Cannabis sativa root has traditionally been used to relieve pain and inflammation, but its pharmacological properties remain underexplored due to low levels of psychoactive cannabinoids. This study aimed to investigate the anti-inflammatory and antinociceptive effects of the ethyl acetate fraction of Cannabis sativa root (CSREA) using in vivo rodent pain models. Mice were subjected to formalin and acetic acid-induced nociceptive tests, while rats were evaluated using a carrageenan-induced paw edema model. CSREA significantly reduced pain-related behaviors in both early (0–10 min) and late phases (15–30 min) of the formalin test and decreased writhing responses in the acetic acid model. Notably, CSREA also improved survival rates following acetic acid injection. Inflammatory markers, including IL-6 and IL-1β, were significantly lowered in serum. Furthermore, CSREA suppressed paw edema and redness in the carrageenan-induced rat model, demonstrating dose-dependent anti-inflammatory efficacy comparable to diclofenac. CSREA also downregulated pain-related gene expression (SCN9A, ASIC1A, TACR1) and regulated key enzymes involved in endocannabinoid metabolism (FAAH, MAGL, DAGL), suggesting its role in the molecular modulation of pain pathways. These effects are likely mediated via modulation of the endocannabinoid system, particularly by rebalancing the CB1R/CB2R ratio. The findings suggest that CSREA holds promise as a natural therapeutic agent for managing pain and inflammation and warrants further investigation into its molecular mechanisms and long-term effects.

Introduction: Long COVID is a persistent symptom that appears in many COVID-19 patients, regardless of whether they are asymptomatic, mild, or severe. At least 10 % of the global population suffers from long COVID. The antiinflammatory effects of six Korean national insurance-covered herbal prescriptions were evaluated in LPS-treated RAW264.7 cells. Additional network pharmacology analysis was conducted to assess their potential for mitigating chronic respiratory tract inflammation. This study aimed to provide an experimental basis for the clinical application of herbal prescriptions and compounds in long COVID patients. Methods: Six candidate prescriptions were selected from the 56 national insurance-covered herbal prescriptions used in Korea. The in vitro efficacies and mechanisms of these six prescriptions on LPS-activated murine macrophages were evaluated. The long COVID-related targets isolated from patients with long COVID and obtained from the GEO online database were identified. These targets were compared with the potential targets of the most potent prescription used in this study. Results: Soshihotang (SST) significantly downregulated iNOS and COX2 expression and reduced the production of nitric oxide and inflammatory cytokines, such as TNF alpha, IL-1 beta, and IL-6. SST also inhibited I kappa B alpha phosphorylation and reduced the nuclear translocation of NF-kappa B. Network pharmacological analysis showed that SST influenced the NF-kappa B signaling pathway. In addition, Saikosaponin D, a major ingredient of Bupleuri Radix (a major herb in SST), showed similar effects and mechanisms to those of SST, including the NF-kappa B and TNF alpha signaling pathways. Thus, SST may modulate the inflammatory pathways associated with long COVID through its active component, saikosaponin D. Conclusion: These findings suggest that the potent anti-inflammatory effects of SST are due to its suppression of inflammatory cytokines by blocking NF-kappa B translocation. In addition, the predicted target was reconfirmed using network pharmacology. Nevertheless, further study on the clinical application of SST as insurance-covered herbal medicine on long COVID patients is needed.

Long COVID is a persistent symptom that appears in many COVID-19 patients, regardless of whether they are asymptomatic, mild, or severe. At least 10% worldwide are suffering from long COVID. The anti-inflammatory effect was evaluated in LPS-treated RAW264.7 cells using six Korean national insurance-covered herbal prescriptions with therapeutic potential for mitigating inflammation in the respiratory tract, and additional network pharmacology analysis was performed. Soshihotang (SST) significantly downregulated iNOS and COX2 expression, reduced the production of NO (nitric oxide) and inflammatory cytokines, such as TNFα, IL-1β, and IL-6, and inhibited IκBα phosphorylation and reduced the nuclear translocation of NF-κB in LPS-treated RAW264.7 cells. Network pharmacological analysis showed that SST was closely related to the NF-κB signaling pathway. Further study on the potential of herbal prescriptions will be needed before they can be used clinically by long COVID patients.

