Dioscorea bulbifera L. (DBL), a traditional herbal medicine used to treat thyroid disorders and tumors, has raised considerable safety concerns due to its potential hepatotoxic effects. Previous research suggests that this liver injury may be associated with the metabolic activation of furan-containing compounds (FCCs) present in DBL. Here, we systematically characterized FCCs and their reactive metabolites in DBL extract through an integrated analytical approach combining ultrahigh performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS) and ultra-high performance liquid chromatography coupled with tandem high-resolution mass spectrometry (UHPLC-HRMS). Employing both targeted and untargeted mass spectrometric analyses, we identified a total of 849 constituents in the aqueous extract of DBL, including 10 furanoditerpenoids and 17 additional FCCs. To elucidate the metabolic activation pathways, we used glutathione (GSH) and 4-bromobenzylamine (BBA) as dual trapping agents within a mouse liver microsomal (MLM) incubation system. Through P450-mediated metabolism, seven corresponding adducts derived from diosbulbin B (DSB), C (DSC), D (DSD), E (DSE), L (DSL), N (DSN), and 2-pentylfuran were successfullytrapped and characterized. This study establishes a sensitive and specific approach for the comprehensive profiling of potentially hepatotoxic furan compounds in DBL, and provides valuable insights into the formation of reactive metabolites and toxicity mechanisms related to furan-containing herbal medicine.

The molecular weight (MW) of dextran involved in glycation is a key factor influencing the structural and functional properties of protein-dextran conjugates. In this study, soy protein isolate (SPI) was glycated with dextran of varying MWs via the Maillard reaction, and the resulting conjugates were evaluated for structural characteristics and flavor-binding capacity. Glycation significantly inhibited SPI thermal aggregation by decreasing surface hydrophobicity (H0) from 296 to 145-217 and free thiol group (R-SH) content from 10.8 to 4.7-7.9 mu mol/g, with both indicators negatively correlated with dextran MW distribution and grafting degree (DG). Multispectral analyses revealed that glycation promoted protein unfolding, induced secondary structure transitions from alpha-helix to beta-sheet, and enhanced overall molecular flexibility. Glycation predominantly occurred at the epsilon-amino groups of lysine and the imidazole groups of arginine, with LYS-(C6H10O5)2 identified as the predominant glycation adduct. Furthermore, glycation with medium- and high-MW dextran reduced SPI binding to off-flavor compounds, with the greatest reduction observed for nonanal (24.7 %), and the extent of reduction was positively correlated with DG. These findings elucidate the MW-dependent effects of glycation and offer mechanistic insights for the targeted design of protein modifications to enhance the flavor quality of plant-based foods.

Risedronate sodium (RIS) is a primary treatment for postmenopausal osteoporosis, but oral administration requires patients to remain upright to prevent side effects like esophagitis, gastritis, and ulcers, which can be difficult for those with fatigue or bone pain. This study developed RIS-loaded Bletilla striata polysaccharide microneedles (RIS-BMNs) with sufficient mechanical strength for effective skin penetration and drug delivery. Franz diffusion cell experiments showed that RIS-BMNs achieved 3 times higher in vitro transdermal absorption than the RIS solution and 2.6 times more than RIS patches. The stimulation test found that RIS-BMNs caused minimal skin irritation, which resolved within 12 h. RIS-BMNs also promoted cell proliferation and wound healing, as shown by in vitro cell viability and migration tests, and improved bone trabeculae and density in osteoporotic rats, according to micro-CT imaging and H&E staining. Biochemical markers (BGP, IL-6, CTX-I, TRAP, OPG, and PINP) indicated that RIS-BMNs had anti-osteoporotic effects comparable to oral RIS but without gastrointestinal side effects, as confirmed by H&E staining. Our new transdermal method for RIS could improve adherence in treating postmenopausal osteoporosis.

In this study, nanoparticles loaded with active components from Polygonum orientale L. (PO), a traditional Chinese herb known for its anti-myocardial ischemic properties, were investigated for cardio-protective properties. Specifically, OVQ-Nanoparticles (OVQ-NPs) with Orientin (Ori), Vitexin (Vit), and Quercetin (Que) was obtained by double emulsion-solvent evaporation method. The OVQ-NPs exhibited a spherical shape, with a uniform size distribution of 136.77 ± 3.88 nm and a stable ζ-potential of -13.40 ± 2.24 mV. Notably, these nanoparticles exhibited a favorable sustained-release characteristic, resulting in an extended circulation time within the living organism. Consequently, the administration of these nanoparticles resulted in significant improvements in electrocardiograms and heart mass index of myocardial ischemic rats induced by isoproterenol, as well as decreased serum levels of CK, LDH, and AST. Furthermore, the results of histopathological examination, such as H&E staining and TUNEL staining, confirmed a reduced level of cardiac tissue pathology and apoptosis. Moreover, the quantification of biochemical indicators (SOD, MDA, GSH, NO, TNF-α, and IL-6) demonstrated that OVQ-NPs effectively mitigated myocardial ischemia by regulating oxidative stress and inflammatory pathways. In conclusion, OVQ-NPs demonstrate promising therapeutic potential as an intervention for myocardial ischemia, providing a new perspective on traditional Chinese medicine treatment in this area.

