Search results for: secret authentication

Authors: Martin Cai, Khadijah B. Hashim, Leng Leo, Edmund F. Tian

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Analysis of traditional Chinese medicinal (TCM) supplements has always been a laborious task, particularly in the case of multi‐ingredient formulations. Traditionally, herbal extracts are analysed using one or few markers compounds. In the recent years, however, pharmaceutical companies are introducing health supplements of TCM active ingredients to cater to the needs of consumers in the fast-paced society in this age. As such, new problems arise in the aspects of composition identification as well as quality analysis. In most cases of products or supplements formulated with multiple TCM herbs, the chemical composition, and nature of each raw material differs greatly from the others in the formulation. This results in a requirement for individual analytical processes in order to identify the marker compounds in the various botanicals. Thin-layer Chromatography (TLC) is a simple, cost effective, yet well-regarded method for the analysis of natural products, both as a Pharmacopeia-approved method for identification and authentication of herbs, and a great analytical tool for the discovery of chemical compositions in herbal extracts. Recent technical advances introduced High-Performance TLC (HPTLC) where, with the help of automated equipment and improvements on the chromatographic materials, both the quality and reproducibility are greatly improved, allowing for highly standardised analysis with greater details. Here we report an industrial consultancy project with ONI Global Pte Ltd for the analysis of LAC Liver Protector, a TCM formulation aimed at improving liver health. The aim of this study was to identify 4 key components of the supplement using HPTLC, following protocols derived from Chinese Pharmacopeia standards. By comparing the TLC profiles of the supplement to the extracts of the herbs reported in the label, this project proposes a simple and cost-effective analysis of the presence of the 4 marker compounds in the multi‐ingredient formulation by using 4 different HPTLC methods. With the increasing trend of small and medium-sized enterprises (SMEs) bringing natural products and health supplements into the market, it is crucial that the qualities of both raw materials and end products be well-assured for the protection of consumers. With the technology of HPTLC, science can be incorporated to help SMEs with their quality control, thereby ensuring product quality.

Keywords: traditional Chinese medicine supplement, high performance thin layer chromatography, active ingredients, product quality

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Authors: Emmanuel Ikechuckwu Onwubuya, Afees Adebayo Oladejo

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Background: The use of medicinal plants in the treatment of chronic diseases especially myocardial infarction, is gaining wide acceptance globally. Andrographis paniculata (Acanthaceae) is a medicinal plant commonly known as the king of bitters in Nigeria and has been acclaimed for several therapeutic activities. Materials and methods: This study investigated the cardio-protective effect of the leaf extract of A. paniculata in isoproterenol-induced myocardial infarction. Fresh green leaves of A paniculata were harvested from the Faculty of Agriculture farmland, Nnamdi Azikiwe University, Awka, Nigeria. Identification and authentication of the plant were carried out at the Department of Botany, Nnamdi Azikiwe University and a voucher specimen was deposited at the herbarium. The plant material was then shredded, air-dried under shade and pulverized. The fine powders obtained were weighed and extraction was done via a solvent combination of water and ethanol (3:7) for 72 hr via maceration. The filtrate gotten was evaporated to dryness to obtain the ethanol extract, which was used for further bioassay study. The bioactive constituents of the plant extract were quantitatively analyzed by Gas chromatography-mass spectrometry (GC-MS). The animals were administered the extract of A. paniculata orally for seven days at a divided dose of 100 mg/kg, 200 mg/kg and 400 mg/kg body weights. On the eighth day, myocardial infarction was induced through subcutaneous administration of isoproterenol at a dose of 150 mg/kg/day diluted in 2 ml of saline on two consecutive days. Subsequently, the blood pressures were monitored and blood was collected for bioassay studies. Results: The results of the study showed that the leaf extract of A. paniculata was rich in Dodecanoic acid (8.261%), 4-Dibenzofuranamine (6.03%), Cyclotrisiloxane (4.679 %). The findings also showed a significant decrease (p>0.05) in the Mean arterial blood pressure, heart rate, aspartate transaminase, alanine transaminase, creatinine kinase and lactate dehydrogenase activities of the treatment group compared with the untreated control group while the antioxidant (superoxide dismutase, catalase and glutathione) activities were significantly increased in the treatment group, compared with the untreated control group. Conclusion: The findings of this work have shown that the leaf of A. paniculata was rich in bioactive compounds, which could be synthesized to produce plant-based products to fight cardiovascular diseases, especially myocardial infarction.

