Structure Elucidation of a Novel Polysaccharide Isolated from Euonymus fortunei and Establishing Its Antioxidant and Anticancer Properties.

Euonymusfortunei polysaccharides (EFPs) have not been extensively investigated yet in terms of their extraction and biological activity. The orthogonal experimental design was employed in this study to evaluate the optimum yield of EFPs. A maximum yield of 2.63 ± 0.23% was attained using material-liquid ratios of 60 mL/g, extraction temperature of 80°C, ultrasonic power of 144 W, and extraction time of 75 mins. The polysaccharide content reached 53.47 ± 0.31% when deproteinized thrice. An analysis of monosaccharide composition revealed that these polysaccharides consist of Gal, Glc, Man, Fuc, and Rha with a molar ratio of 7.14 ∶ 23.99 ∶ 6.29 ∶ 6.55 ∶ 1.00, respectively, in EFPs. Subsequently, the in vitro scavenging capacities of 2,2-diphenylpicrylhydrazyl (DPPH) and ·OH and superoxide anion radicals, along with the reducing power of EFPs, were studied. Results revealed that EFPs have higher antioxidant activity, particularly ·OH scavenging, as well as reducing power, as compared to Astragalus polysaccharides (ASPs) and Lycium barbarum polysaccharides (LBPs). The Cell Counting Kit-8 (CCK-8) method was used to evaluate the effects of different concentrations of polysaccharides on SKOV3 cell proliferation, and the results revealed their inhibition at concentrations in the range of 200-800 µg/mL. In addition, findings from flow cytometry further confirmed that EFPs blocked the cell cycle at G0/G1 and S phases and induced SKOV3 cell apoptosis. In a word, EFPs could be exploited and used further based on the experimental results from this study.

Vegetative cell wall protein OsGP1 regulates cell wall mediated soda saline-alkali stress in rice.

Plant growth and development are inhibited by the high levels of ions and pH due to soda saline-alkali soil, and the cell wall serves as a crucial barrier against external stresses in plant cells. Proteins in the cell wall play important roles in plant cell growth, morphogenesis, pathogen infection and environmental response. In the current study, the full-length coding sequence of the vegetative cell wall protein gene OsGP1 was characterized from Lj11 (Oryza sativa longjing11), it contained 660 bp nucleotides encoding 219 amino acids. Protein-protein interaction network analysis revealed possible interaction between CESA1, TUBB8, and OsJ_01535 proteins, which are related to plant growth and cell wall synthesis. OsGP1 was found to be localized in the cell membrane and cell wall. Furthermore, overexpression of OsGP1 leads to increase in plant height and fresh weight, showing enhanced resistance to saline-alkali stress. The ROS (reactive oxygen species) scavengers were regulated by OsGP1 protein, peroxidase and superoxide dismutase activities were significantly higher, while malondialdehyde was lower in the overexpression line under stress. These results suggest that OsGP1 improves saline-alkali stress tolerance of rice possibly through cell wall-mediated intracellular environmental homeostasis.

Functional Study of Amorpha fruticosa WRKY20 Gene in Response to Drought Stress.

