Cancer is one of the most important problems of modern societies. Recently, studies have reported the anticancer properties of rosiglitazone related to its ability to bind peroxisome proliferator receptor γ (PPARγ), which has various effects on cancer and can inhibit cell proliferation. In this study, we investigated the effect of new 4-thiazolidinone (4-TZD) hybrids Les-4369 and Les-3467 and their effect on reactive oxygen species (ROS) production, metabolic activity, lactate dehydrogenase (LDH) release, caspase-3 activity, and gene and protein expression in human foreskin fibroblast (BJ) cells and lung adenocarcinoma (A549) cells. The ROS production and caspase-3 activity were mainly increased in the micromolar concentrations of the studied compounds in both cell lines. Les-3467 and Les-4369 increased the mRNA expression of PPARG, P53 (tumor protein P53), and ATM (ATM serine/threonine kinase) in the BJ cells, while the mRNA expression of these genes (except PPARG) was mainly decreased in the A549 cells treated with both of the tested compounds. Our results indicate a decrease in the protein expression of AhR, PPARγ, and PARP-1 in the BJ cells exposed to 1 µM Les-3467 and Les-4369. In the A549 cells, the protein expression of AhR, PPARγ, and PARP-1 increased in the treatment with 1 µM Les-3467 and Les-4369. We have also shown the PPARγ modulatory properties of Les-3467 and Les-4369. However, both compounds prove weak anticancer properties evidenced by their action at high concentrations and non-selective effects against BJ and A549 cells. Full article

A high alkaline pH was previously demonstrated to enhance the extraction yield of brewer’s spent grains (BSG) proteins. The effects of extraction pH beyond the extraction yield, however, has not been investigated before. The present work examined the effects of extraction pH (pH 8–12) on BSG proteins’ (1) amino acid compositions, (2) secondary structures, (3) thermal stability, and (4) functionalities (i.e., water/oil holding capacity, emulsifying, and foaming properties). The ideal extraction temperature (60 °C) and BSG-to-solvent ratio (1:20 w/v) for maximizing the extraction yield were first determined to set the conditions for the pH effect study. The results showed that a higher extraction pH led to more balanced compositions between hydrophilic and hydrophobic amino acids and higher proportions of random coils structures indicating increased protein unfolding. This led to superior emulsifying properties of the extracted proteins with more than twofold improvement between pH 8 and a pH larger than 10. The extraction pH, nevertheless, had minimal impact on the water/oil holding capacity, foaming properties, and thermal denaturation propensity of the proteins. The present work demonstrated that a high alkaline pH at pH 11–12 was indeed ideal for both maximizing the extraction yield (37–46 wt.%) and proteins’ functionalities. Full article

Banks have enormous potential to support the achievement of sustainable development goals (SDGs) in accordance with their function as financial intermediaries through sustainable lending. However, the average national financing growth for the sustainable business activity category over the past four years is still 12%. The aim of this research is to identify the conditions of sustainable lending at state-owned conventional banks and analyze the influence of the value chain, economic performance, and ESG performance on sustainable lending. The research was conducted at state-owned commercial banks in Indonesia. The research utilized structural equation modeling (SEM). The SEM results of this study describe value chain, ESG performance (environmental, social, and governance), and economic performance and have direct and positive influences on sustainable lending, of which ESG performance has the biggest influence. As per each individual aspect, social orientation makes the biggest contribution toward sustainable lending. Full article

The safety condition of concrete gravity dams is influenced by multiple factors, and assessing their safety solely based on a single factor is difficult to comprehensively evaluate. Therefore, this paper proposes a comprehensive modeling and analysis approach to assess dam safety by considering long-term temperature, stress, and strain monitoring data of actual concrete gravity dams. Firstly, the K-means clustering algorithm is utilized to classify the data. Then, the study area of the dam is meshed and three indicator evaluation values for all the elements are calculated. The other elements’ evaluation values can be obtained by the Inverse Distance Weighting (IDW) method. Finally, the analytic hierarchy process extended by the D numbers preference relation (D-AHP) method is applied to compute the weights of temperature, stress, and strain and evaluate the dam’s safety comprehensively. The effectiveness of this method is validated through application to specific engineering cases. The results demonstrate that compared to assessing methods considering only single factors, the comprehensive evaluation method proposed in this paper can more comprehensively and accurately reflect the actual safety condition of concrete gravity dams, providing important references for engineering decision-making. Full article

