Enrichment and Evaluation of Antitumor Properties of Total Flavonoids from Juglans mandshurica Maxim.

Flavonoids are important secondary metabolites found in Juglans mandshurica Maxim., which is a precious reservoir of bioactive substances in China. To explore the antitumor actions of flavonoids (JMFs) from the waste branches of J. mandshurica, the following optimized purification parameters of JMFs by macroporous resins were first obtained. The loading concentration, flow rate, and loading volume of raw flavonoid extracts were 1.4 mg/mL, 2.4 BV/h, and 5 BV, respectively, and for desorption, 60% ethanol (4 BV) was selected to elute JMFs-loaded AB-8 resin at a flow rate of 2.4 BV/h. This adsorption behavior can be explained by the pseudo-second-order kinetic model and Langmuir isotherm model. Subsequently, JMFs were identified using Fourier transform infrared combined with high-performance liquid chromatography and tandem mass spectrometry, and a total of 156 flavonoids were identified. Furthermore, the inhibitory potential of JMFs on the proliferation, migration, and invasion of HepG2 cells was demonstrated. The results also show that exposure to JMFs induced apoptotic cell death, which might be associated with extrinsic and intrinsic pathways. Additionally, flow cytometry detection found that JMFs exposure triggered S phase arrest and the generation of reactive oxygen species in HepG2 cells. These findings suggest that the JMFs purified in this study represent great potential for the treatment of liver cancer.

The Carthamus tinctorius L. genome sequence provides insights into synthesis of unsaturated fatty acids.

Domesticated safflower (Carthamus tinctorius L.) is a widely cultivated edible oil crop. However, despite its economic importance, the genetic basis underlying key traits such as oil content, resistance to biotic and abiotic stresses, and flowering time remains poorly understood. Here, we present the genome assembly for C. tinctorius variety Jihong01, which was obtained by integrating Oxford Nanopore Technologies (ONT) and BGI-SEQ500 sequencing results. The assembled genome was 1,061.1 Mb, and consisted of 32,379 protein-coding genes, 97.71% of which were functionally annotated. Safflower had a recent whole genome duplication (WGD) event in evolution history and diverged from sunflower approximately 37.3 million years ago. Through comparative genomic analysis at five seed development stages, we unveiled the pivotal roles of fatty acid desaturase 2 (FAD2) and fatty acid desaturase 6 (FAD6) in linoleic acid (LA) biosynthesis. Similarly, the differential gene expression analysis further reinforced the significance of these genes in regulating LA accumulation. Moreover, our investigation of seed fatty acid composition at different seed developmental stages unveiled the crucial roles of FAD2 and FAD6 in LA biosynthesis. These findings offer important insights into enhancing breeding programs for the improvement of quality traits and provide reference resource for further research on the natural properties of safflower.

Metagenomics reveals the variations in functional metabolism associated with greenhouse gas emissions during legume-vegetable rotation process.

Legume-based rotation is commonly recognized for its mitigation efficiency of greenhouse gas (GHG) emissions. However, variations in GHG emission-associated metabolic functions during the legume-vegetable rotation process remain largely uncharacterized. Accordingly, a soybean-radish rotation field experiment was designed to clarify the responses of microbial communities and their GHG emission-associated functional metabolism through metagenomics. The results showed that the contents of soil organic carbon and total phosphorus significantly decreased during the soybean-radish process (P < 0.05), while soil total potassium content and bacterial richness and diversity significantly increased (P < 0.05). Moreover, the predominant bacterial phyla varied, with a decrease in the relative abundance of Proteobacteria and an increase in the relative abundance of Acidobacteria, Gemmatimonadetes, and Chloroflexi. Metagenomics clarified that bacterial carbohydrate metabolism substantially increased during the rotation process, whereas formaldehyde assimilation, methanogenesis, nitrification, and dissimilatory nitrate reduction decreased (P < 0.05). Specifically, the expression of phosphate acetyltransferase (functional methanogenesis gene, pta) and nitrate reductase gamma subunit (functional dissimilatory nitrate reduction gene, narI) was inhibited, indicating of low methane production and nitrogen metabolism. Additionally, the partial least squares path model revealed that the Shannon diversity index was negatively correlated with methane and nitrogen metabolism (P < 0.01), further demonstrating that the response of the soil bacterial microbiome responses are closely linked with GHG-associated metabolism during the soybean-radish rotation process. Collectively, our findings shed light on the responses of soil microbial communities to functional metabolism associated with GHG emissions and provide important insights to mitigate GHG emissions during the rotational cropping of legumes and vegetables.

