Metabolomics combined with network pharmacology reveals a role for astragaloside IV in inhibiting enterovirus 71 replication via PI3K-AKT signaling.

BACKGROUND: Astragaloside IV (AST-IV), as an effective active ingredient of Astragalus membranaceus (Fisch.) Bunge. It has been found that AST-IV inhibits the replication of dengue virus, hepatitis B virus, adenovirus, and coxsackievirus B3. Enterovirus 71 (EV71) serves as the main pathogen in severe hand-foot-mouth disease (HFMD), but there are no specific drugs available. In this study, we focus on investigating whether AST-IV can inhibit EV71 replication and explore the potential underlying mechanisms. METHODS: The GES-1 or RD cells were infected with EV71, treated with AST-IV, or co-treated with both EV71 and AST-IV. The EV71 structural protein VP1 levels, the viral titers in the supernatant were measured using western blot and 50% tissue culture infective dose (TCID50), respectively. Network pharmacology was used to predict possible pathways and targets for AST-IV to inhibit EV71 replication. Additionally, ultra-high performance liquid chromatography-high resolution mass spectrometry (UHPLC-HRMS) was used to investigate the potential targeted metabolites of AST-IV. Associations between metabolites and apparent indicators were performed via Spearman's algorithm. RESULTS: This study illustrated that AST-IV effectively inhibited EV71 replication. Network pharmacology suggested that AST-IV inhibits EV71 replication by targeting PI3K-AKT. Metabolomics results showed that AST-IV achieved these effects by elevating the levels of hypoxanthine, 2-ketobutyric acid, adenine, nicotinic acid mononucleotide, prostaglandin H2, 6-hydroxy-1 H-indole-3- acetamide, oxypurinol, while reducing the levels of PC (14:0/15:0). Furthermore, AST-IV also mitigated EV71-induced oxidative stress by reducing the levels of MDA, ROS, while increasing the activity of T-AOC, CAT, GSH-Px. The inhibition of EV71 replication was also observed when using the ROS inhibitor N-Acetylcysteine (NAC). Additionally, AST-IV exhibited the ability to activate the PI3K-AKT signaling pathway and suppress EV71-induced apoptosis. CONCLUSION: This study suggests that AST-IV may activate the cAMP and the antioxidant stress response by targeting eight key metabolites, including hypoxanthine, 2-ketobutyric acid, adenine, nicotinic acid mononucleotide, prostaglandin H2, 6-Hydroxy-1 H-indole-3-acetamide, oxypurinol and PC (14:0/15:0). This activation can further stimulate the PI3K-AKT signaling to inhibit EV71-induced apoptosis and EV71 replication.

Two {CuI[P4Mo6]2}-Based Coordination Polymers Incorporating In Situ Converted Tetrapyridyl Ligands for Trace Analysis of Nitrofuran Antibiotics.

Nitrofurans are important synthetic broad-spectrum antibacterial drugs with the basic structure of 5-nitrofuran. Due to their toxicity, it is essential to develop a sensitive sensor with strong anti-interference capabilities for their detection. In this work, two {P4Mo6O31}12--based compounds, [H4(HPTTP)]2{CuI[Mo12O24(OH)6(PO4)3(HPO4)(H2PO4)4]}·xH2O (x = 13 for (1), 7 for (2); HPTTP = 4,4',4″,4‴-(1H-pyrrole-2,3,4,5-tetrayl)tetrapyridine), exhibiting similar coordination but distinct stacking modes. Both compounds were synthesized and used for the electrochemical detection of nitrofuran antibiotics. The tetrapyridine-based ligand was generated in situ during assembly, and its potential mechanism was discussed. Composite electrode materials, formed by mixing graphite powder with compounds 1-2 and physically grinding them, proved to be highly effective in the electrochemical trace detection of furazolidone (FZD) and furaltadone hydrochloride (FTD·HCl) under optimal conditions. Besides, the possible electrochemical detection mechanisms of two nitro-antibiotics were studied.

