BackgroundPolygonatum kingianum, a key species in traditional Chinese medicine, is increasingly valued for its medicinal and nutritional properties. However, wild resources are declining due to over-exploitation, necessitating genomic studies for conservation.ResultsFlow cytometry analysis based on genome size revealed the presence of both putative diploid and tetraploid P. kingianum. By further integrating sequencing data from both Illumina and Oxford Nanopore Technologies (ONT), the organelle genomes were assembled. While the chloroplast genomes of different putative ploidy levels of P. kingianum were highly conserved in structure and features, the mitogenome of putative tetraploid P. kingianum consisted of two chromosomes, whereas that of putative diploid P. kingianum contained only one. Mitochondrial gene annotation showed that putative tetraploid P. kingianum possesses one additional copy each of the nad3 and nad5 genes compared to the putative diploid P. kingianum. Analyses of repetitive sequences indicated that, although no obvious correlation was observed between tandem repeats or dispersed repeats and different putative ploidy levels in the chloroplast genomes or between tandem repeats and different putative ploidy levels in the mitogenomes, simple sequence repeats in the organelle genomes correlated with ploidy variation of P. kingianum. Investigation of RNA editing sites revealed ploidy-dependent variations in the cox2 and nad3 genes of the mitogenomes across different ploidy levels of P. kingianum, suggesting that differences in editing sites may be linked to the adaptability of the different ploidies. Phylogenetic analyses based on mitochondrial genes and nucleotide substitution rate analysis showed no significant differences or distinct variations among different ploidy levels of P. kingianum, supporting that the morphologically highly variable P. kingianum comprises only a single species.ConclusionsIn this study, we successfully assembled the organelle genomes of P. kingianum with different putative ploidy levels and conducted comparative genomic analyses. We found that the mitogenomes of P. kingianum are structurally complex and variable, playing crucial roles in evolution and adaptation. This study not only enhances our understanding of the organelle genome characteristics of different putative ploidy levels within the same species, but also lays the foundation for the subsequent development and utilization of the medicinal value of Polygonatum.

Four new ent-abietane diterpenoids 1-4 were isolated from the methanol extract of the roots of Euphorbia fischeriana. Their structures were elucidated using HRESIMS, NMR spectroscopy, and X-ray diffraction. The biological activities of these compounds were evaluated through cytotoxicity assay against three human breast cancer cell lines (MDA-MB-231, MDA-MB-468, and MCF-7) and anti-inflammatory assay using lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages, respectively. Among them, compound 2 displayed moderate cytotoxicity against MDA-MB-468, with an IC50 of 11.7 mu M, and compound 3 exhibited the most potent antiinflammatory effect, inhibiting NO production with an IC50 of 14.5 mu M.

The inefficient artificial breeding of the endangered largemouth bronze gudgeon (Coreius guichenoti) is partly attributed to compromised immunity and poor hepatic-intestinal health, yet no nutritional immunostimulants have been evaluated in this species. This study evaluated four immunostimulants—curcumin (CU), astaxanthin (AX), chitosan (CH), and Bacillus coagulans CICC 20138 (BC)—on immune function, antioxidant capacity, and intestinal microbiota of C. guichenoti. Six hundred juveniles (8.03±0.31 g) were allocated to five groups (including control, CO) and fed diets supplemented with 200 mg/kg of each immunostimulant, based on previously reported effective levels in other fish species, for 8 weeks. Compared to CO, all immunostimulant groups showed elevated plasma immunoglobulin M, albumin, complement C3/C4, and lysozyme levels. Hepatic glutathione peroxidase (GPX), superoxide dismutase (SOD), and catalase (CAT) activities increased in AX and BC groups, with reduced malondialdehyde (MDA). Intestinal antioxidant enzyme activities (GPX, SOD, CAT) were highest in BC, while MDA decreased in all supplemented groups. BC-fed fish exhibited reduced hepatic vacuolation and improved hepatocyte integrity. Intestinal microbiota analysis revealed higher Chao1 and Faith_pd diversity indices in BC, alongside a marked downregulation of the "enterobacterial common antigen biosynthesis" pathway. This suppression correlated with reduced pathogenic genera (Plesiomonas, Shigella) compared to CO. Collectively, B. coagulans CICC 20138 enhanced immunity, attenuated oxidative stress, mitigated hepatointestinal damage, and suppressed pathogen-associated metabolic pathways. These results identify B. coagulans CICC 20138 as a promising immunostimulant for C. guichenoti, warranting further dose-response optimization.

