Objective: This study aimed to elucidate the temporal characteristics, physiological responses and interactive relationships of Penicillium pathogens in Gonggan mandarin (Citrus reticulata 'Gonggan') fruits, thereby providing a theoretical basis for targeted preservation technologies. Methods: Experiment 1 involved observing storage diseases in Gonggan mandarin fruits, while experiment 2 focused on the physiological responses of the fruits to Penicillium inoculation. In the Penicillium inoculation experiment, three major pathogenic fungi—Penicillium digitatum (Pd), P. italicum (Pi), and P. expansum (Pe)—were isolated and identified from Gonggan mandarin fruits in the Xijiang River Basin of Guangdong Province and used as pathogen material. A total of seven treatments were set up, including single and combined inoculations, along with two control groups (CK1: Fruits immersed in 0.1% NaClO solution for 3 min; CK2: Peel wounded but not inoculated). Disease symptoms were dynamically observed, incidence rate (IR) and disease index (DI) were statistically calculated, fruit storage quality was analyzed, reactive oxygen species metabolism indicators and defense enzyme activities were measured, and a disease-physiology regression model was constructed. Results: During the storage of Gonggan mandarin fruits, green mold was the predominant disease, exhibiting a higher severity than blue mold, with complex infections being commonly observed. In single inoculation treatments, Pi and Pe developed symptoms earlier, but Pd exhibited faster spread and stronger infectivity, with a significantly higher incidence rate (IR) than Pi and Pe just 4 days after inoculation. Following combined inoculation, the DI of the triple mixed inoculation (Pd+Pi+Pe) was lower than that of the (Pd+Pi) combination (P<0.05), indicating that interspecific competition among Penicillium species significantly delayed disease progression. Fruit injury significantly promoted quality deterioration in Gonggan mandarin fruits, resulting in a substantial increase in weight loss rate, and it also induced H₂O₂ accumulation and a significant rise in CAT activity (P<0.05). Pathogen infection induced severe oxidative stress and a disrupted defense enzyme system in Gonggan mandarin fruits, which was manifested by a substantial increase in the generation rate of O₂⁻· and the contents of H₂O₂ and malondialdehyde (MDA); In contrast, the activities of phenylalanine ammonia-lyase (PAL) and catalase (CAT) were significantly inhibited, exhibiting a highly significant negative correlation with disease severity indices IR and DI (P<0.01). Conclusion: During the storage of Gonggan mandarin, green mold was the dominant disease, often accompanied by complex infections. Mechanical damage accelerated fruit decay by facilitating the infection of Penicillium spp. As the predominant pathogen, Penicillium digitatum drove oxidative damage and quality deterioration in the fruit through sustained suppression of PAL activity and induction of CAT activity collapse. Moreover, interspecific competition among Penicillium species mitigated disease development in Gonggan mandarin.

The present study aimed to investigate the synergistic effects of nitric oxide combined with clove essential oil on the postharvest storage quality of peach fruit. The harvested peaches were treated with the sodium nitroprusside and clove essential oil using atomized fumigation. The firmness, ethylene release rate, decay index, titratable acid content, soluble solid content, cell wall polysaccharide contents, cell wall degradation enzymes activity, defense-related enzymes activity, tissue structure and cell wall ultrastructure were analyzed in peaches during the storage period at 0 ℃. The results indicated that the combined atomized fumigation treatment had a better preservation effect on peaches, in comparison with the nitric oxide or clove essential oil alone. The decay index of peach fruit treated with the combined treatment was only 9.25% during the 35 d of storage. The ethylene release rate of combined treatment was reduced by 34.88% while the firmness, titratable acid, and soluble solid contents of peaches fruit was increased by 22.01%, 31.68% and 5.90%, respectively, when compared to the untreated fruit on day 35. Furthermore, the combined treatment inhibited the activities of polygalacturonase, pectin methylesterase, cellulase and β-galactosidase activities. The water-soluble pectin content of combined treatment was reduced by 21.88% compared to the control group. The contents of chelate soluble pectin, sodium carbonate-soluble pectin, hemicellulose and cellulose components increased by 23.40%, 21.31%, 36.62% and 39.89% respectively, thereby maintaining the integrity of the cell wall structure and delaying the fruit softening. We found that combined treatment enhanced the defense enzyme activities of phenylalanine ammonia-lyase, and 4-coumarate coenzyme A ligase activities. Cinnamate-4-hydroxylase activity of combined treatment was 15.71% higher than that of the treatment with nitric oxide alone and 20.98% higher than that of the treatment with clove essential oil alone, enhancing the disease resistance of the fruit. Therefore, the nitric oxide combined with clove essential oil through atomized fumigation can effectively maintain the quality of peaches during low-temperature storage, providing a theoretical basis for the development of quality control technologies for postharvest peaches.

