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.

Functional vinorelbine plus schisandrin B liposomes destroying tumor metastasis in treatment of gastric cancer.

Gastric cancer is one of the leading causes of cancer-related death worldwide with a poor prognosis. Gastric cancer is usually treated with surgery and chemotherapy, accompanied by a high rate of metastasis and recurrence. In this paper, R8 (RRRRRRRR) modified vinorelbine plus schisandrin B liposomes had been successfully constructed for treating gastric cancer. In the liposomes, R8 was used to enhance the intracellular uptake, schisandrin B was incorporated into liposomes for inhibiting tumor cells metastasis, and vinorelbine was encapsulated into liposomes as antitumor drugs. Studies were performed on BGC-823 cells in vitro and were verified in the BGC-823 cell xenografts nude mice in vivo. Results in vitro demonstrated that the targeting liposomes could induce BGC-823 cells apoptosis, inhibit the metastasis of tumor cells, and increase targeting effects to tumor cells. Meanwhile, action mechanism studies showed that the targeting liposomes could down-regulate VEGF, VE-Cad, HIF-1a, PI3K, MMP-2, and FAK to inhibit tumor metastasis. In vivo results exhibited that the targeting liposomes displayed an obvious antitumor efficacy by accumulating selectively in tumor site and induce tumor cell apoptosis. Hence, R8 modified vinorelbine plus schisandrin B liposomes might provide a safe and efficient therapy strategy for gastric cancer.

Characterization and preparation of p(U-MMA-An) interpenetrating polymer network damping and absorbing material.

P(U-MMA-ANI) interpenetrating polymer network (IPN) damping and absorbing material is successfully synthesized by PANI particles served as an absorbing agent with the microemulsion polymerization and P(U-MMA) foam IPN network structure for substrate materials with foaming way. P(U-MMA-ANI) IPN is characterized by the compression mechanical performance testing, TG-DSC, and DSC. The results verify that the P(U-MMA) IPN foam damping material has a good compressive strength and compaction cycle property, and the optimum content of PMMA was 40% (mass) with which the SEM graphs do not present the phase separation on the macro level between PMMA and PU, while the phase separation was observed on the micro level. The DTG curve indicates that because of the formation of P(U-MMA) IPN, the decomposition temperature of PMMA and the carbamate in PU increases, while that of the polyol segment in PU has almost no change. P(U-MMA-ANI) IPN foam damping and absorbing material is obtained by PANI particles served as absorbing agent in the form of filler, and PMMA in the form of micro area in substrate material. When the content of PANI was up to 2.0% (mass), the dissipation factor of composites increased, and with the increasing of frequency the dissipation factor increased in a straight line.

Fabrication of versatile polyvinyl alcohol and carboxymethyl cellulose-based hydrogels for information hiding and flexible sensors: Heat-induced adjustable stiffness and transparency.

With the growing demand for wearable electronics, designing biocompatible hydrogels that combine self-repairability, wide operating temperature and precise sensing ability offers a promising scheme. Herein, by interpenetrating naturally derived carboxymethyl cellulose (CMC) into a polyvinyl alcohol (PVA) gel matrix, a novel hydrogel is successfully developed via simple coordination with calcium chloride (CaCl2). The chelation of CMC and Ca2+ is applied as a second crosslinking mechanism to stabilize the hydrogel at relatively high temperature (95 °C). In particular, it has unique heat-induced healing behavior and unexpected tunable stiffness & transparency. Like the sea cucumber, the gel can transform between a stiffened state and a relaxed state (nearly 23 times modulated stiffness from 453 to 20 kPa) which originates from the reconstruction of the crystallites. The adjustable transparency enables the hydrogel to become an excellent information hiding material. Due to the presence of Ca2+, the hydrogels show favorable conductivity, anti-freezing and long-term stability. Based on the advantages, a self-powered sensor, where chemical energy is converted to electrical energy, is assembled for human motion detection. The low-cost, environmentally friendly strategy, at the same time, complies to the "green" chemistry concept with the full employment of the biopolymers. Therefore, the proposed hydrogel is deemed to find potential use in wearable sensors.

Total astragalosides decrease apoptosis and pyroptosis by inhibiting enterovirus 71 replication in gastric epithelial cells.