Tetrasargltones A and B (1 and 2, respectively), representing the first examples of lindenane sesquiterpenoid (LS) tetramers, were isolated from Sarcandra glabra. Their structures were fully elucidated via HR-MS and NMR, with their authenticity further confirmed by X-ray diffraction. Structurally, 1 and 2 possess two unprecedented fully helically polymerized carbon skeletons, which are formed through plausible sequential Diels-Alder cycloadditions at different sites of the triene LS precursor. Biologically, 1 and 2 exhibit cytotoxic activity against five cancer cell lines, with 2 showing the best activity in HCT116 cells (IC50 = 2.51 ± 0.13 μM) and exerting its cytotoxic effects through NF-κB pathway inhibition.

Herein, we report a novel acyl migration protocol based on a reductive radical-polar crossover process enabled by redox-neutral photocatalysis. Employing tertiary amines or alpha-silylamines as radical sources, modular access to various functionalized alpha-(hetero)aryl-alpha-keto-1,3-diamines could be efficiently realized via reactions of enamides with alpha-aminoalkyl radicals under mild reaction conditions. Additionally, the radical addition/acyl migration cascade process could be extended to the intramolecular version. The synthetic application of acyl migrated products was also demonstrated. Taking advantage of the easy generation of alpha-aminoalkyl radicals and alpha-imido carbanions via a single-electron-transfer process, this new procedure features a broad substrate scope and exogenous reductant-free conditions.

UbiA prenyltransferases (PTs) play an indispensable role in the prenylation of plant metabolites, yielding numerous natural products with enhanced pharmacological activities, such as cannabinoids, polycyclic polyprenylated acylphloroglucinols, prenylated flavonoids, and stilbenoids. These enzymes typically target specific carbon atoms or hydroxy groups of aromatic substrates. Despite the recent identification of dozens of plant‐derived UbiA PTs, their catalytic mechanism remains poorly understood, particularly regarding the precise control of regioselectivity. In this study, we identified and characterized a total of 10 members that catalyzed the regioselective prenylation and geranylation of moracin substrates through comprehensive analysis of the UbiA superfamily in Morus alba . Molecular dockings, dynamics simulations, and quantum chemical calculations revealed the substrate‐induced conformational changes leading to the formation of the hydrophobic reaction pocket, as well as the differential binding between various Ma PTs and moracin M. Additionally, the recognition of prenyl donors by Ma PT27 and the potential mechanism underlying the reversal of regioselectivity induced by different donors are discussed. Finally, structure‐based rational mutation altered the site preference from C7 to C5. These findings suggest that the regioselectivity of plant UbiA PTs is governed by both the inherent chemoselectivity of reaction sites and intricate protein–substrate interactions.

Herein, using reductive radical-polar crossover as a key process, a robust and practical protocol for the N → C acyl migration reaction has been successfully developed. A variety of enamides could react with carboxylic acids for modular access to α-(hetero)aryl-α-aminoketones enabled by redox-neutral photocatalysis. This decarboxylative radical addition/acyl migration cascade process features a broad substrate scope, good functional compatibility, and mild reaction conditions.

Bicyclo[1.1.1]pentane (BCP) heteroaryls make up an important class of BCP derivatives in drug discovery. Herein, we report the visible-light-mediated synthesis of cyanoisopropyl BCP-heteroaryls motifs from N-containing heterocycles, [1.1.1]propellane, and AIBN (2,2 '-azobis(isobutyronitrile)) through three-component cascade reaction. Importantly, this protocol is compatible with pyrazinones, quinoxaline-2(1H)-one, azauracils, quinoline derivatives, and imidazo[1,2-b]pyridazine, as well as various phenyl disulfide derivatives; thus, this operationally simple and general methodology could enable rapid library generation of sought-after BCP derivatives for drug development.