Keywords: cardiovascular disease, myocardial infarction, medicinal plant, andrographis paniculata, isoproterenol

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Authors: Omwoyo Bosire Onyancha

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The Journal Citation Reports (JCR) of the Thomson Reuters has, for decades, provided the data for bibliometrically assessing the impact of journals. In their criticism of the journal impact factor (JIF), a number of scholars such as Priem, Taraborelli, Groth and Neylon (2010) observe that the “JIF is often incorrectly used to assess the impact of individual articles. It is troubling that the exact details of the JIF are a trade secret, and that significant gaming is relatively easy”. The emergence of alternative metrics (Altmetrics) has introduced another dimension of re-assessing how the impact of journals (and other units such as articles and even individual researchers) can be measured. Altmetrics is premised upon the fact that research is increasingly being disseminated through social network sites such as ResearchGate, Mendeley, Twitter, Facebook, LinkedIn, and ImpactStory, among others. This paper adopts informetrics (including altmetrics) techniques to report on the findings of a study conducted to investigate and compare the social media impact of 274 South Africa Post Secondary Education (SAPSE)-accredited journals, which are recognized and accredited by the Department of Higher Education and Training (DHET) of South Africa (SA). We used multiple sources to extract data for the study, namely Altmetric.com and the Thomson Reuters’ Journal Citation Reports. Data was analyzed in order to determine South African journals’ presence and impact on social media as well as contrast the social media impact with Thomson Reuters’ citation impact. The Spearman correlation test was performed to compare the journals’ social media impact and JCR citation impact. Preliminary findings reveal that a total of 6360 articles published in 96 South African journals have received some attention in social media; the most commonly used social media platform was Twitter, followed by Mendeley, Facebook, News outlets, and CiteULike; there were 29 SA journals covered in the JCR in 2008 and this number has grown to 53 journals in 2014; the journals indexed in the Thomson Reuters performed much better, in terms of their altmetrics, than those journals that are not indexed in Thomson Reuters databases; nevertheless, there was high correlation among journals that featured in both datasets; the journals with the highest scores in Altmetric.com included the South African Medical Journal, African Journal of Marine Science, and Transactions of the Royal Society of South Africa while the journals with high impact factors in JCR were South African Medical Journal, Onderstepoort: Journal of Veterinary Research, and Sahara: Journal of Social Aspects of HIV-AIDS; and that Twitter has emerged as a strong avenue of sharing and communicating research published in the South African journals. Implications of the results of the study for the dissemination of research conducted in South Africa are offered. Discussions based on the research findings as well as conclusions and recommendations are offered in the full text paper.

Keywords: altmetrics, citation impact, journal citation reports, journal impact factor, journals, research, scholarly publishing, social media impact, South Africa

