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To investigate the effects and mechanisms of irisin, a newly discovered myokine, in cartilage development, osteoarthritis (OA) pathophysiology and its therapeutic potential for treating OA we applied the following five strategical analyses using (1) murine joint tissues at different developmental stages; (2) human normal and OA pathological tissue samples; (3) experimental OA mouse model; (4) irisin gene knockout (KO) and knock in (KI) mouse lines and their cartilage cells; (5) in vitro mechanistic experiments. We found that Irisin was involved in all stages of cartilage development. Both human and mouse OA tissues showed a decreased expression of irisin. Intra-articular injection of irisin attenuated ACLT-induced OA progression. Irisin knockout mice developed severe OA while irisin overexpression in both irisin KI mice and intraarticular injection of irisin protein attenuated OA progression. Irisin inhibited inflammation and promoted anabolism in chondrogenic ADTC5 cells. Proliferative potential of primary chondrocytes from KI mice was found to be enhanced, while KO mice showed an inhibition under normal or inflammatory conditions. The primary chondrocytes from irisin KI mice showed reduced expression of inflammatory factors and the chondrocytes isolated from KO mice showed an opposite pattern. In conclusion, it is the first time to show that irisin is involved in cartilage development and OA pathogenesis. Irisin has the potential to ameliorate OA progression by decreasing cartilage degradation and inhibiting inflammation, which could lead to the development of a novel therapeutic target for treating bone and cartilage disorders including osteoarthritis.

Background The aim of this study was to investigate a novel method to treat coronary artery aneurysms(CAAs).Methods Three CAAs patients who underwent single or double bare-metal stent implantation were recruited. The CAAs and parent artery diameters were measured for model construction. Single- and double-stent implantation were simulated, and the changes in the CAAs haemodynamics after stenting were analysed. Results In Case 1, the flow velocity in the single-stent model was significantly lower than that in the double-stent model (0.0046±0.013 vs 0.0050±0.011, pConclusion Application of the single or double bare-metal stent technique, according to the neck diameter of the coronary artery aneurysms, can effectively change the flow in vessels and aneurysm haemodynamics to achieve occlusion.Trail registration: Clinicaltrails, NCT04265989. Registered 12 February 2020, https://clinicaltrials.gov/ct2/show/NCT04265989?term=NCT04265989&draw=2&rank=1

TNFα/TNFR signaling plays a critical role in the pathogenesis of various inflammatory and autoimmune diseases, and anti-TNFα therapies have been accepted as the effective approaches for treating several autoimmune diseases. Progranulin (PGRN), a multi-faced growth factor-like molecule, directly binds to TNFR1 and TNFR2, particularly to the latter with higher affinity than TNFα. PGRN derivative Atsttrin is composed of three TNFR-binding domain of PGRN and exhibits even better therapeutic effects than PGRN in several inflammatory disease models, including collagen-induced arthritis. Herein we describe the detailed methodology of using (1) ELISA-based solid phase protein-protein interaction assay to demonstrate the direct binding of Atsttrin to TNFR2 and its inhibition of TNFα/TNFR2 interaction; and (2) tartrate-resistant acid phosphatase (TRAP) staining of in vitro osteoclastogenesis to reveal the cell-based anti-TNFα activity of Atsttrin. Using the protocol described here, the investigators should be able to reproducibly detect the physical inhibition of TNFα binding to TNFR and the functional inhibition of TNFα activity by Atsttrin and various kinds of TNF inhibitors.

The study aims to investigate whether kaemperfol (KAE) inhibits microglia pyroptosis and subsequent neuroinflammatory response to exert neuroprotective effects, along with the underlying mechanisms. The results showed KAE could ameliorate the behavioral deficits of Parkinson's disease (PD) rats, inhibit the activation of microglia and astrocytes, reduce the loss of TH-positive neurons, down-regulate levels of pyroptosis-related NOD-like receptor family pyrin domain containing 3 (NLRP3), GasderminD-N Term (GSDMD-NT), caspase1, apoptosis-associated speck-like protein containing a caspase-recruitment domain (ASC), interleukin (IL)-1β, and IL-18, and decrease the levels of inflammatory molecules (inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2)) and p38 mitogen-activated protein kinase/nuclear factor-kappaB (p38MAPK/NF-κB) signaling pathway molecules (p38MAPK, p-p38MAPK, NF-κB, and p-NF-κB) in the substantia nigra of PD rats. Further in vitro study indicated that KAE reversed the activation of BV2 cells and down-regulated the expressions of pyrolytic proteins, inflammatory mediators and key molecules in p38MAPK/NF-κB signaling pathway. Collectively, KAE inhibits the microglia pyroptosis and subsequent neuroinflammatory response to exert neuroprotective effects on 6-hydroxydopamine (6-OHDA)-induced PD rats and lipopolysaccharide (LPS)-induced BV2 inflammatory cells through inhibiting p38MAPK/NF-κB signaling pathway.

TNFR1 and TNFR2 have received prominent attention because of their dominance in the pathogenesis of inflammation and autoimmunity. TNFR1 has been extensively studied and primarily mediates inflammation. TNFR2 remains far less studied, although emerging evidences demonstrate that TNFR2 plays an anti-inflammatory and immunoregulatory role in various conditions and diseases. Herein, we report that TNFR2 regulates macrophage polarization, a highly dynamic process controlled by largely unidentified intracellular regulators. Using biochemical co-purification and mass spectrometry approaches, we isolated the signaling molecule 14-3-3e as a component of TNFR2 complexes in response to progranulin stimulation in macrophages. In addition, 14-3-3e was essential for TNFR2 signaling-mediated regulation of macrophage polarization and switch. Both global and myeloid-specific deletion of 14-3-3e resulted in exacerbated inflammatory arthritis and counteracted the protective effects of progranulin-mediated TNFR2 activation against inflammation and autoimmunity. TNFR2/14-3-3e signaled through PI3K/Akt/mTOR to restrict NF-κB activation while simultaneously stimulating C/EBPβ activation, thereby instructing macrophage plasticity. Collectively, this study identifies 14-3-3e as a previously-unrecognized vital component of the TNFR2 receptor complex and provides new insights into the TNFR2 signaling, particularly its role in macrophage polarization with therapeutic implications for various inflammatory and autoimmune diseases with activation of the TNFR2/14-3-3e anti-inflammatory pathway.