Objectives: Ephedrae herba (EH) and Coicis semen (CS) has been frequently prescribed for the treatment of obesity.However, effects of combinational extracts of these two herbs on non-alcoholic fatty liver disease are unknown.The aim of the present study was to investigate the effects of EH and CS on lipid accumulation and glucose absorption in free fatty acids (FFAs) or palmitic acid (PA)-treated HepG2 cells.Methods: Five samples of EH and CS were extracted by combination ratios (S1=0:100, S2=25:75, S3=50:50, S4=75:25, S5=100:0).Oil Red O staining was used to measure lipid accumulation in FFAs-induced steatosis cells.Intracellular triglycerides and total cholesterol levels were measured in FFAs-induced steatotic HepG2 cells.In PA-treated cells, intracellular 2-NBDG was detected using a fluorescence microplate reader and flow cytometry.Phosphorylation of key metabolism-related factors of AMP-activated protein kinase and acetyl-CoA carboxylase, expression of key lipid synthesis-related factors carnitine palmitoyltransferase 1 alpha (CPT1α), sterol regulatory element-binding protein 1 (SREBP1), peroxisome proliferator-activated receptor gamma (PPARγ) and CCAAT enhancer binding protein alpha (C/EBPα) were confirmed by western blot.Results: Treatment of EH-CS combination in the FFAs-induced steatotic HepG2 cells significantly reduced lipid accumulation.As the relative ratio of Ephedrae herba increased, the lipid-lowering effects of the combination were increased.However, S1 and S5 of Ephedrae herba and Coicis semen did not significantly reduce triglycerides and total cholesterol induced by FFAs.However, the combination of Ephedrae herba and Coicis semen restored glucose absorption in PA-induced HepG2 cells.Major makers of SREBP1, PPARγ, C/EBPα, and CPT1α expression tended to decrease with EH ratio.Conclusions: The EH-CS combination has advantages over sole EH and CS extracts in improving lipid and glucose metabolism in liver steatosis models.

Massa Medicata Fermentata (MMF) is a naturally fermented product used to treat indigestion and increase stomach activity in traditional medicine. This study examined the ability of the hydrothermal extract of MMF to scavenge free radicals corresponding to biological oxidative stresses, further protecting essential biomolecules. The anti-inflammatory effects of MMF were evaluated in LPS-induced RAW264.7 macrophages and zebrafish. In addition, the effects of MMF on the body mass index (BMI) and cholesterol accumulation in adult zebrafish fed a high-cholesterol diet (HCD) for three weeks were examined. MMF prevented the DNA and lipid damage caused by oxidative stress, inhibited LDL oxidation, and reduced the expression of cytokines and related proteins (MAPK and NFκB), with prominent anti-oxidative pathway (NRF2-HO-1) activation properties. LPS-induced NO production was reduced, and the increase in BMI and TC caused by the HCD diet was suppressed by MMF in zebrafish embryos or adult zebrafish. The bioactive aglycone of quercetin may be contributing to the mechanisms of systemic effects. MMF has excellent antioxidant properties and is useful for improving inflammation status and metabolic profile, thus highlighting its potential as a healthy, functional food.

Objectives: National health insurance herbal prescription of Korean medicine has been serving important role in public healthcare in spite of continuous demand on revision of system. However, the categories of insurance herbal prescriptions are not equally distributed throughout the KCD-based major disease categories. We analyzed statistical database of claimed national health insurance classified as major disease categories by years. We classified all 56 herbal prescriptions as per their total medical indications into 22 major disease categories to analyze their distribution. Significant increase of M and S-T code claims were found, whereas decrease of U code claims by years. We figured out that the 56 prescriptions were unequally distributed along with enrichment of certain codes such as K and J. Meanwhile, the insurance claim of each prescription was positively correlated with number of code types of their indications. As a result, we believe that the reform of national health insurance herbal prescription list is necessary to promote use of it in clinic.