A convenient and efficient reagent system of arylhydrazine salt‐selenium was developed to directly selenizing uracil preparation of 5‐selenium uracil. In the presence of this reagent, a variety of uracil/pyrimidine converts to their corresponding 5‐arylselanyluracils/5‐selenopyrimidine with good yield and high regioselectivity. Elemental selenium of commercially accessible, stable, affordable, and easy to use was used as the selenization reagent. This reaction is attractive and practical since there are no catalysts or ligands needed, and a wide range of functional groups can be tolerated.

A facile method was developed for the selective thioetherification of uracils using sulfonyl hydrazide as the thioetherification reagent.

ETHNOPHARMACOLOGICAL RELEVANCE:As reported in the Ancient Chinese Medicinal Books, Ginkgo biloba L. fruit has been used as a traditional Chinese medicine for the treatment asthma and cough or as a disinfectant. Our previous study demonstrated that G. biloba exocarp extract (GBEE), an extract of a traditional Chinese herb, inhibits the formation of methicillin-resistant Staphylococcus aureus (MRSA) biofilms. However, GBEE is a crude extract that contains many components, and the underlying mechanisms of purified GBEE fractions extracted with solvents of different polarities are unknown.AIM OF THE STUDY:This study aimed to investigate the different components in GBEE fractions extracted with solvents of different polarities and their antibacterial effects and mechanisms against MRSA and Staphylococcus haemolyticus biofilms both in vitro and in vivo.METHODS:The components in different fractions were detected by high-performance liquid chromatography-high resolution mass spectrometry (HPLC-HRMS). Microbroth dilution assays and time growth curves were used to determine the antibacterial effects of the fractions on 15 clinical bacterial isolates. Crystal violet staining, scanning electron microscopy (SEM) and transmission electron microscopy (TEM) were utilized to identify the fractions that affected bacterial biofilm formation. The potential MRSA targets of the GBEE fraction obtained with petroleum ether (PE), denoted GBEE-PE, were screened by transcriptome sequencing, and the gene expression profile was verified by quantitative polymerase chain reaction (qPCR).RESULTS:HPLC-HRMS analysis revealed that the four GBEE fractions (extracted with petroleum ether, ethyl acetate, n-butanol, and water) contained different ginkgo components, and the antibacterial effects decreased as the polarity of the extraction solvent increased. The antibacterial activity of GBEE-PE was greater than that of the GBEE fraction extracted with ethyl acetate (EA). GBEE-PE improved H. illucens survival and reduced MRSA colonization in model mouse organs. Crystal violet staining and SEM and TEM analyses revealed that GBEE-PE inhibited MRSA and S. haemolyticus biofilm formation. Transcriptional analysis revealed that GBEE-PE inhibits MRSA biofilms by altering ion transport, cell wall metabolism and virulence-related gene expression. In addition, the LO2 cell viability and H. illucens toxicity assay data showed that GBEE-PE at 20 mg/kg was nontoxic.CONCLUSION:The GBEE fractions contained different components, and their antibacterial effects decreased with increases in the polarity of the extraction solvent. GBEE-PE limited MRSA growth and biofilm formation by affecting ion transport, cell wall synthesis, and virulence-related pathways. This research provides a more detailed overview of the mechanism by which GBEE-PE inhibits MRSA both in vitro and in vivo and suggests that GBEE-PE is a new prospective antimicrobial with the potential to be used in MRSA therapeutics in the future.

Streptococcus agalactiae is the major cause of invasive neonatal infections and is a recognized pathogen associated with various diseases in nonpregnant adults. The emergence and spread of antibiotic-resistant S. agalactiae necessitate the development of a novel antibacterial agent. Here, the potential antibacterial activities and mechanisms of ginkgolic acid C15:1 (GA (15:1)) from Ginkgo biloba against clinical S. agalactiae are characterized. The MIC50 and MIC90 values for GA (15:1) against 72 clinical S. agalactiae isolates were 6.25 and 12.5 μM, respectively. GA (15:1) showed a strong bactericidal effect against both planktonic bacteria and bacteria embedded in biofilms as well as significant effectiveness in suppressing the growth of S. agalactiae biofilms. Moreover, GA (15:1) possesses intracellular antibacterial activity and could significantly decrease the bacterial burden in the intraperitoneal infection model of S. agalactiae. Mechanistic studies showed that GA (15:1) triggers membrane damage of S. agalactiae through a unique dual-targeting mechanism of action (MoA). First, GA (15:1) targets phospholipids in the bacterial cytoplasmic membrane. Second, by using mass-spectrometry-based drug affinity responsive target stability (DARTS) and molecular docking, lipoprotein signaling peptidase II (lspA) was identified as a target protein of GA (15:1), whose role is crucial for maintaining bacterial membrane depolarization and permeabilization. Our findings suggest a potential therapeutic strategy for developing GA (15:1) to combat S. agalactiae infections.