Polyphenols from Toona sinensis seeds (PTSS) have demonstrated anti-inflammatory effects in various diseases, while the anti-neuroinflammatory effects still remain to be investigated. We aimed to investigate the effects of PTSS on Parkinson's disease and underlying mechanisms using a rat model. We employed 6-hydroxydopamine (6-OHDA) to male Sprague Dawley (SD) rats and PC12 cells to construct the in vivo and vitro models of PD and dopaminergic (DA) neuron injury, respectively. Cell viability was detected by cell counting kit-8 (CCK-8) assay and protein levels of inflammatory mediators and some p38 MAPK pathway molecules were investigated by immunohistochemistry and Western blot analyses. The results showed that 6-OHDA significantly increased protein levels of inflammatory mediators, such as cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), and tumor necrosis factor α (TNF-α), which could be reversed by PTSS through suppressing the p38 MAPK pathway. The anti-inflammatory effects of PTSS were significantly enhanced by the specific p38 inhibitor of SB203580 in vitro. The present work suggests that PTSS can exert anti-inflammatory effects on PD models, which may be attributed to the suppression of p38 MAPK signaling pathway.

Purpose—Image-based computational fluid dynamics (CFD) is widely used to predict intracranial aneurysm wall shear stress (WSS), particularly with the goal of improving rupture risk assessment. Nevertheless, concern has been expressed over the variability of predicted WSS and inconsistent associations with rupture. Previous challenges, and studies from individual groups, have focused on individual aspects of the image-based CFD pipeline. The aim of this Challenge was to quantify the total variability of the whole pipeline. Methods—3D rotational angiography image volumes of five middle cerebral artery aneurysms were provided to participants, who were free to choose their segmentation methods boundary conditions, and CFD solver and settings. Participants were asked to fill out a questionnaire about their solution strategies and experience with aneurysm CFD, and provide surface distributions of WSS magnitude, from which we objectively derived a variety of hemodynamic parameters. Results—A total of 28 datasets were submitted, from 26 teams with varying levels of self-assessed experience. Wide variability of segmentations, CFD model extents, and inflow rates resulted in interquartile ranges of sac average WSS up to 56%, which reduced to < 30% after normalizing by parent artery WSS. Sac-maximum WSS and low shear area were more variable, while rank-ordering of cases by low or high shear showed only modest consensus among teams. Experience was not a significant predictor of variability. Conclusions—Wide variability exists in the prediction of intracranial aneurysm WSS. While segmentation and CFD solver techniques may be difficult to standardize across groups, our findings suggest that some of the variability in image-based CFD could be reduced by establishing guidelines for model extents, inflow rates, and blood properties, and by encouraging the reporting of normalized hemodynamic parameters. Fil: Valen-Sendstad, Kristian. Simula Research Laboratory and Center for Cardiological Innovation; Noruega Fil: Bergersen, Aslak W.. University of Oslo; Noruega Fil: Shimogonya, Yuji. Nihon University; Japón Fil: Goubergrits, Leonid. Charite´ – Universita¨tsmedizin Berlin; Alemania Fil: Bruening, Jan. Charité – Universitätsmedizin Berlin; Alemania Fil: Pallares, Jordi. Universitat Rovira I Virgili; España Fil: Cito, Salvatore. Universitat Rovira I Virgili; España Fil: Piskin, Senol. University Of Texas At San Antonio.; Estados Unidos Fil: Pekkan, Kerem. Koc University; Turquía Fil: Geers, Arjan J.. Universitat Pompeu Fabra; España Fil: Larrabide, Ignacio. Universidad Nacional del Centro de la Provincia de Buenos Aires. Facultad de Ciencias Exactas. Grupo de Plasmas Densos Magnetizados. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Grupo de Plasmas Densos Magnetizados; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Rapaka, Saikiran. Siemens Medical Solutions USA Inc; Estados Unidos Fil: Mihalef, Viorel. Siemens Medical Solutions USA Inc; Estados Unidos Fil: Fu, Wenyu. Beijing Union University; China Fil: Qiao, Aike. Beijing University of Technology; China Fil: Jain, Kartik. Simula Research Laboratory and Center for Cardiological Innovation; Noruega. University of Siegen; Alemania. University of Zürich; Suiza Fil: Roller, Sabine. University of Siegen; Alemania Fil: Mardal, Kent-Andre. Simula Research Laboratory and Center for Cardiological Innovation; Noruega. University of Oslo; Noruega Fil: Kamakoti, Ramji. Dassault Systemes; Francia Fil: Spirka, Thomas. Simpleware Software Solutions; Reino Unido Fil: Ashton, Neil. University of Oxford; Reino Unido Fil: Revell, Alistair. University of Manchester; Reino Unido Fil: Aristokleous, Nicolas. University of Limerick; Irlanda Fil: Houston, J. Graeme. University of Dundee; Reino Unido Fil: Tsuji, Masanori. Mie Chuo Medical Center; Japón Fil: Ishida, Fujimaro. Mie Chuo Medical Center; Japón Fil: Menon, Prahlad G.. University of Pittsburgh; Estados Unidos Fil: Browne, Leonard D.. University of Limerick; Irlanda Fil: Broderick, Stephen. University of Limerick; Irlanda Fil: Shojima, Masaaki. University of Tokyo; Japón Fil: Koizumi, Satoshi. University of Tokyo; Japón Fil: Barbour, Michael. University of Washington; Estados Unidos Fil: Aliseda, Alberto. University of Washington; Estados Unidos Fil: Morales, Hernán G.. Medisys - Philips Research Paris; Francia Fil: Lefèvre, Thierry. Medisys - Philips Research Paris; Francia Fil: Hodis, Simona. Texas A&M University - Kingsville; Estados Unidos Fil: Al-Smadi, Yahia M.. Jordan University of Science and Technology; Jordania Fil: Tran, Justin S.. Stanford University; Estados Unidos Fil: Marsden, Alison L.. Stanford University; Estados Unidos Fil: Vaippummadhom, Sreeja. EinNel Technlogies; India Fil: Einstein, G. Albert. EinNel Technlogies; India Fil: Brown, Alistair G.. Siemens PLM Software; Estados Unidos Fil: Debus, Kristian. Siemens PLM Software; Estados Unidos Fil: Niizuma, Kuniyasu. Tohoku University; Japón Fil: Rashad, Sherif. Tohoku University; Japón Fil: Sugiyama, Shin-ichiro. Kohnan Hospital; Japón Fil: Owais Khan, M.. University of Toronto; Canadá Fil: Updegrove, Adam R.. University of California at Berkeley; Estados Unidos Fil: Shadden, Shawn C.. University of California at Berkeley; Estados Unidos Fil: Cornelissen, Bart M. W.. Academic Medical Center; Países Bajos Fil: Majoie, Charles B. L. M.. Academic Medical Center; Países Bajos Fil: Berg, Philipp. University of Magdeburg; Alemania Fil: Saalfield, Sylvia. University of Magdeburg; Alemania Fil: Kono, Kenichi. Wakayama Rosai Hospital; Japón Fil: Steinman, David A.. University of Toronto; Canadá

