Classification of Sand-Binder Mixtures from the Foundry Industry Using Electrical Impedance Spectroscopy and Support Vector Machines.

Molding sand mixtures used in the foundry industry consist of various sands (quartz sands, chromite sands, etc.) and additives such as bentonite. The optimum control of the processes involved in using the mixtures and in their regeneration after the casting requires an efficient in-line monitoring method that is not available today. We are investigating whether such a method can be based on electrical impedance spectroscopy (EIS). To establish a database, we have characterized various sand mixtures by EIS in the frequency range from 0.5 kHz to 1 MHz under laboratory conditions. Attempts at classifying the different molding sand mixtures by support vector machines (SVM) show encouraging results. Already high assignment accuracies (above 90%) could even be improved with suitable feature selection (sequential feature selection). At the same time, the standard uncertainty of the SVM results is low, i.e., data assigned to a class by the presented SVMs have a high probability of being assigned correctly. The application of EIS with subsequent evaluation by machine learning (machine-learning-enhanced EIS, MLEIS) in the field of bulk material monitoring in the foundry industry appears possible.

Uncoupling protein 2 modulates polarization and metabolism of human primary macrophages via glycolysis and the NF­κB pathway.

Metabolic abnormalities, particularly the M1/M2 macrophage imbalance, play a critical role in the development of various diseases, leading to severe inflammatory responses. The present study aimed to investigate the role of uncoupling protein 2 (UCP2) in regulating macrophage polarization, glycolysis, metabolic reprogramming, reactive oxygen species (ROS) and inflammation. Primary human macrophages were first polarized into M1 and M2 subtypes, and these two subtypes were infected by lentivirus-mediated UCP2 overexpression or knockdown, followed by enzyme-linked immunosorbent assay, reverse transcription-quantitative PCR, western blotting and flow cytometry to analyze the effects of UCP2 on glycolysis, oxidative phosphorylation (OXPHOS), ROS production and cytokine secretion, respectively. The results demonstrated that UCP2 expression was suppressed in M1 macrophages and increased in M2 macrophages, suggesting its regulatory role in macrophage polarization. UCP2 overexpression decreased macrophage glycolysis, increased OXPHOS, decreased ROS production, and led to the conversion of M1 polarization to M2 polarization. This process involved NF-κB signaling to regulate the secretion profile of cytokines and chemokines and affected the expression of key enzymes of glycolysis and a key factor for maintaining mitochondrial homeostasis (nuclear respiratory factor 1). UCP2 knockdown in M2 macrophages exacerbated inflammation and oxidative stress by promoting glycolysis, which was attenuated by the glycolysis inhibitor 2-deoxyglucose. These findings highlight the critical role of UCP2 in regulating macrophage polarization, metabolism, inflammation and oxidative stress through its effects on glycolysis, providing valuable insights into potential therapeutic strategies for macrophage-driven inflammatory and metabolic diseases.

The synergistic effect of trap deactivation and hysteresis suppression at grain boundaries in perovskite interfaces via multifunctional groups.

In spite of the outstanding photoelectric properties of perovskite materials, numerous defects produced in the preparation process eventually result in decomposition of the perovskite layer. To date, the mechanism of defect passivation and hysteresis reduction via additive engineering has still been obscure for perovskite materials, which seriously restricts performance improvement of the devices. Herein, conductive atomic force microscopy (C-AFM) and Kelvin probe force microscopy (KPFM) measurements were applied to probe carbamic acid ethyl ester (EU)-based trap passivation and suppression of hysteresis in perovskite films. The results indicate that the internal interaction between multifunctional bonds ("CO" and "-NH2") of EU and Pb2+ ions of the perovskite may inactivate the trap state and inhibit ion migration within sub-grains and grain boundaries (GBs), resulting in improvement of the long-term stability of the cells. In consequence, the EU-modified champion device prepared in all-air achieved a power conversion efficiency (PCE) of 20.10%, one of the high performances for the devices fabricated in air to date. In short, this work will propose some interesting speculation about ion migration as well as its influence on hysteresis in perovskite materials.