To evaluate the effectiveness of different preservatives on the postharvest quality of ginger, treatments involving chlorine dioxide (ClO₂), ascorbic acid (V_C), cinnamaldehyde (CA), and a combined treatment (ClO₂+V_C+CA) were applied separately. The ginger was stored under refrigerated conditions (13±0.5 ℃, 90% relative humidity), and the impact of these treatments on the physicochemical properties and related enzyme activities of ginger was assessed. Flavour profiles were analyzed using gas chromatography-ion mobility spectrometry (GC-IMS). The results indicated that all preservative treatments significantly delayed the deterioration of ginger quality (P<0.05), with the combined treatment demonstrating a synergistic effect. After 120 days of storage, the water loss rate of ginger was reduced by 57.67% compared to the control (CK). The activities of phenylalanine ammonia-lyase (PAL), superoxide dismutase (SOD), and catalase (CAT) increased by 7.78%, 11.22%, and 28.61%, respectively, while polyphenol oxidase (PPO) and peroxidase (POD) activitie significantly decreased (P<0.05). Retention of gingerol, protein, and polysaccharides improved by 31.65%, 35.47%, and 40.02%, respectively, compared to the control. Moreover, GC-IMS combined with principal component analysis revealed clear differences in volatile profiles among the various treatment groups. The combined treatment promoted the accumulation and retention of important aroma compounds, including green aromas (citral), fruity aromas (β-laurene), and spicy aromas ((E,E)-2,4-hexadienal). In conclusion, the combined preservation approach proved to be an effective strategy for maintaining post-harvest ginger quality by boosting antioxidant enzyme activity and minimizing aroma volatile loss. This research offers a theoretical basis and technical reference for developing ginger preservation technologies.

Objective: To investigate lipid changes during the storage of barley flour, the oxidative patterns of lipids and lipase activity, and their interactions, thereby elucidating the reaction mechanisms underlying oxidative deterioration. Methods: Oxidative stability was evaluated by measuring fatty acid content, lipase activity, and malondialdehyde (MDA) value in barley flour stored for 28 days under high-temperature/high-humidity (HH), normal-temperature/normal-humidity (NN), and low-temperature/low-humidity (LL) conditions. Lipid composition changes were monitored using ultra-performance liquid chromatography-quadrupole/electrostatic field orbitrap high-resolution mass spectrometry (UPLC-Q-Exactive Orbitrap MS/MS) to monitor lipid composition changes. Results: Under LL and NN conditions, changes in fatty acid value, lipase activity, and malondialdehyde value were relatively slow; After 28 days of storage under HH conditions, the fatty acid value increased from 91.08±1.33 mg/100 g to 136.26±2.62 mg/100 g, lipase activity rose from 41.04±0.58 mg/g to 47.91±0.61 mg/g, and malondialdehyde value increased from 45.41±2.10 nmol/g to 79.97±1.37 nmol/g. Lipidomics analysis identified 234 lipid molecules across 23 subclasses, including 176 significantly altered lipids. Correlation analysis revealed positive correlations between lipase activity and sterol lipids (STs), fatty acids (FAs), and glycerolipids (GLs). Principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) indicated that barley lipids under HH conditions underwent significant changes with prolonged storage. Among these, triglycerides (TG) and diglycerides (DG) exhibited the highest susceptibility to oxidation and could serve as biomarkers distinguishing fresh barley from oxidized barley. Conclusion: Significant differences exist in barley flour lipids before and after storage. Enhancing storage stability by reducing lipase activity provides a theoretical basis for establishing barley shelf-life standards.