Background:: Filamentous fungi in the genus Aspergillus are well known for their important roles in production of bioactive secondary metabolites with diversely chemical structures and potential application in pharmaceutical industry. Objective:: The present study aimed to investigate the phenolic bisabolane sesquiterpene (PBS) derivatives from an Arctic marine-derived fungus Aspergillus sydowii MNP-2. Methods:: In this study, antimicrobial activities were carried out according to the broth microdilution assay, nitric oxide (NO) production in mouse macrophages (RAW264.7) and BV2 microglial cells was used to detect the inhibitory effect of compounds in inflammatory reactions, and in vitro inhibitory cell proliferation activity was determined by the cell counting kit-8 (CCK-8) assay. Results:: In this work, chemical investigation of an Arctic marine-derived strain A. sydowii MNP-2 led to the isolation of 11 PBSs (1-11) using various chromatographic methods. Their chemical structures were unambiguously determined by 1H NMR spectroscopy and mass spectrometry analyses as well as comparison with literature data. It is noteworthy that compounds 1, 7 and 11 were firstly obtained from A. sydowii. Antimicrobial assay showed that these chemicals had no potent inhibitory effect on Staphylococcus aureus, Escherichia coli, and Candida albicans with MIC values > 16 μg/mL. Additionally, the inhibition of nitric oxide (NO) production in lipopolysaccharide (LPS)- induced inflammation in mouse macrophages (RAW264.7) and BV2 microglial cells were all below 10% for compounds 4-6 and 8, indicating almost negligible anti-inflammatory efficacy. Among the tested compounds 4-6 and 8 for tumor-cell proliferation inhibition activities, compound 5 demonstrated the strongest inhibitory effect against human acute promyelocytic leukemia cells (HL-6) with a 44.76% inhibition rate. Conclusion:: In the present study, 11 PBS derivatives were purified and characterized from the solidand liquid-state fermentations of the Arctic marine-derived fungus A. sydowii MNP-2. Unfortunately, none of these metabolites had significant antimicrobial, anti-inflammatory, or tumor-cell proliferation inhibition activities.

Helicteres angustifolia L. (H. angustifolia), a well-known traditional Chinese medicine, has been demonstrated to have hypoglycemic activity. We found that the EtOAc extract of H. angustifolia (HAEF) showed stronger α-glucosidase inhibitory activity than that of positive control. Furthermore, the hypoglycemic activity of HAEF was evaluated in streptozotocin (STZ)-induced type 2 diabetes mellitus (T2DM) rats. The results demonstrated that HAEF reduced the drinking quantity, feeding quantity, and controlled weight loss in diabetic rats. Besides, the fasting blood glucose (FBG), viscera index, and the area under time-blood glucose curve (AUC) were significantly decreased, and the oral glucose tolerance was also improved after 5 weeks. Then, the high-performance liquid chromatography with quadrupole time of flight tandem mass spectrometry (HPLC-Q-TOF-MS/MS) method was performed for qualitative analysis of the chemical constituents in HAEF. Twenty-one compounds were identified from in HAEF. Four compounds were further isolated from HAEF and subjected to α-glucosidase inhibition experiments. At the end, molecular docking was empolyed simulate the interaction of three compounds with α-glucosidase. This is the first report on major hypoglycaemic components has been identified in the roots of H. angustifolia. These findings provide a material basis for the use of H. angustifolia in the treatment of diabetes.

This study describes a pioneering visible-light-induced phosphine-catalyzed halogen-atom transfer (XAT) strategy that heralds a new era in the difunctionalization of [1.1.1]propellane.

The traditional Chinese medicine (TCM) formula Ento-PB containing Periplaneta americana (Linnaeus) (Blattidae) and Taraxacum mongolicum Hand.-Mazz. (Compositae) has great potential for treating inflammation. This study explored the effects of Ento-PB on ulcerative colitis (UC). The UC model was induced with 2,4,6-trinitrobenzene sulfonic acid (TNBS) by enema. Male Sprague–Dawley rats (n = 32) were divided into four groups: (1) control group that received 2.5 mL/kg normal saline, (2) TNBS group that received 2.5 mL/kg normal saline, (3) Ento-PB low-dose group that received 100 mg/kg Ento-PB, and (4) Ento-PB high-dose group that received 200 mg/kg Ento-PB. Rats were administered drugs via enema for 14 days after modeling. The disease activity index (DAI), colon mucosa damage index (CMDI), histopathological score (HS), levels of interleukin-8 (IL-8), IL-10, IL-17, tumor necrosis factor-α (TNF-α), and C-reactive protein (CRP) in serum, contents of IL-2, myeloperoxidase (MPO), transforming growth factor-β1 (TGF-β1), and epidermal growth factor (EGF) in the colon, and abundance of Bifidobacterium, Lactobacillus, Enterococcus, Bacteroides, and Escherichia coli were assessed. Ento-PB administration showed a significant reduction in DAI, CMDI, and HS, contents of IL-2, IL-8, IL-17, TNF-α, CRP, and MPO, and a significant increase in the levels of IL-10, TGF-β1, and EGF. Compared with the TNBS-administered group, the abundance of Bifidobacterium, Lactobacillus, Enterococcus, and E. coli decreased, while an obvious increase in the proportion of Bacteroides was found in the Ento-PB-administered groups. Ento-PB alleviated inflammation in UC by regulating the equilibrium of Th1/Th17/Treg cytokines and recovering the imbalance between the gut microbiota. Applying Ento-PB in treating UC could be suggested.