The Modified Exenatide Microspheres: PLGA-PEG-PLGA Gel and Zinc-Exenatide Complex Synergistically Reduce Burst Release and Shorten Platform Stage.

The existing exenatide microspheres have the problem of burst release in the early stage, and minimal release in the middle stage which makes it difficult to achieve effective blood drug concentration (platform period). In this study, the modified exenatide microspheres were constructed to address the aforementioned issues. Poly(D,L-lactic-co-glycolic acid) (PLGA) and triblock copolymer with sol-gel conversion characteristics (PLGA-PEG-PLGA gel) were introduced as carriers to prepare microspheres. The hot gel characteristics and hydrophilicity of PLGA-PEG-PLGA gel were utilized to decline the burst release and shorten the platform period. Simultaneously, zinc acetate and exenatide were combined to generate an insoluble complex to further reduce the burst release. Herein, we prepared three types of exenatide microspheres using the solvent evaporation method and investigated their characterization as well as in vitro and in vivo release. According to the experimental findings, the modified exenatide microspheres, i.e., PLGA-PEG-PLGA gel and PLGA co-loaded zinc-exenatide insoluble complex microspheres (Zn-EXT-Gel-MS), had smooth and rounded surfaces, with a particle size of 24.7 µm, and the encapsulation rate reached 89.43%. And it was released for 40 days in vitro, behaving better than the other two microspheres in terms of release behavior. When this product was administered subcutaneously to rats, it produced a comparatively constant plasma exenatide concentration that lasted for 24 days and superior bioavailability than the exenatide microspheres (EXT-MS). The creation of modified exenatide microspheres may serve as a heuristic method for other long-acting medications. Schematic diagram of the synthesis process and release curves of three types of exenatide microspheres in vitro and in vivo.

3D printed scaffold for repairing bone defects in apical periodontitis.

OBJECTIVES: To investigate the feasibility of the 3D printed scaffold for periapical bone defects. METHODS: In this study, antimicrobial peptide KSL-W-loaded PLGA sustainable-release microspheres (KSL-W@PLGA) were firstly prepared followed by assessing the drug release behavior and bacteriostatic ability against Enterococcus faecalis and Porphyromonas gingivalis. After that, we demonstrated that KSL-W@PLGA/collagen (COL)/silk fibroin (SF)/nano-hydroxyapatite (nHA) (COL/SF/nHA) scaffold via 3D-printing technique exhibited significantly good biocompatibility and osteoconductive property. The scaffold was characterized as to pore size, porosity, water absorption expansion rate and mechanical properties. Moreover, MC3T3-E1 cells were seeded into sterile scaffold materials and investigated by CCK-8, SEM and HE staining. In the animal experiment section, we constructed bone defect models of the mandible and evaluated its effect on bone formation. The Japanese white rabbits were killed at 1 and 2 months after surgery, the cone beam computerized tomography (CBCT) and micro-CT scanning, as well as HE and Masson staining analysis were performed on the samples of the operation area, respectively. Data analysis was done using ANOVA and LSD tests. (α = 0.05). RESULTS: We observed that the KSL-W@PLGA sustainable-release microspheres prepared in the experiment were uniform in morphology and could gradually release the antimicrobial peptide (KSL-W), which had a long-term antibacterial effect for at least up to 10 days. HE staining and SEM showed that the scaffold had good biocompatibility, which was conducive to the adhesion and proliferation of MC3T3-E1 cells. The porosity and water absorption of the scaffold were (81.96 ± 1.83)% and (458.29 ± 29.79)%, respectively. Histological and radiographic studies showed that the bone healing efficacy of the scaffold was satisfactory. CONCLUSIONS: The KSL-W@PLGA/COL/SF/nHA scaffold possessed good biocompatibility and bone repairing ability, and had potential applications in repairing infected bone defects. Clinical significance The 3D printed scaffold not only has an antibacterial effect, but can also promote bone tissue formation, which provides an alternative therapy option in apical periodontitis.