The WRKY gene family in plants regulates the plant's response to drought through regulatory networks and hormone signaling. AfWRKY20 (MT859405) was cloned from Amorpha fruticosa (A. fruticosa) seedlings using RT-PCR. The binding properties of the AfWRKY20 protein and the W-box (a DNA cis-acting element) were verified both in vivo and in vitro using EMSA and Dual-Luciferase activity assays. RT-qPCR detected that the total expression level of AfWRKY20 in leaves and roots was 22 times higher in the 30% PEG6000 simulated drought treatment compared to the untreated group. Under the simulated drought stress treatments of sorbitol and abscisic acid (ABA), the transgenic tobacco with the AfWRKY20 gene showed enhanced drought resistance at the germination stage, with significantly increased germination rate, green leaf rate, fresh weight, and root length compared to the wild-type (WT) tobacco. In addition, the superoxide dismutase (SOD) activity, chlorophyll content, and Fv/Fm ratio of AfWRKY20 transgenic tobacco were significantly higher than those of the WT tobacco under natural drought stress, while the malondialdehyde (MDA) content and 3,3'-diaminobenzidine (DAB) and nitroblue tetrazolium (NBT) staining levels were lower. The expression levels of oxidation kinase genes (NbSOD, NbPOD, and NbCAT) in transgenic tobacco under drought stress were significantly higher than those in WT tobacco. This enhancement in gene expression improved the ability of transgenic tobacco to detoxify reactive oxygen species (ROS). The survival rate of transgenic tobacco after natural drought rehydration was four times higher than that of WT tobacco. In summary, this study revealed the regulatory mechanism of AfWRKY20 in response to drought stress-induced ABA signaling, particularly in relation to ROS. This finding provides a theoretical basis for understanding the pathways of WRKY20 involved in drought stress, and offers genetic resources for molecular plant breeding aimed at enhancing drought resistance.

Beyond adaptive cruise control and lane centering control: drivers' mental model of and trust in emerging ADAS technologies.

Introduction: The potential safety benefits of advanced driver assistance systems (ADAS) highly rely on drivers' appropriate mental models of and trust in ADAS. Current research mainly focused on drivers' mental model of adaptive cruise control (ACC) and lane centering control (LCC), but rarely investigated drivers' understanding of emerging driving automation functions beyond ACC and LCC. Methods: To address this research gap, 287 valid responses from ADAS users in the Chinese market, were collected in a survey study targeted toward state-of-the-art ADAS (e.g., autopilot in Tesla). Through cluster analysis, drivers were clustered into four groups based on their knowledge of traditional ACC and LCC functions, knowledge of functions beyond ACC and LCC, and knowledge of ADAS limitations. Predictors of driver grouping were analyzed, and we further modeled drivers' trust in ADAS. Results: Drivers in general had weak knowledge of LCC functions and functions beyond ACC and LCC, and only 27 (9%) of respondents had a relatively strong mental model of ACC and LCC. At the same time, years of licensure, weekly driving distance, ADAS familiarity, driving style (i.e., planning), and personability (i.e., agreeableness) were associated with drivers' mental model of ADAS. Further, it was found that the mental model of ADAS, vehicle brand, and drivers' age, ADAS experience, driving style (i.e., focus), and personality (i.e., emotional stability) were significant predictors of drivers' trust in ADAS. Discussion: These findings provide valuable insights for the design of driver education and training programs to improve driving safety with ADAS.

Functional Identification and Genetic Transformation of the Ammonium Transporter PtrAMT1;6 in Populus.

The ammonium transporter (AMT) family gene is an important transporter involved in ammonium uptake and transfer in plants and is mainly engaged in the uptake and transport of ammonium from the environment by roots and the reabsorption of ammonium in the aboveground parts. In this study, the expression pattern, functional identification, and genetic transformation of the PtrAMT1;6 gene, a member of the ammonium transporter protein family in P. trichocarpa, were investigated as follows: (1) Fluorescence quantitative PCR demonstrated that the PtrAMT1;6 gene was preferentially expressed in the leaves, with both dark-induced and light-inhibited expression patterns. (2) A functional restoration assay using the yeast ammonium transporter protein mutant strain indicated that the PtrAMT1;6 gene restored the ability of the mutant to transport ammonium with high affinity. (3) Arabidopsis was transformed with pCAMBIA-PtrAMT1;6P, and the transformed lines were stained with GUS, which showed that the rootstock junction, cotyledon petioles, and the leaf veins and pulp near the petioles of the transformed plants could be stained blue, indicating that the promoter of the PtrAMT1;6 gene had promoter activity. (4) The overexpression of the PtrAMT1;6 gene caused an imbalance in carbon and nitrogen metabolism and reduced nitrogen assimilation ability in '84K' poplar and ultimately reduced biomass. The above results suggest that PtrAMT1;6 may be involved in ammonia recycling during nitrogen metabolism in aboveground parts, and overexpression of PtrAMT1;6 may affect the process of carbon and nitrogen metabolism, as well as nitrogen assimilation in plants, resulting in stunted growth of overexpression plants.