Ring-Oven-Assisted In Situ Synthesis of Metal-Organic Frameworks on the Lab-On-Paper Device for Chemiluminescence Detection of Nitrite in Whole Blood.

In situ synthesis of metal-organic frameworks (MOFs) on flexible materials for the fabrication of functional platforms and micro-devices is challenging. The time-/precursor-consuming procedure and uncontrollable assembly are stumbling blocks for constructing this platform. Herein, a novel in situ MOF synthesis method on paper substrates by use of the ring-oven-assisted technique was reported. Utilizing the ring-oven's heating and washing function, MOFs can be synthesized in 30 min on the designated position of paper chips with extremely low-volume precursors. The principle of this method was explained by steam condensation deposition. The MOFs' growth procedure was theoretically calculated by crystal sizes and the results conformed to the Christian equation. As different MOFs (Cu-MOF-74, Cu-BTB, Cu-BTC) can be synthesized successfully on paper-based chips, the ring-oven-assisted in situ synthesis method has great generality. Then, the prepared Cu-MOF-74 loading paper-based chip was applied to the chemiluminescence (CL) detection of nitrite (NO2-), based on the catalysis effect of Cu-MOF-74 on the NO2--H2O2 CL system. Also, by the delicate design of the paper-based chip, NO2- can be detected with the detection limit (DL) of 0.5 nM in whole blood samples without sample pretreatment. This work establishes a distinctive method for the in situ synthesis of MOFs and the application of MOFs on paper-based CL chips.

Evaluation of Electrical Impedance Spectra by Long Short-Term Memory to Estimate Nitrate Concentrations in Soil.

Monitoring the nitrate concentration in soil is crucial to guide the use of nitrate-based fertilizers. This study presents the characteristics of an impedance sensor used to estimate the nitrate concentration in soil based on the sensitivity of the soil dielectric constant to ion conductivity and on electrical double layer effects at electrodes. The impedance of synthetic sandy soil samples with nitrate nitrogen concentrations ranging from 0 to 15 mg/L was measured at frequencies between 20 Hz and 5 kHz and noticeable conductance and susceptance effects were observed. Long short-term memory (LSTM), a variant of recurrent artificial neural networks (RNN), was investigated with respect to its suitability to extract nitrate concentrations from the measured impedance spectra and additional physical properties of the soils, such as mass density and water content. Both random forest and LSTM were tested as feature selection methods. Then, numerous LSTMs were trained to estimate the nitrate concentrations in the soils. To increase estimation accuracy, hyperparameters were optimized with Bayesian optimization. The resulting optimal regression model showed coefficients of determination between true and predicted nitrate concentrations as high as 0.95. Thus, it could be demonstrated that the system has the potential to monitor nitrate concentrations in soils in real time and in situ.

Study on the relationship between genetic polymorphism of reductive folic acid carrier and the risk of neural tube defects.

BACKGROUND: To investigate the association of folate metabolism gene polymorphism with neural tube defects (NTDs) in Chinese population. METHODS: The subjects were divided into two groups, 495 children with NTDs (NTD group) and 255 healthy children (control group). RESULTS: The levels of folic acid, s-adenosine methionine (SAM), and Sam/s-adenosine homocysteine (SAH) in NTD group were lower than those in control group. There were significant differences in hey, SAH, and Sam levels between two groups, but there was no significant difference in folic acid content. High fever in early pregnancy, taking antiepileptic drugs, father's exposure to organic solvents, folic acid deficiency, and mother's diabetes were the important risk factors in NTDs. MTHFR 677C > T gene was a risk factor for NTD in children, while 1298A > C gene was a protective factor. CONCLUSION: Folic acid metabolism markers were different in NTD children and their mothers, and the overall trend showed that folate, SAM, and SAM/SAH levels were decreased, while Hcy and SAH levels were increased; MTHFR 677C > T gene of SNPs was a risk factor for the occurrence of NTDs, and MTHFR 1298A > C gene was a protective factor, and the environmental risk factor had a synergistic effect on occurrence of NTDs.