ObjectiveTumour necrosis factor alpha (TNF-α) signalling plays a central role in the pathogenesis of various autoimmune diseases, particularly inflammatory arthritis. This study aimed to repurpose clinically approved drugs as potential inhibitors of TNF-α signalling in treatment of inflammatory arthritis.MethodsIn vitro and in vivo screening of an Food and Drug Administration (FDA)-approved drug library; in vitro and in vivo assays for examining the blockade of TNF actions by fexofenadine: assays for defining the anti-inflammatory activity of fexofenadine using TNF-α transgenic (TNF-tg) mice and collagen-induced arthritis in DBA/1 mice. Identification and characterisation of the binding of fexofenadine to cytosolic phospholipase A2 (cPLA2) using drug affinity responsive target stability assay, proteomics, cellular thermal shift assay, information field dynamics and molecular dynamics; various assays for examining fexofenadine inhibition of cPLA2 as well as the dependence of fexofenadine’s anti-TNF activity on cPLA2.ResultsSerial screenings of a library composed of FDA-approved drugs led to the identification of fexofenadine as an inhibitor of TNF-α signalling. Fexofenadine potently inhibited TNF/nuclear factor kappa-light-chain-enhancer of activated B cells (NF-ĸB) signalling in vitro and in vivo, and ameliorated disease symptoms in inflammatory arthritis models. cPLA2 was isolated as a novel target of fexofenadine. Fexofenadine blocked TNF-stimulated cPLA2 activity and arachidonic acid production through binding to catalytic domain 2 of cPLA2 and inhibition of its phosphorylation on Ser-505. Further, deletion of cPLA2 abolished fexofenadine’s anti-TNF activity.ConclusionCollectively, these findings not only provide new insights into the understanding of fexofenadine action and underlying mechanisms but also provide new therapeutic interventions for various TNF-α and cPLA2-associated pathologies and conditions, particularly inflammatory rheumatic diseases.

Stem cell transplantation is a promising tool for the treatment of neurodegenerative disorders, including Parkinson's disease (PD); however, the therapeutic routes and mechanisms of mechanical approaches to stem cell transplantation must be explored. This study tests the therapeutic effect of transplantation of rat bone marrow mesenchymal stem cells (MSCs) into the substantia nigra (SN) of the PD rat. 5-Bromo-2-deoxyuridine-labeled rat MSCs were transplanted into the SN of the 6-hydroxydopamine-injected side of PD rat brains. The behavioral changes in PD rats were examined before and 4 and 8 weeks after MSC transplantation. The expression of tyrosine hydroxylase (TH) in the SN and the striatum and the survival and differentiation of MSCs were assessed by immunohistochemical and double immunofluorescence techniques. Abnormal behavior of PD rats was significantly improved by the administration of bone marrow MSCs, and the number of TH-positive cells in the SN and the optical density of TH-positive fibers in the striatum were markedly increased. Transplanted MSCs can survive and migrate in the brain and differentiate into nestin-, neuron-specific enolase-, and GFAP-positive cells. Our findings suggest that transplantation of rat bone marrow MSCs into the SN of PD rats may provide therapeutic effects. © 2016 Wiley Periodicals, Inc.

Background It has been reported that a disintegrin and metalloproteinase with thrombospondin motifs-12 (ADAMTS-12) is a susceptibility gene for rheumatoid arthritis (RA) development, and its level was significantly increased in RA patients. In addition, ADAMTS-12 was also reported to be required for normal inflammation. Objectives This study aims to determine the role of ADAMTS-12 and the underlying mechanisms in the pathogenesis of inflammatory arthritis. Methods Collagen-induced arthritis (CIA) was established in ADAMTS-12-/- mice and their control littermates to determine the role of ADAMTS-12 in vivo. Connective tissue growth factor (CTGF) deficient Raw264.7 were used to determine their functional interplays. Protein-protein interaction assays were performed to detect the interactions of ADAMTS-12 with CTGF Results ADAMTS-12-/- mice are more susceptible to collagen-induced arthritis. Accelerated disease onset, significant increase in the arthritis score and arthritis incidence, were observed in ADAMTS-12-/- mice (figure 1a). Histological analysis of whole ankle joints demonstrated a significant increase in synovitis, destruction of bone and cartilage loss in ADAMTS-12-/- mice. ELISA results indicated that ADAMTS-12-/- CIA mice exhibited enhanced release of pro-inflammatory and reduced secretion of anti-inflammatory cytokine. Collectively, these data demonstrate that ADAMTS-12-/- renders mice highly susceptible to CIA. ADAMTS-12 interacts with and cleaves CTGF, and ADAMTS-12-mediated signalling depends on CTGF during inflammation. It is known that CTGF plays a pro-inflammatory role in the pathogenesis of inflammatory arthritis. We co-transfected CTGF and ADAMTS-12 into 293 T cells and found ADAMTS-12 bound to (figure 1b) and digested CTGF (figure 1c). In vivo studies also demonstrated that CTGF was accumulated in the synovium of ADAMTS-12-/- CIA mice. To further determine whether CTGF is a critical regulator of ADAMTS-12 mediated signalling, we generated CTGF deficient Raw264.7. Overexpression of ADAMTS-12 and CTGF deficiency could decrease the activation of inflammatory signalling markers such as NFkb, p38 and JNK in response to IL-1β at a comparable level. More importantly, overexpression of ADAMTS-12 in CTGF deficient Raw264.7 failed to further inhibit the activation of these signal molecules as compared to CTGF deficient Raw264.7 (figure 1f). Taken together, these results suggest that CTGF is a critical regulator of ADAMTS-12 mediated signalling during inflammation. Blocking CTGF attenuates inflammatory arthritis in ADAMTS12-deficient CIA mice model. To determine whether the accelerated inflammation in ADAMTS-12-/- mice resulted from the accumulated CTGF, we administered CTGF antibody to ADAMTS-12-/- CIA model after disease onset. The arthritis score in ADAMTS-12 deficient mice was significantly reduced in presence of CTGF antibody (figure 1d). Moreover, histological analysis indicated CTGF abrogated further tissue destruction and inflammation (figure 1e). Conclusions ADAMTS-12-mediated regulation of inflammatory arthritis is probably through, at least in part, its interplay with CTGF and blockage of CTGF has been shown to be effective in treating inflammatory arthritis. Disclosure of Interest None declared