Design, synthesis and biological evaluation of structurally new 4-indolyl quinazoline derivatives as highly potent, selective and orally bioavailable EGFR inhibitors.

Targeting the epidermal growth factor receptor (EGFR) has been recognized as an effective strategy for treating non-small-cell lung cancer (NSCLC). Although several representative EGFR inhibitors have been approved for clinical use, it is highly desirable to develop highly potent and selective EGFR inhibitors with novel scaffolds because of the occurrence of acquired resistance after treatment. Here we first demonstrate that the 4-indolyl quinazoline derivatives could potently inhibit EGFR in vitro and in vivo, of which YS-67 effectively and selectively inhibits EGFR[WT] (IC50 = 5.2 nM), EGFR[d746-750] (IC50 = 9.6 nM) and EGFR[L858R] (IC50 = 1.9 nM). The TREEspot™ kinase interaction map further reveals the binding selectivity toward 468 kinases. YS-67 not only potently suppresses p-EGFR and p-AKT, but also effectively inhibits proliferation of A549 (IC50 = 4.1 µM), PC-9 (IC50 = 0.5 µM) and A431 cells (IC50 = 2.1 µM). YS-67 treatment also causes colony formation inhibition, arrests cell cycle progression at G0/G1 phases and induces apoptosis. More importantly, YS-67 is well tolerated in A431 xenograft model after oral administration, showing effective tumor growth suppression and low toxicity. Collectively, YS-67 represents an underexplored scaffold for developing new EGFR inhibitors.

Inhibition of LSD1 induces ferroptosis through the ATF4-xCT pathway and shows enhanced anti-tumor effects with ferroptosis inducers in NSCLC.

Lysine-specific demethylase 1 (LSD1) has been identified as an important epigenetic target, and recent advances in lung cancer therapy have highlighted the importance of targeting ferroptosis. However, the precise mechanisms by which LSD1 regulates ferroptosis remain elusive. In this study, we report that the inhibition of LSD1 induces ferroptosis by enhancing lipid peroxidation and reactive oxygen species (ROS) accumulation. Mechanistically, LSD1 inhibition downregulates the expression of activating transcription factor 4 (ATF4) through epigenetic modification of histone H3 lysine 9 dimethyl (H3K9me2), which sequentially inhibits the expression of the cystine-glutamate antiporter (xCT) and decreases glutathione (GSH) production. Furthermore, LSD1 inhibition transcriptionally upregulates the expression of transferrin receptor (TFRC) and acyl-CoA synthetase long chain family member 4 (ACSL4) by enhancing the binding of histone H3 lysine 4 dimethyl (H3K4me2) to their promoter sequences. Importantly, the combination of an LSD1 inhibitor and a ferroptosis inducer demonstrates an enhanced anti-tumor effect in a xenograft model of non-small cell lung cancer (NSCLC), surpassing the efficacy of either agent alone. These findings reveal new insights into the mechanisms by which LSD1 inhibition induces ferroptosis, offering potential guidance for the development of new strategies in the treatment of NSCLC.

Discovery of YS-363 as a highly potent, selective, and orally efficacious EGFR inhibitor.