This study aims to examine the impact of the microfluidic preparation process on the quality of poly (lactic-co-glycolic acid) (PLGA) nanoparticles (NPs) co-delivered with scutellarin (SCU) and paeoniflorin (PAE) in comparison to a conventional emulsification method and to evaluatethe potential cardio-protective effect of SCU-PAE PLGA NPs produced through emulsification method. As compared with microfluidics, the nanoparticles prepared by emulsification method exhibited a smaller size, higher encapsulation efficiency, higher drug loading and lower viscosity for injection. Subsequently, a rat myocardial ischemia (MI) was established using male Sprague-Dawley (SD) rats (250 ± 20 g) subcutaneously injected with 85 mg/kg isoproterenol (ISO) for two consecutive days. The pharmacokinetic findings demonstrated that our SCU-PAE PLGA NPs exhibited prolonged blood circulation time in MI rats, leading to increased levels of SCU and PAE in the heart. This resulted in significant improvements in electrocardiogram and cardiac index, as well as reduced serum levels of CK, LDH, AST. Histopathological analysis using H&E and TUNEL staining provided further evidence of improved cardiac function and decreased apoptosis. Additionally, experiments measuring SOD, MDA, GSH, NO, TNF-α and IL-6 levels indicated that SCU-PAE PLGA NPs may effectively treat MI through oxidative stress and inflammatory pathways, thereby establishing it as a promising therapeutic intervention.

Although the potent antibacterial ability of radezolid against Staphylococcus aureus has been widely reported worldwide, its antibacterial and anti-biofilm activity against the S. aureus clinical isolates from China remains elusive. In this study, the minimum inhibitory concentration (MIC) of radezolid was determined in S. aureus clinical isolates from China using the agar dilution method, and the relationship between radezolid susceptibility and ST distribution was also investigated. The anti-biofilm activity of radezolid against S. aureus was determined by a crystal violet assay and compared with that of linezolid and contezolid. The quantitative proteomics of S. aureus treated with radezolid was analyzed, and the genetic mutations in radezolid-induced resistant S. aureus were determined by whole-genome sequencing. The dynamic changes in transcriptional expression levels of several biofilm-related genes were analyzed by quantitative RT-PCR. Our data showed that radezolid MIC ranged from ≤0.125 to 0.5 mg/L, which was almost 1/4 × MIC of linezolid against S. aureus, indicating the greater antibacterial activity of radezolid than linezolid. The S. aureus clinical isolates with radezolid MICs of 0.5 mg/L were most widely distributed in ST239 of MRSA and ST7 of MSSA. Moreover, the more robust anti-biofilm activity of radezolid with subinhibitory concentrations (1/8 × MIC and 1/16 × MIC) was demonstrated against S. aureus when compared with that of contezolid and linezolid. Genetic mutations were found in glmS, 23S rRNA, and DUF1542 domain-containing protein in radezolid-induced resistant S. aureus selected by in vitro induction of drug exposure. Quantitative proteomic analysis of S. aureus indicated that the global expression of some biofilm-related and virulence-related proteins was downregulated. Quantitative RT-PCR further confirmed that the expressions of some downregulated biofilm-related proteins, including sdrD, carA, sraP, hlgC, sasG, spa, sspP, fnbA, and oatA, were decreased after 12 h and 24 h of exposure to radezolid. Conclusively, radezolid shows robust antibacterial and anti-biofilm activity against S. aureus clinical isolates from China when compared with contezolid and linezolid.

Gastric ulcers are a common clinical presentation affecting anyone, regardless of their age or gender. Nanoparticles (NPs) containing Bletilla striata polysaccharide (BSP) and omeprazole (OME) were investigated in the study for their therapeutic effect on gastric ulcers. Ethanol-induced gastric ulcers in rats (240 ± 30 g) were established. Our OME-BSP NPs were more stable than free OME in the acidic environment and can increase the absorption of OME in rat stomach, which was confirmed by in situ gastric absorption and distribution experiments. The extended blood circulation of OME-BSP NPs was also observed in rats with gastric ulcer. More importantly, OME-BSP NPs not only decreased the area of gastric ulcer and inhibited gastric acid secretion but also reversed gastric tissue damage and cell apoptosis, as revealed by HE and TUNEL staining. Subsequent SOD, MDA, PGE2, IL-6, and TNF-α tests further verified the superiority of OME-BSP NPs against rat gastric ulcer, which properly originated from superior antioxidant and anti-inflammatory effects. As a result, our OME-BSP NPs' drug delivery system improved the stability and absorption of OME in the rat stomach and achieved targeted treatment of gastric ulcers.

