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Rotundic acid (RA), an ursane-type pentacyclic triterpene acid isolated from the dried barks of Ilex rotunda Thunb. (Aquifoliaceae), possesses diverse bioactivities. To further study its pharmacokinetics, a simple and sensitive liquid chromatography with triple quadrupole mass spectrometry (LC-QqQ-MS/MS) method was developed and validated to quantify RA concentration in rat plasma and tissue using etofesalamide as an internal standard (IS). Plasma and tissue samples were subjected to one-step protein precipitation. Chromatographic separation was achieved on a ZORBAX Eclipse XDB-C18 column (4.6 mm × 50 mm, 5 μm) under gradient conditions with eluents of methanol:acetonitrile (1:1, V/V) and 5 mM ammonium formate:methanol (9:1, V/V) at 0.5 mL/min. Multiple reaction monitoring transitions were performed at m/z 487.30 → 437.30 for RA and m/z 256.10 → 227.10 for IS in the negative mode. The developed LC-QqQ-MS/MS method exhibited good linearity (2–500 ng/mL) and was fully validated in accordance with U.S. Food and Drug Administration bioanalytical guidelines. Dose proportionality and bioavailability in rats were determined by comparing pharmacokinetic data after single oral (10, 20, and 40 mg/kg) and intravenous (10 mg/kg) administration of RA. Tissue distribution was studied following oral administration at 20 mg/kg. The results showed that the absolute bioavailability of RA after administration at different doses ranged from 16.1% to 19.4%. RA was rapidly absorbed in a dose-dependent manner and highly distributed in the liver. In conclusion, this study is the first to systematically elucidate the absorption and distribution characteristics of RA in rats, which could provide additional information for further development and evaluation of RA in drug metabolism and pharmacokinetic studies.

Background Head and neck squamous cell carcinoma (HNSCC) remains an unmet medical challenge. Metabolic reprogramming is a hallmark of diverse cancers, including HNSCC. Methods We investigated the metabolic profile in HNSCC by using The Cancer Genome Atlas (TCGA) (n = 481) and Gene Expression Omnibus (GEO) (n = 97) databases. The metabolic stratification of HNSCC samples was identified by using unsupervised k-means clustering. We analyzed the correlations of the metabolic subtypes in HNSCC with featured genomic alterations and known HNSCC subtypes. We further validated the metabolism-related subtypes based on features of ENO1, PFKFB3, NSDHL and SQLE expression in HNSCC by Immunohistochemistry. In addition, genomic characteristics of tumor metabolism that varied among different cancer types were confirmed. Results Based on the median expression of coexpressed cholesterogenic and glycolytic genes, HNSCC subtypes were identified, including glycolytic, cholesterogenic, quiescent and mixed subtypes. The quiescent subtype was associated with the longest survival and was distributed in stage I and G1 HNSCC. Mutation analysis of HNSCC genes indicated that TP53 has the highest mutation frequency. The CDKN2A mutation frequency has the most significant differences amongst these four subtypes. There is good overlap between our metabolic subtypes and the HNSCC subtype. Conclusion The four metabolic subtypes were successfully determined in HNSCC. Compared to the quiescent subtype, glycolytic, cholesterogenic and mixed subtypes had significantly worse outcome, which might offer guidelines for developing a novel treatment strategy for HNSCC.

This article focuses on the different administration methods of lipid-based nanoparticles in cancer treatment.

Macrophage-mediated inflammation compromises bone repair in diabetic patients. Electrical signaling cues are known to regulate macrophage functions. However, the biological effects of electrical microenvironment from charged biomaterials on the immune response for regulating osteogenesis under diabetic conditions remain to be elucidated. Herein the endogeneous electrical microenvironment of native bone tissue was recapitulated by fabricating a ferroelectric BaTiO3/poly (vinylidene fluoridetrifluoroethylene) (BTO/P(VDF-TrFE)) nanocomposite membrane. In vitro, the polarized BaTiO3/poly (vinylidene fluoridetrifluoroethylene) (BTO/P(VDF-TrFE)) nanocomposite membranes inhibited high glucose-induced M1-type inflammation, by effecting changes in cell morphology, M1 marker expression and pro-inflammatory cytokine secretion in macrophages. This led to enhanced osteogenic differentiation of human bone marrow mesenchymal stem cells (BM-MSCs). In vivo, the biomimetic electrical microenvironment recapitulated by the polarized nanocomposite membranes switched macrophage phenotype from the pro-inflammatory (M1) into the pro-healing (M2) phenotype, which in turn enhanced bone regeneration in rats with type 2 diabetes mellitus. Mechanistic studies revealed that the biomimetic electrical microenvironment attenuated pro-inflammatory M1 macrophage polarization under hyperglycemic conditions by suppressing expression of AKT2 and IRF5 within the PI3K-AKT signaling pathway, thereby inducing favorable osteo-immunomodulatory effects. Our study thus provides fundamental insights into the biological effects of restoring the electrical microenvironment conducive for osteogenesis under DM conditions, and offers an effective strategy to design functionalized biomaterials for bone regeneration therapy in diabetic patients., Graphical abstract Within the restored electrical microenvironment mediated by polarized ferroelectric nanocomposite membrane, macrophages exhibit suppressed M1 polarization and enhanced M2 polarization. The restored electrical microenvironment downregulates AKT2-IRF5 signaling that induces macrophages to the M2 phenotype, which indicates a favorable osteo-immunomodulatory effects. Hence mimicking the endogeneous electrical microenvironment promotes osteogenic differentiation of BM-MSCs mediated by macrophages, thereby improving bone regeneration.Image 1, Highlights • Electrical microenvironment recapitulated by BTO membranes switched pro-inflammatory M1 into pro-healing M2 phenotype. • The macrophage phenotype transformation from M1 to M2 promotes bone regeneration in rats with type 2 diabetes mellitus. • Restored electrical microenvironment attenuated M1 macrophage polarization via downregulation of AKT2-IRF5.