The Epidermal Growth Factor Receptor (EGFR) tyrosine kinase inhibitors (TKIs) are the standard first-line therapy for EGFR-mutated NSCLC. However, long-term clinical treatment often leads to acquired drug resistance, making NSCLC refractory. Therefore, it is essential to design new EGFR inhibitors as potential drugs against NSCLC. This study reports on a novel quinazoline-based compound called YS-363 that acts as a new EGFR inhibitor. YS-363 demonstrated potent inhibition against both wild-type and L858R mutant forms of EGFR with IC50 values of 0.96 nM and 0.67 nM, respectively. Additionally, YS-363 had a reversible inhibitory effect on cellular EGFR signaling, had excellent inhibitory activity on cell proliferation and migration, and induced G0/G1 cell cycle arrest and apoptosis. In xenograft models dependent on EGFR signaling, oral administration of YS-363 substantially suppressed tumor growth by inhibiting this pathway. In summary, YS-363 is a promising selective reversible inhibitor with a novel quinazoline scaffold that can potentially develop more effective anti-lung cancer agents targeting EGFR in patients who have developed resistance to current therapies such as TKIs like gefitinib or erlotinib.

Multiple Metabolites Derived from Mushrooms and Their Beneficial Effect on Alzheimer's Diseases.

Mushrooms with edible and medicinal potential have received widespread attention because of their diverse biological functions, nutritional value, and delicious taste, which are closely related to their rich active components. To date, many bioactive substances have been identified and purified from mushrooms, including proteins, carbohydrates, phenols, and vitamins. More importantly, molecules derived from mushrooms show great potential to alleviate the pathological manifestations of Alzheimer's disease (AD), which seriously affects the health of elderly people. Compared with current therapeutic strategies aimed at symptomatic improvement, it is particularly important to identify natural products from resource-rich mushrooms that can modify the progression of AD. This review summarizes recent investigations of multiple constituents (carbohydrates, peptides, phenols, etc.) isolated from mushrooms to combat AD. In addition, the underlying molecular mechanisms of mushroom metabolites against AD are discussed. The various mechanisms involved in the antiAD activities of mushroom metabolites include antioxidant and anti-neuroinflammatory effects, apoptosis inhibition, and stimulation of neurite outgrowth, etc. This information will facilitate the application of mushroom-derived products in the treatment of AD. However, isolation of new metabolites from multiple types of mushrooms and further in vivo exploration of the molecular mechanisms underlying their antiAD effect are still required.

Combined effects of microplastics and chlortetracycline on the intestinal barrier, gut microbiota, and antibiotic resistome of Muscovy ducks (Cairina moschata).

Antibiotics and microplastics (MPs) have become critical concerns worldwide because of their increasing amount and ecological risks to ecosystems. However, how MPs exposure affects the bioaccumulation and risks of antibiotics in waterfowls remains poorly understood. In this study, Muscovy ducks were exposed to single and combined contamination with polystyrene MPs and chlortetracycline (CTC) for 56 days, and the effects of MPs on CTC bioaccumulation and their risks in duck intestines were investigated. MPs exposure reduced the bioaccumulation of CTC in the intestine and liver of ducks and increased their fecal CTC excretion. MPs exposure caused severe oxidative stress, inflammatory response, and intestinal barrier damages. Microbiome analysis showed that MPs exposure induced microbiota dysbiosis by increasing the abundance of Streptococcus and Helicobacter, the increase of which may exacerbate intestinal damages. Co-exposure to MPs and CTC alleviated the intestinal damage by regulating the gut microbiome. Metagenomic sequencing revealed that the combined exposure to MPs and CTC increased the abundance of Prevotella, Faecalibacterium, and Megamonas and incidence of total antibiotic resistance genes (ARGs), especially tetracycline ARGs subtypes in the gut microbiota. The results obtained herein provide new insights into the potential risks of polystyrene MPs and antibiotics in waterfowls living in aquatic environments.

Transcriptomic responses of the fast-growing bacterium Vibrio natriegens during cold-induced loss of culturability.