Tetrakis(4-pyridylphenyl)ethylene-based Zinc Metal-Organic Framework with Aggregation-Induced Chemiluminescence Emission on a Paper Platform for Formaldehyde Detection in Breath.

Volatile formaldehyde (FA) in exhaled breath (EB) is considered as a biomarker for lung cancer (LC). On-the-spot selective and sensitive detection of gaseous FA is rather important for LC screening and diagnosis. Herein, a tetrakis(4-pyridylphenyl)ethylene (Py-TPE)-based zinc metal-organic framework (MOF) with excellent aggregation-induced emission (AIE) property was utilized for absorption and selective detection of FA in EB. The porous Zn-Py-TPE served as a gaseous confinement cavity for the adsorption of FA in EB. Interestingly, Zn-Py-TPE was aggregated on paper, and then aggregation-induced chemiluminescence (CL) emission can be triggered by only adding bis(2,4,6-trichlorophenyl)oxalate (TCPO). Though without H2O2, the CL of Zn-Py-TPE-TCPO was enhanced greatly by FA. FA promoted the aggregation of Zn-Py-TPE on paper by forming halogen bonding between FA and Zn-Py-TPE, which contributed to the better selectivity. FA can also stimulate the production of more singlet oxygen (1O2) in the Zn-Py-TPE-TCPO CL system. Hence, FA could be detected via the proposed Zn-Py-TPE-TCPO system with a quantification linear range of 1.0-100.0 ppb and detection limit of 0.3 ppb. This portable, low-cost, and sensitive paper-based platform can achieve trace FA detection in EB and is expected to provide an on-the-spot screening platform for lung cancer.

VdPT1 Encoding a Neutral Trehalase of Verticillium dahliae Is Required for Growth and Virulence of the Pathogen.

Verticillum dahliae is a soil-borne phytopathogenic fungus causing destructive Verticillium wilt disease. We previously found a trehalase-encoding gene (VdPT1) in V. dahliae being significantly up-regulated after sensing root exudates from a susceptible cotton variety. In this study, we characterized the function of VdPT1 in the growth and virulence of V. dahliae using its deletion-mutant strains. The VdPT1 deletion mutants (ΔVdPT1) displayed slow colony expansion and mycelial growth, reduced conidial production and germination rate, and decreased mycelial penetration ability and virulence on cotton, but exhibited enhanced stress resistance, suggesting that VdPT1 is involved in the growth, pathogenesis, and stress resistance of V. dahliae. Host-induced silencing of VdPT1 in cotton reduced fungal biomass and enhanced cotton resistance against V. dahliae. Comparative transcriptome analysis between wild-type and mutant identified 1480 up-regulated and 1650 down-regulated genes in the ΔVdPT1 strain. Several down-regulated genes encode plant cell wall-degrading enzymes required for full virulence of V. dahliae to cotton, and down-regulated genes related to carbon metabolism, DNA replication, and amino acid biosynthesis seemed to be responsible for the decreased growth of the ΔVdPT1 strain. In contrast, up-regulation of several genes related to glycerophospholipid metabolism in the ΔVdPT1 strain enhanced the stress resistance of the mutated strain.

A metal-organic framework loaded paper-based chemiluminescence sensor for trace acetone detection in exhaled breath.

Trace acetone determination in breath can be regarded as a noninvasive method for diagnosis of diabetes. Here, a paper-based CL gas sensor combined with UiO-66 as the preconcentrator was established for sensitive detection of trace acetone in exhaled breath. UiO-66 with excellent adsorption performance and unique water stability was used for the adsorption and enrichment of acetone gas under high humidity conditions in exhaled breath. As acetone can remarkably increase the chemiluminescence (CL) of the 2,4-dinitrophenylhydrazine (DNPH)-potassium permanganate (KMnO4) system, a sensitive CL device based on a paper substrate for trace acetone detection was established and the detection limit was 0.03 ppm. The fabricated method was used to assess the content of trace acetone in exhaled breath with satisfactory recoveries of 90-110%. It is expected to realize the noninvasive determination of acetone for diabetic patients in exhaled breath.

IRF4 suppresses osteogenic differentiation of BM-MSCs by transcriptionally activating miR-636/DOCK9 axis.