Successful implant-bone integration remains a formidable challenge in osteoporotic patients, because of excessive inflammatory reactions and osteoclastogenesis around the peri-implant bone tissue. This study designed biomimetic micro/sub-micro hierarchical surfaces on titanium implants based on natural bone hierarchical structures to mitigate macrophage-mediated inflammatory reactions, osteoclastogenesis, and osteogenesis

As one of the world’s five terminally ills, tumours can cause important genetic dysfunction. However, some current medicines for tumours usually have strong toxic side effects and are prone to drug resistance. Studies have found that farnesyltransferase inhibitors (FTIs) extracted from natural materials have a good inhibiting ability on tumours with fewer side effects. This article describes several FTIs extracted from natural materials and clarifies the current research progress, which provides a new choice for the treatment of tumours., Executive summary Farnesyltransferase (FTase) When Ras gene is activated, it becomes an oncogene with oncogenic activity. Ras protein plays a crucial role in cancer cells. FTase is the first step to activate Ras protein. FTIs not only have anti-tumour effects, but also makes great contribution to the treatment of plasmodium falciparum, parasitic diseases and progeria. There are still some problems with FTIs, but natural products FTIs are worthy of further research as a new class of low-toxic, safe and effective anticancer drugs. Summarised natural product-derived FTase inhibitors This review summarised several FTIs extracted from natural materials and clarifies their anti-tumour activity (IC50 value) and structure, providing a reference for further research on tumour therapy.

Purpose: The combination of a bone graft with a barrier membrane is the classic method for guided bone regeneration (GBR) treatment. However, the insufficient osteoinductivity of currently-available barrier membranes and the consequent limited bone regeneration often inhibit the efficacy of bone repair. In this study, we utilized the piezoelectric properties of biomaterials to enhance the osteoinductivity of barrier membranes. Methods: A flexible nanocomposite membrane mimicking the piezoelectric properties of natural bone was utilized as the barrier membrane. Its therapeutic efficacy in repairing critical-sized rabbit mandible defects in combination with xenogenic grafts of deproteinized bovine bone (DBB) was explored. The nanocomposite membranes were fabricated with a homogeneous distribution of piezoelectric BaTiO3 nanoparticles (BTO NPs) embedded within a poly(vinylidene fluoridetrifluoroethylene) (P(VDF-TrFE)) matrix. Results: The piezoelectric coefficient of the polarized nanocomposite membranes was close to that of human bone. The piezoelectric coefficient of the polarized nanocomposite membranes was highly stable, with more than 90% of the original piezoelectric coefficient (d33) remaining up to 28 days after immersion in culture medium. Compared with commercially-available polytetrafluoroethylene (PTFE) membranes, the polarized BTO/P(VDF-TrFE) nanocomposite membranes exhibited higher osteoinductivity (assessed by immunofluorescence staining for runt-related transcription factor 2 (RUNX-2) expression) and induced significantly earlier neovascularization and complete mature bone-structure formation within the rabbit mandible critical-sized defects after implantation with DBB Bio-Oss® granules. Conclusion: Our findings thus demonstrated that the piezoelectric BTO/P(VDF-TrFE) nanocomposite membranes might be suitable for enhancing the clinical efficacy of GBR.

In the last decade, numerous circRNAs were discovered by virtue of the RNA-Seq technique. With the deepening of experimental research, circRNAs have brought to light the key biological functions and progression of human diseases. CircRNA ITCH has been demonstrated to be a tumor suppressor in numerous cancers, and recently it was found to play an important role in bone diseases, diabetes mellitus, and cardiovascular diseases. However, the functions of circ-ITCH have not been completely understood. In this review, we comprehensively provide a conceptual framework to elucidate circ-ITCH biological functions of cell proliferation, apoptosis and differentiation, and the pathological mechanisms of inflammation, drug resistance/toxicity, and tumorigenesis. Finally, we summarize its clinical applications in various diseases. This research aimed at clarifying the role of circ-ITCH, which could be a promising therapeutic target.