The network pharmacology (NP) approach is a valuable novel methodology for understanding the complex pharmacological mechanisms of medicinal herbs. In addition, various in silico analysis techniques combined with the NP can improve the understanding of various issues used in natural product research. This study assessed the therapeutic effects of Arum ternata (AT), Poria cocos (PC), and Zingiber officinale (ZO) on hyperlipidemia after network pharmacologic analysis. A protein-protein interaction (PPI) network of forty-one key targets was analyzed to discover core functional clusters of the herbal compounds. The Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway and gene ontology (GO) term enrichment analysis identified significant categories of hypolipidemic mechanisms. The STITCH database indicated a high connection with several statin drugs, deduced by the similarity in targets. AT, PC, and ZO regulated the genes related to the energy metabolism and lipogenesis in HepG2 cells loaded with free fatty acids (FFAs). Furthermore, the mixture of three herbs had a combinational effect. The herbal combination exerted superior efficacy compared to a single herb, particularly in regulating acetyl-CoA carboxylase (ACC) and carnitine palmitoyltransferase 1 (CPT-1). In conclusion, the network pharmacologic approach was used to assess potential targets of the herbal combination for treatment. Experimental data from FFA-induced HepG2 cells suggested that the combination of AT, PC, and ZO might attenuate hyperlipidemia and its associated hepatic steatosis.

The enhanced therapeutic effects and mechanisms of certain herbal combination in various herbal prescriptions are mostly unclear. A combination of two herbs, namely Ephedrae herba (EH) and Coicis semen (CS), has been commonly prescribed for obesity. In our previous work, the combination of EH and CS was studied using network pharmacological approach to predict its pharmacological targets and in vitro experiments to evaluate its efficacy on obesity. Although we demonstrated enhanced anti-adiposity effects of the combination on matured adipocytes, the molecular mechanisms and contributing compounds underlying the effects of EH-CS combination on adiposity or adipogenesis were not fully elucidated. The current study adopted integrated bioinformatics analysis to precisely validate potential targets of EH-CS by screening differentially expressed genes (DEGs) of morbid obesity patients from NCBI gene expression omnibus (GEO). Based on the functional cluster analysis of down-regulated DEGs, the anti-adipogenesis mechanism of EH-CS combination was speculated with KEGG enrichment analysis. Furthermore, we investigated the combinational effects of EH and coixol, or stigmasterol, the two compounds in CS which were expected to have main beneficial effects in metabolic diseases. Moreover, distinct effect of the combination on transcriptional activity of glucocorticoid receptor (GR) was investigated using electrophoretic mobility shift assay (EMSA). The EH-CS combination was predicted to modulate down-regulated genes which are involved in KEGG pathways crucial to metabolic disease in morbidly obese individuals. The combination of EH with CS compounds significantly increased the phosphorylation of acetyl-coA carboxylase (ACC), AMP-activated protein kinase (AMPK), and protein kinase B (AKT) in 3T3-L1 cells and decreased intracellular lipid accumulation. The two CS compounds significantly increased the anti-adipogenesis/lipogenesis effects of EH by inhibiting the gene expression levels. Finally, the combination of EH and coixol inhibited dexamethasone-induced GR translocation to the nucleus and transcriptional binding activity in adipocytes. The combination of EH and CS could be considered a therapeutic strategy for treating metabolic diseases, including obesity.