Phellinotus (Polyporales) is a common genus of wood-decay fungi in tropical and subtropical areas, endemic to the Seasonally Dry Tropical Forest (SDTF) biome. However, Phellinotus diversity remains unexplored, despite being a major threat to living trees. Therefore, this study is aimed at confirming and characterizing through morphological and molecular data the first isolates of Phellinotus teixeirae in Pithecellobium dulce (Fabaceae) trees (locally referred to as ‘Chiminango’) from the endangered Colombian SDTF biome. Fifteen fungal specimens were recovered from living P. dulce trees, in the urban area and at the Universidad del Valle campus, and classified as P. teixeirae based on taxonomical descriptors. Phylogenetic relationships were inferred from a four-loci dataset (ribosomal and gene-coding regions), including 82 taxa covering 3991 nucleotide positions. The analysis recovered seven highly supported (>90% bootstrapping) monophyletic taxa of the ‘Phellinotus Clade’, and confirmed the new distribution range of P. teixeirae (100% bootstrap support), which extends approx. 1000 km north in the Neotropics. Hierarchical stratified Analysis of MOlecular VAriance (AMOVA) provided a clear genetic distinction between species (70% of variation, p-value = 0.001) and low differentiation among country of origin within species (11%, p-value = 0.044). Discriminant Analysis for Principal Components (DAPC) indicated complex clustering including closely related species, probably a signal of recent radiation and weak species boundaries. Median-joining haplotype network analysis identified unique haplotypes, which may correlate with new host colonization and population expansion (Tajima’s D ≤ −0.5). In conclusion, this study provides the first assessment of the genetic diversity of P. teixeirae in a novel geography (SDTP) and host tree (P. dulce). However, increasing the number of isolates remains critical to understand further the genus’ distribution patterns and drivers of genetic diversity. Full article

Bovine serum albumin (BSA) hydrogels are non-immunogenic, low-cost, biocompatible, and biodegradable. In order to avoid toxic cross-linking agents, gellan was oxidized with NaIO₄ to obtain new functional groups like dialdehydes for protein-based hydrogel cross-linking. The formed dialdehyde groups were highlighted with FT-IR and NMR spectroscopy. This paper aims to investigate hydrogel films for biomedical applications obtained by cross-linking BSA with oxidized gellan (OxG) containing immobilized β-cyclodextrin–curcumin inclusion complex (β-CD–Curc) The β-CD–Curc improved the bioavailability and solubility of Curc and was prepared at a molar ratio of 2:1. The film’s structure and morphology were evaluated using FT-IR spectroscopy and SEM. The swelling degree (Q%) values of hydrogel films depend on hydrophilicity and pH, with higher values at pH = 7.4. Additionally, the conversion index of -NH₂ groups into Schiff bases increases with an increase in OxG amount. The polymeric matrix provides protection for Curc, is non-cytotoxic, and enhances antioxidant activity. At pH = 5.5, the skin permeability and release efficiency of encapsulated curcumin were higher than at pH = 7.4 because of the interaction of free aldehyde and carboxylic groups from hydrogels with amine groups from proteins present in the skin membrane, resulting in a better film adhesion and more efficient curcumin release. Full article

This paper introduces a new type of flexible rudder surface based on the cosine-type zero Poisson’s ratio honeycomb to enhance the adaptive capabilities of aircraft and enable multi-condition, rudderless flight. The zero Poisson’s ratio honeycomb structure exhibits exceptional in-plane and out-of-plane deformation capacities, as well as a high load-bearing capability. To investigate the deformation characteristics of flexible rudder surfaces utilizing cosine honeycomb structures, this study undertakes a comprehensive investigation through finite element simulation and 3D printing experiments. Moreover, this study analyzed the impact of honeycomb parameters and layout on the deflection performance and weight. The flexible rudder surface, fabricated from nylon, achieves smooth and consistent chordwise bending deformation, as well as uniform spanwise deformation within a tolerance of ±25°, and the maximum equivalent stress observed was 31.99 MPa, which is within the material’s allowable stress limits (50 MPa). Finite element simulation results indicate that once the deflection angle of the rocker exceeds 15°, a discernible deviation arises between the actual deflection angle of the flexible control surface and that of the rocker. Furthermore, this deviation escalates with increasing rocker rotation angles, and this discrepancy can be mitigated by augmenting the number of cosine honeycomb cells within the flexible rudder surface. Finally, a prototype of the flexible rudder surface was successfully produced using 3D printing technology, and the experimental results confirmed the deformation behavior, aligning with simulation outcomes with a deviation of less than 20%. These findings confirm the effective deflection performance of the designed flexible rudder surface, highlighting its potential application in small unmanned aerial vehicles. Full article