Actinomyces qiguomingii sp. nov., isolated from the Pantholops hodgsonii.

Two previously undescribed, Gram-stain-positive, rod-shaped strains, 410T and 553, were isolated from faeces of the Tibetan antelope (Pantholops hodgsonii) from the Tibet-Qinghai Plateau, PR China. The optimum growth conditions of the two novel strains were 1 % (w/v) NaCl, 37 °C and pH 7. The end products from glucose fermentation included ethanol and lactic acid. Based on results of 16S rRNA gene sequence comparison and phylogenetic and phylogenomic analyses, strains 410T and 553 were classified into the genus Actinomyces, and were closely related to Actinomyces ruminicola (97.6 %), Actinomyces oricola (93.5 %) and Actinomyces dentalis (90.8 %). The genomic G+C content of strain 410T was 67.4 mol%. Digital DNA-DNA hybridization values between strain 410T and each of the closely related species were under 70 %. The respiratory quinones were MK-10 (68 %) and MK-9 (32 %). The main cellular fatty acids of the isolates were C16 : 0, followed by C18 : 1 ω9c. The major polar lipids were diphosphatidylglycerol and phosphatidylinositol-mannoside. The whole-cell sugars contained rhamnose, ribose and glucose. The diagnostic amino acids of cell-wall peptidoglycan included alanine, glutamic acid, lysine and ornithine. The results of biochemical, chemotaxonomic and genotypic analyses revealed that the two novel strains represent a novel species of genus Actinomyces, for which the name Actinomyces qiguomingii sp. nov. is proposed. The type strain is 410T (=CGMCC 1.16361T= DSM 106201T).

Enhanced antitumor effects of follicle-stimulating hormone receptor-mediated hexokinase-2 depletion on ovarian cancer mediated by a shift in glucose metabolism.

BACKGROUND: Most cancers favor glycolytic-based glucose metabolism. Hexokinase-2 (HK2), the first glycolytic rate-limiting enzyme, shows limited expression in normal adult tissues but is overexpressed in many tumor tissues, including ovarian cancer. HK2 has been shown to be correlated with the progression and chemoresistance of ovarian cancer and could be a therapeutic target. However, the systemic toxicity of HK2 inhibitors has limited their clinical use. Since follicle-stimulating hormone (FSH) receptor (FSHR) is overexpressed in ovarian cancer but not in nonovarian healthy tissues, we designed FSHR-mediated nanocarriers for HK2 shRNA delivery to increase tumor specificity and decrease toxicity. RESULTS: HK2 shRNA was encapsulated in a polyethylene glycol-polyethylenimine copolymer modified with the FSH ß 33-53 or retro-inverso FSH ß 33-53 peptide. The nanoparticle complex with FSH peptides modification effectively depleted HK2 expression and facilitated a shift towards oxidative glucose metabolism, with evidence of increased oxygen consumption rates, decreased extracellular acidification rates, and decreased extracellular lactate and glucose consumption in A2780 ovarian cancer cells and cisplatin-resistant A2780CP counterpart cells. Consequently, cell proliferation, invasion and migration were significantly inhibited, and tumor growth was suppressed even in cisplatin-resistant ovarian cancer. No obvious systemic toxicity was observed in mice. Moreover, the nanoparticle complex modified with retro-inverso FSH peptides exhibited the strongest antitumor effects and effectively improved cisplatin sensitivity by regulating cisplatin transport proteins and increasing apoptosis through the mitochondrial pathway. CONCLUSIONS: These results established HK2 as an effective therapeutic target even for cisplatin-resistant ovarian cancer and suggested a promising targeted therapeutic approach.

Near-Infrared Afterglow Luminescent Aggregation-Induced Emission Dots with Ultrahigh Tumor-to-Liver Signal Ratio for Promoted Image-Guided Cancer Surgery.