BackgroundHerbal combinations are regarded as basic strategy in oriental medicine with various purposes. Ephedrae herba (EH) and Coicis semen (CS) are two herbal medicines used to treat obesity in many herbal prescriptions, yet the effect and significance of this herbal pair have not been evaluated.PurposeThis study is to elucidate the effect of a novel herbal pair, EH-CS, on obesity and identify the key synergistic mechanism underlying it.MethodsWe investigated the network of herbs comprising the anti-obesity herbal prescriptions. Using the tools of network pharmacology, we investigated the compound-target interactions of EH and CS in combination to predict their effects in combination. Five EH-CS samples with different EH to CS ratios were prepared to investigate their efficacies in adipocytes.Results1-mode network analysis of herbs in prescriptions based on literature review revealed the importance of EH-CS in anti-obesity prescriptions. The herbal combination comprised of equivalent weights (1:1) of EH and CS most potently reduced mature adipocyte adiposity, although several markers of adipogenesis and lipid synthesis were more suppressed by pure EH. PTGS2 (COX-2 gene) expression, a common target of EH and CS as deduced by compound-target network analysis, was affected by EH-CS extract treatments. However, EH at high concentration (25 μg/ml) notably increased PTGS2 expression without adversely affecting cell viability. However, EH-CS combination of the same concentration markedly decreased PTGS2 gene expression.ConclusionThese results show that the compounds in CS and EH act in concert to enhance the pharmacological effect of EH, but control unexpected effects of EH treatment.

This study aimed to evaluate the infection control rate of palliative arthroscopic debridement, antibiotics, and implant retention (DAIR) for the high mortality risk or terminal cancer stage patients. From March 2018 to August 2021, 21 patients met the following inclusion criteria: old age of more than 80, diagnosed as a terminal stage of cancer, high risk of mortality and morbidity representing as Charlson comorbidity index (CCI) ≥ 5, low daily activity with disabled extremity, and re-infection after two-stage revision. Each patient underwent arthroscopic DAIR and additional continuous irrigation for 48 hours. The need for subsequent re-arthroscopic DAIR or two-stage revision was determined by the post-operative trends of C-reactive protein (CRP) levels. Infection control was defined as continuing controlled status of infection based on clinical and laboratory results by one or two times of arthroscopic DAIR within initial two months. Treatment failure was defined as more than three times arthroscopic debridement, two-stage revision surgery, or expired due to uncontrolled infection. Arthroscopic DAIR controlled the infection in 19 (90.5%) of the 21 cases. The other knee underwent a total of three times of re-arthroscopic DAIR and the other one underwent two-stage revision. Although five patients expired during the follow-up period due to worsening medical problems or terminal cancer, there were no deaths from uncontrolled infection, sepsis, or surgery-related complications. Arthroscopic debridement with continuous irrigation for the infection TKA with high mortality risk or terminal cancer patients showed a 90.5% infection control rate. For high-risk patients, arthroscopic debridement with continuous irrigation can be an alternative treatment to improve the quality of life during survival.

Targeting Toll-like receptor 4/myeloid differentiation factor 2 (TLR4/MD2) signaling is regarded as a potential strategy for treating inflammatory diseases. Saponaria officinalis L. is rich in saponin, which include quillaic acid, gypsogenin, saponarin, and hederagenin. We evaluated the pharmacological activity of a Saponaria officinalis extract in THP-1 derived macrophages and RAW264.7 macrophages. TLR4/MyD88 complex formation and downstream signals were investigated by co-immunoprecipitation (Co-IP). In silico docking simulation was conducted to predict binding scores and perform 3D modeling of saponarin-TLR4/MD2 complex. A hexane fraction of Saponaria officinalis (SH) and fr.1 (a sub-fraction 1 of SH) inhibited mitogen-activated protein kinase (MAPK) signaling, nuclear factor kappa b (NF-kappa B) activity, cytokine production, and the expressions of marker genes specific for M1 polarization. The inhibitory effects of fr.1 and saponarin on TLR4/MyD88 complex formation were observed by western blotting TLR4 co-immunoprecipitated proteins. Saponarin and fr.1 markedly attenuated LPS-induced inflammatory cytokines, thus reducing mortality and morphological abnormality in zebrafish larvae. Finally, docking simulation revealed that saponarin can directly interact with TLR4/MD2 complex to inhibit downstream signalings. Our findings suggest that saponarin reduces downstream inflammatory response by disrupting TLR4/MD2 complex and blocking MyD88-dependent inflammatory signaling.

