, several nonconventional yeast species including Hansenula polymorpha, Pichia pastoris, and Yarrowia lipolytica have gained increasing attention as alternative hosts for the industrial production of recombinant proteins. In this review, we address the established and emerging genetic tools and host strains suitable for recombinant protein production in various yeast expression systems, particularly focusing on current efforts toward synthetic biology approaches in developing yeast cell factories for the production of therapeutic recombinant proteins.

To evaluate the efficacy of intravitreal anti-vascular endothelial growth factor (VEGF) monotherapy for patients diagnosed with exudative age-related macular degeneration (AMD) accompanied by submacular hemorrhage.Retrospective, observational case series.Ninety-one eyes of 91 patients who initially presented with submacular hemorrhage associated with exudative AMD from October 2009 to September 2012. Patients were followed up for at least 6 months after treatment.Best-corrected visual acuity (BCVA) was measured at diagnosis and at 1, 3, and 6 months after treatment. The duration of symptoms was estimated. The extent of hemorrhage was estimated using fundus photography, and central foveal thickness was measured using optical coherence tomography. Change in BCVA during 6 months after treatment was estimated. The correlation of BCVA at 6 months with duration of symptoms, extent of hemorrhage, and central foveal thickness was evaluated.The BCVA, duration of symptoms, extent of hemorrhage, and central foveal thickness.The mean duration of symptoms was 27.6±39.5 days. The mean extent of hemorrhage was 7.8±5.6 disc areas, and the mean central foveal thickness was 610.1±249.6 μm. All eyes were treated with 3.2±0.8 (range, 1-5) monthly intravitreal anti-VEGF injections during the 6-month follow-up period. The logarithm of the minimum angle of resolution BCVA at diagnosis and at 1, 3, and 6 months after the initial diagnosis was 1.38±0.53 (Snellen equivalent, 20/479), 1.27±0.57, 1.05±0.58, and 0.96±0.65 (Snellen equivalent, 20/182), respectively. The BCVA at 6 months significantly improved from baseline (P 0.001). Poor BCVA at 6 months correlated with a longer duration of symptoms, greater extent of hemorrhage, and greater central foveal thickness (P = 0.008, P = 0.004, and P = 0.014, respectively).Anti-VEGF monotherapy was found to be a useful treatment option for exudative AMD accompanied by submacular hemorrhage. However, the limited efficacy in eyes with large hemorrhage may suggest the need for more aggressive treatment in these cases.