Background Functional heterogeneity is a hallmark of macrophages, which can classified into 2 major phenotypes with opposite role in inflammation termed M1 (inflammatory or classically activated) macrophages and M2 (alternatively activated) macrophages. In addition, M1-M2 polarisation of macrophages is a highly dynamic process and the phenotype of polarised macrophages can be switched under physiological and pathological conditions. Progranulin (PGRN), a multiple functional growth factor, binds to TNF receptor 2 (TNFR2) and activates the protective and anti-inflammatory pathway in inflammatory arthritis. In addition, 14–3–3e was identified as a component of PGRN/TNFR2 complexes in Raw264.7 macrophages. Objectives In this study, we examined whether 14–3–3e regulated macrophage polarisation and if so, whether this was important for PGRN’s anti-inflammatory action in inflammatory arthritis. Methods LysMCre and14–3–3e F/F mouse line was obtained from Jackson Laboratory. Results : 14–3–3e regulates macrophage polarisationin vitro. We found that 14–3–3e deficiency enhanced M1 while inhibited M2 polarisation (figure 1a, b). Interestingly, PGRN showed reverse effects on macrophage polarisation. In addition, PGRN’s effects were largely lost in 14–3–3e deficient BMDMs (figure 1a, b). Together, these data indicate that 14–3–3e is a critical downstream mediator of PGRN regulation of macrophage polarisation. Macrophage-specific 14–3–3e contributes to control of inflammatory arthritis and is critical for PGRN’s anti-inflammatory action. We then explored the role of macrophage-specific 14–3–3e in inflammatory arthritis and whether PGRN’s anti-inflammatory activity depended on 14–3–3e in vivo. We established CIA in 14–3–3eF/F (serve as WT) and 14–3–3eΔ/Δ mice, followed by i.p. injection of recombinant PGRN. The clinical arthritis score demonstrated that 14–3–3eΔ/Δ mice displayed increased severity of CIA compared with WT CIA. In addition, PGRN’s protective effects against inflammatory arthritis was compromised in 14–3–3eΔ/Δ mice (figure 1c), suggesting that 14–3–3e is critical downstream mediator of PGRN’s anti-inflammatory effects. In addition, FACS analysis showed that total numbers of F4/80 cells were not different in all WT and knockout CIA mice. However, analysis of CD45 +CD11b+cell population in spleen demonstrated a significant increase in mean fluorescence intensity (MFI) of iNOS and a significant decrease of CD206 +cells in PBS treated 14–3–3eΔ/Δ CIA mice as compared with PBS treated WT CIA mice; moreover, there was a significant decrease of iNOS MFI while a dramatic increase of CD206 +cells in PGRN-treated WT mice compared to that in PBS-treated mice. Further PGRN-mediated effects on macrophage polarisation was lost in 14–3–3eΔ/Δ CIA mice (Fig 1d, e). Cellectively, these results indicate that PGRN skews macrophage toward M2 polarisation to resolve inflammation and this effect depends on 14–3–3e. Conclusions Both in vitro and in vivo results indicate that 14–3–3e is a key molecule regulating macrophage polarisation which plays an important role in inflammatory arthritis, and it is an essential component for PGRN/TNFR2 mediated protective effect against inflammatory arthritis. Disclosure of Interest None declared

Background Braided stents, such as Pipeline Embolization Device (PED; ev3 Neurovascular, Irvine, CA, USA), are commonly used to treat cerebral aneurysms. However, little information is available on the compression and bending characteristics of such stents. Objective This paper investigates how geometrical parameters of braided stents influence their radial compression and bending characteristics. Methods Six groups of braided stent models with different braiding angles, numbers of wires and wire diameters are constructed. Parametric analyses of these models are conducted using Abaqus/Explicit software. The numerical results of a finite element analysis are validated by comparison with data of theoretical analysis. Results The results show that the radial stiffness is not uniform along the longitudinal direction of the stent. When the braiding angle increases from 30° to 75°, the minimum radial deformation decreases from 0.85 mm to 0.0325 mm (at a pressure of 500 Pa, for 24 braided wires). When the wire diameter increases from 0.026 mm to 0.052 mm, the minimum radial deformation decreases from 0.65 mm to 0.055 mm (at a pressure of 500 Pa and a braiding angle of 60°, for 24 braided wires). Frictions don't affect stent diameter and its axial length when braided stent is crimping, but the friction must be considered when it is related to the radial pressure required for compression the braided stent. Conclusions Compared with commonly used intracranial stents, a braided stent with geometrical parameters close to PED stent has a smaller radial stiffness but a considerably greater longitudinal flexibility. The results of this analysis of braided stents can help in the design and selection of flow diverter stents for clinical treatment of cerebral aneurysms.