Afterglow imaging through the collection of persistent luminescence after the stopping of light excitation holds enormous promise for advanced biomedical uses. However, efficient near-infrared (NIR)-emitting afterglow luminescent materials and probes (particularly the organic and polymeric ones) are still very limited, and their in-depth biomedical applications such as precise image-guided cancer surgery are rarely reported. Here, we design and synthesize a NIR afterglow luminescent nanoparticle with aggregation-induced emission (AIE) characteristics (named AGL AIE dots). It is demonstrated that the AGL AIE dots emit rather-high NIR afterglow luminescence persisting over 10 days after the stopping of a single excitation through a series of processes occurring in the AIE dots, including singlet oxygen production by AIE luminogens (AIEgens), Schaap's dioxetane formation, chemiexcitation by dioxetane decomposition, and energy transfer to NIR-emitting AIEgens. The animal studies reveal that the AGL AIE dots have the innate property of fast afterglow signal quenching in normal tissues, including the liver, spleen, and kidney. After the intravenous injection of AGL AIE dots into peritoneal carcinomatosis bearing mice, the tumor-to-liver ratio of afterglow imaging is nearly 100-fold larger than that for fluorescence imaging. The ultrahigh tumor-to-liver signal ratio, together with low afterglow background noise, enables AGL AIE dots to give excellent performance in precise image-guided cancer surgery.

Autophagy induced by enterovirus 71 regulates the production of IL-6 through the p38MAPK and ERK signaling pathways.

Enterovirus 71 (EV71) is the main causative agent of hand, foot, and mouth disease (HFMD), which has high morbidity and mortality. It mainly threatens children under six years of age. Because of a poor understanding of its pathogenesis, there are no effective drugs to control EV71 infection. Previous studies showed that EV71 infection induced autophagy and the production of cytokine IL-6. However, the underlying mechanisms between autophagy and the production of IL-6 induced by EV71 remain unclear. This study aimed to reveal the regulatory mechanisms between autophagy and the expression of IL-6 induced by EV71 infection. Our results showed that the proliferation of human gastric epithelial (GES-1) cells was inhibited by EV71 in a time- and dose-dependent manner. In addition, EV71 induced autophagy in GES-1 cells. EV71 infection promoted the expression and the release of IL-6 to the extracellular space, although the expression and release were inhibited by autophagy inhibitors 3-methyladenine (3-MA) and chloroquine (CQ) in GES-1 cells. The phosphorylated levels of p38MAPK and ERK proteins in GES-1 cells also increased after infection with EV71, and these changes were also reversed by 3-MA and CQ treatment. Our findings suggested that EV71-induced autophagy regulated the production of IL-6 through the p38MAPK and ERK signaling pathways.

Polymeric Encapsulation of Turmeric Extract for Bioimaging and Antimicrobial Applications.

As a herb of the ginger family, the turmeric plant has been used as spice and colorant in the Oriental countries. The rhizome part of the plant is rich in curcumin, which has been proven to be the main ingredient responsible for turmeric's biological effects. Most research endeavors have been upon the investigation of pharmaceutical activities of curcumin, yet the fluorescence of curcumin is a bit far from well-studied. The major drawbacks associated with curcumin are its poor aqueous solubility and low stability. In this communication, the encapsulation of fluorescent turmeric extract into polymeric nanoparticles (NPs) for bioimaging and antibacterial applications is reported. Through poly(d,l-lactic-co-glycolic acid) (PLGA) encapsulation, solubility of curcumin is greatly increased, and the biodegradable nature of PLGA further enhances the biocompatibility of curcumin. These Cur-PLGA NPs are successfully demonstrated to be efficient fluorescence probes for bioimaging, and promising for antibacterial application.

Enhanced human enterovirus 71 infection by endocytosis inhibitors reveals multiple entry pathways by enterovirus causing hand-foot-and-mouth diseases.