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Authors: Abdul Basit Kiani

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Web development, the art of creating and maintaining websites, has witnessed remarkable advancements. The aim is to provide an overview of some of the cutting-edge developments in the field. Firstly, the rise of responsive web design has revolutionized user experiences across devices. With the increasing prevalence of smartphones and tablets, web developers have adapted to ensure seamless browsing experiences, regardless of screen size. This progress has greatly enhanced accessibility and usability, catering to the diverse needs of users worldwide. Additionally, the evolution of web frameworks and libraries has significantly streamlined the development process. Tools such as React, Angular, and Vue.js have empowered developers to build dynamic and interactive web applications with ease. These frameworks not only enhance efficiency but also bolster scalability, allowing for the creation of complex and feature-rich web solutions. Furthermore, the emergence of progressive web applications (PWAs) has bridged the gap between native mobile apps and web development. PWAs leverage modern web technologies to deliver app-like experiences, including offline functionality, push notifications, and seamless installation. This innovation has transformed the way users interact with websites, blurring the boundaries between traditional web and mobile applications. Moreover, the integration of artificial intelligence (AI) and machine learning (ML) has opened new horizons in web development. Chatbots, intelligent recommendation systems, and personalization algorithms have become integral components of modern websites. These AI-powered features enhance user engagement, provide personalized experiences, and streamline customer support processes, revolutionizing the way businesses interact with their audiences. Lastly, the emphasis on web security and privacy has been a pivotal area of progress. With the increasing incidents of cyber threats, web developers have implemented robust security measures to safeguard user data and ensure secure transactions. Innovations such as HTTPS protocol, two-factor authentication, and advanced encryption techniques have bolstered the overall security of web applications, fostering trust and confidence among users. Hence, recent progress in web development has propelled the industry forward, enabling developers to craft innovative and immersive digital experiences. From responsive design to AI integration and enhanced security, the landscape of web development continues to evolve, promising a future filled with endless possibilities.

Keywords: progressive web applications (PWAs), web security, machine learning (ML), web frameworks, advancement responsive web design

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Authors: Triveni Gogoi, Rima Chatterjee

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Shear wave velocity (Vs) estimation is an important approach in the seismic exploration and characterization of a hydrocarbon reservoir. There are varying methods for prediction of S-wave velocity, if recorded S-wave log is not available. But all the available methods for Vs prediction are empirical mathematical models. Shear wave velocity can be estimated using P-wave velocity by applying Castagna’s equation, which is the most common approach. The constants used in Castagna’s equation vary for different lithologies and geological set-ups. In this study, multiple regression analysis has been used for estimation of S-wave velocity. The EMERGE module from Hampson-Russel software has been used here for generation of S-wave log. Both single attribute and multi attributes analysis have been carried out for generation of synthetic S-wave log in Upper Assam basin. Upper Assam basin situated in North Eastern India is one of the most important petroleum provinces of India. The present study was carried out using four wells of the study area. Out of these wells, S-wave velocity was available for three wells. The main objective of the present study is a prediction of shear wave velocities for wells where S-wave velocity information is not available. The three wells having S-wave velocity were first used to test the reliability of the method and the generated S-wave log was compared with actual S-wave log. Single attribute analysis has been carried out for these three wells within the depth range 1700-2100m, which corresponds to Barail group of Oligocene age. The Barail Group is the main target zone in this study, which is the primary producing reservoir of the basin. A system generated list of attributes with varying degrees of correlation appeared and the attribute with the highest correlation was concerned for the single attribute analysis. Crossplot between the attributes shows the variation of points from line of best fit. The final result of the analysis was compared with the available S-wave log, which shows a good visual fit with a correlation of 72%. Next multi-attribute analysis has been carried out for the same data using all the wells within the same analysis window. A high correlation of 85% has been observed between the output log from the analysis and the recorded S-wave. The almost perfect fit between the synthetic S-wave and the recorded S-wave log validates the reliability of the method. For further authentication, the generated S-wave data from the wells have been tied to the seismic and correlated them. Synthetic share wave log has been generated for the well M2 where S-wave is not available and it shows a good correlation with the seismic. Neutron porosity, density, AI and P-wave velocity are proved to be the most significant variables in this statistical method for S-wave generation. Multilinear regression method thus can be considered as a reliable technique for generation of shear wave velocity log in this study.