Water-based antimicrobial agents, used in environmentally friendly applications, are widely used in wood protection industries. Furthermore, nanomaterials as antimicrobial agents, because of their biocidal component, huge specific surface area, and unique nanoscale effect, have attracted attention in the field of biodurability. We employed aqueous dispersed nano-silver with a diameter of 10 nm~20 nm to treat poplar wood and evaluated its leaching resistance and anti-mold effect on the wood surface. The results revealed that the higher the retention of the nano-silver, the stronger the protection efficiency of the wood surface against three molds (Aspergillus niger V. Tiegh, Penicillium citrinum Thom, and Trichoderma viride Pers. ex Fr); and the leachability of the nano-silver presented a slowly growing trend with the increase in the retention. When the wood surface attained a silver retention of 0.324 g·m−2, its anti-mold efficiency against Aspergillus niger V. Tiegh, Penicillium citrinum Thom, and Trichoderma viride Pers. ex Fr reached 80, 75, and 80%, respectively, which achieved or even exceeded the required standard value of effective mold inhibition (75%). Notably, the nano-silver leaching rate at this retention attained merely 4.75 %. The nanoparticle, well distributed on a wood surface, may promote sufficient contact with fungi as well as strong interaction with wood cell wall components, which probably contributed to the effective anti-mold efficiency and the leaching resistance. This study provided positive evidence for the anti-mold effect of nano-silver on wood surface.

Cancer is a long-known incurable disease, and the medical use of cisplatin has been a significant discovery. However, the side-effects of cisplatin necessitate the development of new and improved drug. Therefore, in this study, we focused on the photoactivatable Pt(IV) compounds Pt[(X1)(X2)(Y1)(Y2)(N3)2], which have a completely novel mechanism of action. Pt(IV) can efficiently overcome the side-effects of cisplatin and other drugs. Here, we have demonstrated, summarized and discussed the effects and mechanism of these compounds. Compared to the relevant articles in the literature, we have provided a more detailed introduction and a made comprehensive classification of these compounds. We believe that our results can effectively provide a reference for the development of these drugs.

Petroleum-based plastics, such as PP, PE, PVC, etc., have become an important source of environmental pollution due to their hard degradation, posing a serious threat to the human health. Isolating nanocellulose from abundant biomass waste resources and further integrating the nanocellulose into hydrophobic transparent film (i.e., nanopaper), to replace the traditional nondegradable plastic film, is of great significance for solving the problem of environmental pollution and achieving sustainable development of society. This study respectively extracted nanocellulose from the branches of Amorpha fruticosa Linn., wheat straw, and poplar residues via combined mechanical treatments of grinding and high-pressure homogenization. Among them, the nanocellulose derived from the Amorpha fruticosa has a finer structure, with diameter of about 10 nm and an aspect ratio of more than 500. With the nanocellulose as building block, we constructed hydrophilic nanopaper with high light transmittance (up to 90%) and high mechanical strength (tensile strength up to 110 MPa). After further hybridization by incorporating nano-silica into the nanopaper, followed by hydrophobic treatment, we built hydrophobic nanopaper with transmittance over 82% and a water contact angle of about 102&deg, that could potentially replace transparent plastic film and has wide applications in food packaging, agricultural film, electronic device, and other fields.

As a lightweight and highly insulating composite material, epoxy resin syntactic foam is increasingly widely used for insulation filling in electrical equipment. To avoid core burning and cracking, which are prone to occur during the casting process, the epoxy resin-based syntactic foam insulation materials with high thermal conductivity and low coefficient of thermal expansion are required for composite insulation equipment. The review is divided into three sections concentrating on the two main aspects of modifying the thermal properties of syntactic foam. The mechanism and models, from the aspects of thermal conductivity and coefficient of thermal expansion, are presented in the first part. The second part aims to better understand the methods for modifying the thermal properties of syntactic foam by adding functional fillers, including the addition of thermally conductive particles, hollow glass microspheres, negative thermal expansion filler and fibers, etc. The third part concludes by describing the existing challenges in this research field and expanding the applicable areas of epoxy resin-based syntactic foam insulation materials, especially cross-arm composite insulation.

It has been proven that the surface topographic cues of fiber arrangement can induce osteogenic differentiation of mesenchymal stem cells. However, this effect alone is weak and insufficient to meet the needs of regenerative medicine. In this work, electroactivity concept was introduced to enhance the osteoinductivity of fibrous scaffolds. The randomly oriented and aligned electroactive fibrous scaffolds of poly-(l-lactic acid) (PLLA) with incorporation of ferroelectric ceramic BaTiO3 (BTO) nanoparticles (NPs) were fabricated by electrospinning. Physicochemical properties, including fiber morphology, microstructure, composition, thermal stability, surface roughness, and surface wettability, of these fibrous scaffolds were studied. The dielectric properties of the scaffolds were evaluated. The results showed that the randomly oriented BTO/PLLA composite fibrous scaffolds had the highest dielectric permittivity of 1.19, which is of the same order of magnitude as the natural bone. The combined effects of fiber orientation and electrical activity on the osteogenic responses of bone marrow mesenchymal stem cells (BM-MSCs) were specifically investigated. Randomly oriented composite fibrous scaffolds significantly promoted polygonal spreading and encouraged early osteogenic differentiation in BM-MSCs, whereas aligned composite fibrous scaffolds promoted cell elongation and discouraged osteogenic differentiation. These results evidenced that randomly fiber orientation and biomimetic electric activity have combining effects on osteogenic differentiation of BM-MSCs. Our findings indicate that coupling effects of multi-physical properties should be paid more attention to mimic the microenvironment for enhancing osteogenic differentiation of BM-MSCs.