OBJECTIVES: Osteoblasts are derived from Bone Marrow-derived Mesenchymal Stem Cells (BM-MSCs), which play an indispensable role in bone formation. In this study, the authors aim to investigate the role of IRF4 in the osteogenic differentiation of BM-MSCs and its potential molecular mechanism. METHODS: The authors used lentivirus infection to overexpress IRF4 in BM-MSCs. The expression of IRF4 and osteogenesis-related genes were detected by qRT-PCR and western blot analysis. The osteogenic differentiation of BM-MSCs was evaluated by Alkaline Phosphatase (ALP) activity, Alizarin red staining, and Alkaline Phosphatase (ALP) staining. Chromatin Immunoprecipitation (ChIP), Dual-Luciferase reporter assay and RNA Immunoprecipitation Assay were applied to confirm the regulatory mechanism between IRF4, miR-636 and DOCK9. RESULTS: The authors found IRF4 was down-regulated during the osteogenic differentiation of BM-MSCs, and IRF4 overexpression could decrease the osteogenic differentiation of BM-MSCs by specifically promoting the reduction of Alkaline Phosphatase (ALP) activity and down-regulating osteogenic indicators, including OCN, OPN, Runx2 and CollA1. Mechanistically, IRF4 activated microRNA-636 (miR-636) expression via binding to its promoter region, and Dedicator of Cytokinesis 9 (DOCK9) was identified as the target of miR-636 in BM-MSCs. Moreover, the damage in the capacity of osteogenic differentiation of BM-MSCs induced by IRF4 overexpression could be rescued by miR-636 inhibition. CONCLUSIONS: In summary, this paper proposed that IRF4/miR-636/DOCK9 may be considered as targets for the treatment of osteoporosis (OP).

Regional Homogeneity Alterations in Patients with Impaired Consciousness. An Observational Resting-State fMRI Study.

PURPOSE: It is always challenging to correctly differentiate between minimally conscious state (MCS) and vegetative state/unresponsive wakefulness syndrome (VS/UWS) among disorders of consciousness (DOC) patients. However, the underlying neural mechanisms of awareness identification remain incompletely understood. METHODS: Using regional homogeneity (ReHo) analysis, we evaluated how regional connectivity of brain regions is disrupted in MCS and VS/UWS patients. Resting-state functional magnetic resonance imaging was conducted in 14 MCS patients, 25 VS/UWS patients, and 30 age-matched healthy individuals. RESULTS: We found that MCS and VS/UWS patients demonstrated DOC-dependent reduced ReHo within widespread brain regions including posterior cingulate cortices (PCC), medial prefrontal cortices (mPFC), and bilateral fronto-parieto-temporal cortices and showed increased ReHo in limbic structures. Moreover, a positive correlation between Coma Recovery Scale-Revised (CRS-R) total scores and reduced ReHo in the left precuneus was observed in VS/UWS patients, despite the linear trend was not found in MCS patients. In addition, ReHo were also observed reduced in three mainly intrinsic connectivity networks (ICNs), including default mode network (DMN), executive control network (ECN), and salience network (SN). Notably, as the clinical symptoms of consciousness disorders worsen from MCS to VS/UWS, ReHo in dorsal DMN, left ECN, and posterior SN became significantly reduced. CONCLUSION: These findings make a further understanding of the underlying neural mechanism of regional connectivity among DOC patients and provide additional neuroimaging-based biomarkers for the clinical diagnosis of MCS and VS/UWS patients.

Recent Advances and Applications in Paper-Based Devices for Point-of-Care Testing.

Point-of-care testing (POCT), as a portable and user-friendly technology, can obtain accurate test results immediately at the sampling point. Nowadays, microfluidic paper-based analysis devices (µPads) have attracted the eye of the public and accelerated the development of POCT. A variety of detection methods are combined with µPads to realize precise, rapid and sensitive POCT. This article mainly introduced the development of electrochemistry and optical detection methods on µPads for POCT and their applications on disease analysis, environmental monitoring and food control in the past 5 years. Finally, the challenges and future development prospects of µPads for POCT were discussed.

