•Total steroid saponins from Dioscorea zingiberensis C.H. Wright reveal bioactivities.•The sub-acute and chronic toxicity studies of TSSN were investigated on dog animals.•The LD50 was calculated greater than 3000 mg/Kg.•No toxic appearance was observable after examining the corresponding parameters.Sub-acute and chronic toxic effects of total steroidal saponins (TSSN) extracts from Dioscorea zingiberensis C.H. Wright on various internal organs and biochemical indicators have never been studied before and this study is the first of its kind to demonstrate sub-acute and chronic toxicities of TSSN on dogs. Administration of TSSN extracts at doses up to 3000 mg/Kg daily for 14 days, no biochemical and organ changes were observed on the experimental groups of dogs. Further, chronic toxicity study through oral administration of TSSN extracts at the gradual doses of 50, 250 and 500 mg/Kg for 90 days followed by a 2-week recovery assay revealed absence of significant architectural and morphological changes in internal organs which were confirmed through histopathological examination and merely no significant alteration in the biochemical indicators including hematologic and urine analysis and electrocardiogram compared to the control dogs. This toxicological evaluation came across with the finding that the herbal preparation can be considered as nontoxic and animals could tolerate the extracts at doses up to 500 mg/Kg with LD50 greater than 3000 mg/Kg. It may serve as a preliminary scientific evidence for further therapeutic investigations.Download high-res image (424KB)Download full-size image

•A facile and general approach to imprint glycoprotein in mild conditions.•Combination of boronate affinity and surface imprinting technique to prepare polymers with synergistic selectivity.•Directly aldehyde-functionalized magnetic nanoparticles utilized as supports to simplify the preparation procedure.In light of the significance of glycoprotein biomarkers for early clinical diagnostics and treatments of diseases, it is essential to develop efficient and selective enrichment platforms for glycoproteins. In this study, we present a facile and general strategy to prepare the boronate affinity-based magnetic imprinted nanoparticles. Boronic acid ligands were first grafted on the directly aldehyde-functionalized magnetic nanoparticles through amidation reaction. Then, template glycoproteins were immobilized on the boronic acid-modified magnetic nanoparticles via boronate affinity binding. Subsequently, a thin layer of dopamine was formed to coat the surface of magnetic nanoparticles through self-polymerization. After the template glycoproteins were removed, the cavities that can specific bind the template glycoproteins were fabricated. Adopting horseradish peroxidase as model template, the effects of imprinting conditions as well as the properties and performance of the obtained products were investigated. The resultant imprinted materials exhibit highly favorable features, including uniform surface morphology with thin imprinted shell of about 8 nm, super-paramagnetic property, fast kinetics of 40 min, high adsorption capacity of 60.3 mg g−1, and satisfactory reusability for at least five cycles of adsorption-desorption without obvious deterioration. Meanwhile, the obtained magnetic imprinted nanoparticles could capture target glycoprotein from nonglycoproteins, but also from other glycoproteins because the synergistic selectivity of boronate affinity and imprinting effect. In addition, the facile preparation method shows feasibility in the imprinting of different glycoproteins.

•Gelatin with three functions in synthesis process.•Preparation conditions optimized by CCD-RSM.•High extraction efficiency and favorable selectivity to E2 in water matrices.•Easy and rapid separation by an external magnetic field.•Application for extraction and determination of trace E2 in environmental water samples.Endocrine disrupting compounds (EDCs) are a potential risk for wildlife and humans for their existence in water. The efficient extraction and clean-up steps are required before detection of low concentration levels of EDCs. In this work, a novel water-compatible magnetic molecularly imprinted nanoparticles is synthesized for the selective extraction of 17β-estradiol (E2) in environmental water samples. The preparation is carried out by introducing aldehyde groups to the surface of amino-functionalized magnetic nanoparticles through a simple one-step modification, followed by copolymerization of functional monomer gelatin and template E2 via surface imprinting technique. The gelatin with abundant active groups could not only act as functional monomer reacting with template, but also assemble covalently at the surface of magnetic nanoparticles. At the same time, gelatin would improve the water-compatibility of imprinted materials for attaining high extraction efficiency. To obtain high imprinting effect, the preparation conditions are optimized in detail using Central composite design-response surface methodology. The resultant polymers have uniform spherical shape with a shell thickness of about 8 nm, stable crystalline form, and super-paramagnetic property. Meanwhile, the obtained polymers have high capacity of 12.87 mg g−1 and satisfactory selectivity to template molecule. To testify the feasibility of the magnetic imprinted polymers in sample pretreatment, a method for determination of trace E2 in environmental water samples was set up by combination of solid-phase extraction (SPE) using the prepared polymers as sorbents and HPLC for rapid isolation and determination of E2. The limit of detection of proposed method is 0.04 ng mL−1, the intra- and inter-day relative standard deviations (RSDs) are less than 4.6% and 5.7%, respectively. The recoveries of E2 from environmental water samples are in the range from 88.3% to 99.1% with the RSDs less than 7.2%.