Objective We aimed to investigate how systemic bone metabolism was affected after 1 year of treatment with tumour necrosis factor (TNF) inhibitors in rheumatoid arthritis (RA) patients. Methods A total of 29 seropositive RA patients not treated for osteoporosis were enrolled and TNF inhibitors were administered for a year. Bone mineral density (BMD) at the lumbar spine, femur neck, and total hip was measured at baseline and 12 months after anti-TNF treatment. Blood samples were collected at baseline and 6 and 12 months after anti-TNF treatment and osteoclasts were cultured on hone slices. Weight was the strongest factor influencing systemic bone loss. Patients were categorised into two groups: obese (body mass index (BM!) >= 25 kg/m(2)) and non-obese (BM! <25 kg/m(2)). Results All patients showed decreased BMD at all sites. The obese group showed relatively little change in BMD, although the non-obese group showed significant decreases in BMD at all sites after 1 year of treatment with TNF inhibitors. Resorption pits created by osteoclasts decreased at 6 months and increased at 12 months in the non-obese group, while the obese group presented with steadily decreasing sizes of resorption pits at all-time points. Levels of receptor activator of nuclear factor kappa B ligand were significantly decreased at 12 months compared to baseline in the obese group, while they were increased in the non-obese group. Conclusion One year of treatment with TNF inhibitors failed to halt systemic bone loss in RA patients, but obesity may have protective effects against bone loss.

Network pharmacology (NP) is a useful, emerging means of understanding the complex pharmacological mechanisms of traditional herbal medicines. Sochehwan (SCH) is a candidate herbal prescription for drug repurposing as it has been suggested to have beneficial effects on metabolic syndrome. In this study, NP was adopted to complement the shortcomings of literature-based drug repurposing strategies in traditional herbal medicine. We conducted in vitro studies to confirm the effects of SCH on potential pharmacological targets identified by NP analysis. Herbal compounds and molecular targets of SCH were explored and screened from a traditional Chinese medicine systems pharmacology database and analysis platform (TCMSP) and an oriental medicine advanced searching integrated system (OASIS). Forty-seven key targets selected from a protein-protein interaction (PPI) network were analyzed with gene ontology (GO) term enrichment and KEGG pathway enrichment analysis to identify relevant categories. The tumor necrosis factor (TNF) and mitogen-activated protein kinase (MAPK) signaling pathways were presented as significant signaling pathways with lowest p-values by NP analysis, which were downregulated by SCH treatment. The signal transducer and activator of transcription 3 (STAT3) was identified as a core key target by NP analysis, and its phosphorylation ratio was confirmed to be significantly suppressed by SCH. In conclusion, the NP-based approach used for target prediction and experimental data obtained from Raw 264.7 cells strongly suggested that SCH can attenuate inflammatory status by modulating the phosphorylation status of STAT3.

Sochehwan (SCH) is an herbal prescription from traditional oriental medicine and is currently used to treat digestive ailments. In a previous study, SCH was found to have the potential to attenuate metabolic syndrome (MetS) by activating AMPK and downstream signaling. From the view of drug repurposing, the efficacy of SCH on alcoholic liver injury is implied in classic medical texts but is yet to be proven. C57BL/6J mice were pre-treated with SCH orally for 5 days and challenged by providing a pair-fed Lieber DeCarli diet containing alcohol for 20 days. Hepatic enzyme and triglyceride levels and endoplasmic reticulum (ER) stress-related markers were analyzed. Moreover, mitogen-activated protein kinases (MAPKs) and cytochrome P450 2E1 (CYP2E1) levels were determined. CYP2E1-transfected HepG2 cells were used to test the cytoprotective efficacy of SCH against the adverse effects of alcohol in vitro. In mice, SCH administration notably reduced hepatic enzyme activity and neural lipid levels. Furthermore, ER-stress markers and MAPK phosphorylation were reduced due to ROS suppression, which was attributed to decreased CYP2E1 expression in liver tissue. In addition, SCH successfully protected CYP2E1-transfected HepG2 cells against ethanol. Our findings suggest SCH attenuated alcohol-induced liver injury by inhibiting CYP2E1 expression and indicate drug repurposing should be considered as a valuable option for drug development in traditional herbal medicines.