With the rapid emergence of microbial infections induced by "superbugs", public health and the global economy are threatened by the lack of effective and biocompatible antibacterial agents. Herein, we systematically design a series of secondary ammonium-based hyperbranched poly(amidoamine) (SAHBP) with different alkyl chain lengths for probing high-efficacy antibacterial agents. SAHBP modified with alkyl tails at the hyperbranched core could efficiently kill Escherichia coli and Staphylococcus aureus, two types of clinically important bacteria worldwide. The best SAHBP with 12-carbon-long alkyl tails (SAHBP-12) also showed high activity against problematic multidrug-resistant bacteria, including Pseudomonas aeruginosa and methicillin-resistant S. aureus (MRSA). Based on ζ potential, isothermal titration microcalorimetry (ITC), and membrane integrity assays, it is found that SAHBP-12 could attach to the cell membrane via electrostatic adsorption and hydrophobic interactions, following which the integrity of the bacterial cell wall and the cell membrane is disrupted, resulting in severe cell membrane damage and the leakage of cytoplasmic contents, finally causing bacterial cell death. Impressively, benefiting from excellent membrane-active property, SAHBP-12 exhibited robust therapeutic efficacy in MRSA-infected mice wounds. Moreover, SAHBP-12 also showed excellent biosafety in vitro and in vivo, which undoubtedly distinguished it as a potent weapon in combating the growing threat of problematic multidrug-resistant bacterial infections.

Natural deep eutectic solvent (NaDES) is widely applied in the extraction of nutrients from natural resources as a greener alternative for fossil solvent. In the present work, 27 different NaDESs were screened for the extraction of paeoniflorin (PF) and galloyl paeoniflorin (GPF) from Radix Paeoniae Rubra (RPR). After screening and extraction parameter optimization, the extraction yields of PF and GPF reached up to 182.8 mg/g and 77.4 mg/g with the selected NaDES, ChCl-Sor. Furthermore, the antioxidant activity in vitro and neuroprotectivity in vivo of the 'ready-to-use' extracts were evaluated comprehensively. Especially in vivo, the cerebral ischemic/ reperfusion injury model was established in rats and the protective effects of the RPR extracts were determined. The results not only proved that NaDES is a valuable green extraction media, but also indicated the safety and potential pharmaceutical application of NaDES based 'ready-to-use' extracts from medical plants.

With the continuous emergence of drug-resistant pathogens, new strategies with high antibacterial efficacy are urgently needed. Herein, five cationic nano-sized hyperbranched polymers (CNHBPs) with cationic functional groups have been constructed, and their antibacterial mechanism has been studied in detail. CNHBPs bearing secondary ammonium salt groups and long alkyl chains (S12-CNHBP) exhibited weak antibacterial and antibiofilm ability, while CNHBPs bearing quaternary ammonium salt groups and long alkyl chains (Q12-CNHBP) showed the highest antimicrobial and strongest antibiofilm activities. ζ potential and isothermal titration microcalorimetry (ITC) results suggest that the negatively charged surfaces of bacterial cells provided Q12-CNHBP with a higher intrinsic electrostatic driving force for bacterial killing than that with S12-CNHBP. Fluorescent tracing and morphological observations indicate that the bacterial genome might be another antibacterial target for S12-CNHBP in addition to the cell wall and membrane, which are mainly antibacterial targets for Q12-CNHBP, making it less likely to induce bacterial resistance. Surprisingly, Q12-CNHBP exhibited superior in vivo therapeutic efficacy in a mouse wound model of methicillin-resistant Staphylococcus aureus (MRSA) infection with low toxicity during treatment. These advantages and ease of preparation will undoubtedly distinguish Q12-CNHBP as a new class of suitable candidates to combat multidrug-resistant pathogen infections. This study opens up a new avenue for exploiting antibacterial biomaterials to treat infections caused by drug-resistant bacteria.