Smart buildings play a key role in the complex ecosystem of cities and are often subject to barriers that limit their development. Although identifying these barriers is fundamental to creating an enabling environment for this segment’s expansion, few works aim to identify these challenges. This work has two main objectives: (1) to research the main barriers limiting the development of new smart building projects and (2) to prioritize these barriers from the perspective of professionals with experience in the field. We adopted an exploratory approach common in research that focuses on identifying and prioritizing variables related to a phenomenon, which is based on two main actions: obtaining information through a careful literature review and consulting professionals who work in the concerned field. The results showed that professionals assessed the 23 barriers identified through bibliographic research as important, with the most important being related to lack of qualified professionals, shortage of government policies, higher initial and construction costs, macroeconomic barriers and access to financing, high cost of intelligent systems and technologies, regulatory barriers, lack of knowledge about the current and potential benefits of smart buildings, and more complex design and construction. Full article

Numerous studies have reported a correlation between gut microbiota and influenza A virus (IAV) infection and disease severity. However, the causal relationship between these factors remains inadequately explored. This investigation aimed to assess the influence of gut microbiota on susceptibility to human infection with H7N9 avian IAV and the severity of influenza A (H1N1)pdm09 infection. A two-sample Mendelian randomization analysis was conducted, integrating our in-house genome-wide association study (GWAS) on H7N9 susceptibility and H1N1pdm09 severity with a metagenomics GWAS dataset from a Chinese population. Twelve and fifteen gut microbiotas were causally associated with H7N9 susceptibility or H1N1pdm09 severity, separately. Notably, Clostridium hylemonae and Faecalibacterium prausnitzii were negative associated with H7N9 susceptibility and H1N1pdm09 severity, respectively. Moreover, Streptococcus peroris and Streptococcus sanguinis were associated with H7N9 susceptibility, while Streptococcus parasanguini and Streptococcus suis were correlated with H1N1pdm09 severity. These results provide novel insights into the interplay between gut microbiota and IAV pathogenesis as well as new clues for mechanism research regarding therapeutic interventions or IAV infections. Future studies should concentrate on clarifying the regulatory mechanisms of gut microbiota and developing efficacious approaches to reduce the incidence of IAV infections, which could improve strategy for preventing and treating IAV infection worldwide. Full article

Hydrogen energy is now a crucial technological option for decarbonizing energy systems. Comprehensive utilization is a typical mode of hydrogen energy deployment, leveraging its excellent conversion capabilities. Hydrogen is often used in combination with electrical and thermal energy. However, current hydrogen utilization modes are relatively singular, resulting in low energy utilization efficiency and high wind curtailment rates. To improve energy utilization efficiency and promote the development of hydrogen energy, we discuss three utilization modes of hydrogen energy, including hydrogen storage, integration into a fuel cell and gas turbine hybrid power generation system, and hydrogen methanation. We propose a hydrogen energy system with multimodal utilization and integrate it into an electrolytic hydrogen–thermal integrated energy system (EHT-IES). A mixed-integer linear programming (MILP) optimization scheduling model for the EHT-IES is developed and solved using the Cplex solver to improve the operational feasibility of the EHT-IES, focusing on minimizing economic costs and reducing wind curtailment rates. Case studies in northwest China verify the effectiveness of the proposed model. By comparing various utilization modes, energy storage methods, and scenarios, this study demonstrated that integrating a hydrogen energy system with multimodal utilization into the EHT-IES offers significant technical benefits. It enhances energy utilization efficiency and promotes the absorption of wind energy, thereby increasing the flexibility of the EHT-IES. Full article

The process integration and electrification concept has significant potential to support the industrial transition to low- and net-zero-carbon process heating. This increasingly essential concept requires an expanded set of process analysis tools to fully comprehend the interplay of heat recovery and process electrification (e.g., heat pumping). In this paper, new Exergy Pinch Analysis tools and methods are proposed that can set lower bound work targets by acutely balancing process heat recovery and heat pumping. As part of the analysis, net energy and exergy load curves enable visualization of energy and exergy surpluses and deficits. As extensions to the grand composite curve in conventional Pinch Analysis, these curves enable examination of different pocket-cutting strategies, revealing their distinct impacts on heat, exergy, and work targets. Demonstrated via case studies on a spray dryer and an evaporator, the exergy analysis targets net shaft-work correctly. In the evaporator case study, the analysis points to the heat recovery pockets playing an essential role in reducing the work target by 25.7%. The findings offer substantial potential for improved industrial energy management, providing a robust framework for engineers to enhance industrial process and energy sustainability. Full article

