Study of Icaritin Films by Low-Energy Electron Beam Deposition

In this paper, icaritin film was prepared by low-energy beam electron beam deposition (EBD). The material test showed that the structure and composition of icaritin were not changed after electron beam deposition. Then, the film was sliced and immersed in simulated body fluids, it can be seen that the film was released quickly in the first 7 days. With the extension of soaking time, the release rate gradually slowed down, and the release amount exceeded 90% in about 20 days. In vitro cytotoxicity test showed that the relative cell viability rate of the film was still 92.32±1.30% (p<0.05), indicating that the film possessed excellent cytocompatibility.

Synthesis of Yellow-Fluorescent Carbon Nano-dots by Microplasma for Imaging and Photocatalytic Inactivation of Cancer Cells

In recent years, multifunctional nanoparticles with combined diagnostic and therapeutic functions show great promise in nanomedicine. In this study, we report the environmentally friendly synthesis of fluorescent carbon nano-dots such as carbon quantum dots (CQDs) by microplasma using o-phenylenediamine. The produced CQDs exhibited a wide absorption peaks at 380–500 nm and emitted bright yellow fluorescence with a peak at 550 nm. The CQDs were rapidly taken up by HeLa cancer cells. When excited under blue light, a bright yellow fluorescence signal and intense reactive oxygen species (ROS) were efficiently produced, enabling simultaneous fluorescent cancer cell imaging and photodynamic inactivation, with a 40% decrease in relative cell viability. Furthermore, about 98% cells were active after the incubation with 400 μg mL−1 CQDs in the dark, which revealed the excellent biocompatibility of CQDs. Hence, the newly prepared CQDs are thus demonstrated to be materials which might be effective and safe to use for in vivo bioimaging and imaging-guided cancer therapy.

Green Synthesis of Yellow-fluorescent Carbon Nano-Dots for Cancer-Cell Image and Photocatalytic Inactivation by Using Microplasma

In recent years, multifunctional nanoparticles with combined diagnostic and therapeutic functions show great promise in nanomedicine. In this work, we report the environmentally friendly synthesis of fluorescent carbon nano-dots such as carbon quantum dots (CQDs) by microplasma using o-phenylenediamine. The produced CQDs exhibited a wide absorption peaks at 380-500 nm and emitted bright yellow fluorescence with a peak at 550 nm. The CQDs were rapidly taken up by HeLa cancer cells. When excited under blue light, a bright yellow fluorescence signal and intense reactive oxygen species (ROS) were efficiently produced, enabling simultaneous fluorescent cancer cell imaging and photodynamic inactivation, with a 40% decrease in relative cell viability. Furthermore, the CQDs has 98% cell viability at concentrations of 400 μg·mL-1 in the dark demonstrating excellent biocompatibility. The newlyprepared CQDs are thus demonstrated to be materials that can be effectively for imaging-guided cancer therapy.

Simonkolleite Coating on Poly(Amino Acids) to Improve Osteogenesis and Suppress Osteoclast Formation in Vitro

Zinc can enhance osteoblastic bone formation and stimulate osteogenic differentiation, suppress the differentiation of osteoclast precursor cells into osteoclasts, and inhibit pathogenic bacterial growth in a dose-dependent manner. In this study, simonkolleite, as a novel zinc resource, was coated on poly (amino acids) (PAA) via suspending PAA powder in different concentrations of zinc chloride (ZnCl2) solution, and the simonkolleite-coated PAA (Zn–PAA) was characterized by SEM, XRD, FT-IR and XPS. Zinc ions were continuously released from the coating, and the release behavior was dependent on both the concentration of the ZnCl2 immersing solution and the type of soak solutions (SBF, PBS and DMEM). The Zn–PAA was cultured with mouse bone marrow stem cells (BMSCs) through TranswellTM plates, and the results indicated that the relative cell viability, alkaline phosphatase (ALP) activity and mineralization of BMSCs were significantly higher with Zn–PAA as compared to PAA. Moreover, the Zn–PAA was cultured with RAW264.7 cells, and the results suggested an inhibiting effect of Zn–PAA on the cell differentiation into osteoclasts. In addition, Zn–PAA exhibited an antibacterial activity against both S. aureus and E. coli. These findings suggest that simonkolleite coating with certain contents could promote osteogenesis, suppress osteoclast formation and inhibit bacteria, indicating a novel way of enhancing the functionality of synthetic bone graft material and identifying the underline principles for designing zinc-containing bone grafts.