Natural deep eutectic solvents (NaDESs) are promising green alternatives to conventional solvents widely applied in the extraction of natural products due to their physical and chemical superiorities. In present study, 22 NaDESs consisted from food grade ingredients were screened in ultrasonic assisted extraction (UAE) of bioactive compounds from safflower. The oral bioavailabilities of hydroxysafflor yellow A (HSYA) and anhydrosafflor yellow B (ASYB) in the extracts were then investigated in SD rats with the help of HPLC-MS technique. The results revealed that l-proline-acetamide (l-Pro-Am) was an effective solvent with the yields of HSYA and ASYB at 32.83 and 8.80 mg/g. Pharmacokinetic studies revealed that the blood level of HSYA and ASYB were significantly higher after oral administration of l-Pro-Am extract than that of aqueous extract. Especially, the relative bioavailabilities (to aqueous extract) of HSYA and ASYB were calculated 183.5% and 429.8%.

Natural deep eutectic solvent (NaDES) is generally considered as a greener alternative to fossil solvent, with great potential in various areas. In the present work, 25 different NaDESs were screened for the extraction of puerarin (PUE) and its two natural derivatives from Radix Pueraria (RP). As the main isoflavone in RP, PUE has a wide range of biological activities. However, its application is restricted due to its poor solubility in water and low oral bioavailability. In this study, the extraction of PUE with NaDESs showed significant advantages compared with traditional solvents. While using L-Pro-Maa (L-proline-malic acid) under optimal conditions, the optimized yields of PUE, 3-MPR and PRX were 98.7 mg/g, 16.3 mg/g and 9.9 mg/g, respectively, which were 2.2-, 2.9- and 3.4-fold higher than that of water. Furthermore, the oral bioavailability of PUE in NaDES extracts was comparatively investigated in rats with HPLC-MS technique. Pharmacokinetic analysis revealed that the relative bioavailability of PUE in L-Pro-Maa extract is 323%. The result indicated that NaDES is not only a sustainable ionic liquid with higher extraction efficiency, but also an enhancer of oral bioavailability of specific natural products.

BackgroundThromboembolic events are leading causes of mortality and morbidity all over the world. Tongmai (TM) is a botanical drug with valid clinical efficacy and safety in the management of thrombosis and ischemic cardiovascular diseases, however, its active compounds and underlying mechanism are largely unclear.PurposeTo investigate the endogenous effects, therapeutic mechanism and active compounds of TM in thrombus formation.Study designCombined with transgenic zebrafish models and high-content imaging system, this study evaluated the endogenous antithrombotic effects of TM and screened for the active compounds.MethodsThe PHZ-induced thrombotic model in erythrocytes or platelets labeled transgenic zebrafish were established, to dynamically evaluate the antithrombotic effects of TM. The oxidative damage levels were analyzed by specific fluorescent probes, and the expression levels of key factors in coagulation cascades and platelet activation were examined by QPCR. TM were dissected into fractions by reverse phase chromatography and subsequently screened for their antithrombotic effects in the transgenic fish models. The compounds of the active TM fraction were then analyzed by UPLC-Q-TOF analysis and further verified for their antithrombotic effects and mechanisms.ResultsIn PHZ-induced zebrafish thrombotic model, TM incubation markedly increased cardiac blood flow, decreased peripheral erythrocytes aggregation, and recovered peripheral platelet circulation. Besides, the levels of oxidative stress and lipid peroxidation were increased in the PHZ-induced thrombotic fish, which were greatly decreased by TM treatment. Moreover, TM significantly reduced the expression of coagulation factor II (thrombin) and the downstream fibrinogen. In order to identify the active compounds of TM, four fractions were separated from the extract by reverse phase chromatography, which were subsequently screened for their antithrombotic effects in the fish model. As a result, fraction 4 showed the strongest effect in inhibiting thrombosis. Finally, through UPLC-Q-TOF analysis and endogenous screening, cryptotanshione was identified as the main active compound with antithrombotic effects.ConclusionOur study demonstrated the endogenous antithrombotic effects of TM, which is possibly mediated by inhibiting oxidative stress and coagulation cascade. Cryptotanshione was identified as a major compound with antithrombotic activity and is a promising candidate for novel antithrombotic therapy.

This article presents a new label-free fluorescence assay based on supramolecular self-assembly of cucurbit[7]uril and specific peptide Gly-Pro-Phe-Gly for monitoring DPP4 activity in clinical samples. It also displays a good potential application in high-throughput screening of DPP4 inhibitors.