Enterovirus 71 (EV71) is one of the primary pathogens involved in severe hand, foot and mouth disease in children. EV71 infection causes various types of programmed cell death. However, there are currently no clinically approved specific antiviral drugs for control of EV71 infection. Astragalus membranaceus (AM), a Traditional Chinese medicine, has been used in antiviral therapy in China. The aim of the present study was to determine whether total astragalosides (ASTs), bioactive components of AM, protect against EV. DAPI nuclear staining was used to observe morphological changes of the nucleus and the protective effect of ASTs, which revealed that the nucleus shrank following EV71 infection, while ASTs reversed it. Cell Counting Kit-8 assay found that human normal gastric epithelial cell (GES-1 cell) viability decreased following EV71 infection, while lactate dehydrogenase (LDH) assay showed that EV71 infection induced GES-1 cell damage. Western blotting was used to measure the expression levels of apoptosis and pyroptosis marker protein to determine whether EV71 infection induced apoptosis and pyroptosis in GES-1 cells. Reverse transcription-quantitative PCR was used to determine the anti-EV71 effect of ASTs. The results showed that ASTs protected GES-1 cells from EV71-induced cell apoptosis and pyroptosis. Furthermore, the present data demonstrated that the protective effect of ASTs was exerted by suppressing EV71 replication and release. These findings suggested that ASTs may represent a potential antiviral agent for the treatment of EV71 infection.

[FTIR spectra study on the film of polyurethane coated urea controlled-release fertilizer].

The polyurethane films were prepared to wrap the urea in order to achieve a desirable release rate by mixing isocyanate, polyols and wax. The effect of wax, urea and isocyanate on the structure and properties of the films was investigated by FTIR. The structural changes were monitored as the polyurethane films together with the wrapped urea were immersed into ammonia water for 28 days, which is used to model soil conditions. The FTIR results showed that the width and intensity of the NH-free band increased remarkably with time, and all kinds of carbonly bands shifted to high wavenumber and their intensity increased obviously. The results suggest that the structure of the polyurethane films was destroyed more heavily in soil than in water, and this explains the relatively fast release rate of urea in soil. It was observed that the increase in the chemical crosslinking density in the polyurethane films can effectively decrease the release rate of the urea nitrogen in soil.

Retention of prefabricated titanium dowels cemented with three luting resins.

PURPOSE: to evaluate the effects of three luting resins and roughening of the dentinal walls on the long-term retention of tapered titanium dowels. MATERIALS AND METHODS: ninety-six single-rooted teeth were endodontically treated. All root specimens were divided into three groups (n = 32). Dowel spaces were then prepared with ISO 90 drills 10 mm deep. Corresponding prefabricated tapered titanium dowels were air abraded and luted with Panavia 21 (P21), Super Bond C&B (SB) or with Chemiace II (CH) according to the manufacturers' instructions. In the subgroups of each luting agent, the dentinal walls were either left in their original state as prepared by the drills (n = 16), or roughened with a handheld diamondcoated ISO 90 cutting instrument (n = 16). The bonded specimens were stored in water at 37°C for 3 days (n = 8) or 150 days with simulated aging conditions of 37,500 thermal cycles (5°C/55°C) and 300,000 mechanical loading cycles with 30 N (n = 8). Dowel retention in N was measured using a universal testing machine with a crosshead speed of 2 mm/min. Statistical analysis was performed with ANOVAs followed by Scheffé post-hoc tests. RESULTS: the dowel retention with P21 and SB was significantly higher than that of CH (p ≤ 0.05). Roughening the dentinal walls significantly increased the retention for all cements (p ≤ 0.01). Storage for 150 days with thermal cycling and mechanical loading caused a significant decrease in dowel retention (p ≤ 0.05). CONCLUSION: P21 and SB luting resins provided a durable retention of prefabricated titanium dowels in root canals, especially for root canals with roughened dentinal walls, as the combined result of sliding friction, micromechanical interlocking and adhesive dentin bonding.

Local penetration of doxorubicin via intrahepatic implantation of PLGA based doxorubicin-loaded implants.

Doxorubicin (DOX) is widely used in the chemotherapy of a wide range of cancers. However, intravenous administration of DOX causes toxicity to most major organs which limits its clinical application. DOX-loaded drug delivery system could provide a continuous sustained-release of drugs and enables high drug concentrations at the target site, while reducing systemic toxicity. Additionally, local chemotherapy with DOX may be a promising approach for lowering post-surgical recurrence of cancer. In this study, the sustained-release DOX-loaded implants were prepared by melt-molding method. The implants were characterized with regards to drug content uniformity, micromorphology and drug release profiles. Furthermore, differential scanning calorimetry (DSC) and Fourier transform infrared spectroscopy (FTIR) analyses were carried out to investigate the drug-excipient compatibility. To determine the local penetration of DOX in liver, the minipigs received intrahepatic implantation of DOX-loaded implants by abdominal surgery. UPLC-MS/MS method was used to detect the concentration of DOX in liver tissues. Our results suggested that DOX-loaded implants delivered high doses of drug at the implantation site for a prolonged period and provided valuable information for the future clinical applications of the DOX-loaded implants.

[Separation and purification process of total flavones and total tannins from Apocynum venetum leaves with macroreticular resin].