By analyzing students’ understanding of a certain subject’s knowledge and learning process, and evaluating their learning level, we can formulate students’ learning plans and teachers’ curricula. However, the large amount of data processing consumes a lot of manpower and time resources, which increases the burden on educators. Therefore, this study aims to use a machine learning model to build a model to evaluate students’ learning levels for art education. To improve the prediction accuracy of the model, SVM was adopted as the basic model in this study, and was combined with SSA, ISSA, and KPCA-ISSA algorithms in turn to form a composite model. Through the experimental analysis of prediction accuracy, we found that the prediction accuracy of the KPCA-ISSA-SVMM model reached the highest, at 96.7213%, while that of the SVM model was only 91.8033%. Moreover, by putting the prediction results of the four models into the confusion matrix, it can be found that with an increase in the complexity of the composite model, the probability of classification errors in model prediction gradually decreases. It can be seen from the importance experiment that the students’ achievements in target subjects (PEG) have the greatest influence on the model prediction effect, and the importance score is 9.5958. Therefore, we should pay more attention to this characteristic value when evaluating students’ learning levels. Full article

COVID-19 is an airborne respiratory disease that mainly affects the lungs. To date, COVID-19 has infected 580 million people with a mortality of approximately 7 million people worldwide. The emergence of COVID-19 has also affected the infectivity, diagnosis, and disease outcomes of existing diseases such as influenza, TB, and asthma in human populations. These are airborne respiratory diseases with symptoms and mode of transmission similar to those of COVID-19. It was speculated that the protracted nature of the COVID-19 pandemic coupled with vaccination could impact other respiratory diseases and mortality. In this study, we analyzed the impact of COVID-19 on flu, tuberculosis (TB), and asthma. Our analyses suggest that COVID-19 has a potential impact on the mortality of flu, TB, and asthma. These impacts vary across before the COVID-19 era, during the peak period of the pandemic, and after vaccinations/preventive measures were implemented, as well as across different regions of the world. Overall, the spread of flu generally reduced during the pandemic, resulting in a reduced expenditure on flu-related hospitalizations, although there were sporadic spikes at setting times. In contrast, TB deaths generally increased perhaps due to the disruption in access to TB services and reduction in resources. Asthma deaths, on the other hand, only marginally varied. Collectively, the emergence of COVID-19 added extra cost to the overall expenditure on some respiratory infectious diseases, while the cost for other infectious diseases was either reduced or somewhat unaffected. Full article

Three polysaccharides (SnNG, SnFS and SnFG) were purified from the body wall of Stichopus naso. The physicochemical properties, including monosaccharide composition, molecular weight, sulfate content, and optical rotation, were analyzed, confirming that SnFS and SnFG are sulfated polysaccharides commonly found in sea cucumbers. The highly regular structure {3)-L-Fuc_2S-(α1,}_n of SnFS was determined via a detailed NMR analysis of its oxidative degradation product. By employing β-elimination depolymerization of SnFG, tri-, penta-, octa-, hendeca-, tetradeca-, and heptadeca-saccharides were obtained from the low-molecular-weight product. Their well-defined structures confirmed that SnFG possessed the backbone of {D-GalNAc_4S6S-β(1,4)-D-GlcA}, and each GlcA residue was branched with Fuc_2S4S. SnFS and SnFG are both structurally the simplest version of natural fucan sulfate and fucosylated glycosaminoglycan, facilitating the application of low-value sea cucumbers S. naso. Bioactivity assays showed that SnFG and its derived oligosaccharides exhibited potent anticoagulation and intrinsic factor Xase (iXase) inhibition. Moreover, a comparative analysis with the series of oligosaccharides solely branched with Fuc_3S4S showed that in oligosaccharides with lower degrees of polymerization, such as octasaccharides, Fuc_2S4S led to a greater increase in APTT prolongation and iXase inhibition. As the degree of polymerization increases, the influence from the sulfation pattern diminishes, until it is overshadowed by the effects of molecular weight. Full article