Objective: To investigate the regulatory effects of GPA on serum differential metabolites and elucidate the underlying mechanisms in D-Gal-induced aging mice. Method: The aging model was established through injection with D-Gal, and the administration group given GPA treatment simultaneously. The spatial cognition and memory ability was determined by Morris water maze test (MWM). Then detect mice serum antioxidant enzyme activity and lipid peroxide levels with the kit method. And Non-targeted metabolomics analysis of mice serum using UPLC-QTOF/MS. Results: Compared with the model group, GPA could shorten the escape latency of aging mice with various degrees, and the mice of GPA group spent significantly more time in the target quadrant. GPA (50 mg/kg) could increase mice serum activities of SOD (P<0.01), GSH-Px (P<0.01), and CAT (P<0.01) and reduce the level of MDA (P<0.01) in the serum of aging mice. In the serum of mice in the GPA group, 18 classes of differential metabolites were upregulated, and 6 classes were downregulated. KEGG pathway enrichment analysis indicated that the differential metabolites were mainly enriched in inositol phosphate metabolism, steroid hormone biosynthesis, glutathione metabolism and primary bile acid biosynthesis metabolism pathways, etc. Conclusion: Oxidized glutathione, 18-hydroxycorticosterone, inositol-1,3,4,5-tetraphosphate, and D-inositol-1,4,5,6-tetraphosphate which may be important metabolites related to improve oxidative damage. This confirms that GPA could alleviate oxidative damage in aging mice, which may be related to enhancing the endogenous antioxidant system and the regulation of inositol phosphate metabolism, steroid hormone biosynthesis, glutathione metabolism and primary bile acid biosynthesis metabolism pathways, etc.

To elucidate the composition of chlorogenic acid homologs and anthocyanins in blueberry extracts and their protective effects on human L-02 hepatocytes, the present study utilized organic solvent extraction and macroporous adsorption resin separation to prepare chlorogenic acid extracts (CGAE) and anthocyanin extracts (ANTE) from 15 blueberry samples of distinct cultivars, origins, and geographical locations. In vitro experiments were conducted to assess free radical scavenging capacity and evaluate the activity of antioxidant enzymes in L-02 hepatocytes. The results showed that chlorogenic acid>neochlorogenic acid>cryptochlorogenic acid>isochlorogenic acid B>isochlorogenic acid A>isochlorogenic acid C and delphinidin>petunidin>peonidin>malvidin>cyanidin in the average content of chlorogenic acids and anthocyanins in 15 blueberry samples. Correlation analysis showed that the contents of chlorogenic acid, neochlorogenic acid, and cryptochlorogenic acid were significantly negatively correlated with DPPH and ABTS⁺ radical scavenging rates, and significantly positively correlated with FRAP and RP. The contents of delphinidin-3-galactoside and delphinidin-3-arabinoside were significantly positively correlated with FRAP and RP, and negatively correlated with ABTS⁺ radical scavenging rate, but only the correlation for delphinidin-3-arabinoside was significant. In contrast, delphinidin-3-glucoside was significantly positively correlated with ABTS⁺ radical scavenging rate, and significantly negatively correlated with FRAP and RP. Among cyanidin, petunidin, peonidin, and malvidin, only cyanidin-3-glucoside was significantly negatively correlated with FRAP and RP, while petunidin-3-galactoside and petunidin-3-arabinoside were significantly positively correlated with FRAP and RP. The results indicated that the active component content and antioxidant activity of blueberries varied significantly depending on the different cultivars, growing regions, and geographical origins. Principal component analysis (PCA) was employed to statistically validate the distinct chemotypic profiles among different varieties. A functional validation model at the cellular level was established, and the optimal induction conditions were determined as treatment with 350 μmol/L t-BHP for 4 hours. Using this optimized model, the protective effects of both CGAE and ANTE extracts were rigorously evaluated. In summary, 20~60 μg/mL CGAE and 15~25 μg/mL ANTE significantly reduced ROS levels while significantly increasing the activities of GSH-Px and SOD, and the antioxidant effect of ANTE was significantly higher than that of CGAE. This study screened blueberry extracts with high antioxidant activity, providing a reference for research on the antioxidant applications of blueberry extracts.