BACKGROUND: Human enterovirus 71 (EV71) was previously known to enter cells through clathrin or caveolar mediated endocytic pathways. However, we observed chlorpromazine (CPZ) or dynasore (DNS), which inhibit clathrin and dynamin mediated endocytosis, did not suppress EV71 cell entry in particular cell types. So the current knowledge of entry mechanisms by EV71 is not complete. METHODS: Viral infection was examined by flow cytometry or end-point dilution assays. Viral entry was monitored by immunofluorescence or pseudoviral infections. Various inhibitors were utilized for manipulating endocytic pathways. Cellular proteins were knockdown by siRNA. RESULTS: CPZ and DNS did not inhibit but rather enhance viral infection in A549 cells, while they inhibited infections in other cells tested. We further found CPZ did not affect EV71 binding to target cells and failed to affect viral translation and replication, but enhanced viral entry in A549 cells. Immunofluorescence microscopy further confirmed this increased entry. Using siRNA experiment, we found that the enhancement of EV71 infection by CPZ did not require the components of clathrin mediated endocytosis. Finally, CPZ also enhanced infection by Coxackivirus A16 in A549 cells. CONCLUSIONS: CPZ and DNS, previously reported as EV71 entry inhibitors, may rather lead to increased viral infection in particular cell types. CPZ and DNS increased viral entry and not other steps of viral life cycles. Therefore, our study indicated an unknown dynamin-independent entry pathway utilized by enteroviruses that cause Hand-Foot-and-Mouth Diseases.

Bioinspired polymer vesicles and membranes for biological and medical applications.

Biological membranes play an essential role in living organisms by providing stable and functional compartments, preserving cell architecture, whilst supporting signalling and selective transport that are mediated by a variety of proteins embedded in the membrane. However, mimicking cell membranes - to be applied in artificial systems - is very challenging because of the vast complexity of biological structures. In this respect a highly promising strategy to designing multifunctional hybrid materials/systems is to combine biological molecules with polymer membranes or to design membranes with intrinsic stimuli-responsive properties. Here we present supramolecular polymer assemblies resulting from self-assembly of mostly amphiphilic copolymers either as 3D compartments (polymersomes, PICsomes, peptosomes), or as planar membranes (free-standing films, solid-supported membranes, membrane-mimetic brushes). In a bioinspired strategy, such synthetic assemblies decorated with biomolecules by insertion/encapsulation/attachment, serve for development of multifunctional systems. In addition, when the assemblies are stimuli-responsive, their architecture and properties change in the presence of stimuli, and release a cargo or allow "on demand" a specific in situ reaction. Relevant examples are included for an overview of bioinspired polymer compartments with nanometre sizes and membranes as candidates in applications ranging from drug delivery systems, up to artificial organelles, or active surfaces. Both the advantages of using polymer supramolecular assemblies and their present limitations are included to serve as a basis for future improvements.

Enterovirus 71 protease 2Apro targets MAVS to inhibit anti-viral type I interferon responses.

Enterovirus 71 (EV71) is the major causative pathogen of hand, foot, and mouth disease (HFMD). Its pathogenicity is not fully understood, but innate immune evasion is likely a key factor. Strategies to circumvent the initiation and effector phases of anti-viral innate immunity are well known; less well known is whether EV71 evades the signal transduction phase regulated by a sophisticated interplay of cellular and viral proteins. Here, we show that EV71 inhibits anti-viral type I interferon (IFN) responses by targeting the mitochondrial anti-viral signaling (MAVS) protein--a unique adaptor molecule activated upon retinoic acid induced gene-I (RIG-I) and melanoma differentiation associated gene (MDA-5) viral recognition receptor signaling--upstream of type I interferon production. MAVS was cleaved and released from mitochondria during EV71 infection. An in vitro cleavage assay demonstrated that the viral 2A protease (2A(pro)), but not the mutant 2A(pro) (2A(pro)-110) containing an inactivated catalytic site, cleaved MAVS. The Protease-Glo assay revealed that MAVS was cleaved at 3 residues between the proline-rich and transmembrane domains, and the resulting fragmentation effectively inactivated downstream signaling. In addition to MAVS cleavage, we found that EV71 infection also induced morphologic and functional changes to the mitochondria. The EV71 structural protein VP1 was detected on purified mitochondria, suggesting not only a novel role for mitochondria in the EV71 replication cycle but also an explanation of how EV71-derived 2A(pro) could approach MAVS. Taken together, our findings reveal a novel strategy employed by EV71 to escape host anti-viral innate immunity that complements the known EV71-mediated immune-evasion mechanisms.