Objective It has been reported that ADAMTS-12 is a susceptibility gene for rheumatoid arthritis (RA) development, and its level is significantly increased in RA patients. In addition, ADAMTS-12 is reported to be required for inflammation in otherwise healthy subjects. This study was undertaken to determine the role of ADAMTS-12 and the underlying mechanisms in the pathogenesis of inflammatory arthritis. Methods The collagen-induced arthritis (CIA) model was established in ADAMTS-12-deficient mice and their control littermates to determine the role of ADAMTS-12 in vivo. Micro-computed tomography scanning was used to demonstrate the destruction of the ankle joint; histologic analysis illustrated synovitis, pannus formation, and bone and cartilage destruction; enzyme-linked immunosorbent assay was performed to measure serum levels of inflammatory cytokines; and protein-protein interaction assays were performed to detect the interactions of ADAMTS-12 and its various deletion mutants with connective tissue growth factor (CTGF). Results Deficiency of ADAMTS-12 led to accelerated inflammatory arthritis in the CIA mouse model. Loss of ADAMTS-12 caused enhanced osteoclastogenesis. In vitro and in vivo protein-protein interaction assays demonstrated that ADAMTS-12 bound and processed CTGF, a previously unrecognized substrate of ADAMTS-12. In addition, deletion of ADAMTS-12 enhanced, while overexpression of ADMATS-12 reduced, CTGF-mediated inflammation. Furthermore, ADAMTS-12 regulation of inflammation was largely lost in CTGF-deficient macrophages. Importantly, blocking of CTGF attenuated elevated inflammatory arthritis seen in the ADAMTS-12-deficient CIA mouse model. Conclusion This study provides evidence that ADAMTS-12 is a critical regulator of inflammatory arthritis and that this is mediated, at least in part, through control of CTGF turnover.

Background Atsttrin, an engineered protein composed of three tumor necrosis factor receptor (TNFR)-binding fragments of progranulin (PGRN), shows therapeutic effect in multiple murine models of inflammatory arthritis . Additionally, intra-articular delivery of PGRN protects against osteoarthritis (OA) progression. The purpose of this study is to determine whether Atsttrin also has therapeutic effects in OA and the molecular mechanisms involved. Methods Surgically induced and noninvasive rupture OA models were established in mouse and rat, respectively. Cartilage degradation and OA were evaluated using Safranin O staining, immunohistochemistry, and ELISA. Additionally, expressions of pain-related markers, degenerative factors, and anabolic and catabolic markers known to be involved in OA were analyzed. Furthermore, the anabolic and anti-catabolic effects and underlying mechanisms of Atsttrin were determined using in-vitro assays with primary chondrocytes. Results Herein, we found Atsttrin effectively prevented the accelerated OA phenotype associated with PGRN deficiency. Additionally, Atsttrin exhibited a preventative effect in OA by protecting articular cartilage and reducing OA-associated pain in both nonsurgically induced rat and surgically induced murine OA models. Mechanistic studies revealed that Atsttrin stimulated TNFR2-Akt-Erk1/2-dependent chondrocyte anabolism, while inhibiting TNFα/TNFR1-mediated inflammatory catabolism. Conclusions These findings not only provide new insights into the role of PGRN and its derived engineered protein Atsttrin in cartilage homeostasis as well as OA in vivo, but may also lead to new therapeutic alternatives for OA as well as other relative degenerative joint diseases. Electronic supplementary material The online version of this article (doi:10.1186/s13075-017-1485-8) contains supplementary material, which is available to authorized users.

To evaluate the influence of deployment strategy on the mechanical interaction between braided stent and parent artery of intracranial aneurysm (the elasticity of the arterial wall is considered), finite-element analyses are carried out by referring to computational models of flow-diverter device and arterial wall. Two implantation strategies are used to virtually implant the braided stent into the ideal intracranial aneurysm model. One is the noncompacted implantation method, and the other is the implantation method of using push-pull technique. During the process of the implantation, the changes of the arterial shape around the aneurysm and the changes of the wall pressure at the contact area between the braided stent and the inner wall of the artery are analyzed. The results indicate that the average contact pressure in the area of low porosity is 57 mmHg using the push-pull technique, and the average contact pressure of the parent artery is 10.45 mmHg using the non-push-pull technique. The diameter of the parent artery at the aneurismal orifice increased about 0.2 mm when using the push-pull technique, so the elasticity of the vessel should be considered in the mechanical analysis of interaction between stent and vessel.

Progranulin (PGRN) restrains inflammation and is therapeutic against inflammatory arthritis; however, the underlying immunological mechanism remains unknown. In this study, we demonstrated that anti-inflammatory cytokine IL-10 was a critical mediator for PGRN-mediated anti-inflammation in collagen-induced arthritis by using PGRN and IL-10 genetically modified mouse models. IL-10 green fluorescent protein reporter mice revealed that regulatory T (Treg) cells were the predominant source of IL-10 in response to PGRN. In addition, PGRN-mediated expansion and activation of Treg cells, as well as IL-10 production, depends on JNK signaling, but not on known PGRN-activated ERK and PI3K pathways. Furthermore, microarray and chromatin immunoprecipitation sequencing screens led to the discovery of forkhead box protein O4 and signal transducer and activator of transcription 3 as the transcription factors required for PGRN induction of IL-10 in Treg cells. These findings define a previously unrecognized signaling pathway that underlies IL-10 production by PGRN in Treg cells and present new insights into the mechanisms by which PGRN resolves inflammation in inflammatory conditions and autoimmune diseases, particularly inflammatory arthritis.-Fu, W., Hu, W., Shi, L., Mundra, J. J. Xiao, G., Dustin, M. L., Liu, C. Foxo4- and Stat3-dependent IL-10 production by progranulin in regulatory T cells restrains inflammatory arthritis.

The pigmentation of mammalian skin and hair develops through the interaction of two basic cell types — pigment donors and recipients. The pigment donors are melanocytes, which produce and distribute melanin through specialized structures. The pigment recipients are epithelial cells, which acquire melanin and put it to use, collectively yielding the pigmentation visible to the eye. This review will focus on the pigment recipients, the historically less understood cell type. These end-users of pigment are now known to exert a specialized control over the patterning of pigmentation, as they identify themselves as melanocyte targets, recruit pigment donors, and stimulate the transfer of melanin. As such, this review will discuss the evidence that the skin is like a coloring book: the pigment recipients create a “picture,” a blueprint for pigmentation, which is colorless initially but outlines where pigment should be placed. Melanocytes then melanize the recipients and “color in” the picture.

Computational fluid dynamics analyses were performed on three models which have a giant aneurysm with or without a stenosis. The first is a model with an aneurysm (no stenosis and no stent), the second is a model with a preaneurysm stenosis, and the third is a model with an aneurysm implanted with a stent. The increase in pressure in aneurismal sac caused by a 50% stenosis is about 10.3[Formula: see text]mmHg at peak systole (comparison between the second model and the first model). It must pay attention to the increase of the pressure for the patient which has an aneurysm accompanied by a stenosis when making the treatment plan. Otherwise, it may cause the aneurysm rupture.