Development of a Novel Cell-Based Assay System for High-Throughput Screening of Compounds Acting on Background Two-Pore Domain K+ Channels

Two-pore domain K+ (K2P) channels are thought to be druggable targets. However, only a few agents specific for K2P channels have been identified, presumably due to the lack of an efficient screening system. To develop a new high-throughput screening (HTS) system targeting these channels, we have established a HEK293-based “test cell” expressing a mutated Na+ channel (Nav1.5) with markedly slowed inactivation, as well as a K+ channel (Kir2.1) that sets the membrane potential quite negative, close to K+ equilibrium potential. We found in this system that Kir2.1 block by 100 μM Ba2+ application consistently elicited a large depolarization like a long-lasting action potential. This maneuver resulted in cell death, presumably due to the sustained Na+ influx. When either the TWIK-related acid-sensitive K+ (TASK)-1 or TASK-3 channel was expressed in the test cells, Ba2+-induced cell death was markedly weakened. Stronger activation of TASK-1 by extracellular acidification further decreased the cell death. In contrast, the presence of K2P channel blockers enhanced cell death. IC50 values for TASK-1 and/or TASK-3 blockers acquired by measurements of relative cell viability were comparable to those obtained using patch-clamp recordings. Both blockers and openers of K2P channels can be accurately assessed with high efficiency and throughput by this novel HTS system.

The in vitro Antiviral Mechanisms of Stronger Neo-Minophafen C, an Established Formulation of Compound Glycyrrhizin

Background: Glycyrrhiza glabra (liquorice) has been extensively used since ancient Egypt, Greek and Roman, and is an important herb in traditional Chinese medicine. Its major compound, Glycyrrhizin (GL) possesses multiple pharmacological activities, such as anti-virus waiting for exploration indepth. Objective: The aim of this research is determining the antiviral mechanisms of Stronger Neo-Minophafen C (SNMC), an established formulation of compound GL based on Interferon (IFN) system, an important cytokine family best known for its antiviral ability. Methods: Four cell lines, A549, Hela, SMMC-7721 and TC-1, were recruited. The relative cell viability (RCV) was measured with 3(4, 5 dimethylthiazol) 2, 5 diphenyltetrazolium bromide (MTT). The gene transcription of key elements on IFN system, such as IFN-β1, IRF3 and ISG15 were evaluated using realtime RT-PCR. The expressions of key enzymes on IFN system were measured by Western blot. The concentrations of IFN-γ and IRF1, representative members of type II interferon, were detected by ELISA. Results: SNMC reduces RCV with concurrent induction of antiviral genes majorly belong to type I IFN pathway, focusing on IRF3-IFN-β1- ISG15 axis. The expression of IFN-β1, IRF3 and ISG15 genes in A549 and Hela cells peak at 12 h post-SNMC incubation, in a time- and dosage- dependent manner. The expression of IFN-β1 protein reaches a peak at 24 h in A549 and SMMC-7721 cells, and peaks at 48 h in Hela and TC-1 cells. The expression of ISG15 reaches a peak at 24 h in A549, Hela and TC-1 cells, and at 48 h in SNMC-7721 cells. The expression of Mx reaches a peak at 24 h in A549 and Hela cells, and at 48 h in SMMC-7721 and TC-1 cells. However, SNMC could not induce the expression of type II IFN signal pathway. Conclusion: We demonstrated that SNMC can induce the expression of important anti-viral genes in type I interferon pathway and indicate the existence of a key pathway response for the anti-viral effects of SNMC.