Vibrio natriegens has massive biotechnological potential owing to its fast growth rate. However, this bacterium rapidly loses its culturability during low-temperature preservation (LTP), the reason for which is still unknown. To reveal the metabolic responses of V. natriegens during LTP, we analyzed and compared the transcriptome before and after 8 days of preservation at 4 or 25 °C (room-temperature preservation (RTP)) in liquid culture medium. Most genes exhibited significant transcriptional responses to LTP. Using gene set enrichment analysis, we compared the transcriptional responses of different V. natriegens Gene Ontology (GO) sets during LTP or RTP. The enrichment of the GO set "SOS response" during LTP, but not RTP, indicated the occurrence of DNA damage during LTP. The GO set "respiratory electron transport chain" was suppressed during LTP and RTP. Although the GO set "response to oxidative stress" was not significantly altered, we observed an increase in reactive oxygen species (ROS) during LTP, suggesting a relationship between ROS and cold-induced loss of culturability (CILC) in V. natriegens. The faster loss of culturability and accumulation of ROS in 20 mL compared to 100 mL of liquid culture medium further suggested a relationship between CILC and oxygen availability. Furthermore, we showed that the deletion of Na+-translocating NADH-ubiquinone oxidoreductase, but not type-II NADH dehydrogenase, accelerated CILC and increased intracellular ROS levels in V. natriegens. These findings will help to understand the cause of CILC which may lead to improving the stability of V. natriegens at low temperatures.

Quantification of the antagonistic and synergistic effects of Pb2+, Cu2+, and Zn2+ bioaccumulation by living Bacillus subtilis biomass using XGBoost and SHAP.

Bioaccumulation and adsorption are efficient methods for removing heavy metal ions (HMIs) from aqueous environments. However, methods to quantifiably characterize the removal selectivity for co-existing HMIs are limited. In this study, we applied Shapley additive explanations (SHAP) following extreme gradient boosting (XGBoost) modeling, to generate SHAP values. We used these values to create an affinity interference index (AII) that quantitatively represented the interference between metal ions in a multi-metal bioaccumulation system. The selectivity for simultaneous bioaccumulation of Pb2+, Cu2+, and Zn2+ by living Bacillus subtilis biomass was then characterized as a proof of concept. The AII indicated that the bioaccumulation of Zn2+ was more strongly inhibited by Pb2+/Cu2+ (AII = 1) than that of Pb2+/Cu2+ by Zn2+. Moreover, the presence of Zn2+ promoted the bioaccumulation of Pb2+ (AII = 0.39), which was confirmed in further experiments where the bioaccumulation of Pb2+ (300 µM) was increased by 38% with Zn2+ (300 µM). This study demonstrated that the combination of XGBoost and SHAP is effective in the quantifiable characterization of the antagonistic and synergistic effects in a multi-metal simultaneous bioaccumulation system. This method could also be generalized to similar tasks for analyzing the selectivity effects in a multi-component system.

Genome-wide analysis of CtNF-YB and lipid synthesis regulation of CtNF-YB12 in Carthamus tinctorius L.

KEY MESSAGE: The nuclear Factor YB of Carthamus tinctorius L. increased the content of unsaturated fatty acids by regulating the expression of genes involved in fatty acid synthesis and oil accumulation. Safflower (Carthamus tinctorius L.) seed oil is rich in linoleic acid and is widely used in food and medicine. Therefore, key genes regulating oil synthesis were mined through genetic engineering to provide genetic resources for improving oil content. Based on the conserved domain of the NF-YB, we screened and identified 14 CtNF-YB transcription factors in the safflower genome and divided them into three subfamilies through phylogenetic analysis. Regulatory motif analysis of the CtNF-YB promoter revealed specific cis-regulatory elements related to abiotic stress, growth, and development. Expression analysis of CtNF-YB family genes showed that non-Leafy Cotyledon 1(non-LEC1) genes were highly expressed in roots, leaves, and flowers; Leafy Cotyledon 1(LEC1) genes were highly expressed during early seed development; and Dr1-like genes were highly expressed in roots, stems, and leaves. CtNF-YB12 was identified as a LEC1 transcription factor based on phylogeny and BLAST alignment. Heterologous CtNF-YB12 expression in Arabidopsis thaliana increased seed pod length and seed size. Moreover, CtNF-YB12 overexpression increased the oil content of seeds, upregulated genes involved in fatty acid biosynthesis and glycolysis, and altered the content of unsaturated fatty acids, including oleic acid (C18:1), linoleic acid (C18:2), and linolenic acid (C18:3), as well as of sucrose, fructose, and glucose. CtNF-YB12 may increase the oil content by regulating key enzyme genes of oil synthesis, so it can be used as a reliable target.