In-vitro and in-vivo evaluation of ciprofloxacin liposomes for pulmonary administration.

OBJECTIVE: The aim of this study was to investigate the ciprofloxacin liposome of high encapsulation efficiency with optimal physical properties for pulmonary administration and to test its in-vivo potential in rats. METHODS: Ciprofloxacin-loaded liposome was prepared by gradient of ammonium sulfate method. The particle size and morphology were determined using a NANOPHOX particle size analyzer and a transmission electron microscope, respectively. Encapsulation efficiency was calculated by UV spectrophotometry. Ciprofloxacin liposome released in vitro was performed using simulated lung fluid. In-vivo studies, pharmacokinetics and pulmonary distribution, HPLC method was established to determine the concentration of ciprofloxacin in rat plasma and lung tissue. The pulmonary pathological section was used to observe the change of pulmonary pathology. RESULTS: The optimized ciprofloxacin liposome, which had a high encapsulation efficiency of 93.96%, and an average particle size of 349.6 nm with a span of 0.42, showed sustained in-vitro release. The optimized ciprofloxacin liposome was further examined in the in-vivo study in rats. The concentration of ciprofloxacin in lung and blood was simultaneously determined in each rat. The ratio of the AUClung value between ciprofloxacin liposome and ciprofloxacin solution was 288.33, whereas the relative bioavailability was 72.42%, and the drug targeting efficiency of ciprofloxacin liposome and ciprofloxacin solution by intratracheal administration were 799.71 and 2.01, respectively. CONCLUSION: Ciprofloxacin liposome for pulmonary administration offered an attractive alternative that was able to deliver high concentrations of antibiotic directly to the chosen target site while minimizing the local irritation.

[Recent advances in Sphingobium sp. SYK-6 for lignin aromatic compounds degradation--a review].

Lignin is complex heteropolymer produced from hydroxycinnamyl alcohols through radical coupling. In nature, white-rot fungi are assumed initially to attack native lignin and release lignin-derived-low-molecular-weight compounds, and soil bacteria play an importent role for completely degradation of these compounds. Study on the soil bacteria degrading lignin-derived-low-molecular-weight compounds will give way to understand how aromatic compounds recycle in nature, and to utilize lignin compounds as the renewable materials for valuable materials production. Sphingobium sp. SYK-6 that grows on lignin biphenyl (5,5'-dehydrodivanillate) had been isolated from pulp effluent in 1987. We have researched this bacterium more than 25 years, a serious aromatic metabolic pathway has been determined, and related genes have been isolated. As the complete genome sequence of SYK-6 has been opened to the public in 2012, the entire aromatic compounds degradation mechanisms become more clear. Main contents in our review cover: (1) genome information; (2) aryl metabolism; (3) biphenyl metabolism; (4) ferulate metabolism; (5) tetrahydrofolate-dependent O-demethylation system for lignin compound degrdation; (6) protocatechuate 4,5-cleavage pathway; (7) multiple pathways for 3-O-methylgallate metabolism.

Surgery Outcomes of Prolene Suture Gonioscopy-Assisted Transluminal Trabeculotomy (GATT): Up to 4 Years Follow-Up and Prognostic Factors.