With the enhancement of people&rsquo, s environmental awareness, waterborne polyurethane (PU) paint&mdash, with its advantages of low release of volatile organic compounds (VOCs), low temperature flexibility, acid and alkali resistance, excellent solvent resistance and superior weather resistance&mdash, has made its application for wood furniture favored by the industry. However, due to its lower solid content and weak intermolecular force, the mechanical properties of waterborne PU paint are normally less than those of the traditional solvent-based polyurethane paint, which has become the key bottleneck restricting its wide applications. To this end, this study explores nanocellulose derived from biomass resources by the 2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO) oxidation method to reinforce and thus improve the mechanical properties of waterborne PU paint. Two methods of adding nanocellulose to waterborne PU&mdash, chemical addition and physical blending&mdash, are explored. Results show that, compared to the physical blending method, the chemical grafting method at 0.1 wt% nanocellulose addition results in the maximum improvement of the comprehensive properties of the PU coating. With this method, the tensile strength, elongation at break, hardness and abrasion resistance of the waterborne PU paint increase by up to 58.7%, ~55%, 6.9% and 3.45%, respectively, compared to the control PU, while the glossiness and surface drying time were hardly affected. Such exploration provides an effective way for wide applications of water PU in the wood industry and nanocellulose in waterborne wood coating.

Early osseointegration plays a crucial role in determining the therapeutic efficacy of orthopedic implants. Some factors responsible for early osseointegration, such as inherent mechanical properties and surface topology of implants, are well-characterized. However, the synergistic effects of elastic modulus and surface topology of implants on the osteogenic differentiation of stem cells and early osseointegration have not been thoroughly investigated. In this study, the titanium (Ti) and β-titanium alloy Ti–24Nb–4Zr–8Sn (TNZS) were used to evaluate the synergistic effects of elastic modulus and surface topology on the biological performance of these materials in vitro and in vivo. Scanning electron microscopy imaging confirmed the presence of a micro-scale porous oxide layer on the Ti and TNZS surfaces upon treatment with microarc oxidation (MAO), which resulted in increased surface roughness, enhanced surface wettability and favourable mechanical properties. As compared with Ti-MAO, the TNZS-MAO samples with lower elastic modulus displayed increased cell attachment, alkaline phosphatase activity, collagen secretion, osteogenic marker expression, and mineralization of rat bone marrow mesenchymal stem cells. Upon implantation in rat femoral condylar defects, an inherently low elastic modulus could cooperatively accelerate early osseointegration and maturation of trabecular bone after 4 weeks implantation with the MAO modified surface. These results demonstrated that elastic modulus and the surface micro-scale topographical structure of Ti alloy implants have a synergistic effect on their osseointegration.

Wood-based panels covered by melamine-impregnated paper are widely used in floors and furniture, due to its good surface texture, hardness, wear resistance, and waterproof function. However, there are still some problems, such as formaldehyde release from the impregnated resin, non-wood touch, and complex preparation processes. Therefore, this study designed glycidyl methacrylate (GMA) and ethyleneglycol dimethacrylate (EGDMA), combined with maleic anhydride (MAN) as a reactive catalyst, to build an active monomers system. It was first impregnated into poplar veneers, and then in-situ polymerized within the veneer using a hot pressing process, which realized the gluing of the veneer onto the wood-based panel substrate, synchronously. Such treatment aims to obtain wood-based panel composites decorated by the modified veneer, with real solid wood touch feeling, satisfied surface properties, and environment friendly glue bonding. The results indicated that the optimized reaction ratio of the active monomers (GMA:EGDMA) was 2:1 (molar ratio), and the maleic anhydride addition accounted for 6 wt.% of the total monomers. Under the optimized hot pressing condition, the modified veneer closely bonded to the wood-based panel substrate without obvious interfacial gaps. The hardness, abrasion resistance, modulus of rupture, and water resistance of the composites were significantly improved. Such results indicate that the treatment realized the perfect merging of solid wood touch feeling, environment friendly feature, and excellent properties of the composite. It was highly expected to replace the traditional melamine-impregnated paper to decorate wood-based panels, and could be potentially applied as surface decorating materials in wide areas of desktop, floor, cupboard, wardrobe, and so on.