To investigate the biochemical changes in striatum after rat bone marrow mesenchymal stem cells (MSCs) were transplanted into hemiparkinsonian rats and to further confirm the therapeutic effects of rat MSCs for Parkinson's disease (PD).5-bromo-2-deoxyuridine (BrdU)-labeled MSCs were transplanted into the corpus striatum of the 6-hydroxydopamine (6-OHDA)-injected side of six PD model rats. Before and 8 weeks after MSC transplantation, ethological changes in PD rats were assessed. The expression of tyrosine hydroxylase (TH) in substantia nigra (SN) and striatum were measured using immunohistochemical methods. The differentiation of MSCs was detected by double immunofluorescence techniques. The concentrations of neural metabolites of N-acetylaspartate (NAA), choline (Cho) and creatine (Cr) were measured by ¹H-magnetic resonance spectroscopy (MRS). Relative concentrations of NAA/Cr and Cho/Cr were calculated.The behavior of PD rats in rotarod tests improved, and there were statistical differences in TH-positive cells in SN and TH-positive terminals in striatum after the transplantation of BrdU-labeled MSCs. Transplanted MSCs differentiated into MAP-2-positive neurons. Especially compared with pre-MSC transplantation, the neural metabolite NAA/Cr ratio of the 6-OHDA-injected side of the striatum increased (P0.05) and the Cho/Cr ratio decreased (P0.05).MSCs transplantation apparently improves neuronal function in the striatum of PD rats.

There is still a considerable lack of quantitative information concerning the effects of stent structures on blood flow in an aneurismal cavity. In this paper, five virtual stents with different structures and wire cross-sections were designed for incorporation into the same patient-specific aneurysm model. Computational fluid dynamics simulations were performed so as to study how these five types of stents modified hemodynamic parameters. Numerical results demonstrated that the mean flow rate in the aneurismal cavity decreased the most in the model that used a stent with a rectangular wire cross-section, and that the wall shear stresses at the dome and neck of the aneurysm decreased more in models that used a stent with a circular wire cross-section or a spiral stent with a rectangular wire cross-section compared to other models. In addition, the wall pressure on the aneurysm increased slightly after implantation of the stent in all five models. This result differs from that previously published, and may help guide the design and assist clinicians in selecting an appropriate stent for treating cerebral aneurysms.

Oxidative stress is considered as a possible molecular mechanism involved in lead toxicity. This study was carried out to investigate whether lead acetate could induce oxidative stress in mice, and the following damages as well. Lead acetate was given orally to mice for 4 weeks at doses of 0, 10, 50, 100mg/kg body weight every other day, respectively. Production of reactive oxygen species (ROS) and malondialdehyde (MDA) were measured as indicators of oxidative stress. DNA damage in peripheral blood lymphocytes was determined by comet assay. Ultrastructure alteration was detected using transmission electron microscopy. The alterations of p53, Bax, and Bcl-2 expression were determined by western blotting. The results showed that lead acetate significantly increased the levels of ROS and MDA in mice. Meanwhile, severe DNA damage and ultrastructure alterations were obviously observed. In addition, p53 and Bax expressions increased and the imbalance of Bax/Bcl-2 occurred. Therefore, it strongly suggests that lead may induce oxidative stress and change the expressions of apoptosis-related proteins in mouse liver.

PmrA/PmrB and PhoP/PhoQ are a pair of two-component systems (TCSs) that allow the Gram-negative bacteria to survive the cationic antimicrobial peptide polymyxin B. The two TCSs are linked by the polymyxin resistance protein, PmrD. The PhoP-activated PmrD protects the phosphorylated response regulator PmrA from dephosphorylation, and promotes the transcription of PmrA-activated genes responsible for polymyxin resistance. PmrD is the first protein identified to mediate the connectivity between two TCSs by protecting the phosphorylated response regulator of the downstream TCS. PmrD shows no homology to proteins with known structures. We present here the solution structure of PmrD from Escherichia coli, the first three-dimensional structure of the PmrD family. Our study provides the structural basis of the novel interacting mechanism of bacterial two-component signal-transduction systems.

Braided stents are commonly used to treat cerebral aneurysm, but there is little information about the bending characteristic of braided stent used for cerebral aneurysm. This paper investigates how geometrical parameters of braided stent influence its flexibility. Eight groups of braided stent models with different geometries (i.e., nominal diameter, length, braiding angle, number of wires, diameter of wire, frictional coefficient among wires and porosity) were constructed. Parametric analyses of these models were carried out by using Abaqus/Explicit. When the nominal diameter varied from 2[Formula: see text]mm to 5.5[Formula: see text]mm, the forces required for flexural deformation decrease from [Formula: see text][Formula: see text]N to [Formula: see text][Formula: see text]N; when the axial length varied from 10[Formula: see text]mm to 40[Formula: see text]mm, the forces required for flexural deformation decrease from [Formula: see text][Formula: see text]N to [Formula: see text][Formula: see text]N; when the braiding angle increases from 30[Formula: see text] to 75[Formula: see text] (the number of wires is 48 and the diameter of the wire is 0.026[Formula: see text]mm), the forces required for bending deformation decrease from [Formula: see text][Formula: see text]N to [Formula: see text][Formula: see text]N; when the diameter of wires increases from 0.026[Formula: see text]mm to 0.052[Formula: see text]mm (the number of wires is 24 and the braiding angle is 60[Formula: see text]), the forces required for flexural deformation increase from [Formula: see text][Formula: see text]N to [Formula: see text][Formula: see text]N; and when the number of wires increases from 14 to 48 (the braiding angle is 75[Formula: see text] and the diameter of the wire is 0.026[Formula: see text]mm), the forces required for flexural deformation increase from [Formula: see text][Formula: see text]N to [Formula: see text][Formula: see text]N. From the data above it can be seen that the diameter of wires, the number of wires and braiding angle have a larger impact on bending characteristics of braided stent; and the axial length and nominal diameter have a smaller impact on bending characteristics of braided stent. Results of the present study may provide theoretical guidance for the design of self-expanding braided stent and its clinical practice.