Plastisphere microbiome: Methodology, diversity, and functionality.

Broad topics of the plastisphere in various environments are reviewed, including its methodologies, diversity, functionality, and outlook.

Beneficial Effects of Oleosomes Fused with Human Fibroblast Growth Factor 1 on Wound Healing via the Promotion of Angiogenesis.

In our previous study, human fibroblast growth factor 1 was successfully fused with oleosomes, energy-storing organelles of seeds, which are considered to be excellent "expression carriers" for substances with a convenient purification process. The present work aimed to explore the beneficial effects of oleosomes fused with human fibroblast growth factor 1 (OLAF) on wound healing. The data showed marked improvements in terms of the angiogenesis, vascular integrity, collagen and inflammation on the wound sites of rats with a full-thickness skin defect. Moreover, the positive role of OLAF in promoting angiogenesis and its possible pathways were clarified in vivo and in vitro. The results showed that the number, length and branches of the blood vessels of the chick embryo chorioallantoic membrane were markedly increased after OLAF treatment. Meanwhile, the in vitro results also revealed that 100 ng/mL OLAF exhibited a promoting effect on the proliferation, migration and tube formation of human umbilical vein endothelial cells. In addition, the potential of OLAF to improve wound angiogenesis was demonstrated to be associated with an up-regulated PI3K/Akt pathway by transcriptome sequencing analysis and the introduction of a PI3K/Akt pathway inhibitor (LY294002). These findings suggest that OLAF has many prospects in the development of drugs for wound healing.

Xanthan gum/oil body-microgel emulsions with enhanced transdermal absorption for accelerating wound healing.

The oil body comprises lipid droplets surrounded by a surface embedded with oil body-related proteins. To form a drug delivery system, an oleosin can be fused with foreign proteins and bound to the oil body surface. Here, safflower oil bodies carrying oleosin-human epidermal growth factor (hEGF) were mixed with xanthan gum to form self-assembled polymers, referred as an oil body microgel emulsion (OBEME) without any chemical crosslinking agent. The physicochemical properties of OBEME were evaluated and compared with those of natural lipid droplets. The electrostatic interaction between xanthan gum and oil bodies prevents excessive cross-linking and forms a uniform network structure. The basic properties of OBEME were characterized by scanning electron microscopy, cryo-scanning electron microscopy, rheology, and thermogravimetric analysis. The OBEME is an interconnected network and presents a smooth surface without any pores; it remains stable at room temperature for 90 days, and is not affected by low-speed centrifugation and repeated freeze-thaw cycles as indicated by particle size, potential, and fluorescence microscopy analyses. The OBEME enlarges the skin tissue gap, enhances skin permeability, and shows a good slow-release effect in the transdermal absorption test in vivo. It demonstrates a wound healing effect; further, it regulates the inflammatory response of full-layer skin wounds in rats, as well as accelerate angiogenesis, and promote re-epithelialization and remodeling. The OBEME as a bioactive molecule-carbohydrate complex can effectively accelerate skin regeneration and has great translational potential to provide low-cost alternative wound care treatments.

Exploration of Strategies for Organic Integration of Labor Education and Occupational Therapy for College Students.