Effects of Gypenoside XLIX on fatty liver cell gene expression in vitro: a genome-wide analysis.

OBJECTIVE: To perform Genome-wide analysis of Gypenoside XLIX (Gyp-XLIX) in the treatment of fatty liver cells. METHODS: The gene profiles of 3 normal liver cells, 3 fatty liver cells, and 3 fatty liver cells treated with Gyp-XLIX were detected by high-throughput sequencing to identify the differentially expressed genes (DEGs) in fatty liver treated by Gyp-XLIX. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis were used to explore the biological functions of DEGs. By constructing lncRNA-mRNA co-expression network of DEGs, network node genes were mined. Possible target genes of differentially expressed lncRNA were predicted by cis regulation. RESULTS: 782 DEGs were screened out; that is, 172 genes were highly expressed in fatty liver cells, and the expression decreased to the level of normal liver cells after Gyp-XLIX treatment; 610 genes were under expressed in fatty liver cells, and the expression increased to the level of normal liver cells after Gyp-XLIX treatment. Functional analysis of KEGG and GO showed that DEGs process DNA-binding transcription factor activity and ion transmembrane transporter activity in the plasma membrane region. This mediates glycerophospholipid metabolism, bile secretion, fatty acid degradation and other signaling pathways. lncRNA analysis showed that the expression of 16 lncRNAs was low in fatty liver cells, and the expression was increased to the level of normal liver cells after Gyp-XLIX treatment. Target gene prediction showed that 16 differentially expressed lncRNAs had cis potential to regulate target genes, among which lncRNA RPARP-AS1 had a high degree of relationship with other genes. lncRNA-mRNA co-expression network results showed that lncRNA RPARP-AS1 may acted on NFKB2. CONCLUSION: LncRNA was differentially expressed in fatty liver cells and Gyp-XLIX treated fatty liver cells, and lncRNA RPARP-AS1 may be a regulatory gene in Gyp-XLIX treated fatty liver.

Iron-Catalyzed Sulfonylthiocyanation of α,ß-Unsaturated Amides/Esters via the Insertion of Sulfur Dioxide.

An iron-catalyzed four-component sulfonylthiocyanation between α,ß-unsaturated amides/esters, TMSNCS, aryldiazonium tetrafluoroborates, and sulfur dioxide (from SOgen) is demonstrated. This protocol is characterized by mild reaction conditions, good functional group compatibility, broad substrate scope, and good to excellent yields, providing a feasible method for the preparation of ß-thiocyanated sulfone compounds. The preliminary mechanism investigation shows that a radical pathway may be involved in the process.

Thermally Activated n-Doping of Organic Semiconductors Achieved by N-Heterocyclic Carbene Based Dopant.

Molecular doping plays an important role in the modification of carrier density of organic semiconductors thus enhancing their optoelectronic performance. However, efficient n-doping remains challenging, especially owing to the lack of strongly reducing and air-stable n-dopants. Herein, an N-heterocyclic carbene (NHC) precursor, DMImC, is developed as a thermally activated n-dopant with the excellent stability in air. Its thermolysis in situ regenerates free NHC and subsequently dopes typical organic semiconductors. In sequentially doped FBDPPV films, DMImC does not disturb the π-π packing of the polymer and achieves good miscibility with the polymer. As a result, a high electrical conductivity of up to 8.4 S cm-1 is obtained. Additionally, the thermally activated doping and the excellent air stability permit DMImC to be noninteractively co-processed with polymers in air. Our results reveal that DMImC can be served as an efficient n-dopant suitable for various organic semiconductors.

Development and Validation of a Wearable Inertial Sensors-Based Automated System for Assessing Work-Related Musculoskeletal Disorders in the Workspace.