IRF4 suppresses osteogenic differentiation of BM-MSCs by transcriptionally activating miR-636/DOCK9 axis

Abstract Objectives Osteoblasts are derived from Bone Marrow-derived Mesenchymal Stem Cells (BM-MSCs), which play an indispensable role in bone formation. In this study, the authors aim to investigate the role of IRF4 in the osteogenic differentiation of BM-MSCs and its potential molecular mechanism. Methods The authors used lentivirus infection to overexpress IRF4 in BM-MSCs. The expression of IRF4 and osteogenesis-related genes were detected by qRT-PCR and western blot analysis. The osteogenic differentiation of BM-MSCs was evaluated by Alkaline Phosphatase (ALP) activity, Alizarin red staining, and Alkaline Phosphatase (ALP) staining. Chromatin Immunoprecipitation (ChIP), Dual-Luciferase reporter assay and RNA Immunoprecipitation Assay were applied to confirm the regulatory mechanism between IRF4, miR-636 and DOCK9. Results The authors found IRF4 was down-regulated during the osteogenic differentiation of BM-MSCs, and IRF4 overexpression could decrease the osteogenic differentiation of BM-MSCs by specifically promoting the reduction of Alkaline Phosphatase (ALP) activity and down-regulating osteogenic indicators, including OCN, OPN, Runx2 and CollA1. Mechanistically, IRF4 activated microRNA-636 (miR-636) expression via binding to its promoter region, and Dedicator of Cytokinesis 9 (DOCK9) was identified as the target of miR-636 in BM-MSCs. Moreover, the damage in the capacity of osteogenic differentiation of BM-MSCs induced by IRF4 overexpression could be rescued by miR-636 inhibition. Conclusions In summary, this paper proposed that IRF4/miR-636/DOCK9 may be considered as targets for the treatment of osteoporosis (OP).

A Controlled Study of Continuous Lumbar Drainage of Fluid and Lumbar Puncture Drainage for Aneurysmal SAH after Intracranial Aneurysm Clipping.

Objective: To analyze the different effects of Continuous Lumbar Drainage of fluid and lumbar puncture drainage for aneurysmal subarachnoid hemorrhage (SAH) after intracranial aneurysm clipping. Method: Seventy-five patients with aneurysmal SAH who underwent aneurysm clipping were retrospectively analyzed and were divided into two groups according to the different postoperative drainage methods. The lumbar spine group received lumbar puncture drainage, and the lumbar cistern group received lumbar pool continuous drainage to compare the efficacy. Result: The time to normalize intracranial pressure and headache relief after drainage treatment in the lumbar cistern group was shorter than that in the lumbar spine group. The GOS score was higher than that in the lumbar spine group, and the cerebral artery flow velocity and NIHSS score were significantly lower than those in the lumbar spine group (P < 0.05). The total effective rate of drainage treatment was 76.32% in the lumbar cistern group, which was higher than that in the lumbar spine group (54.05%) (P < 0.05). The total complication rate was 18.42% in the lumbar cistern group, which was lower than that in the lumbar spine group (40.54%) (P < 0.05). Conclusion: Continuous Lumbar Drainage of fluid after intracranial aneurysm clipping for aneurysmal SAH can control symptoms more rapidly, reduce neurological deficits, and improve prognosis than lumbar puncture. Also, the drainage is safer and more widely used.

Neferine Inhibits Expression of Inflammatory Mediators and Matrix Degrading Enzymes in IL-1ß-Treated Rat Chondrocytes via Suppressing MAPK and NF-κB Signaling Pathways.