Mitochondrial Respiratory Chain Inhibitors Involved in ROS Production Induced by Acute High Concentrations of Iodide and the Effects of SOD as a Protective Factor

A major source of reactive oxygen species (ROS) generation is the mitochondria. By using flow cytometry of the mitochondrial fluorescent probe, MitoSOX Red, western blot of mitochondrial ROS scavenger Peroxiredoxin (Prx) 3 and fluorescence immunostaining, ELISA of cleaved caspases 3 and 9, and TUNEL staining, we demonstrated that exposure to 100 μM KI for 2 hours significantly increased mitochondrial superoxide production and Prx 3 protein expression with increased expressions of cleaved caspases 3 and 9. Besides, we indicated that superoxide dismutase (SOD) at 1000 unit/mL attenuated the increase in mitochondrial superoxide production, Prx 3 protein expression, and lactate dehydrogenase (LDH) release and improved the relative cell viability at 100 μM KI exposure. However, SOD inhibitor diethyldithiocarbamic acid (DETC) (2 mM), Rotenone (0.5 μM), a mitochondrial complex I inhibitor, and Antimycin A (10 μM), a complex III inhibitor, caused an increase in mitochondrial superoxide production, Prx 3 protein expression, and LDH release and decreased the relative cell viability. We conclude that the inhibitors of mitochondrial respiratory chain complex I or III may be involved in oxidative stress caused by elevated concentrations of iodide, and SOD demonstrates its protective effect on the Fischer rat thyroid cell line (FRTL) cells.

Biocompatibility of Silver-Containing Calcium-Phosphate Cements with Anti-Bacterial Properties

We have previously synthesized silver-containing hydroxyapatite (Ag-HAp) powders by an ultrasonic spray-pyrolysis (USSP) technique. On the other hand, we have successfully fabricated novel calcium-phosphate cements (CPCs) composed of mainly β-tricalcium phosphate (β-TCP) phase with anti-washout property (hereafter, β-TCP cement), which was set on the basis of chelate-bonding ability of inositol phosphate (IP6). In this study, we developed novel CPCs with both anti-bacterial and anti-washout properties by adding the Ag-HAp powder into the above β-TCP cements, and examined their anti-bacterial property and cytotoxicity. The Ag-HAp powders with Ag contents of 0, 2, and 5 mol% as a nominal composition were synthesized by an USSP technique. The raw powder for β-TCP cement was prepared by ball-milling the commercially-available β-TCP powder in the IP6 solution. The Ag-HAp/β-TCP powders were prepared by mixing Ag-HAp powder and β-TCP cement powder at a ratio of 25:75 in mass. The Ag-HAp/β-TCP cement was fabricated by mixing the above-mentioned Ag-HAp/β-TCP powder and 2.5 mass% Na2HPO4 solution at a powder/liquid ratio of 1/0.3 [g/cm3]. The anti-bacterial property of resulting cements was evaluated using Staphylococcus aureus by biofilm formation test. The Ag-HAp/β-TCP cements containing 2 and 5 mol% Ag showed strong anti-bacterial property among examined specimens. Furthermore, the cytotoxicity of Ag+ ion eluted from these cements was also examined using osteoblastic MC3T3-E1 cells and Transwell® kit. The relative cell viability cultured on each Ag-containing cement specimen was over 80 %, compared with the control (polystyrene plate). These results demonstrate that the present Ag-HAp/β-TCP cements containing 2 mol% Ag are promising one of the candidates as CPCs with both anti-bacterial property and biocompatibility.

The role of photosynthetic activity in the response of isolated Glycine max mesophyll cells to ozone

The hypothesis that active photosynthesis results in a more severe cellular response to ozone (O3) was tested. Using suspensions of isolated soybean (Glycine max (L.) Merr. cv. Chippewa 64) mesophyll cells, we measured photosynthetic carbon fixation rates and cell viability, as determined by fluorescein diacetate staining, before and after treatment with O3 in the light and dark in the presence or absence of NaHCO3. Ozone reduced the photosynthesis rates of isolated mesophyll cells to a greater degree than cell viability, suggesting greater sensitivity of photosynthesis. Posttreatment photosynthesis rates were higher in cells that were fumigated in media containing NaHCO3 than in cells fumigated in its absence. The only interaction detected was between the gaseous treatments and light, in which relative cell viability was reduced more by O3 in the light than in the dark, in the second experiment. The interaction was, at least in part, a reflection of experimental error. Of greater significance was the observation that the photosynthetic function could be affected equally by O3 in the dark and in the light.