Wood is susceptible to swelling deformation and decay fungi due to moisture adsorption that originates from the dynamic nanopores of the cell wall and the abundant hydroxyl groups in wood components. This study employed as a modifier maleic anhydride (MAn), with the help of acetone as solvent, to diffuse into the wood cell wall, bulk nanopores, and further chemically bond to the hydroxyl groups of wood components, reducing the numbers of free hydroxyl groups and weakening the diffusion of water molecules into the wood cell wall. The derived MAn-bulked wood, compared to the control wood, presented a reduction in water absorptivity (RWA) of ~23% as well as an anti-swelling efficiency (ASE) of ~39% after immersion in water for 228 h, and showed an improvement in decay resistance of 81.42% against white-rot fungus and 69.79% against brown-rot fungus, respectively. The method of combined cell wall bulking and hydroxyl group bonding could effectively improve the dimensional stability and decay resistance with lower doses of modifier, providing a new strategy for wood durability improvement.

Immune responses can determine thein vivofate of implanted materials. The strategy for developing implants has shifted towards using materials with immunomodulatory activity. However, the immunoregulatory effect of hydrophilicity of titanium surface on the macrophage behavior and its underlying mechanism remain poorly understood. Here, the Ti surface hydrophilicity-dependent behavior of murine RAW264.7 macrophages was investigatedin vitro. Two laboratory models with significantly different surface hydrophilicity and similar roughness were established with Ti-polished and Ti-H2O2surfaces. The results of cell morphology observation showed that the Ti-H2O2surface yielded enhanced cell adhesion and less multinucleated cell formation. CCK-8 assay indicated that the growth rate of macrophage on Ti-H2O2surface is higher than that of Ti-polished. ELISA assay result revealed lower level of proinflammatory factor TNF-αand higher level of anti-inflammatory factor IL-10 on the Ti-H2O2surface compared to Ti-polished. Subsequently, immunofluorescence and western blotting analysis showed that activation of the NF-κB-TNF-αpathway might be involved in the modulation of the immune response by surface hydrophilicity. Together, these results suggested that relative high hydrophilic Ti surface might attenuate the immune response of macrophage by activating NF-κB signaling. These findings could provide new insights into designing implant devices for orthopedic applications.

The design and fabrication of biodegradable barrier membranes with satisfactory structure and composition remain a considerable challenge for periodontal tissue regeneration. We have developed a biomimetic nanofibrous membrane made from a composite of gelatin andβ-tricalcium phosphate (β-TCP). We previously confirmed thein vitrobiological performance of the membrane material, but the efficacy of the membranes in promoting bone repairin situhas not yet been examined. Gelatin/β-TCP composite nanofibers were fabricated by incorporation of 20 wt.%β-TCP nanoparticles into electrospun gelatin nanofibers. Electron microscopy showed that the composite membranes presented a nonwoven structure with an interconnected porous network and had a rough surface due to theβ-TCP nanoparticles, which were distributed widely and uniformly throughout the gelatin-fiber matrix. The repair efficacy of rabbit mandible defects implanted with bone substitute (Bio-Oss) and covered with the gelatin/β-TCP composite nanofibrous membrane was evaluated in comparison with pure gelatin nanofibrous membrane. Gross observation, histological examination, and immunohistochemical analysis showed that new bone formation and defect closure were significantly enhanced by the composite membranes compared to the pure gelatin ones. From these results, we conclude that nanofibrous gelatin/β-TCP composite membranes could serve as effective barrier membranes for guided tissue regeneration.

Staphylococcus aureus is a significant Gram-positive bacterium that is associated with a broad spectrum of diseases ranging from minor skin infections to lethal pneumonia, endocarditis, and toxinoses. α-Hemolysin is one of the most important exotoxins that contribute to the pathogenesis of S. aureus infections. Liquiritigenin is one of the most significant active components in licorice. In this study, hemolysis, western blot, and real-time reverse transcription-PCR assays were performed to investigate the impact of liquiritigenin on the production of S. aureus α-hemolysin. The results showed that low concentrations of liquiritigenin remarkably decreased S. aureus α-hemolysin production in a dose-dependent manner. Using live/dead cell staining and lactate dehydrogenase assays, we found that liquiritigenin could protect human lung cells (A549) from α-hemolysin-mediated injury. The data indicated that this compound could potentially be useful in developing drugs aiming at staphylococcal α-hemolysin.

In this study, hemolysis, Western blot, and real-time RT-PCR assays were performed to evaluate silibinin's activity against S. aureus α-toxin secretion. In addition, live/dead cell staining and lactate dehydrogenase activity assays were introduced to examine the influence of silibinin on α-toxin-induced cell injury in human alveolar epithelial cells. Furthermore, we tested the influence of silibinin on S. aureus pneumonia in a mouse model. We show that silibinin inhibits the expression of α-toxin in a dose-dependent manner and alleviates α-toxin-induced lung cell injury. The IC50 of silibinin that inhibits the hemolytic activity of S. aureus was 14.33 µg/mL for strain 8325-4. Moreover, this compound provides effective protection on the lung injury of staphylococcal pneumonia.