Candida albicans causes mastitis in dairy cows, which reduces milk production, causes milk quality decline and increases the risk of foodborne infections. Antimicrobial peptides (AMPs) play important roles in insects due to their ability to inhibit the growth of bacteria, fungi and viruses. Moreover, sorbic acid and benzoic acid are widely used preservatives in the food industry. In this study, one of the housefly larval AMPs, Phormicin C-NS, modified with sorbic acid or benzoic acid, was synthesized, and the anti-C. albicans activity was evaluated. In addition, the potential mechanism of action of Phormicin C-NS against C. albicans was revealed by transcriptomic analysis. Phormicin C-NS inhibited the C. albicans bud-to-hyphal transition and biofilm formation by downregulating the hypha-specific genes HWP1, Ece1p and Als3. Moreover, it inhibited C. albicans propagation in pasteurized milk. The MIC of Phormicin C-NS for C. albicans was nearly 30 times lower than that of sorbic acid. Furthermore, Phormicin C-NS will significantly reduce sorbic acid usage in the milk industry as an alternative peptide. Eventually, the peptide is degraded into amino acids. In summary, Phormicin C-NS has good biosafety, enhances milk amino acid levels, and has great potential as a new preservative for use in the milk industry.

Ethnopharmacological relevance: The fruit of Ginkgo biloba L. (Ginkgo nuts) has been used for a long time as a critical Chinese medicine material to treat cough and asthma, as well as a disinfectant. Similar records were written in the Compendium of Materia Medica (Ben Cao Gang Mu, pinyin in Chinese) and Sheng Nong's herbal classic (Shen Nong Ben Cao Jing, pinyin in Chinese). Recent research has shown that Ginkgo biloba exocarp extract (GBEE) has the functions of unblocking blood vessels and improving brain function, as well as antitumour activity and antibacterial activity. GBEE was shown to inhibit methicillin-resistant Staphylococcus aureus (MRSA) biofilm formation as a traditional Chinese herb in our previous report in this journal. Aim of the stud: yThe antibiotic resistance of clinical bacteria has recently become increasingly serious. Thus, this study aimed to investigate the Ginkgo biloba exocarp extract (GBEE) antibacterial lineage, as well as its effect and mechanism on S. haemolyticus biofilms. This study will provide a new perspective on clinical multidrug resistant (MDR) treatment with ethnopharmacology herbs.Methods: The microbroth dilution assay was carried out to measure the antibacterial effect of GBEE on 13 types of clinical bacteria. Bacterial growth curves with or without GBEE treatment were drawn at different time points. The potential targets of GBEE against S. haemolyticus were screened by transcriptome sequencing. The effects of GBEE on bacterial biofilm formation and mature biofilm disruption were determined by crystal violet staining and scanning electron microscopy. The metabolic activity of bacteria inside the biofilm was assessed by colony -forming unit (CFU) counting and (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2HY-tetrazolium bromide (MTT) assay. Quantitative polymerase chain reaction (qPCR) was used to measure the gene expression profile of GBEE on S. haemolyticus biofilm-related factors. Results: The results showed that GBEE has bacteriostatic effects on 3 g-positive (G thorn ) and 2 g-negative (G-) bacteria among 13 species of clinical bacteria. The antibacterial effect of GBEE supernatant liquid was stronger than the antibacterial effect of GBEE supernviaould-like liquid. GBEE supernatant liquid inhibited the growth of S. epidermidis, S. haemolyticus, and E. faecium at shallow concentrations with minimum inhibitory concentrations (MICs) of 2 mu g/ml, 4 mu g/ml and 8 mu g/ml, respectively. Genes involved in quorum sensing, two-component systems, folate biosynthesis, and ATP-binding cassette (ABC) transporters were differentially expressed in GBEE-treated groups compared with controls. Crystal violet, scanning electron microscopy (SEM) and MTT as-says showed that GBEE suppressed S. haemolyticus biofilm formation in a dose-dependent manner. Moreover, GBEE supernatant liquid downregulated cidA, cidB and atl, which are involved in cell lysis and extracellular DNA (eDNA) release, as well as downregulated the cbp, ebp and fbp participation in encoding cell-surface binding proteins. Conclusions: GBEE has an excellent antibacterial effect on gram-positive bacteria and also inhibits the growth of gram-negative bacteria, such as A. baumannii (carbapenem-resistant Acinetobacter baumannii) CRABA and S. maltophilia. GBEE inhibits the biofilm formation of S. haemolyticus by altering the regulation and biofilm material-related genes, including the release of eDNA and cell-surface binding proteins.