This paper analyzes the actual functions of the labor education and occupational therapy phase through an in-depth study of labor education and occupational therapy for college students and explores the strategies of organic integration of labor education and occupational therapy for college students. This paper carries out a holistic theoretical construction of labor education in schools and systematically analyzes the components of labor education objectives in colleges and universities, labor education contents in schools, labor education implementation in schools, and labor education evaluation in schools. The main goal of labor education in colleges and universities is to guide students in establishing a scientific concept of labor and constructing a labor education resource system. It focuses on strengthening the construction and improvement of the labor education theoretical system, promoting the construction of labor education disciplines and professional courses, and attaching importance to the training of labor education teachers in the new era to contribute to the perfection of labor education. The system of labor education practice should be innovated. Of course, the labor practice activities based on institutionalization should be implemented. The cycle of labor activities based on the principle of continuity should be strengthened, and the effectiveness of labor education should be improved totally from explicit and implicit instructions. The core of occupational therapy is to help college students adapt to social work, give full play to their self-worth, obtain economic resources, and improve self-confidence. The organic integration of labor education and occupational therapy shows that it can help students ease their social adaptability and adapt to social functions, improve the reemployment rate, and improve students' overall health. Therefore, the practical research on labor education and occupational therapy should be strengthened.

Ultrasonic-Assisted Extraction of Flavonoids from Juglans mandshurica Maxim.: Artificial Intelligence-Based Optimization, Kinetics Estimation, and Antioxidant Potential.

Ultrasonic-assisted extraction (UAE) of flavonoids (JMBF) from Juglans mandshurica Maxim., an important industrial crop in China, was investigated in the present study. To improve the extraction efficiency of JMBF, suitable UAE was proposed after optimization using a hybrid response surface methodology-artificial neural network-genetic algorithm approach (RSM-ANN-GA). The maximum extraction yield (6.28 mg·g-1) of JMBF was achieved using the following optimum UAE conditions: ethanol concentration, 62%; solid-liquid ratio, 1:20 g·mL-1; ultrasonic power, 228 W; extraction temperature, 60 °C; extraction time, 40 min; total number of extractions, 1. Through the investigation of extraction kinetics, UAE offered a higher saturated concentration (Cs) for JMBF in comparison to traditional solvent extraction (TSE). Scanning electron microscopy (SEM) images showed that deeper holes were generated in J. mandshurica powder under the action of ultrasound, indicating that ultrasound significantly changed the structure of the plant materials to facilitate the dissolution of active substances. Extracts obtained using UAE and TSE were compared by Fourier-transform infrared spectroscopy analysis, the results of which revealed that the functional group of bioactive compounds in the extract was unaffected by the ultrasonication process. Moreover, JMBF was further shown to exhibit significant antioxidant properties in vitro. This study provides a basis for the application of JMBF as a natural antioxidant.

Toxicity alleviation and metabolism enhancement of nonylphenol in green algae Dictyosphaerium sp. by NaHCO3.

Nonylphenol (NP) toxicity limits the improvements in its algal remediation efficiency. This study comprehensively investigated the performance and mechanism of NaHCO3-driving effects on NP-exposed algae. The results showed that NaHCO3 enhanced algal resistance to NP and the corresponding EC50 values increased 1.31-4.25 times. Further, the toxicological effects of NP reduced with increasing pyrenoid volume and chlorophyll and carotenoids production, and decreasing cellular damage degree. Moreover, the concentration of extracellular polymeric substances was enhanced and more NP adsorption sites were formed. Consistently, RNA-seq demonstrated significant expression alterations in genes related to energy metabolism, cellular synthesis, photosynthesis, and carbon fixation. Besides, NP biodegradation rate was increased by 15.2 % and 11.1 % in the 1, and 4 mg/L NP treatments, respectively. Identification of degradation intermediates and their toxicity via Ecological Structure Activity Relationship program showed that NaHCO3 accelerated sequential α-C removal from NP in algae with faster generation of less toxic metabolites, namely, 4-ethylphenol, 4-cresol and 4-hydroxybenzoic acid. This study provides new insights into the role of NaHCO3 in toxicity alleviation and metabolism enhancement of NP in algae and can assist NP bioremediation efforts in aquatic environment.