The formation of nitrogen-containing organic interstellar molecules is of great importance to reveal chemical processes and the origin of life on Earth. Benzonitrile (BN) is one of the simplest nitrogen-containing aromatic molecules in the interstellar medium (ISM) that has been detected in recent years. Methanol (CH₃OH) exists widely in interstellar space with high reactivity. Herein, we measured the infrared (IR) spectra of neutral and cationic BN–CH₃OH clusters by vacuum ultraviolet (VUV) photoionization combined with time-of-flight mass spectrometry. Combining IR spectra with the density functional theory calculations, we reveal that the BN–CH₃OH intends to form a cyclic H-bonded structure in neutral clusters. However, after the ionization of BN–CH₃OH clusters, proton-shared N···H···O and N···H···C structures are confirmed to form between BN and CH₃OH, with the minor coexistence of H-bond and O-π structures. The formation of the proton-shared structure expands our knowledge of the evolution of the life-related nitrogen-containing molecules in the universe and provides a possible pathway to the further study of biorelevant aromatic organic macromolecules. Full article

In the European Union, salmonellosis is one of the most important zoonoses reported. Poultry meat and egg products are the most common food matrices associated with Salmonella presence. Moreover, wild and domestic animals could represent an important reservoir that could favour the direct and indirect transmission of pathogens to humans. Salmonella spp. can infect carnivorous or omnivorous wild birds that regularly ingest food and water exposed to faecal contamination. Birds kept in captivity can act as reservoirs of Salmonella spp. following ingestion of infected prey or feed. In this paper, we describe the isolation of different Salmonella serovars in several species of raptors hosted in aviaries in an Italian wildlife centre and in the raw chicken necks used as their feed but intended for human consumption. Characterisations of strains were carried out by integrating classical methods and whole genome sequencing analysis. The strains of S. bredeney isolated in poultry meat and birds belonged to the same cluster, with some of them being multidrug-resistant (MDR) and carrying the Col(pHAD28) plasmid-borne qnrB19 (fluoro)quinolone resistance gene, thus confirming the source of infection. Differently, the S. infantis found in feed and raptors were all MDR, carried a plasmid of emerging S. infantis (pESI)-like plasmid and belonged to different clusters, possibly suggesting a long-lasting infection or the presence of additional undetected sources. Due to the high risk of fuelling a reservoir of human pathogens, the control and treatment of feed for captive species are crucial. Full article

Dry-powder inhalers (DPIs) are valued for their stability but formulating them is challenging due to powder aggregation and limited flowability, which affects drug delivery and uniformity. In this study, the incorporation of L-leucine (LEU) into hot-melt extrusion (HME) was proposed to enhance dispersibility while simultaneously maintaining the high aerodynamic performance of inhalable microparticles. This study explored using LEU in HME to improve dispersibility and maintain the high aerodynamic performance of inhalable microparticles. Formulations with crystalline itraconazole (ITZ) and LEU were made via co-jet milling and HME followed by jet milling. The LEU ratio varied, comparing solubility, homogenization, and aerodynamic performance enhancements. In HME, ITZ solubility increased, and crystallinity decreased. Higher LEU ratios in HME formulations reduced the contact angle, enhancing mass median aerodynamic diameter (MMAD) size and aerodynamic performance synergistically. Achieving a maximum extra fine particle fraction of 33.68 ± 1.31% enabled stable deep lung delivery. This study shows that HME combined with LEU effectively produces inhalable particles, which is promising for improved drug dispersion and delivery. Full article

Background and Objectives: New investigations have detected an enhanced probability for women to develop menstrual cycle alterations after anti-COVID-19 vaccination. Moreover, given that the protective immunity provided by anti-COVID-19 vaccination appears to wane quickly, booster vaccination has been recommended. Nonetheless, whether adverse events arise from such repeated immunization has not been studied. Materials and Methods: We studied the incidence of menstrual cycle alterations, the quantity of menstrual cycle alterations per subject, and of altered menstrual cycles in nonpregnant women of fertile age after anti-COVID-19 vaccination in a cohort of vaccinated female subjects by the means of a standardized questionary that was applied via telephone calls each month. Subjects that received up to four doses were studied for 6 months after each dose. We calculated the odds ratio for enhanced incidence, as well as quadratic functions for the tendencies. A sensitivity analysis excluding subjects taking hormonal birth control and those with polycystic ovary syndrome was performed. Results: Anti-COVID-19 vaccination enhanced the probability to develop menstrual cycle alterations (OR 1.52, CI at 95% 1.2–1.8, p < 0.0001) and, interestingly, such a tendency was enhanced when subjects received more doses (R² = 0.91). Furthermore, the same trends repeated for the quantity of alterations per subject, and of altered cycles. Such an effect was further demonstrated to be independent upon the vaccine brand being applied, the birth control status, and the diagnosis of polycystic ovary syndrome. Conclusions: Vaccination is the most cost-effective measure for primary prevention and is considered to be safe. Nonetheless, in this article, we show data that suggest that repeated vaccination of adult female subjects may lead to an enhanced incidence of menstrual cycle–related adverse events, quantity of alterations per subject, and altered cycles. We therefore think that the development of new vaccine formulations that produce longer-lasting immunity is of paramount importance to reduce the potential for dose accumulation–dependent enhanced risk. Full article

