Liubao tea (LBT), a traditional dark tea, is recognized for its health-promoting properties, particularly its potential to mitigate obesity. However, differences in functional efficacy among distinct aroma types remain unexplored. In this study, we systematically compared the anti-obesity activities and phytochemical characteristics of three aroma-type LBTs, namely, ginseng (GA), betelnut (BA), and stale (SA), to clarify the compositional and functional distinctions. Phytochemical analysis showed that all three teas were rich in polyphenols, flavonoids, organic acids, and other compounds, with GA-type LBT containing the highest polyphenol (477.32 mg/g) and flavonoid (240.83 mg/g) contents. In vitro, all LBT extracts displayed antioxidant and anti-inflammatory activities. Among them, GA exhibited the strongest effects, scavenging over 60% of free radicals, inhibiting pancreatic lipase by 89.27%, and reducing nitric oxide production in macrophages by 67.95%. In a high-fat diet-induced obese mouse model, GA-type LBT also showed the most pronounced anti-obesity outcomes, reducing weight gain by 52.98%, alleviating hepatic steatosis and systemic inflammation, and improving serum lipid profiles. Mechanistic studies revealed that LBT extracts modulated the gut microbiota by increasing beneficial microbes and decreasing harmful ones, while also regulating lipid metabolism pathways related to glycerophospholipid, glycerolipid, and arachidonic acid. In conclusion, although all three aroma types possess anti-obesity potential, GA-type LBT stands out due to its unique bioactive composition and consistently strong efficacy both in vitro and in vivo. This study identifies GA-type LBT as a promising candidate for anti-obesity dietary supplements and provides a scientific basis for the aroma-specific selection of LBT.

Cassava (Manihot esculenta Crantz) storage roots exhibit significant variation in starch content among cultivars, yet the metabolic and molecular mechanisms governing carbon allocation between storage and structural components remain poorly understood. Here, we investigated carbon partitioning in two cassava cultivars with distinct starch phenotypes: high-starch FX01 and low-starch SC16. Using 13C isotope labeling coupled with metabolomic analysis, we traced the pathway of carbohydrates through primary and secondary metabolism. The results revealed that SC16 exhibits enhanced photosynthetic capacity and elevated soluble sugar content in storage roots, whereas FX01 demonstrates superior starch synthesis due to its efficient glucose and fructose phosphorylation. Conversely, SC16 exhibits a faster conversion of 13C-labeled ferulic acid, directing carbon flow towards lignin biosynthesis via the phenylpropanoid pathway. Further, by silencing the MeCOMT8 gene, encoding a key enzyme in ferulic acid biosynthesis, we observed a reduction in lignin content and an increase in ADP-glucose levels in the MeCOMT8-silenced cassava plants, suggesting a regulatory link between these competing pathways. Our research elucidated that the variations in carbon allocation between starch and lignin biosynthesis among different cultivars are finely orchestrated through the specific-step alteration of metabolic flux. These findings provide potential candidate targeted points and valuable insights for high-starch breeding in cassava. A study shows that variations in carbon allocation between starch and lignin biosynthetic pathways among cassava cultivars are regulated by specific metabolic flux step alterations and demonstrates MeCOMT8’s critical role in lignin synthesis.

Chilli veinal mottle virus (ChiVMV) is the most important virus known to threaten chilli pepper (Capsicum annuum) growth and yield in Asia and Africa. Here, we identified the class III peroxidase gene CaPOD49 as a key regulator of chilli pepper immunity against ChiVMV. The CaPOD49 expression was strongly induced in response to ChiVMV infection at 48 h post-inoculation in both resistant and susceptible pepper cultivars. Silencing CaPOD49 via virus-induced gene silencing (VIGS) significantly increased disease severity, reduced plant survival rates and increased the disease index of ChiVMV infection. This susceptibility was associated with the elevated accumulation of reactive oxygen species (ROS), while the treatment with the ROS scavenger diphenyleneiodonium (DPI) recovered pepper resistance. Additionally, the defence-related genes CaPR2, CaPR5 and CaPR10 were significantly suppressed in CaPOD49-silenced plants, as well as increasing its lipid peroxidation level. Heterologous expression of CaPOD49 in yeast enhanced its oxidative stress tolerance, consistent with the protective effects demonstrated by its peroxidase activity in vitro assays. Our findings demonstrate that CaPOD49 positively regulates chilli pepper immunity by fine-tuning ROS levels and activating defence responses, offering potential genetic targets for disease-resistant chilli pepper breeding.

Cassava (Manihot esculenta Crantz) is a tropical crop with storage roots containing large amounts of starch, but excessive lignin deposition negatively affects processing properties and quality. Developing cassava germplasm with enhanced starch accumulation and reduced lignin content is therefore an important breeding objective. Nevertheless, it remains unclear whether induced genetic variation can modulate starch and lignin accumulation and produce genotypes with improved compositional traits. In this study, an in vitro mutagenesis approach combining atmospheric and room temperature plasma (ARTP) and ethyl methanesulfonate (EMS) was established in cassava cultivar SC205. A total of 184 regenerated lines were obtained, among which 16 lines exhibiting vigorous growth and stable storage root formation were selected for further evaluation. Considerable variation in storage root composition was observed among the mutant lines, and starch accumulation was consistently negatively correlated with lignin content across two consecutive planting years. Notably, mutant line A18 exhibited increased starch accumulation and reduced lignin content compared with the wild type (WT), whereas E27 displayed the opposite phenotype. To investigate the molecular basis underlying the differences in starch and lignin accumulation, metabolomic profiling and qPCR analyses were conducted using storage roots from A18, E27, and WT plants. Metabolomic and transcriptional analyses revealed that A18 exhibited enhanced starch and sucrose metabolism and increased expression of starch biosynthesis-related genes, whereas E27 showed activation of phenylpropanoid metabolism and elevated expression of lignin biosynthesis-related genes. This study demonstrates that ARTP- and EMS-induced mutagenesis generated substantial compositional diversity while maintaining the negative association between starch accumulation and lignification. The identified high-starch/low-lignin mutant A18 provides a valuable genetic resource for elucidating the regulatory relationship between starch accumulation and lignin biosynthesis in cassava storage roots.