The industrial societies face difficulty applying traditional work-related musculoskeletal disorder (WMSD) risk assessment methods in practical applications due to in-situ task dynamics, complex data processing, and the need of ergonomics professionals. This study aims to develop and validate a wearable inertial sensors-based automated system for assessing WMSD risks in the workspace conveniently, in order to enhance workspace safety and improve workers' health. Both postural ergonomic analysis (RULA/REBA) and two-dimensional static biomechanical analysis were automatized as two toolboxes in the proposed system to provide comprehensive WMSD risk assessment based on the kinematic data acquired from wearable inertial sensors. The effectiveness of the developed system was validated through a follow-up experiment among 20 young subjects when performing representative tasks in the heavy industry. The RULA/REBA scores derived from our system achieved high consistency with experts' ratings (intraclass correlation coefficient ≥0.83, classification accuracy >88%), and good agreement was also found between low-back compression force from the developed system and the reference system (mean intersystem coefficient of multiple correlation >0.89 and relative error <9.5%). These findings suggested that the wearable inertial sensors-based automated system could be effectively used for WMSD risk assessment of workers when performing tasks in the workspace.

A thermally activated and highly miscible dopant for n-type organic thermoelectrics.

N-doping plays an irreplaceable role in controlling the electron concentration of organic semiconductors thus to improve performance of organic semiconductor devices. However, compared with many mature p-doping methods, n-doping of organic semiconductor is still of challenges. In particular, dopant stability/processability, counterion-semiconductor immiscibility and doping induced microstructure non-uniformity have restricted the application of n-doping in high-performance devices. Here, we report a computer-assisted screening approach to rationally design of a triaminomethane-type dopant, which exhibit extremely high stability and strong hydride donating property due to its thermally activated doping mechanism. This triaminomethane derivative shows excellent counterion-semiconductor miscibility (counter cations stay with the polymer side chains), high doping efficiency and uniformity. By using triaminomethane, we realize a record n-type conductivity of up to 21 S cm-1 and power factors as high as 51 µW m-1 K-2 even in films with thicknesses over 10 µm, and we demonstrate the first reported all-polymer thermoelectric generator.

Identification of novel and potent small-molecule inhibitors of tubulin with antitumor activities by virtual screening and biological evaluations.

Microtubules (made up of α and ß-tubulin subunits) play an essential role in the process of mitosis and cell proliferation, thus making them an ideal target for anticancer drugs discovery. Microtubule-targeted drugs, including taxanes and vinca alkaloids, can suppress microtubule dynamics, cause mitotic block and apoptosis, which have been widely used in the treatment of various cancers. There are three unique binding sites (taxanes, vinca alkaloids, and colchicine) in tubulin can be targeted to develop tubulin inhibitors. In this study, we selected the colchicine binding site in tubulin as our target. By performing molecular docking-based virtual screening combined with in vitro tubulin polymerization inhibition assay, we identified two novel and potent tubulin inhibitors (9 and 19). These two compounds arrested cell cycle progression at the G2/M phase and induced apoptosis at sub µM concentrations. In addition, they displayed potent antiproliferative activity with IC50 values in the nM range. Finally, the probable binding modes of 9 and 19 were probed by molecular docking. These two compounds with novel scaffold will shed new light on the lead molecules discovery and the design of new microtubule-targeting agents (MTAs).

Micheliolide alleviates hepatic steatosis in db/db mice by inhibiting inflammation and promoting autophagy via PPAR-γ-mediated NF-кB and AMPK/mTOR signaling.

