Identification of Protein Quality Markers in Toad Venom from Bufo gargarizans.

Toad venom is a traditional Chinese medicine with high medicinal value. The existing quality evaluation standards of toad venom have obvious limitations because of the lack of research on proteins. Thus, it is necessary to screen suitable quality markers and establish appropriate quality evaluation methods for toad venom proteins to guarantee their safety and efficacy in clinical applications. SDS-PAGE, HPLC, and cytotoxicity assays were used to analyze differences in protein components of toad venom from different areas. Functional proteins were screened as potential quality markers by proteomic and bioinformatic analyses. The protein components and small molecular components of toad venom were not correlated in content. Additionally, the protein component had strong cytotoxicity. Proteomics analysis showed that 13 antimicrobial proteins, four anti-inflammatory and analgesic proteins, and 20 antitumor proteins were differentially expressed extracellular proteins. A candidate list of functional proteins was coded as potential quality markers. Moreover, Lysozyme C-1, which has antimicrobial activity, and Neuropeptide B (NPB), which has anti-inflammatory and analgesic activity, were identified as potential quality markers for toad venom proteins. Quality markers can be used as the basis of quality studies of toad venom proteins and help to construct and improve safe, scientific, and comprehensive quality evaluation methods.

Impaired barrier integrity of endothelial cells induced by PEGylated black phosphorus nanosheets.

PEGylated black phosphorus nanosheets (PEG-BPNSs) have shown promising applications in biomedicine and potentially interact with the vasculature following iatrogenic exposures. Whether the exposure to PEG-BPNSs could induce toxic effects on endothelial cells that line the blood vessels remains largely unknown. Herein, we investigate the cellular response and transcriptional profiling of human umbilical vein endothelial cells (HUVECs) after the exposure to BPNSs and PEG-BPNSs. BPNSs and PEG-BPNSs induce cellular elongation and cause significant cytotoxicity to HUVECs at 0.8 µg/mL, with viabilities of 87.8% and 87.7% respectively. The transcriptome analysis indicates that BPNSs and PEG-BPNSs at 0.4 µg/mL cause marked alterations in the expression of genes associated with detection of stimulus, ion transmembrane transport and components of plasma membrane. BPNSs and PEG-BPNSs at 0.4 µg/mL decrease the transendothelial electrical resistance (TEER) across monolayers of HUVECs by 22.8% and 20.3% compared to the control, respectively. The disturbance of tight junctions (TJs) after 24 h exposure to 0.4 µg/mL BPNSs and PEG-BPNSs is indicated with the downregulated mRNA expression of zona occluden-1 (ZO-1) by respective 16.5% and 29.9%, which may be involved in the impairment of endothelial barrier integrity. Overall, the response of HUVECs to PEG-BPNSs and BPNSs has no statistical difference, suggesting that PEGylation does not attenuate the BPNSs-induced endothelial injury. This study demonstrates the detrimental effects of BPNSs and PEG-BPNSs on barrier integrity of HUVECs, contributing to our understanding on the potential toxicological mechanisms.

Preparation of multiresponsive hydrophilic molecularly imprinted microspheres for rapid separation of gardenia yellow and geniposide from gardenia fruit.

In this work, a robust method for the separation of gardenia yellow and geniposide from gardenia fruit was developed based on a molecularly imprinted solid phase extraction (MISPE) procedure. First, hydrophilic molecularly imprinted microspheres (HMIMs) were prepared using gardenia yellow as the template via reversible addition fragmentation chain transfer (RAFT) precipitation polymerization. The resultant HMIMs demonstrated the multiresponsiveness to pH, temperature, and magnetism, achieving controllable uptake and release of gardenia yellow and easy recovery by external magnets. Meanwhile, the HMIMs possessed high adsorption capacity, fast binding kinetics, specific recognition, and reusability. Finally, a MISPE approach using HMIMs as adsorbent was developed for extraction of gardenia yellow and purification of geniposide after optimization of the adsorption and elution conditions. Thus, efficient separation of gardenia yellow and geniposide with relative purities of 99.77 ± 0.05% (94.04 ± 0.10% recovered) and 94.50 ± 0.62% (95.40 ± 0.86% recovered), respectively, was achieved.

Biobased Epoxies Derived from Myrcene and Plant Oil: Design and Properties of Their Cured Products.

Two biobased epoxy resin monomers derived from myrcene and plant oil are synthesized without using petroleum-based bisphenol A. To obtain material with balanced strength and toughness, the two epoxy monomers are cured together in different weight proportions. Properties of cured epoxy resin are tested by different techniques. Tensile and impact tests indicate that when the content of myrcene-based epoxy is 50-75 wt %, the cured sample has a high strain of 32.30-161.47%, and a moderate tensile strength of 9.57-15.96 MPa. Dynamic mechanical analysis suggests that the glass transition temperature (T g) of cured samples increases from 17 to 71 °C with the increasing content of myrcene-based epoxy. Morphology of fracture surface indicates that the cured sample containing plant oil-based epoxy resin shows obvious plastic deformation. The curing kinetics of the two epoxies resin is studied by differential scanning calorimetry. Also, the calculated activation energy is 70.49 kJ/mol for myrcene-based epoxy and 64.02 kJ/mol for poly-fatty acid-derived epoxy resin. The thermogravimetric analysis indicates that the main degradation temperature of all cured samples is above 300 °C. The sustainable biobased epoxy has some potential in preparing flexible epoxy materials and can be used to toughen conventional petroleum-based epoxy.