•Protodioscin was obtained with combined traditional and modern separation methods.•Protodioscin alleviated ischemia-reperfusion (I/R) cerebral injury on animal mode.•Anti-inflammatory and anti-apoptosis were its underlying mechanisms.•The neuroprotection of protodioscin was mediated by inactivating NF-κB pathways.The aim of the current research is to investigate the cerebral-protection of protodioscin on a transient cerebral ischemia-reperfusion (I/R) model and to explore its possible underlying mechanisms. The rats were preconditioned with protodioscin at the doses of 25 and 50 mg kg−1 prior to surgery. Then the animals were subjected to right middle cerebral artery occlusion (MCAO) using an intraluminal method by inserting a thread (90 min surgery). After the blood flow was restored in 24 h via withdrawing the thread, some representative indicators for the cerebral injury were evaluated by various methods including TTC-staining, TUNEL, immunohistochemistry, and Western blotting. As compared with the operated rats without drug intervening, treatment with protodioscin apparently lowered the death rate and improved motor coordination abilities through reducing the deficit scores and cerebral infarct volume. What’s more, an apparent decrease in neuron apoptosis detected in hippocampus CA1 and cortex of the ipsilateral hemisphere might attribute to alleviate the increase in Caspase-3 and Bax/Bcl-2 ratio. Meanwhile, concentrations of several main pro-inflammatory cytokines (TNF-α, IL-1β and IL-6) in the serum were also significantly suppressed. Finally, the NF-κB and IκBa protein expressions in the cytoplasm of right injured brain were remarkably up-regulated, while NF-κB in nucleus was down-regulated. Therefore, these observed findings demonstrated that protodioscin appeared to reveal potential neuroprotection against the I/R injury due to its anti-inflammatory and anti-apoptosis properties. This therapeutic effect was probably mediated by the inactivation of NF-κB signal pathways.Download full-size image

•The gracillin was isolated from the rhizome of Dioscorea zingiberensis C.H. Wright.•HPLC–MS/MS was established to detect the concentration of gracillin in rat plasma.•This method was used to evaluate the pharmacokinetic analysis after validation.•This method was accurate, simple and reproducible to analyze the bio-samples.A sensitive and credible high performance liquid chromatography hyphenated to mass spectrometry (HPLC–MS/MS) was established to quantify the concentration of gracillin in rat plasma. The plasma samples were subjected to a direct protein precipitation process with acetonitrile as a precipitant in a single-step. Ginsenoside Rb1 was selected as an internal standard (IS). The chromatographic separation of analyte and IS were carried out on an Inersil ODS-3 C18 column (250 × 4.6 mm, 5 μm) with a binary solvent system containing acetonitrile and 0.1% formic acid in water at a flow rate of 1 mL min−1 under a gradient elution mode. Mass spectrometric detection was performed on a triple quadrupole tandem mass spectrometer by the multiple reaction monitoring (MRM) mode to examine the precursor-to-daughter ion transitions of 1110.3 → 948.2 for IS and 886.1 → 739.9 for gracillin, respectively, in a positive electrospray ionization mode. The calibration curve showed a promising linearity over a concentration range of 0.065–800 ng mL−1 with a better regression coefficient of r2 = 0.9960. The intra- and inter-day precisions (as relative standard deviation) of the assay at three quality control levels were all less than 3.48%, while the intra- and inter-day accuracies (as relative error) ranged from −8.43% to 9.74%, whose data were within the acceptable limits. The mean extraction recoveries of analyte from rat plasma were all more than 74.11%, and no notable matrix effect was observed. Stability experiments revealed that gracillin remained stable throughout the analytical procedure under various stored conditions. The above validated method was successfully used to investigate the pharmacokinetic behaviors of gracillin orally administrated to rats at three proportion doses. The pharmacokinetic analysis would pave the way for understanding the pharmacological actions and provide a meaningful foundation for further development and application in preclinical and clinical use of gracillin in the near future.Download full-size image

•Diosgenin alleviated ischemia-reperfusion (I/R) cerebral injury on animal mode.•Diosgenin suppressed the pro-inflammatory cytokines in animal subjected to MCAO.•Diosgenin ameliorated the imbalance between the pro- and anti-apoptotic factors.•Its potential mechanisms were related with anti-inflammation and anti-apoptosis.•The NF-κB pathways was inactivated after preliminary administration with diosgenin.The aim of the present study is to explore the potential cerebroprotection of diosgenin against the transient focal cerebral ischemia-reperfusion (I/R) injury and its possible underlying mechanisms. The diosgenin at two dose levels, namely 100 and 200 mg kg−1, was intragastrically administrated once daily for 7-day period prior to the surgery. Then, the rats were subjected to middle cerebral artery occlusion (MCAO) using the intraluminal thread for 90 min. After 24 h reperfusion, several diagnostic indicators were evaluated and all animals were sacrificed to harvest their brains and blood for subsequent biochemical analyses. The results indicated that diosgenin treatment significantly inhibited the death rate and improved the impaired neurological functions along with neurological deficit scores and cerebral infarct size as compared with the rats exposed to I/R insult without agents administration. The increase in the number of apoptotic cells determined by TUNEL in the hippocampus CA1 and cortex was also apparently attenuated in the diosgenin treatment group, which was closely correlated with suppression of Caspase-3 activity and Bax/Bcl-2 ratio. In addition the elevated concentrations of pro-inflammatory cytokines (TNF-α, IL-1β, and IL-6) in blood serum of the I/R treated rats were reduced almost to their normal level. Further results obtained from the Western blotting analysis revealed that the protein expression of IκBα in the injured brain was up-regulated, while the p65 subunit of NF-κB was down-regulated in nucleus after the treatment. Collectively, this neuroprotection of diosgenin against I/R injury may be attained through its anti-apoptosis, anti-inflammation and intervening the NF-κB signal pathway properties. Due to the satisfactory findings, diosgenin might be a powerful therapeutic agent to combat the similar disease in future clinic.Download full-size image