Keywords: Castagna's equation, multi linear regression, multi attribute analysis, shear wave logs

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Authors: Jessie Rapoza, Petr Moroshkin, Jimmy Xu

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Chirality is omnipresent, in nature, in life, and in the field of physics. One intriguing example is the homochirality that has remained a great secret of life. Another is the pairs of mirror-image molecules – enantiomers. They are identical in atomic composition and therefore indistinguishable in the scalar physical properties. Yet, they can be either therapeutic or toxic, depending on their chirality. Recent studies suggest a potential link between abnormal levels of certain D-amino acids and some serious health impairments, including schizophrenia, amyotrophic lateral sclerosis, and potentially cancer. Although indistinguishable in their scalar properties, the chirality of a molecule reveals itself in interaction with the surrounding of a certain chirality, or more generally, a broken mirror-symmetry. In this work, we report on a system for chiral molecule detection, in which the mirror-symmetry is doubly broken, first by asymmetric structuring a nanopatterned plasmonic surface than by the incidence of circularly polarized light (CPL). In this system, the incident circularly-polarized light induces a surface plasmon polariton (SPP) wave, propagating along the asymmetric plasmonic surface. This SPP field itself is chiral, evanescently bound to a near-field zone on the surface (~10nm thick), but with an amplitude greatly intensified (by up to 104) over that of the incident light. It hence probes just the molecules on the surface instead of those in the volume. In coupling to molecules along its path on the surface, the chiral SPP wave favors one chirality over the other, allowing for chirality detection via the change in an optical rectification current measured at the edges of the sample. The asymmetrically structured surface converts the high-frequency electron plasmonic-oscillations in the SPP wave into a net DC drift current that can be measured at the edge of the sample via the mechanism of optical rectification. The measured results validate these design concepts and principles. The observed optical rectification current exhibits a clear differentiation between a pair of enantiomers. Experiments were performed by focusing a 1064nm CW laser light at the sample - a gold grating microchip submerged in an approximately 1.82M solution of either L-arabinose or D-arabinose and water. A measurement of the current output was then recorded under both rights and left circularly polarized lights. Measurements were recorded at various angles of incidence to optimize the coupling between the spin-momentums of the incident light and that of the SPP, that is, spin-momentum locking. In order to suppress the background, the values of the photocurrent for the right CPL are subtracted from those for the left CPL. Comparison between the two arabinose enantiomers reveals a preferential signal response of one enantiomer to left CPL and the other enantiomer to right CPL. In sum, this work reports on the first experimental evidence of the feasibility of chiral molecule detection via optical rectification in a metal meta-grating. This nanoscale interfaced electrical detection technology is advantageous over other detection methods due to its size, cost, ease of use, and integration ability with read-out electronic circuits for data processing and interpretation.

Keywords: Chirality, detection, molecule, spin

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Authors: Bander Alzahrani, Mohammed Alreshoodi

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Information-centric networking (ICN) using architectures such as the Publish-Subscribe Internet Technology (PURSUIT) has been proposed as a new networking model that aims at replacing the current used end-centric networking model of the Internet. This emerged model focuses on what is being exchanged rather than which network entities are exchanging information, which gives the control plane functions such as routing and host location the ability to be specified according to the content items. The forwarding plane of the PURSUIT ICN architecture uses a simple and light mechanism based on Bloom filter technologies to forward the packets. Although this forwarding scheme solve many problems of the today’s Internet such as the growth of the routing table and the scalability issues, it is vulnerable to brute force attacks which are starting point to distributed- denial-of-service (DDoS) attacks. In this work, we design and analyze a novel source-routing and information delivery technique that keeps the simplicity of using Bloom filter-based forwarding while being able to deter different attacks such as denial of service attacks at the ingress of the network. To achieve this, special forwarding nodes called Edge-FW are directly attached to end user nodes and used to perform a security test for malicious injected random packets at the ingress of the path to prevent any possible attack brute force attacks at early stage. In this technique, a core entity of the PURSUIT ICN architecture called topology manager, that is responsible for finding shortest path and creating a forwarding identifiers (FId), uses a cryptographically secure hash function to create a 64-bit hash, h, over the formed FId for authentication purpose to be included in the packet. Our proposal restricts the attacker from injecting packets carrying random FIds with a high amount of filling factor ρ, by optimizing and reducing the maximum allowed filling factor ρm in the network. We optimize the FId to the minimum possible filling factor where ρ ≤ ρm, while it supports longer delivery trees, so the network scalability is not affected by the chosen ρm. With this scheme, the filling factor of any legitimate FId never exceeds the ρm while the filling factor of illegitimate FIds cannot exceed the chosen small value of ρm. Therefore, injecting a packet containing an FId with a large value of filling factor, to achieve higher attack probability, is not possible anymore. The preliminary analysis of this proposal indicates that with the designed scheme, the forwarding function can detect and prevent malicious activities such DDoS attacks at early stage and with very high probability.