Background:CT-P13, an infliximab biosimilar, is effective for treating ankylosing spondylitis (AS) at a dose of 5 mg/kg infused once every 6–8 weeks. Evidence suggests that patients with AS may benefit from a lower dose, and individualised dose/interval adjustments should be based on treatment response.Objectives:To analyse real-world treatment patterns (doses and infusion intervals) and outcomes for CT-P13-treated patients with AS over 5 years.Methods:The RAAS study collected medical record data for adults with AS treated with CT-P13 at five referral hospitals in the Republic of Korea (2012–2017). Patients were infliximab naïve at CT-P13 initiation (‘naïve’) or had switched to CT-P13 from reference infliximab (‘switched’). Patients were analysed by baseline dose (BD) (<4 mg/kg; ≥4–<5 mg/kg; ≥5 mg/kg), defined as the third (naïve) or first (switched) infusion dose. Baseline infusion intervals were the average of the three infusion intervals after BD. Over time, patients with both constant dose and infusion interval were compared with those with changes in dose and/or infusion interval. Data were analysed by Kruskal–Wallis test, chi-squared test and one-way analysis of variance, and drug survival by log-rank test.Results:Overall, 337 patients (219 naïve; 118 switched) were identified. Of those with BD data, 71, 117 and 82 patients had BDs of <4 mg/kg, ≥4–<5 mg/kg and ≥5 mg/kg, respectively. Most patients were male (74.8%). Patients with higher BDs tended to have higher Bath Ankylosing Spondylitis Disease Activity Index (BASDAI) scores; switched patients had lower scores than naïve patients. Of 186 evaluable patients (118 naïve; 68 switched), 85 (46 naïve; 39 switched) did not have dose and/or interval changes (‘combined constant’ group). More naïve (n=72; 61.0%) versus switched (n=29; 42.6%) patients had dose and/or interval changes (‘combined changed’ group). Considering dose and interval separately, 18/235 evaluable patients (152 naïve; 83 switched) had dose changes (12 increased; 6 decreased) and 110/224 evaluable patients (140 naïve; 84 switched) had interval changes (79 increased; 31 decreased). Cumulative annual doses were similar between naïve and switched patients; switched patients had longer infusion intervals than naïve patients (Figure 1). There were no significant differences in drug survival between BD groups overall or for naïve and switched patients. BASDAI scores over time showed that disease activity was well controlled (Table 1). Patients in the combined changed versus combined constant group had greater improvements in BASDAI score.Table 1.BASDAI scoresGroupStatisticW0W54W102W156W210Combined constantTotal (N=85)n7273574231Mean (SD)5.50 (3.12)2.50 (1.71)2.35 (1.66)2.42 (1.66)2.36 (1.68)Median6.322.602.402.552.60Naïve (n=46)n3939261812Mean (SD)7.86 (1.45)2.49 (1.82)2.41 (1.75)2.34 (1.70)1.94 (1.58)Median7.802.201.751.901.44Switched (n=39)n3334312419Mean (SD)2.71 (2.07)2.50 (1.60)2.31 (1.60)2.48 (1.67)2.63 (1.73)Median2.582.752.502.802.70Combined changedTotal (N=101)n8785765335Mean (SD)5.68 (2.89)1.81 (1.45)1.58 (1.27)1.49 (1.34)1.40 (1.24)Median6.701.321.191.201.00Naïve (n=72)n6360543418Mean (SD)7.18 (1.37)2.01 (1.44)1.65 (1.22)1.54 (1.26)1.52 (0.97)Median7.301.831.351.311.35Switched (n=29)n2425221917Mean (SD)1.74 (1.94)1.33 (1.36)1.42 (1.41)1.39 (1.50)1.28 (1.50)Median0.920.800.850.700.58SD, standard deviation; W, WeekConclusion:These real-world data demonstrate that adjusting dose and infusion interval can improve clinical outcomes for CT-P13-treated patients with AS. Drug survival and BASDAI results show that patients with lower baseline BASDAI receiving low CT-P13 doses can achieve the same outcomes as those dosed with ≥5 mg/kg. Findings support the lack of impact of switching from reference infliximab to CT-P13 on efficacy, underlining conclusions previously drawn for efficacy and safety.1References:[1]Kim T-H, et al. Clin Drug Investig 2020;40:541–53.Acknowledgements:Funding: This study was supported by Celltrion Healthcare Co., Ltd. (Incheon, Republic of Korea). Medical writing support was provided by Beatrice Tyrrell, DPhil (Aspire Scientific, Bollington, UK), and funded by Celltrion Healthcare Co., Ltd. (Incheon, Republic of Korea).Disclosure of Interests:Shin-Seok Lee: None declared, Tae-Hwan Kim: None declared, Won Park Consultant of: Celltrion, Inc., Yeong Wook Song: None declared, Chang-Hee Suh Speakers bureau: AbbVie Inc., Astellas Pharma Inc., Samsung Bioepis Co., Ltd, Consultant of: Celltrion Healthcare Co., Ltd., Eli Lilly and Company, GlaxoSmithKline plc, Janssen Pharmaceuticals, Yungjin Pharmaceutical, Co., Ltd, SooKyoung Kim Shareholder of: Celltrion Healthcare Co., Ltd., Employee of: Celltrion Healthcare Co., Ltd., DaeHyun Yoo Speakers bureau: Celltrion, Consultant of: Celltrion, Grant/research support from: Celltrion