Chrysin is hypothesized to possess the ability to prevent different illnesses, such as diabetes, cancer, and neurodegenerative disorders. Nonetheless, chrysin has a low solubility under physiological conditions, resulting in limited bioavailability. In a previous study, we utilized an oil-in-water emulsion system (chrysin-ES or chrysin-NE) to encapsulate chrysin, thereby increasing its bioaccessibility and preserving its antioxidant and anti-Alzheimer’s properties. To promote the chrysin-ES as a supplementary and functional food, it was obligatory to carry out a safety assessment. Cytotoxicity testing showed that chrysin-ES was harmless, with no killing effect on 3T3-L1 (adipocytes), RAW 264.7 (macrophages), HEK293 (kidney cells), and LX-2 (hepatic stellate cells). The acute toxicity evaluation demonstrated that the 50% lethal dose (LD₅₀) for chrysin-ES was greater than 2000 mg/kg BW. Genotoxicity assessments found that chrysin-ES did not induce DNA mutations in vitro or in vivo. Furthermore, chrysin and chrysin-ES exhibited anti-mutagenic properties against PhIP-induced and IQ-induced mutagenesis in the Ames test, while they inhibited urethane-, ethyl methanesulfonate-, mitomycin C-, and N-nitrosomethylurea-mediated mutations in Drosophila. The present study illustrates the safety and anti-genotoxicity properties of chrysin-ES, allowing for the further development of chrysin-based food supplements and nutraceuticals. Full article

States in the U.S. are newly implementing universal school meal (USM) policies, yet little is known about the facilitators of their success and the challenges they confront. This study evaluated the challenges and facilitators faced by school food authorities (SFAs) implementing California’s universal school meal (USM) policy during its inaugural year (2022–2023) using an online survey. In March 2023, 430 SFAs reported many benefits, including increased meal participation (64.2% of SFAs) and revenues (65.7%), reduced meal debt (41.8%) and stigma (30.9%), and improved meal quality (44.3%) and staff salaries (36.9%). Reported challenges include product/ingredient availability (80.9%), staffing shortages (77.0%), vendor/distributor logistics issues (75.9%), and administrative burden (74.9%). Top facilitators included state funding (78.2%) and increased federal reimbursement (77.2%). SFAs with fewer students eligible for free or reduced-price meals (as opposed to SFAs with more) reported greater increases in meal participation and reductions in stigma but also more administrative burdens. Larger SFAs reported greater increases in revenues, staff salaries, and improvements in meal quality than smaller SFAs but also more challenges. Overall, California’s USM policy has enhanced student access to healthy meals while mitigating social and financial barriers. Understanding California’s experience can inform other jurisdictions considering or implementing similar policies. Full article

In most cases, the number of honeybee stings received by the body is generally small, but honeybee stings can still cause serious allergic reactions. This study fully simulated bee stings under natural conditions and used ¹H Nuclear Magnetic Resonance (¹H NMR) to analyze the changes in the serum metabolome of Sprague–Dawley (SD) rats stung once or twice by honeybees to verify the impact of this mild sting on the body and its underlying mechanism. The differentially abundant metabolites between the blank control rats and the rats stung by honeybees included four amino acids (aspartate, glutamate, glutamine, and valine) and four organic acids (ascorbic acid, lactate, malate, and pyruvate). There was no separation between the sting groups, indicating that the impact of stinging once or twice on the serum metabolome was similar. Using the Principal Component Discriminant Analysis ( PCA-DA) and Variable Importance in Projection (VIP) methods, glucose, lactate, and pyruvate were identified to help distinguish between sting groups and non-sting groups. Metabolic pathway analysis revealed that four metabolic pathways, namely, the tricarboxylic acid cycle, pyruvate metabolism, glutamate metabolism, and alanine, aspartate, and glutamate metabolism, were significantly affected by bee stings. The above results can provide a theoretical basis for future epidemiological studies of bee stings and medical treatment of patients stung by honeybees. Full article