OBJECTIVE: To investigate the purification process of total flavones and total tannins from Apocynum venetum leaves with macroreticular resin. METHOD: The static capacity absorption, dynamic absorption ratio and dynamic elution ratio of different types of macroreticular resins were studied and compared in order to find the best one among the eight macroreticular resins, and the technical process of the type of HPD-400 type macroreticular resin was studied in detail. RESULT: The type of HPD-400 type macroreticular resin showed the best comprehensive absorption property with the following technological conditions: the current velocity was 1 BV x h(-1), the volume of distilled water was 2 BV, the eluting reagent was 60% ethanol, and the volume of 60% ethanol was 3 BV. CONCLUSION: The purity of total flavones and total tannins in the product is up to 80% after purified with HPD-400 macroreticular resin. Therefore, this purification process is feasible.

Flowable materials as an intermediate layer could improve the marginal and internal adaptation of composite restorations in Class-V-cavities.

OBJECTIVES: The purpose of this in vitro study was to evaluate the marginal and internal adaptation of restorative systems in combination with flowable materials as an intermediate layer in Class-V-cavities. METHODS: Thirty Class-V-cavities with occlusal margins in enamel and gingival margins in dentin/cementum were prepared and randomly assigned to five groups. The following five restorative systems were used: compomer, composite E, flowable compomer/composite E, composite RF and flowable composite/composite RF. The flowable materials were added between the restorative composites and the cervical margins of the cavities. Marginal and internal adaptation were quantitatively evaluated before and after thermal (2,500 times between 5 and 55 degrees C) and mechanical load cycling (25,000 times 100 N) using standard SEM procedures. Statistical analysis was performed using the Kruskal-Wallis and Mann-Whitney U-Test. The morphology of the internal restorative interfaces was also evaluated. RESULTS: The present study revealed that the best marginal adaptation in dentin was attained with the compomer restoration (95.8% perfect margin). The marginal adaptation of composite restorative systems was improved by the use of a flowable compomer (45.5% vs. 68.2% perfect margin in dentin) or a flowable composite (46.7% vs. 80.7% perfect margin in dentin, p<0.05) as an intermediate layer. No differences were observed between the used materials for the parameter internal adaptation. SIGNIFICANCE: In Class-V-cavities compomers showed the best marginal adaptation and the marginal adaptation of composite fillings could be improved by the use of flowable materials as an intermediate layer.

Two new limonoids isolated from the fuits of Melia toosendan.

In the present study, two new limonoids, 1α, 7α-dihydroxyl-3α-acetoxyl-12α-ethoxylnimbolinin (1) and 1α-tigloyloxy-3α-acetoxyl-7α-hydroxyl-12ß-ethoxylnimbolinin (2), together with other four known limonoids (3-6), were isolated from the fruits of Melia toosendan. Their structures were elucidated by means of extensive spectroscopic analyses (NMR and ESI-MS) and comparisons with the data reported in the literature. The isolated compounds were evaluated for their antibacterial activities. Compound 4 exhibited significant antibacterial activity against an oral pathogen, Porphyromonas gingivalis ATCC 33277, with an MIC value of 15.2 µg·mL(-1). Compound 2 was also active against P. gingivalis ATCC 33277, with an MIC value of 31.25 µg·mL(-1). In conlcusion, our resutls indicate that these compounds may provide a basis for future development of novel antibiotics.

Effects of incorporation of nano-fluorapatite or nano-fluorohydroxyapatite on a resin-modified glass ionomer cement.

This study aimed to investigate the fluoride release properties and the effect on bond strength of two experimental adhesive cements. Synthesized particles of nano-fluorapatite (nano-FA) or nano-fluorohydroxyapatite (nano-FHA) were incorporated into a resin-modified glass ionomer cement (Fuji Ortho LC) and characterized using X-ray diffraction and scanning electron microscopy. Blocks with six different concentrations of nano-FA or nano-FHA were manufactured and their fluoride release properties evaluated by ultraviolet spectrophotometry. The unaltered glass ionomer cement Fuji Ortho LC (GC, control) and the two experimental cements with the highest fluoride release capacities (nano-FA+Fuji Ortho LC (GFA) and nano-FHA+Fuji Ortho LC (GFHA)) were used to bond composite blocks and orthodontic brackets to human enamel. After 24 h water storage all specimens were debonded, measuring the micro-tensile bond strength (µTBS) and the shear bond strength (SBS), respectively. The optimal concentration of added nano-FA and nano-FHA for maximum fluoride release was 25 wt.%, which nearly tripled fluoride release after 70 days compared with the control group. GC exhibited a significantly higher SBS than GFHA/GFA, with GFHA and GFA not differing significantly (P>0.05). The µTBS of GC and GFA were significantly higher than that of GFHA (P≤0.05). The results seem to indicate that the fluoride release properties of Fuji Ortho LC are improved by incorporating nano-FA or nano-FHA, simultaneously maintaining a clinically sufficient bond strength when nano-FA was added.