Flavodoxins are a family of small electron transferases widely distributed in prokaryotes. They utilize a noncovalently bound flavin mononucleotide (FMN) molecule as the redox center, and are able to switch between three different redox states, namely the oxidized (ox) state, the one-electron reduced semiquinone (sq) state, and the twoelectron reduced hydroquinone (hq) state (Knight and Hardy 1967). Since flavodoxins display two redox potentials and can transfer either one or two electrons at a time, they are functionally versatile. Flavodoxins have been identified to play important roles in various biological processes, including photosynthesis, methionine synthesis, biotin synthesis, and the activation of important enzymes such as pyruvate-formate lyase and ribonucleotide reductase (Sancho 2006). In eukaryotes, flavodoxins are integrated into multi-domain proteins and carry out similar redox functions. Flavodoxins can be further classified into the long-chain and short-chain subfamilies, which are distinguished based on the presence or absence of a 20-amino acid extra segment. The flavodoxin-like domains in eukaryotic multidomain proteins are more closely related to the short-chain subfamily. Previous investigations on the extra sequence suggested it is not directly involved in cofactor binding but may play a role in protein partner recognition and interactions, and it was proposed that the short-chain may derive from the long-chain group during evolution (LopezLlano et al. 2004a, b). However, the underlying molecular mechanism is yet unclear and the structural determinants for the distinct biological functions of different flavodoxins remain elusive. The Escherichia coli genome harbors several genes encoding flavodoxin proteins. Among them, the fldA gene encodes the well-characterized flavodoxin 1 protein which belongs to the long-chain subfamily and is essential for bacteria survival (Gaudu and Weiss 2000). The fldB gene encodes another long-chain flavodoxin which shares over 40 % sequence identity with FldA yet is functionally distinct. Insertion mutation of the fldB gene is not lethal to E. coli, and over expression of FldB could not substitute FldA (Gaudu and Weiss 2000). Evidence suggested that the fldB gene is a member of the superoxide response soxRS regulon, and its expression level was induced by paraquat (methyl viologen) (Gaudu and Weiss 2000). However, the exact function of FldB protein remains unclear. We have previously carried out structural and dynamic studies of E. coli flavodoxins including the short-chain MioC and YqcA proteins as well as the long-chain FldA by solution NMR spectroscopy (Hu et al. 2006; Ye et al. 2014). The FldA protein exhibit global conformational exchanges without the presence of FMN cofactor, rendering one third of its backbone amide signals missing in the NMR spectra, and we were unable to determine its solution structures in the apo-form. In contrast, the FldB protein Q. Ye W. Fu Y. Hu (&) C. Jin (&) Beijing Nuclear Magnetic Resonance Center, Peking University, Beijing 100871, China e-mail: yunfei@pku.edu.cn

Validation studies are prerequisites for computational fluid dynamics (CFD) simulations to be accepted as part of clinical decision-making. This paper reports on the 2011 edition of the Virtual Intracranial Stenting Challenge. The challenge aimed to assess the reproducibility with which research groups can simulate the velocity field in an intracranial aneurysm, both untreated and treated with five different configurations of high-porosity stents. Particle imaging velocimetry (PIV) measurements were obtained to validate the untreated velocity field. Six participants, totaling three CFD solvers, were provided with surface meshes of the vascular geometry and the deployed stent geometries, and flow rate boundary conditions for all inlets and outlets. As output, they were invited to submit an abstract to the 8th International Interdisciplinary Cerebrovascular Symposium 2011 (ICS'11), outlining their methods and giving their interpretation of the performance of each stent configuration. After the challenge, all CFD solutions were collected and analyzed. To quantitatively analyze the data, we calculated the root-mean-square error (RMSE) over uniformly distributed nodes on a plane slicing the main flow jet along its axis and normalized it with the maximum velocity on the slice of the untreated case (NRMSE). Good agreement was found between CFD and PIV with a NRMSE of 7.28%. Excellent agreement was found between CFD solutions, both untreated and treated. The maximum difference between any two groups (along a line perpendicular to the main flow jet) was 4.0 mm/s, i.e. 4.1% of the maximum velocity of the untreated case, and the average NRMSE was 0.47% (range 0.28-1.03%). In conclusion, given geometry and flow rates, research groups can accurately simulate the velocity field inside an intracranial aneurysm-as assessed by comparison with in vitro measurements-and find excellent agreement on the hemodynamic effect of different stent configurations.

In order to assess the variability in the calculation of the pressure gradient through a moderate thoracic aortic coarctation (MTAC), a 3D finite element model of MTAC was constructed, which includes the ascending aorta, the aortic arch, the descending aorta, and the three large branches (the innominate artery, the left common carotid artery, and the left subclavian artery), as well as with a coarctation in the descending aorta. The surface model of MTAC in STL format was imported into ANSYS ICEM CFD12.1 to generate volume mesh.A finite element model suitable for hemodynamics analysis of patient-specific MTAC was established.Numerical simulation of hemodynamics in this model was performed by means of Computational fluid dynamics (CFD) using ANSYS CFX12.1. The temporal distributions of homodynamic variables such as streamlines, wall pressure, velocity vector and wall shear stress in the arteries were analyzed during a cardiac cycle. The maximum and the average of pressure gradient in a cardiac cycle through a MTAC are 13 mmHg and 2.84 mmHg respectively. The pressure difference between the systolic and the diastolic in a cardiac cycle proximal to the coarctation is about 38 mmHg, which is smaller than the difference between the recorded systolic and diastolic pressures of 115 and 65 mmHg (i.e. the difference is 115-65=50). Similarly, the pressure gradient through the coarctation under exercise conditions could be predicted via modifying the inflow and outflow boundary conditions under resting conditions. CFD techniques make it possible to obtain information (such as pressure when the patient is under exercise condition) which is difficult to get in clinic practice or in experiment based on patient-specific data.

The objective of this study is to investigate the hemodynamics in patient-specific internal carotid aneurysm and discuss the reason for rupture of aneurysm. A 3-Dimensional pulsatile blood flow in internal carotid with a sidewall aneurysm was studied numerically with the average Reynolds number of 704. Patient-specific model whose parent artery has a large "S" bending and a sidewall aneurysm was constructed from CT data. Unsteady, incompressible, 3-Dimensional Navier-Stokes equations were employed to solve the flow field. The temporal distributions of hemodynamic variables during the cardiac cycle such as streamlines, wall pressure and wall shear stress (WSS) in the arteries and aneurysm were analyzed. From streamlines it can be found that there is an obvious vortex flow in aneurismal cavity in a cardiac cycle. The type of this vortex flow is not changed in a cardiac cycle. As far as Wall Shear Stress, there is a region in aneurismal neck where the value of WSS is relatively high. Growth and rupture mechanism of internal carotid aneurysm in patient can be analyzed based on patient-specific model and hemodynamics simulation.