To investigate the absorption pathways and efficacy of Gold Butterfly Collagen Peptides in mice, Gold Butterfly Collagen Peptides were extracted and purified using an in vitro digestion method, and their amino acid composition was analyzed. Subsequently, isotope-labeled peptide segments [¹³C₅-¹⁵N₁-Pro] were prepared via solid-phase synthesis to replace all homologous amino acids in the target peptide segments, and peptides were administered to C57BL/6 mice via gavage. Ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was employed to dynamically monitor the metabolic distribution of labeled proline in various tissues and organs over 24 hours. The characteristic predominant amino acids of Gold Butterfly Collagen Peptides were glycine (20.80%), proline (10.72%), and alanine (10.60%). The small intestine was the primary organ absorbing Gold Butterfly Collagen Peptides, with uptake exceeding 50% across all time points. Skin absorption increased to 11.71% at 12 hours, suggesting potential involvement of Gold Butterfly Collagen Peptides in skin tissue repair. Uptake into the stomach, cecum, and colon exhibited dynamic fluctuations, while uptake by the heart and lungs was relatively low. This study demonstrates that Gold Butterfly Collagen Peptide digestion and absorption processes exhibit prominent temporal and organ-specific characteristics, providing theoretical foundation for their nutritional applications.

Objective: This study evaluated the efficacy of donkey-hide gelatin (Ejiao) in alleviating microcirculatory disorders in mice and explored its underlying mechanisms. Methods: Female ICR mice were randomly divided into four groups: Healthy control, model, Ejiao mid-dose (1.5 g/kg), and Ejiao high-dose (3 g/kg). A microcirculatory disorder model was established by injecting dextran into the tail vein. Real-time blood perfusion in the auricles was continuously monitored using a laser speckle flowmetry system. The number of functional capillaries was assessed under a stereomicroscope, and hemorheological parameters were quantified using a hemorheometer. Enzyme-linked immunosorbent assay was used to measure levels of vascular activity- and metabolism-related factors, neurotransmitters, hormones, and inflammatory cytokines to elucidate the potential mechanisms. Additionally, histopathological changes in the liver and kidneys were assessed. Results: In the mid-dose Ejiao group, auricular blood perfusion levels at 15 and 30 minutes increased significantly by 36.76% and 36.02%, respectively, compared to that at baseline (0 min) (P<0.01). Furthermore, at 30 minutes, both the mid- and high-dose Ejiao groups exhibited significant increases in the number of open capillaries by 34.61% and 38.46%, respectively, compared to that in the model group (P<0.01). Hemorheological analysis revealed that the model group had significantly elevated levels of whole-blood and plasma viscosity, hematocrit, and fibrinogen (P<0.05 or P<0.01), along with a reduced erythrocyte sedimentation rate (P<0.05). In contrast, the mid-dose Ejiao group showed significant improvement in these parameters (P<0.05). Both mid- and high-dose Ejiao treatments significantly decreased the level of plasminogen activator inhibitor-1 (PAI-1) (P<0.01) and markedly increased the level of 6-keto-prostaglandin F1α, a stable metabolite of prostacyclin (P<0.05), compared to that in the model group, suggesting a potential role of Ejiao in modulating vascular function. Additionally, both Ejiao-treated groups showed increased levels of Ca²⁺/Mg²⁺ ATPase and cyclic guanosine monophosphate, indicating that Ejiao might alleviate microcirculatory disorders by enhancing energy metabolism. The levels of the neurotransmitters acetylcholine and norepinephrine, as well as the hormones estradiol and luteinizing hormone, were closer to those of the healthy group in Ejiao-treated mice, indicating regulatory effects on neuroendocrine imbalances. Inflammatory factor analysis demonstrated that mid-dose Ejiao significantly reduced the levels of tumor necrosis factor-α and interleukin-1β (P<0.05), thereby mitigating inflammation associated with microcirculatory disorders. Histopathological observations further confirmed that Ejiao treatment alleviated tissue damage and promoted structural recovery in the liver and kidneys. Conclusion: Mid-dose Ejiao effectively alleviated microcirculatory disorders by modulating vascular activity, correcting metabolic imbalances, restoring neuroendocrine homeostasis, and suppressing inflammatory responses. These findings provide strong scientific evidence supporting the therapeutic potential of Ejiao for the treatment of microcirculatory disorders.