Staphylococcus aureus is a common human pathogenic bacteria that can cause serious infections, including lethal staphylococcal pneumonia. The development of antimicrobial resistance has limited treatment options for this pathogen; consequently, novel antibiotics and strategies are urgently desired to combat these infections. In recent years, virulence factors secreted by pathogenic microorganisms have been developed as targets for drug discovery. Alpha-hemolysin, a pore-forming cytotoxin that is secreted by most S. aureus strains, is essential for the pathogenesis of S. aureus pneumonia. In this study, we report that apigenin, a compound extracted from parsley that has no antimicrobial activity vs. S. aureus in vitro, can remarkably decrease the production of α-hemolysin at low concentrations. When added to the A549 cells and S. aureus co-culture system, apigenin protected A549 cells from α-hemolysin-mediated injury. Furthermore, in vivo tests indicated that apigenin alleviated injury of the lung tissue and decreased cytokine levels in the bronchoalveolar lavage fluid in the mouse model of S. aureus pneumonia.

An unmet need in engineered bone regeneration is to develop scaffolds capable of manipulating stem cells osteogenesis. Graphene oxide (GO) has been widely used as a biomaterial for various biomedical applications. However, it remains challenging to functionalize GO as ideal platform for specifically directing stem cell osteogenesis. Herein, we report facile functionalization of GO with dopamine and subsequent bioactive glass (BG) to enhance stem cell adhesion, spreading, and osteogenic differentiation. On the basis of graphene, we obtained dopamine functionalized graphene oxide/bioactive glass (DGO/BG) hybrid scaffolds containing different content of DGO by loading BG nanoparticles on graphene oxide surface using sol-gel method. To enhance the dispersion stability and facilitate subsequent nucleation of BG in GO, firstly, dopamine (DA) was used to modify GO. Then, the modified GO was functionalized with bioactive glass (BG) using sol-gel method. The adhesion, spreading, and osteoinductive effects of DGO/BG scaffold on rat bone marrow mesenchymal stem cells (rBMSCs) were evaluated. DGO/BG hybrid scaffolds with different content of DGO could influence rBMSCs’ behavior. The highest expression level of osteogenic markers suggests that the DGO/BG hybrid scaffolds have great potential or elicit desired bone reparative outcome.

In this study, we aimed to evaluate the effect of α- cyperone on S. aureus. We used a hemolysin test to examine the hemolytic activity in supernatants of S. aureus cultured with increasing concentrations of α- cyperone. In addition, we evaluated the production of α- hemolysin (Hla) by Western blotting. Real-time RT-PCR was performed to test the expression of hla (the gene encoding Hla) and agr (accessory gene regulator). Furthermore, we investigated the protective effect of α- cyperone on Hla-induced injury of A549 lung cells by live/ dead and cytotoxicity assays. We showed that in the presence of subinhibitory concentrations of α-cyperone, Hla production was markedly inhibited. Moreover, α- cyperone protected lung cells from Hla-induced injury. These findings indicate that α-cyperone is a promising inhibitor of Hla production by S. aureus and protects lung cells from this bacterium. Thus, α-cyperone may provide the basis for a new strategy to combat S. aureus pneumonia.

In the present study, the antimicrobial activity of glycyrrhetinic acid (GA) against Staphylococcus aureus, and its influence on the production of S. aureus alpha-haemolysin (Hla) were investigated, along with the in vivo activity of GA against S. aureus-induced pneumonia. GA could not inhibit the growth of S. aureus, but the secretion of Hla by S. aureus was significantly inhibited by low concentrations of GA in a dose-dependent manner. Furthermore, in vivo data show that GA provides protection against staphylococcal pneumonia in a murine model system.

Aim: To elucidate the effect of chrysin on α-haemolysin production by Staphylococcus aureus and protection against pneumonia in a murine model. Methods and Results: Haemolysis, Western blot and real-time RT-PCR assays were performed to evaluate the effect of chrysin on α-haemolysin secretion by Staph. aureus. The efficacy of chrysin against human alveolar epithelial cell injury by α-haemolysin was tested using live/dead staining or by measuring lactate dehydrogenase activity. Furthermore, we determined the protective effect of chrysin against Staph. aureus pneumonia through histopathology experiments in a mouse model. The production of α-haemolysin by Staph. aureus was inhibited when presented with an increasing subinhibitory concentration of chrysin in vitro. Consistent with this result, chrysin prevented α-haemolysin-mediated cell injury and protected mice from Staph. aureus pneumonia. Conclusions: Chrysin is a potent inhibitor of α-haemolysin expression by Staph. aureus, and it conferred a significant degree of protection against Staph. aureus pneumonia. Significance and Impact of Study: The chrysin-mediated inhibition of α-haemolysin production and protection against Staph. aureus pneumonia may offer a new strategy in combating pathogen infections.