Keywords: forwarding identifier, filling factor, information centric network, topology manager

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Authors: Saeid H. Mobini, Monika Lulsdorf, Thomas D. Warkentin

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The length of the breeding cycle from seed to seed is a limiting factor in the development of improved homozygous lines for breeding or recombinant inbred lines (RILs) for genetic analysis. The objective of this research was to accelerate the production of field pea RILs through application of rapid generation technology (RGT). RGT is based on the principle of growing miniature plants in an artificial medium under controlled conditions, and allowing them to produce a few flowers which develop seeds that are harvested prior to normal seed maturity. We aimed to maintain population size and genetic diversity in regeneration cycles. The effects of flurprimidol (a gibberellin synthesis inhibitor), plant density, hydroponic system, scheduled fertilizer applications, artificial light spectrum, photoperiod, and light/dark temperature were evaluated in the development of RILs from a cross between cultivars CDC Dakota and CDC Amarillo. The main goal was to accelerate flowering while reducing maintenance and space costs. In addition, embryo rescue of immature seeds was tested for shortening the seed fill period. Data collected over seven generations included plant height, the percentage of plant survival, flowering rate, seed setting rate, the number of seeds per plant, and time from seed to seed. Applying 0.6 µM flurprimidol reduced the internode length. Plant height was decreased to approximately 32 cm allowing for higher plant density without a delay in flowering and seed setting rate. The three light systems (T5 fluorescent bulbs, LEDs, and High Pressure Sodium +Metal-halide lamp) evaluated did not differ significantly in terms of flowering time in field pea. Collectively, the combination of 0.6 µM flurprimidol, 217 plant. m-2, 20 h photoperiod, 21/16 oC light/dark temperature in a hydroponic system with vermiculite substrate, applying scheduled fertilizer application based on growth stage, and 500 µmole.m-2.s-1 light intensity using T5 bulbs resulted in 100% of plants flowering within 34 ± 3 days and 96.5% of plants completed seed setting in 68.2 ± 3.6 days, i.e., 30-45 days/generation faster than conventional single seed descent (SSD) methods. These regeneration cycles were reproducible consistently. Hence, RGT could double (5.3) generations per year, using 3% occupying space, compared to SSD (2-3 generation/year). Embryo rescue of immature seeds at 7-8 mm stage, using commercial fertilizer solutions (Holland’s Secret™) showed seed setting rate of 95%, while younger embryos had lower germination rate. Mature embryos had a seed setting rate of 96.5% without either hormones or sugar added. So, considering the higher cost of embryo rescue using a procedure which requires skill, additional materials, and expenses, it could be removed from RGT with a further cost saving, and the process could be stopped between generations if required.

Keywords: field pea, flowering, rapid regeneration, recombinant inbred lines, single seed descent

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Authors: Z. Giagkazoglou, D. Loukovitis, C. Gubili, A. Imsiridou