Objective Epidemiological studies on inflammatory myopathies (IMs) show widely variable results, and studies on Asians are lacking. Despite emerging interest in the cardiovascular disease (CVD) risk associated with IMs, the prevalence of CVD in IM patients and its impact on mortality remain unclear. We conducted a nationwide, population-based study on the incidence, mortality, and associated major CVD events of IMs in the Republic of Korea over 11 years. Method Using the nationwide, population-based National Health Insurance claims database and the Rare Intractable Disease registration programme, we estimated incidence, mortality, and CVD occurrence. Survival was examined using the Kaplan-Meier method. Mortality rate in IMs with CVD was analysed by Cox proportional hazards regression. Results There were 3014 incident cases, 640 of whom died during the study period. The mean annual incidence was 7.16/10(6). Dermatomyositis (DM) and polymyositis (PM) had 5 year survival rates of 76.8% and 79.3%, respectively. Cardiovascular events occurred in 155 patients and 40.6% of IM patients with CVD died. Acute myocardial infarction in men had the highest risk of any CVD event in both DM [standardized incidence ratio (SIR) 4.2, 95% confidence interval (95% CI) 2.4-7.2] and PM (SIR 3.5, 95% CI 1.8-7.0). Haemorrhagic stroke had the highest hazard ratio (HR) in both DM (HR 2.31, 95% CI 1.13-4.70) and PM patients (HR 2.10, 95% CI 1.03-4.27) compared with the general population with CVD. Conclusion We found persistently low incidence, poor survival, and high major CVD incidence in IMs, and increased mortality in IMs with CVD.

Relatively high proportions of proinflammatory M1-like macrophages in tissues may lead to vascular impairment and trigger numerous diseases including atherosclerosis-related cardiovascular disease (CVD). Jisil Haebaek Gyeji-tang (JHGT), a polyherbal decoction, is traditionally used to treat various human ailments including chest pain, angina, and myocardial infarction. In the present study, we investigated the anti-inflammatory effects of JHGT on lipopolysaccharide- (LPS-) stimulated M1 macrophage polarization generated via the mitogen-activated protein kinases (MAPKs) pathway in RAW 264.7 mouse macrophages. The reducing power of JHGT was also investigated using DAF-FA DA in a zebrafish model. JHGT  significantly reduced inflammatory mediator levels, including iNOS, COX2, TNF-α, IL-6, and IL-1β, as compared with LPS-stimulated controls in vitro and ex vivo. Furthermore, JHGT suppressed the ERK1/2, JNK, and p38 MAPK pathways and reduced p-IκBα levels and the nuclear translocation of NF-κB in RAW 264.7 cells. In addition, treatment with JHGT significantly reduced the NO levels in LPS-treated zebrafish larva ex vivo. Our findings show the potent anti-inflammatory properties of JHGT are due to its suppression of MAPK signaling, NF-κB translocation, and M1 macrophage polarization.