Staphylococcus aureus is a versatile pathogen that can cause life-threatening infections. The growing emergence of methicillin-resistant S. aureus strains and a decrease in the discovery of new antibiotics warrant the search for new therapeutic targets to combat infections. Staphylococcus aureus produces many extracellular virulence factors that contribute to its pathogenicity. Therefore, targeting bacterial virulence as an alternative strategy to the development of new antimicrobials has gained great interest. α-Toxin is a 33.2-kDa, water-soluble, pore-forming toxin that is secreted by most S. aureus strains. α-Toxin is essential for the pathogenesis of pneumonia, as strains lacking α-toxin display a profound defect in virulence. In this report, we demonstrate that isoalantolactone (IAL), a naturally occurring compound found in Inula helenium (Compositae), has no anti-S. aureus activity as per MIC evaluation in vitro. However, IAL can markedly inhibit the expression of α-toxin in S. aureus at very low concentrations. Furthermore, the in vivo data indicate that treatment with IAL protects mice from S. aureus pneumonia.

Nano-β-tricalcium phosphate/collagen (n-β-TCP/Col) is considered with good osteoconductivity. However, the therapeutic effectiveness of n-β-TCP/Col scaffolds in combination with autologous bone marrow stromal cells (BMSCs) remains to be elucidated for the repair of critical-sized bone defects. In this study, we found that n-β-TCP/Col scaffolds exhibited high biocompatibilityin vitro. The introduction of BMSCs expandedin vitroto the scaffolds dramatically enhanced their efficiency to restore critical-sized bone defects, especially during the initial stage after implantation. Collectively, these results suggest that autologous BMSCs in n-β-TCP/Col scaffolds have the potential to be applied in bone tissue engineering.

The protein α-hemolysin (α-HL) is a self-assembling exotoxin that binds to the membrane of a susceptible host cell. In this paper, experimental studies show that cyrtominetin (CTM) can inhibit the hemolytic activity of α-HL. To understand how CTM can affect hemolytic activity, molecular dynamics simulations were carried out for α-HL-CTM complex and these results were compared with the crystal structure of monomeric α-HL. With this approach, the analysis revealed that the inhibition of CTM involves CTM directly binding to α-HL. Due to the binding of CTM, the conformation of the critical "Loop" region was restrained. This mechanism was confirmed by the experimental data. These findings indicate that CTM hinders the lysis activity of α-HL through a novel mechanism.

Staphylococcal pneumonia provoked by methicillin-resistant Staphylococcus aureus (MRSA) is a life-threatening infection in which α-toxin is an essential virulence factor. In this study, we investigate the influence of naringenin on α-toxin production and further assess its therapeutic performance in the treatment of staphylococcal pneumonia. Remarkably, the expression of α-toxin was significantly inhibited when the organism was treated with 16 μg/ml of naringenin. When studied in a mouse model of S. aureus pneumonia, naringenin could attenuate the symptoms of lung injury and inflammation in infected mice. These results suggest that naringenin is a promising agent for treatment of S. aureus infection.

α-Hemolysin (α-HL) is secreted by Staphylococcus aureus as a water-soluble monomer that assembles in the lipid bilayers of susceptible host cells to form cylindrical heptameric pores. In this study, we found that baicalein (BAE), a natural compound with weak anti-S. aureus activity, could inhibit the hemolytic activity of α-HL at low concentrations. Further, molecular dynamics simulations, free energy calculations, and mutagenesis assays were carried out to determine the potential mode of action. The data revealed that BAE could directly bind to α-HL, restricting the conformation of the critical Loop region in α-HL and preventing the formation of α-HL heptameric transmembrane pore, thereby inhibiting the hemolytic activity of α-HL. This mechanism was confirmed by experimental data based on a deoxycholate-induced oligomerization assay. Moreover, in vivo tests indicated that BAE provides protection against lethal S. aureus pneumonia in a mouse model system. The mortality, bacterial load and histopathologic manifestation in lung tissue, percent neutrophils and cytokine levels in the bronchoalveolar lavage fluids were significantly decreased following treatment with BAE. These findings indicate that BAE hinders the lysis activity of α-HL through a novel mechanism, which should facilitate the development of novel therapeutics that target S. aureus α-HL.

Staphylococcus aureus causes a broad range of life-threatening diseases in humans. The pathogenicity of this micro-organism is largely dependent upon its virulence factors. One of the most extensively studied virulence factors is the extracellular protein α-toxin. In this study, we show that allicin, an organosulfur compound, was active against S. aureus with MICs ranged from 32 to 64 μg/mL. Haemolysis, Western blot and real-time RT-PCR assays were used to evaluate the effects of allicin on S. aureus α-toxin production and on the levels of gene expression, respectively. The results of our study indicated that sub-inhibitory concentrations of allicin decreased the production of α-toxin in both methicillin-sensitive S. aureus (MSSA) and methicillin-resistant S. aureus (MRSA) in a dose-dependent manner. Furthermore, the transcriptional levels of agr (accessory gene regulator) in S. aureus were inhibited by allicin. Therefore, allicin may be useful in the treatment of α-toxin-producing S. aureus infections.