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Along with the increase in human population, demand for seafood has increased. Despite the strict labeling regulations that exist for most marketed species in the European Union, seafood substitution remains a persistent global issue. Food fraud occurs when food products are traded in a false or misleading way. When one species is traded under the name of another, it is considered mislabeling, and it can be intentional or unintentional. Crustaceans are one of the most regularly consumed seafood in Greece. Shrimps, prawns, lobsters, crayfish, and crabs are considered a delicacy and can be encountered in a variety of market presentations (fresh, frozen, pre-cooked, peeled, etc.). With most of the external traits removed, products as such are susceptible to species substitution. DNA barcoding has proven to be the most accurate method for the detection of fraudulent seafood products. This study aims, for the first time in Greece, to use the DNA barcoding methodology in order to investigate the labeling practices for crustacean products available in the market. A total of 100 tissue samples were collected from various retailers and markets across four Greek cities. In an effort to cover the highest range of products possible, different market presentations were targeted (fresh, frozen and cooked). Genomic DNA was extracted using the DNeasy Blood & Tissue Kit, according to the manufacturer's instructions. The mitochondrial gene selected as the target region of the analysis was the cytochrome c oxidase subunit I (COI). PCR products were purified and sequenced using an ABI 3500 Genetic Analyzer. Sequences were manually checked and edited using BioEdit software and compared against the ones available in GenBank and BOLD databases. Statistical analyses were conducted in R and PAST software. For most samples, COI amplification was successful, and species-level identification was possible. The preliminary results estimate moderate mislabeling rates (25%) in the identified samples. Mislabeling was most commonly detected in fresh products, with 50% of the samples in this category labeled incorrectly. Overall, the mislabeling rates detected by our study probably relate to some degree of unintentional misidentification and lack of knowledge surrounding the legal designations by both retailers and consumers. For some species of crustaceans (i.e., Squila mantis), the mislabeling appears to be also affected by the local labeling practices. Across Greece, S. mantis is sold in the market under two common names, but only one is recognized by the country's legislation, and therefore, any mislabeling is probably not profit-motivated. However, the substitution of the speckled shrimp (Metapenaus monoceros) for the distinct, giant river prawn (Macrobranchium rosenbergii) is a clear example of deliberate fraudulent substitution, aiming for profit. Until now, no scientific study investigating substitution and mislabeling rates in crustaceans has been conducted in Greece. For a better understanding of Greece's seafood market, similar DNA barcoding studies in other regions with increased touristic importance (e.g., the Greek islands) should be conducted. Regardless, the expansion of the list of species-specific designations for crustaceans in the country is advised.