PRCIS: Long-term success was achievable after GATT. GATT performed at early stage of glaucoma had better surgery outcomes. Trabeculoplasty may compromise surgery success. PURPOSE: To evaluate the long-term effectiveness of prolene suture gonioscopy-assisted transluminal trabeculotomy (GATT) and identify factors that may affect surgical outcomes. PATIENTS AND METHODS: This is a retrospective cohort study of adult patients with prolene suture GATT performed by a single surgeon at 1 medical center. RESULTS: Of the 145 eyes from 124 patients studied, intraocular pressure was reduced from 22.1±7.8 to 15.1±3.2 and 15.1±3.5 mm Hg, and the number of glaucoma medications was reduced from 3.2±1.1 to 1.3±1.4 and 1.4±1.5 at postoperative years 3 and 4, respectively. Ninety-three and 71 eyes completed a 3- and 4-year follow-up, with 44% of the eyes at year 4 remaining medication free. Compared with eyes with combined GATT/cataract extraction (CE), eyes with GATT alone had significantly more preoperative medications and a higher reoperation rate (31% vs. 16.5%). Eyes with prior trabeculoplasty had a higher reoperation rate (28.8%) than those without (16.1%). Kaplan-Meier survival analysis revealed that GATT/CE eyes without trabeculoplasty had a longer median time to failure (48 mo) than GATT/CE eyes with trabeculoplasty (18 mo), and GATT eyes with or without trabeculoplasty (9 and 12 mo, respectively). CONCLUSION: Prolene suture GATT successfully reduced IOP. Eyes with more preoperative medications responded less well to GATT. Prior laser trabeculoplasty was associated with poorer outcomes. Further study is needed to verify these findings.

TLR4 signalling: the key to controlling EV71 replication and inflammatory response.

Hand, foot, and mouth disease (HFMD) is a common infectious disease caused by enterovirus 71 (EV71) that frequently affects children, leading to severe infections in some cases. In general, when infection occurs, the body upregulates inflammatory responses to eliminate pathogenic microorganisms to protect the host from infection. However, EV71 may inhibit host's innate immunity to promote virus infection. At present, it is not fully understood how EV71 hijack the host cells for its own replication. Toll-like receptor 4 (TLR4), a natural immune receptor, historically associated with bacterial endotoxin-induced inflammatory responses. However, it is still unclear whether and how TLR4 is altered during EV71 infection. In this study, we observed a reduction in both TLR4 protein and gene transcript levels in RD, GES-1, and Vero cells following EV71 infection, as detected by RT-qPCR, immunofluorescence staining and western blot. Furthermore, we observed that the TLR4 downstream molecules of MYD88, p-NF-κB p65, p-TBK1 and related inflammatory cytokines were also reduced, suggesting that antiviral innate immune and inflammatory response were suppressed. To determine the impact of TLR4 changes on EV71 infection, we interfered EV71-infected RD cells with TLR4 agonist or inhibitor and the results showed that activation of TLR4 inhibited EV71 replication, while inhibition of TLR4 promote EV71 replication. Besides, EV71 replication was also promoted in TLR4 siRNA-transfected and EV71-infected RD cells. This suggests that down-regulation the expression of TLR4 by EV71 can inhibit host immune defense to promote EV71 self-replication. This novel mechanism may be a strategy for EV71 to evade host immunity.

CuFe2O4/lignin-based carbon composited photothermal and photodynamic agents for synergetic antibacterial therapy.

Bacterial infection has become the second leading cause of death in the world. Exploring a new highly antibacterial catalyst to replace traditional antibacterial agent is crucial for the society development of human beings. In this study, CuFe2O4/Lg-based carbon composited catalysts were rationally constructed by facile hydrothermal method. Lignin-derived carbon with enormous oxygen-containing functional group was beneficial to anchor CuFe2O4 nanoparticles. The close contact interface between CuFe2O4 and Lignin-based carbon material was expected to extend the range of optical absorption and promote the separation and transportation of photogenerated carriers. Under NIR (980 nm, 1.5 W/cm2) light irradiation, the as-prepared CuFe2O4/Lg (20 µg/mL) exhibited excellent photo/photothermal synergetic in vitro (against Escherichia coli and Staphylococcus aureus) and in vivo (against Staphylococcus aureus-infected mouse wound model) antibacterial performance. Furthermore, the cell count assay kit 8 (CCK-8 kit) demonstrated the good biocompatibility of this material. On the basis of the experimental results, a possible antibacterial mechanism based on the synergetic photothermal and photodynamic therapies was proposed. This work presented a lignin- derived carbon-based highly efficient antibacterial disinfection agent with desirable biosafety.