Osteoarthritis (OA), in which inflammation plays a crucial role, is the most common joint disease characterized by cartilage degradation. Neferine (Nef), a dibenzyl isoquinoline alkaloid, has shown its anti-inflammatory effects on other inflammatory diseases. Therefore, we hypothesized that Nef might also have an anti-inflammatory effect on OA and explored its effect on IL-1ß-treated rat chondrocytes. Sprague Dawley (SD) rat chondrocytes were stimulated with IL-1ß (10 ng/ml) and Nef (1, 5, and 10 µM) or IL-1ß (10 ng/ml) alone for 24 h. Expression of inducible nitric oxide synthase (iNOS), cyclooxygenase 2 (COX-2), matrix metalloproteinases (MMPs), and thrombospondin motifs-5 (ADAMTS5) was determined by quantitative real-time PCR and Western blotting. Expression of collagen II and aggrecan was examined by Western blotting, immunofluorescence, and safranin O staining. In addition, activation of MAPK and NF-κB signaling pathway was examined by Western blotting, and p65 nuclear translocation was evaluated by immunofluorescence. Nef reduced expression of inflammatory regulators (iNOS and COX-2) in IL-1ß-treated chondrocytes. Expression of IL-1ß-induced major catabolic enzymes (MMP3, MMP13, and ADAMTS5) was inhibited by Nef. Meanwhile, downregulation of collagen II and aggrecan expression was also ameliorated. Furthermore, Nef dampened abnormal activation of MAPK and NF-κB signaling pathway triggered by IL-1ß. Overall, the results above showed that Nef inhibited IL-1ß-induced excess production of inflammatory and catabolic factors in rat chondrocytes via inhibiting the MAPK and NF-κB pathways, suggesting a promising pharmacotherapy for OA.

Protective effects of Erdosteine on interleukin-1ß-stimulated inflammation via inhibiting the activation of MAPK, NF-κB, and Wnt/ß-catenin signaling pathways in rat osteoarthritis.

Osteoarthritis (OA), a degenerative arthropathy, is featured with progressive degradation of cartilage and a chondrocyte inflammatory response. Erdosteine (ER) showed the anti-oxidant properties and various anti-inflammatory effects in various diseases. However, whether it protects against OA remains unknown. In this study, we explore the potential therapeutic properties of ER on IL-1ß-stimulated rat chondrocytes and its underlying mechanism in vitro and vivo. Cell viability, pro-inflammatory cytokines and the degradation of ECM biomarkers were tested to determine the effects of ER at 10, 20, and 40 µM doses on IL-1ß-induced rat chondrocytes for 24 h in virto. In vivo, intra-articular injections of 50 µl of 100 mg/ml ER twice a week for 8 weeks. The results showed ER significantly suppressed the expressions of IL-1ß-induced the production of inflammatory factors in a dose-dependent pattern (4.30-fold decrease in COX-2, p < 0.05; 4.77-fold decrease in iNOS, p < 0.05 at 40 µM in protein levels). Moreover, ER could attenuate the degradation of ECM in IL-1ß-induced rat chondrocytes by repressing the expression of OA-related factors (2.40-fold decrease in ADAMTS-5, p < 0.05; 3.12-fold decrease in MMP1, p < 0.05; 3.97-fold decrease in MMP3, p < 0.05; and 2.62-fold decrease in MMP-13, p < 0.05 at 40 µM in protein levels). Furthermore, our study revealed that ER could inhibit the activations of IL-1ß-induced MAPK and Wnt/ß-catenin. Besides, ER could suppress the process of IL-1ß-induced P65 from the cytoplasm into the nucleus. In vivo, ER delaied the osteoarthritis progression in rat OA models. Collectively, ER might become a new therapeutic agent for OA.

NOV/CCN3 induces cartilage protection by inhibiting PI3K/AKT/mTOR pathway.