Keywords: COI gene, food fraud, labelling control, molecular identification

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Authors: Hosein Maghsoudi

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Rheumatois arthritis (RA) is progressive inflammatory autoimmune diseases that primarily affect the joints, characterized by synovial hyperplasia and inflammatory cell infiltration, deformed and painful joints, which can lead tissue destruction, functional disability systemic complications, and early dead and socioeconomic costs. The cause of rheumatoid arthritis is unknown, but genetic and environmental factors are contributory and the prognosis is guarded. However, advances in understanding the pathogenesis of the disease have fostered the development of new therapeutics, with improved outcomes. The current treatment strategy, which reflects this progress, is to initiate aggressive therapy soon after diagnosis and to escalate the therapy, guided by an assessment of disease activity, in pursuit of clinical remission. The pathobiology of RA is multifaceted and involves T cells, B cells, fibroblast-like synoviocyte (FLSc) and the complex interaction of many pro-inflammatory cytokine. Novel biologic agents that target tumor necrosis or interlukin (IL)-1 and Il-6, in addition T- and B-cells inhibitors, have resulted in favorable clinical outcomes in patients with RA. Despite this, at least 30% of RA patients are résistance to available therapies, suggesting novel mediators should be identified that can target other disease-specific pathway or cell lineage. Among the inflammatory cell population that might participated in RA pathogenesis, FLSc are crucial in initiaing and driving RA in concert of cartilage and bone by secreting metalloproteinase (MMPs) into the synovial fluid and by direct invasion into extracellular matrix (ECM), further exacerbating joint damage. Invasion of fibroblast-like synoviocytes (FLSc) is critical in the pathogenesis of rheumatoid-arthritis. The metalloproteinase (MMPs) and activator of Toll-like receptor 4 (TLR4)/nuclear factor- κB pthway play a critical role in RA-FLS invasion induced by lipopolysaccharide (LPS). The present study aimed to explore the anti-invasion activity of Glycyrrhizic Acid as a pharmacologically safe phytochemical agent with potent anti-inflammatory properties on IL-1beta and TNF-alpha signalling pathways in Bovine fibroblast-like synoviocyte ex- vitro, on LPS-stimulated bovine FLS migration and invasion as well as MMP expression and explored the upstream signal transduction. Results showed that Glycyrrhizic Acid suppressed LPS-stimulated bovine FLS migration and invasion by inhibition MMP-9 expression and activity. In addition our results revealed that Glycyrrhizic Acid inhibited the transcriptional activity of MMP-9 by suppression the nbinding activity of NF- κB in the MMP-9 promoter pathway. The extract of licorice (Glycyrrhiza glabra L.) has been widely used for many centuries in the traditional Chinese medicine as native anti-allergic agent. Glycyrrhizin (GL), a triterpenoidsaponin, extracted from the roots of licorice is the most effective compound for inflammation and allergic diseases in human body. The biological and pharmacological studies revealed that GL possesses many pharmacological effects, such as anti-inflammatory, anti-viral and liver protective effects, and the biological effects, such as induction of cytokines (interferon-γ and IL-12), chemokines as well as extrathymic T and anti-type 2 T cells. GL is known in the traditional Chinese medicine for its anti-inflammatory effect, which is originally described by Finney in 1959. The mechanism of the GL-induced anti-inflammatory effect is based on different pathways of the GL-induced selective inhibition of the prostaglandin E2 production, the CK-II- mediated activation of both GL-binding lipoxygenas (gbLOX; 17) and PLA2, an anti-thrombin action of GL and production of the reactive oxygen species (ROS; GL exerts liver protection properties by inhibiting PLA2 or by the hydroxyl radical trapping action, leading to the lowering of serum alanine and aspartate transaminase levels. The present study was undertaken to examine the possible mechanism of anti-inflammatory properties GL on IL-1beta and TNF-alpha signalling pathways in bovine fibroblast-like synoviocyte ex-vivo, on LPS-stimulated bovine FLS migration and invasion as well as MMP expression and explored the upstream signal transduction. Our results clearly showed that treatment of bovine fibroblast-like synoviocyte with GL suppressed LPS-induced cell migration and invasion. Furthermore, it revealed that GL inhibited the transcription activity of MMP-9 by suppressing the binding activity of NF-κB in the MM-9 promoter. MMP-9 is an important ECM-degrading enzyme and overexpression of MMPs in important of RA-FLSs. LPS can stimulate bovine FLS to secret MMPs, and this induction is regulated at the transcription and translational levels. In this study, LPS treatment of bovine FLS caused an increase in MMP-2 and MMP-9 levels. The increase in MMP-9 expression and secretion was inhibited by ex- vitro. Furthermore, these effects were mimicked by MMP-9 siRNA. These result therefore indicate the the inhibition of LPS-induced bovine FLS invasion by GL occurs primarily by inhibiting MMP-9 expression and activity. Next we analyzed the functional significance of NF-κB transcription of MMP-9 activation in Bovine FLSs. Results from EMSA showed that GL suppressed LPS-induced NF-κB binding to the MMP-9 promotor, as NF-κB regulates transcriptional activation of multiple inflammatory cytokines, we predicted that GL might target NF-κB to suppress MMP-9 transcription by LPS. Myeloid differentiation-factor 88 (MyD88) and TIR-domain containing adaptor protein (TIRAP) are critical proteins in the LPS-induced NF-κB and apoptotic signaling pathways, GL inhibited the expression of TLR4 and MYD88. These results demonstrated that GL suppress LPS-induced MMP-9 expression through the inhibition of the induced TLR4/NFκB signaling pathway. Taken together, our results provide evidence that GL exerts anti-inflammatory effects by inhibition LPS-induced bovine FLSs migration and invasion, and the mechanisms may involve the suppression of TLR4/NFκB –mediated MMP-9 expression. Although further work is needed to clarify the complicated mechanism of GL-induced anti-invasion of bovine FLSs, GL might be used as a further anti-invasion drug with therapeutic efficacy in the treatment of immune-mediated inflammatory disease such as RA.

Keywords: glycyrrhizic acid, bovine fibroblast-like synoviocyte, tlr4/nf-κb, metalloproteinase-9

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