The anti-inflammatory, immunomodulatory, and anticancer effects of micheliolide (MCL) isolated from Michelia champaca were previously reported, but its role and underlying mechanisms in relieving liver steatosis remain unclear. Herein, we investigated the effects of MCL on hepatic steatosis using a db/db mouse model and lipid mixture (LM)-induced AML12 and LO2 cells. The body and liver weights, food consumption, lipid content and liver aminotransferase levels in serum, the lipid content and inflammatory cytokine levels in liver tissue, and the extent of hepatic steatosis in db/db mice were increased compared with those in db/m mice, and these increases were reversed by MCL treatment. Similarly, MCL also attenuated the inflammatory responses and lipid accumulation in LM-treated AML12 and L02 cells by upregulating PPAR-γ and decreasing p-IкBα and p-NF-κB/p65, thereby inhibiting the NF-κB pathway and reducing lipotoxicity. Furthermore, MCL administration increased LC3B, Atg7 and Beclin-1 expression and the LC3B-II/I ratio in db/db mouse livers and LM-treated AML12 and L02 cells, and these MCL-induced increases were mediated by the activation of PPAR-γ and p-AMPK and inhibition of p-mTOR and induce autophagy. These effects were blocked by PPAR-γ and AMPK inhibitors. Our findings suggest that MCL ameliorates liver steatosis by upregulating PPAR-γ expression, thereby inhibiting NF-κB-mediated inflammation and activating AMPK/mTOR-dependent autophagy.

Dietary taurine impairs intestinal growth and mucosal structure of broiler chickens by increasing toxic bile acid concentrations in the intestine.

Three experiments were conducted to determine the effect of taurine on the intestinal development, bile acid concentrations, and hormonal status of chickens. In experiment 1, a total of 250 one-day-old broilers were randomly allocated to 5 treatments and supplemented with 0, 0.25, 0.50, 1.00, and 2.00 g/kg of taurine, respectively. Growth performance, weight and length of the small intestine, and intestinal morphology were measured on d 7, 22, and 44. The gene expression levels of several hormones, including epidermal growth factor and cholecystokinin, were also evaluated. In experiment 2, 60 one-day-old broilers were supplemented with 0, 1.0, and 5.0 g/kg of taurine to assess cell proliferation in the jenunal crypt. In experiment 3, 100 newly hatched broilers were assigned randomly to 5 treatments (0, 0.10, 0.50, 2.00, 8.00 g/kg of taurine) to evaluate the bile acid concentrations in the jejunal mucosa. Our results indicated that dietary taurine decreased the length and weight of small intestine, the villus width, surface area, and crypt depth in the duodenum and jejunum (P < 0.05). Taurine also increased the expression of cholecystokinin and epidermal growth factor on the jejunal mucosa (P < 0.001). Taurine has little effect on stimulating the proliferation of intestinal crypt cells, except for 5 g/kg of taurine supplementation on d 14 (P < 0.05). Additionally, a linear increase in the jejunal concentrations of taurocholic acid, taurochenodeoxycholic acid, and taurolithocholic acid was observed on d 7 in broilers fed increasing levels of taurine. In conclusion, we suggested that taurine impairs intestinal mucosal development partly through generation of toxic bile acids.

[The inhibitory effect of compound liuyuexue on DHBV DNA].

OBJECTIVE: To study the inhibitory effect of traditional Chinese medicine Compound Liuyuxeue(CLYX) on duck hepatitis B virus (DHBV) DNA, and provide experimental basis for developing a new drug for the clinical treatment. METHODS: One - day old Guangxi brown spotted ducks infected with DHBV were used as the hepatitis B virus infected animal model. Positive ducks were detected by PCR at 13 days after the infection of DHBV, and were randomly divided into five groups: the high dose group, middle dose group and low dose group of Compound Liuyexue( CLYX) , model group, positive control group. Every group had 10 ducks and CLYX was given for 14 days. The content of DHBV DNA in serum were measured by Fluoresceence Quantitative PCR( FQ-PCR). RESULTS: The serum DHBV DNA content was decreased significantly by the treatment with CLYX. The high dose group and middle dose group of CLYX could significantly inhibit DHBV DNA replication in vivo( P <0. 01). DHBV DNA content in serum in high dose group and middle dose group did not return significantly 3 days after stopping treatment, and its inhibitory effects were dose-and tim-dependents. CONCLUSION: CLYX can inhibit significantly DHBV DNA.