3D printing of fish-scale derived hydroxyapatite/chitosan/PCL scaffold for bone tissue engineering.

In the field of bone tissue repair, the treatment of bone defects has always posed a significant challenge. In recent years, the advancement of bone tissue engineering and regenerative medicine has sparked great interest in the development of innovative bone grafting materials. In this study, a novel hydroxyapatite (HA) material was successfully prepared and comprehensively characterized. Antimicrobial experiments and biological evaluations were conducted to determine its efficacy. Based on the aforementioned research findings, 3D printing technology was employed to fabricate HA/chitosan (CS)/ polycaprolactone (PCL) scaffolds. The composition of the scaffold materials was confirmed through X-ray diffractometer (XRD) and Fourier Transform Infrared Spectroscopy (FT-IR) tests, while the influence of different HA ratios on the scaffold surface morphology was observed. Additionally, antimicrobial experiments demonstrated the favorable antimicrobial activity of the scaffolds containing 30%HA + 5%CS + PCL. Furthermore, the water contact angle measurements confirmed the superhydrophilicity of the scaffolds. Finally, the excellent bioactivity and ability to promote tissue regeneration of the scaffolds were further confirmed by in vitro and in vivo experiments. This study provides new options for future repair and regeneration of bone tissue and clinical applications.

Fabricating active Egg Albumin/Sodium Alginate/Sodium Lignosulfonate Nanoparticles film with significantly improved multifunctional characteristics for food packing.

In food packaging, sodium lignosulfonate nanoparticles (SLS NPs) showed significant antibacterial properties, antioxidant and UV barrier activities. Herein, the SLS NPs were synthesized via a sustainable green method and were added into egg albumin/sodium alginate mixture (EA/SA) to fabricate a safe, edible EA/SA/SNPs food packaging. A composite film EA/SA/SNP was examined microstructurally and physicochemically. The mechanical characteristics, UV protection, water resistance, and the composite film's thermal stability were all enhanced by the inclusion of SLS NPs, and water vapor permeability reduced by 44 %. This composite film exhibited robust antioxidative properties with DPPH and ABTS free radical scavenging rates reaching 76.84 % and 92.56 %, and effective antimicrobial activity against Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) with antibacterial rates reaching 98.25 % and 97.13 % for the positively charged nanoparticles interacting with the cell membrane. Freshness tests showed that the EA/SA/SNPs packaging film could delay the quality deterioration of fresh tomatoes. This composite film can slow down spoilage bacteria proliferation and prolongs food's preservation period by eight days at ambient temperature.

Isolated fracture of the coronoid process following a molar extraction: A rare case report.

Isolated coronoid process fractures are uncommon, and iatrogenic isolated fractures are extremely rare. This case describes a displaced fracture of an isolated coronoid process thought to be due to excessive force applied by a dentist that had been overlooked and left untreated for about a month. The patient was a woman in her late 50's and she had undergone a molar extraction. Her dentist had confused her symptoms of trismus, pain, and facial oedema with the complex tooth extraction procedure. Following a cone-beam computed tomography (CBCT) scan we showed that the mandibular coronoid process on her right side had suffered a longitudinal fracture, and the fractured fragment had rotated upwards and inwards. Following successful surgical elimination of the fragmented coronoid process, the patient received targeted physiotherapy sessions that yielded excellent results. At the five-month follow-up, the ability of the patient to open her mouth had improved enormously, and her facial appearance almost recovered to its original state.