Inorganic bone xenograft materials have recently found extensive surgical application in the clinic. Previously we have demonstrated that calcinated antler cancellous bone (CACB) has great potential for bone defect repair, due to the similar structure and composition compared with human bone. However, the effect of intrinsic material characteristics, particularly deer age, on the physicochemical and biological properties of CACB scaffolds has not been clarified. The aim of this study is to investigate the structure, composition and in vitro solubility of CACB scaffolds derived from deer of varying ages, including young (CACB-Y), middle-aged (CACB-M), and old (CACB-O) deer, and to determine subsequent biological performance. Microstructural analyses showed looser crystal arrangement and lower porosity in CACB-M compared to CACB-Y and CACB-O. Phase-structure analysis showed that CACB-M had the largest crystal size. Component characterization results showed that CACB-M had the most carbonated substitute and the highest content of trace elements (Na, Fe). The in vitro solubility test showed that CACB-M had the fastest dissolution and apatite deposition rates with new crystalline phases. In addition, CACB-M could be conducive for attachment, proliferation and osteogenic differentiation of rat bone marrow mesenchymal stem cells in vitro, as well as conducive for bone regeneration in vivo. These findings indicate that animal age should be seriously considered as a key parameter in optimizing the physicochemical and biological properties of deproteinized antler cancellous bone substitutes for bone regeneration applications.

Alpha-hemolysin (α-HL) is a self-assembling, channel-forming toxin produced by most Staphylococcus aureus strains as a 33.2-kDa soluble monomer. Upon binding to a susceptible cell membrane, the monomer self-assembles to form a 232.4-kDa heptamer that ultimately causes host cell lysis and death. Consequently, α-HL plays a significant role in the pathogenesis of S. aureus infections, such as pneumonia, mastitis, keratitis and arthritis. In this paper, experimental studies show that oroxylin A (ORO), a natural compound without anti-S. aureus activity, can inhibit the hemolytic activity of α-HL. Molecular dynamics simulations, free energy calculations, and mutagenesis assays were performed to understand the formation of the α-HL-ORO complex. This combined approach revealed that the catalytic mechanism of inhibition involves the direct binding of ORO to α-HL, which blocks the conformational transition of the critical “Loop” region of the α-HL protein thereby inhibiting its hemolytic activity. This mechanism was confirmed by experimental data obtained from a deoxycholate-induced oligomerization assay. It was also found that, in a co-culture system with S. aureus and human alveolar epithelial (A549) cells, ORO could protect against α-HL-mediated injury. These findings indicate that ORO hinders the lytic activity of α-HL through a novel mechanism, which should facilitate the design of new and more effective antibacterial agents against S. aureus., Author Summary The mechanism controlling protein-ligand interactions is one of the most important processes in rational drug design. X-ray crystallography is a traditional tool used to investigate the interaction of ligands and proteins in a complex. However, protein crystallography is inefficient, and the development of crystal technology and research remains unequally distributed. Thus, it seems impractical to explore the structure of the α-hemolysin-ORO monomer complex by crystallography. Therefore, we used molecular dynamics simulations to investigate the receptor-ligand interaction in the α-HL-ORO monomer complex. In this study, we found that oroxylin A (ORO), a natural compound with little anti-S. aureus activity, can inhibit the hemolytic activity of α-HL at low concentrations. Through molecular docking and molecular dynamics simulations, we determined the potential binding mode of the protein-ligand interaction. The data revealed that ORO directly binds to α-HL, an interaction that blacks the conformational transition of the critical “Loop” region in α-HL and thus prevents the formation of the α-HL heptameric transmembrane pore, which ultimately inhibits the hemolytic activity of α-HL. This mechanism was confirmed by experimental data. Furthermore, we demonstrated that ORO could protect against α-HL-mediated injury in human alveolar epithelial (A549) cells.

BACKGROUND: α-toxin is one of the major virulence factors secreted by most Staphylococcus aureus strains, which played a central role in the pathogenesis of S. aureus pneumonia. The aim of this study was to investigate the impact of capsaicin on the production of α-toxin by community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) strain USA 300 and to further assess its performance in the treatment of CA-MRSA pneumonia in a mouse model. METHODOLOGY/PRINCIPAL FINDINGS: The in vitro effects of capsaicin on α-toxin production by S. aureus USA 300 were determined using hemolysis, western blot, and real-time RT-PCR assays. The influence of capsaicin on the α-toxin-mediated injury of human alveolar epithelial cells was determined using viability and cytotoxicity assays. Mice were infected intranasally with S. aureus USA300; the in vivo protective effects of capsaicin against S. aureus pneumonia were assessed by monitoring the mortality, histopathological changes and cytokine levels. Low concentrations of capsaicin substantially decreased the production of α-toxin by S. aureus USA 300 without affecting the bacterial viability. The addition of capsaicin prevented α-toxin-mediated human alveolar cell (A549) injury in co-culture with S. aureus. Furthermore, the in vivo experiments indicated that capsaicin protected mice from CA-MRSA pneumonia caused by strain USA 300. CONCLUSIONS/SIGNIFICANCE: Capsaicin inhibits the production of α-toxin by CA-MRSA strain USA 300 in vitro and protects mice from CA-MRSA pneumonia in vivo. However, the results need further confirmation with other CA-MRSA lineages. This study supports the views of anti-virulence as a new antibacterial approach for chemotherapy.