Objective: To utilize extracellular vesicles (EVs) derived from Lactobacillus paracasei as delivery carriers to construct an astaxanthin (AST)-encapsulated system (AST@EVs), thereby improving the water solubility, stability, and bioavailability of AST. Methods: EVs were isolated via differential centrifugation and ultracentrifugation, and systematic evaluations of the physicochemical properties, digestive stability, antioxidant activity, and anti-fatigue effect of AST@EVs were conducted using characterization analysis, in vitro simulated digestion, free radical scavenging assays, and the C2C12 cell model. Results: EVs with a typical spherical structure and an average particle size of 85.97 nm were successfully prepared. After optimizing the encapsulation ratio (2:1), the encapsulation efficiency and protein loading capacity of AST@EVs reached 84.94% and 12.74%, respectively, with the particle size increasing to 109.37 nm. Additionally, Fourier transform infrared spectroscopy (FTIR) and electron microscopy results confirmed the effective encapsulation of AST in EVs. In vitro digestion, the retention rate of AST in AST@EVs was significantly higher than that in the free AST group and the physical mixture group. The scavenging rates of AST@EVs against 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) cation radical (ABTS⁺) were 83.92% and 62.81%, respectively, indicating a significant enhancement in antioxidant capacity. Cellular experiments showed that AST@EVs could promote the uptake of AST by C2C12 cells (reaching 5.27±0.21 μg/mL), alleviate oxidative stress, regulate the activities of superoxide dismutase (SOD) and catalase (CAT), and reduce the level of malondialdehyde (MDA). Meanwhile, AST@EVs inhibited the increase in lactate dehydrogenase (LDH) and creatine kinase (CK) activities, enhanced glycogen storage, and restored the activities of enzymes related to mitochondrial function. Conclusion: The EV-based encapsulation strategy can significantly enhance the stability, antioxidant capacity, and anti-fatigue efficacy of AST. This study provides a new idea for the development of functional foods and anti-fatigue drugs.

To evaluate the nutritional value of different edible parts of Fuyong Fatty Crab (Scylla serrata) as well as its varietal advantages, the nutritional components of three of its core edible tissues (ovary, hepatopancreas, and muscle) were systematically analyzed and compared with previously reported nutritional data on Suifenhe Mitten Crab (Eriocheir hepuensis, a representative freshwater crab) and Swimming Crab (Portunus trituberculatus, a representative marine crab). The results showed that the average body weight, carapace length and width, and ovary index (10.18%, Grade I premium roe crab) of Fuyong Fatty Crab were higher than those of the two control crab species. Among the three tissues of Fuyong Fatty Crab, the ovary was low in fat, high in protein, and rich in five carotenoid species (total content higher than that of Portunus trituberculatus); The muscle had the highest essential amino acid and total amino acid contents and optimal average amino acid score, and the hepatopancreas had the highest contents of fatty acids such as DHA and EPA (P<0.05) and the lowest n-3/n-6 ratio (P<0.05) significantly. All three tissues were high-quality mineral sources, with differences in mineral composition. This study indicated that the overall nutritional value of Fuyong Fatty Crab was superior to those of Suifenhe Mitten Crab and Portunus trituberculatus, and had the various tissue parts differed considerably in nutritional composition and quality. These results provide a scientific basis for the dietary selection and deep processing of Fuyong Fatty Crab.