Osteoarthritis (OA), an age-related degenerative joint disease, is pathologically characterized by articular cartilage degeneration and synovial inflammation. Nephroblastoma overexpressed (NOV or CCN3), a matricellular protein, is a primary member of the CCN family (Cyr61, Ctgf, NOV) of proteins and is involved in various inflammatory disorders. Previous studies reported that CCN3 might play a therapeutic role in OA. However, the underlying mechanism remains unclear. In this study, we confirmed the expression of CCN3 was decreased in human and rat OA articular cartilage. Recombinant CCN3 ameliorated the IL-1ß-induced matrix catabolism, as demonstrated by MMP1, MMP3, MMP13, ADAMTS5 and iNOS expression, in vitro. In addition, the degradation of cartilage matrix such as collagen 2 and aggrecan could be reversed by CCN3. Furthermore, we found CCN3 promoted autophagy as Atg5, Beclin1 and LC3-II expression were increased. High-mobility group box 1 was negatively correlated with CCN3 in IL-1ß-induced osteoarthritis responses, and HMGB1 is involved in the protective effect of CCN3 in OA. Moreover, CCN3 overexpression decreased the expression of HMGB1 and reversed the IL-1ß induced MMPs production. Additionally, recombinant CCN3 or CCN3 overexpression attenuated the activation of PI3K/AKT/mTOR pathway induced by IL-1ß. Our study presents new mechanisms of CCN3 in osteoarthritis and indicates that CCN3 can serve as a novel potential therapeutic target for osteoarthritis.

Vanillic acid attenuates cartilage degeneration by regulating the MAPK and PI3K/AKT/NF-κB pathways.

Osteoarthritis (OA) is a frequently seen arthropathy that features cartilage loss and destruction. Vanillic acid (VA), is a well-known flavonoid, which possesses various pharmacological activities. However, the effects of Vanillic acid on articular cartilage destruction and the pathogenesis of OA remain unknown. The present study observed that VA attenuated OA progression in vivo and vitro. VA inhibited the expression of inflammation responses, including cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), matrix metalloproteinases (MMPs) and aggrecanase -2(ADAMTS5). Moreover, the major markers of hypertrophic chondrocytes such as Collagen X, Runt related transcription factor 2 (Runx2) and Vascular endothelial growth factor (VEGFA) were also reduced by VA. In addition, the interleukin-1ß (IL-1ß) -stimulated collagen 2 and aggrecan destruction were suppressed by VA. Moreover, VA concentration -dependently inhibited IL-1ß induced production of High-mobility group box 1 (HMGB1), a classic damage-associated molecular pattern (DAMP) molecule with strong pro-inflammatory effects in OA. Meanwhile, we revealed that VA suppressed the IL-1ß induced activation of MAPK and PI3K/AKT/NF-κB pathways. In vivo, VA alleviated osteoarthritis progression in a rat OA model. Collectively, our results demonstrate that VA could potentially be a new candidate for OA therapy.

Pravastatin alleviates interleukin 1ß-induced cartilage degradation by restoring impaired autophagy associated with MAPK pathway inhibition.

BACKGROUND: Osteoarthritis (OA) is a chronic degenerative joint disease characterized by cartilage degradation driven by proinflammatory cytokines; meanwhile, statins display anti-inflammatory effects. Here we assessed the effects of pravastatin on inflammatory rat chondrocytes and explored the underlying mechanism. METHODS: Rat articular chondrocytes were pretreated with pravastatin and subsequently stimulated with IL-1ß. Then, the expression levels of OA- and autophagy-related effectors, at the mRNA and protein levels, were examined by real-time polymerase chain reaction (RT-PCR) and Western blotting, respectively. Autophagic flux in chondrocytes in different treatment groups was monitored via GFP-mRFP-LC3 adenovirus transfection and confocal microscopy. Activation of MAPK, PI3K/Akt, and NF-κB pathways in chondrocytes with or without pravastatin treatment during IL-1ß stimulation was examined by Western blotting. RESULTS: Our results showed that pravastatin downregulated the degradation related genes MMP3, MMP13 and ADAMTS5, as well as extracellular matrix degeneration induced by IL-1ß. In addition, pravastatin upregulated the autophagy related genes atg7, atg12, Beclin1, and LC3 II in IL-1ß stimulated chondrocytes. GFP-mRFP-LC3 adenovirus transfection also indicated that pravastatin restored impaired autophagy in OA chondrocytes. Furthermore, we demonstrated that regulation of the MAPK signaling pathway may be responsible for autophagy regulation in the articular cartilage. CONCLUSIONS: Taken together, these findings suggested that pravastatin restores impaired autophagic flux by inhibiting MAPK activation and protects the cartilage against inflammatory responses, suggesting a potential role for autophagic flux in pravastatin-mediated cartilage protection.