An intracranial aneurysm (IA) is a pathological dilatation of the cerebral artery wall. Blood flow and pressure inside IA are not steady. Interaction between blood flow and vessel wall reveals stress and deformation of vessel wall in vivo. It also provides more true and reliable information about blood flow. So analysis of Fluid Structure Interaction (FSI) is favor of proper explanation for formation and development of aneuris-mal disease. Patient-specific model whose parent artery has a large “S” bending and a side wall aneurysm was constructed from CT data. Pulsatile calculation of FSI for this constructed model was carried out. From streamlines it can be found that there is an obvious vortex flow in aneurismal cavity in a cardiac cycle. The type of this vortex flow is not changed in a cardiac cycle. As far as Wall Shear Stress (WSS), there is a region in aneurismal neck where the value of WSS is relatively high. At 0.27s in a cardiac cycle, deformation of the vessel wall is the biggest. There are two regions in aneurismal dome where the value of Von Mises Stress is the locally biggest.

Endovascular stents with different porosities and wire cross-section shapes may create different effects on the hemodynamics of aneurysm treated with interventional therapy because stents with different wire cross-section shapes induce different flow resistances through the stent strut. In order to analyze the flow resistance of the new stent with triangular cross-section by numerical simulation method, and to provide a reference basis for the structural design and optimization of endovascular stent, four kinds of models of the triangular cross-section bare stent were constructed using SoildWorks 2007 software. Numerical simulations of steady and transient blood flows in the four models were performed respectively according to CFD method using finite element software ANSYS 11. Hemodynamics data in the four models were collected, such as the flow patterns, the distribution of pressure and the flow resistance. The results show that the stent with triangular wire cross-section can produce large resistance of blood inflow to aneurysm and small resistance of outflow from aneurysm. Thus, outflow is easy and inflow is difficult. Thereby blood perfusion and flow of aneurysm cavity is restrained, and the pressure of aneurysm cavity is reduced, which plays certain effect on the intervention treatment of aneurysm. This result provides some instructions for the design of stent structure.

An intracranial aneurysm (IA) is a pathological dilatation of the cerebral artery wall. In clinical practice it is currently widely accepted that maximum diameter of IA is an indicator for patient whether to treat or not. But studies have demonstrated that maximum diameter is not a reliable determinant of IA rupture. According to the theory of strength, if the stress value exceeds the strength limit of material, it will result in failure. So wall stress may be a better indicator to predict aneurismal rupture (failure). Study about wall stress calculated from solid stress model using static pressure has been performed. But blood flow and pressure inside IA are not steady. The dynamic interaction between the unsteady flow and wall may influence the distribution and magnitude of wall stress. The objective of the present study was to compare static and dynamic wall stress analysis of patient specific IA. Patient-specific model of cerebral aneurysm whose host artery has large bending was created from CT data. Simulations about fluid structure interaction model and static structural model were carried out respectively. Commercial software Ansys 11 was used for simulation. The results demonstrated that FSI can change local wall stresses slightly. However, as far as the peak wall stress is concerned, the change of wall stress is negligible. Therefore static structural simulation can be used to predict rupture risk of aneurysm if wall stress was used as an indicator of aneurismal rupture and computational cost can be reduced greatly.

The bacterial chemotaxis adaptor protein CheW physically links the chemoreceptors (MCPs) and the histidine kinase CheA. Extensive investigations using bacterium Escherichia coli have established the central role of CheW in the MCP-modulated activation of CheA. Here we report the solution structure of CheW from E. coli determined by NMR spectroscopy. The results show that E. coli CheW shares an overall fold with previously reported structure of CheW from Thermotoga maritima, whereas local conformational deviations are observed. In particular, the C-terminal alpha-helix is considerably longer in E. coli CheW and appears to shrink the active binding pocket with CheA. Our study provides the structural basis for further investigations in E. coli chemotaxis.

Tributyltin (TBT) is one of the environmental pollutants, which is mostly accumulated in marine animals. The toxic effects of TBT have been extensively documented in several types of cells, but the molecular mechanisms responsible for TBT-induced cell damage are still not fully elucidated. The present study was undertaken to evaluate the apoptotic related biochemical changes in human amnion cells induced by TBT. After cells were exposed to TBT at the concentrations of 1–4 μM for 2 h, the results suggested that TBT could induce an early and typical apoptosis, moreover caspase-3, the modifications of cytoskeletal structure and the Bcl-2 family were involved in this process. The results will deepen our understanding about the toxic mechanism of TBT on human amnion cells.

Previous studies have shown that ultraviolet (UV) A light and the polycyclic aromatic hydrocarbon benzo[ a ]pyrene (BaP) can synergistically enhance the formation of 8-hydroxy-2 ′ deoxyguanosine (8-OHdG) in living cells. It has been postulated that the underlying mechanism is production of reactive oxygen species (ROS) via photosensitization, but direct evidence supporting this hypothesis has been lacking. This study examined intracellular ROS production in living cells co-exposed to UV-A and BaP as well as the relationship between intracellular production of ROS and formation of 8-OHdG. KB cells were exposed to BaP for 24 h, followed by exposure to UV-A (365 nm) or UV-B (312 nm). The levels of intracellular ROS were directly measured by use of the fluorescent probe dihydrorhodamine 123 (DHR-123) in flow cytometry. Levels of 8-OHdG were measured by high performance liquid chromatography coupled with electrochemical detection (HPLC-ECD). The results demonstrated that UV-B itself induced a much greater level of intracellular ROS than did UV-A alone under the same dose of energy (0.10 mW/cm 2 , 20 min). The presence of BaP (13.3 μM) substantially increased ROS production in UV-A-treated cells (2.9-fold), but only slightly enhanced ROS production in UV-B-treated cells (1.3-fold). These results show that BaP acts mainly as a photosensitizer of UV-A, but not UV-B. Furthermore, greater intracellular ROS production was proportional to both BaP concentration and UV-A dosage. There was a linear relationship between ROS production and 8-OHdG formation in cells co-exposed to BaP and UV-A. Results of this study suggest that UV-A and BaP act synergistically to enhance ROS production and formation of 8-OHdG, resulting in increased DNA damage.