Background: Vascular calcification is highly correlated with atherosclerosis. Ankylosing spondylitis (AS) is associated with a process of accelerated atherosclerosis. Wnt signaling plays an important role in the pathogenesis of vascular calcification. However, there has been no study of the role of Wnt signaling in vascular calcification in patients with AS. Objectives: We investigated the relationship between vascular calcification and Wnt signaling in patients with AS. Methods: Sixteen male patients aged over 20 years with AS were enrolled. They fulfilled the modified New York criteria and each of their ankylosing spondylitis disease activity score was more than 2.1. Sex and age matched nineteen healthy controls were also recruited. Mouse MOVAS vascular smooth muscle cell line (American Type Culture Collection, ATCC® CRL-2797™) were stabilized in maintain media for 24 hours. Then media were exchanged for the 10% serum of patients with AS or controls in maintain media. Cells were stimulated for another 72hours. We exchanged this medium with calcification medium. Cells were cultured until 2 weeks then stained with Alizarin Red S and the optical density (OD) was measured. For Western blotting and RT-qPCR, cells were stabilized for 24 hours and stimulated for another 72 hours through the same procedure as that of Alizarin Red S staining. After cell stimulation, the level of mRNA and protein were measured by RT-qPCR and western blot, respectably. We measure the level of Low-density lipoprotein receptor-related protein (LRP)5, LRP6, Dickkopf-related protein 1, Wnt3a, matrix metalloproteinase-7(MMP-7), beta-catenin for canonical Wnt signaling; Receptor Tyrosine Kinase Like Orphan Receptor 2, Wnt5a, Runt-related transcription factor 2 (RUNX2) for non-canonical Wnt signaling. We also checked the level of Alkaline phosphatase (ALP), IL-17, IL-23 and TNF-a. Results: The level of OD of MOVAS cells treated with serum from AS patients (19.503 ± 6.422, mean ± SD) was significantly higher than that from controls (10.994 ± 4.291) (P=0.000, Mann-Whitney test). The protein level of MMP-7 and beta-catenin of MOVAS cells treated with serum from AS patients (1.881 ± 0.687; 1.301 ± 0.342) was significantly higher than that from controls (0.779 ± 0.48; 0.854 ± 0.285) respectively (P=0.005,: P=0.002, Mann-Whitney test). The mRNA level of RUNX2, ALP, IL-17 and IL-23 of serum from AS patients (2.697 ± 1.46; 2.687 ± 1.753; 2.253 ± 1.128; 2.574 ± 1.142) was significantly higher than that from controls (1.396 ± 0.587; 1.696 ± 0.637; 1.358 ± 0.473; 1.386 ± 0.714) respectively (P=0.000; P=0.037; P=0.044; P=0.007, Mann-Whitney test). There was positive correlation between the mRNA level of WNT5a and RUNX2 (rho=-0.705, p=0.002, Spearman rank correlation coefficient) and the protein level of WNT5a and ALP, MMP-7 and TNF-a, MMP-7 and IL-17, MMP-7 and IL-23 (rho=-0.601, p=0.039; rho=-0.769, p=0.026; rho=-0.828, p=0.011; rho=-0.777, p=0.003), respectively. Conclusion: 1. Vascular smooth muscle cell calcification was increased in patients with ankylosing spondylitis than those of the control group. 2. The level of several molecules(i.e. Beta-catenin, RUNX2, MMP-7) related to Wnt signaling of vascular smooth muscle cells treated with serum of patients with AS was elevated significantly compared to those of controls and positively related. 3. Wnt signaling can play an important role in vascular calcification in patients with ankylosing spondylitis. Acknowledgments: This research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (NRF-2017R1D1A1B03030825) Disclosure of Interests: None declared