Caffeic acid O-methyltransferase (COMT) is a pivotal enzyme in phenylpropanoid biosynthesis. We identified 39 COMT genes in cassava. Among these, the proximal duplicate gene pair MeCOMT8a and MeCOMT8b (MeCOMT8a/b) was a key candidate associated with lignin and flavonoids. Co-silencing of MeCOMT8a and MeCOMT8b promoted adventitious root (AR) growth but reduced lignin and flavonoid content. Multiomics analyses revealed downregulation of phenylpropanoid- and flavonoid-related genes, and decreased O-methylated flavonoids, such as laricitrin. In vitro enzymatic assays confirmed that both MeCOMT8a and MeCOMT8b efficiently catalyzed the conversion of myricetin to laricitrin. Silencing of MeCOMT8a/b also led to elevated auxin levels and upregulation of auxin-responsive genes, providing a mechanistic explanation for the enhanced AR phenotype. Consequently, MeCOMT8a/b-silenced plants exhibited enhanced drought tolerance, attributable to a more robust root system and reduced oxidative damage. Thus, silencing of MeCOMT8a/b enhances adventitious root development and drought tolerance in cassava by modulating O-methylated flavonoid metabolism.

Background: Flavonoid secondary metabolites accumulate in xylem during heartwood formation and critically influence wood color and economic value. However, the distribution patterns of individual flavonoids across sapwood, transition zone, and heartwood remain poorly characterized in Castanopsis hystrix, limiting mechanistic understanding of heartwood formation and rational resource utilization. Methods: Sapwood, transition zone, and heartwood samples were collected and analyzed using ultra-performance liquid chromatography coupled with Q-Exactive Orbitrap mass spectrometry (UPLC-QE-MS) for qualitative and quantitative flavonoid determination. Multivariate statistical approaches, including hierarchical clustering, principal component analysis (PCA), and orthogonal partial least-squares discriminant analysis (OPLS-DA), were applied to compare regional metabolic profiles and to identify characteristic differential markers. Results: A total of 26 flavonoids were identified, covering flavones, flavonols, flavanones, dihydroflavonols, and flavanols. Quantitative and cluster analyses revealed distinct region-preferential accumulation: dihydrorobinetin, dihydromyricetin, morin, and kaempferol were highly enriched in heartwood; fustin, taxifolin, and cyanidin predominated in the transition zone, whereas afzelin and astilbin were more abundant in sapwood. PCA and OPLS-DA further selected four characteristic markers, namely dihydromyricetin, dihydrorobinetin, kaempferol and (−)-epicatechin, that effectively discriminated the three regions. Conclusions: This study establishes a clear radial gradient of flavonoid accumulation in C. hystrix xylem, demonstrating selective enrichment of specific metabolites from sapwood to heartwood. These findings provide essential data for deciphering the biochemical mechanisms of heartwood formation and offer a metabolic basis for quality assessment and high-value utilization of this timber species.

Metabolic dyshomeostasis of bile acid (BA) and microbiota-derived short-chain fatty acid (SCFA) is a critical driver of obesity pathogenesis. Liupao tea (LPT) has garnered attention for its anti-obesity potential, owing to potent lipid-lowering, microbiota- and BA metabolism-regulatory properties. However, whether LPT coordinately modulates BA and SCFA homeostasis remains unclear. This study investigated the therapeutic potential of LPT extract (LPTe) in alleviating obesity by focusing on the host-microbial re-establishment of SCFA and BA homeostasis. Results showed that five potential bioactive LPTe-derived constituents were identified in vivo. LPTe intervention effectively attenuated body weight gain, dyslipidemia, hepatic steatosis, intestinal barrier dysfunction, and systemic inflammation in HFD mice. LPTe treatment was associated with a shift in BA biosynthesis toward a chenodeoxycholic acid (CDCA)-enriched profile and with altered enterohepatic BA circulation. This shift coincided with increased fecal BA excretion and diminished lithocholic acid (LCA) accumulation, ultimately mitigating hepatic inflammation and lipotoxicity. LPTe altered gut microbial composition, reducing the relative abundances of genera potentially associated with secondary BA metabolism, in parallel with lower LCA levels. Simultaneously, increased SCFA levels were accompanied by higher colonic mRNA expression of GPR41 and GPR109A, increased ZO-1 and Occludin protein expression, and improved mucosal barrier-related indices. Crucially, antibiotic-mediated microbiota depletion abolished these metabolic benefits. Collectively, LPTe alleviated HFD-induced obesity through microbiota-dependent remodeling of the gut-liver axis, driven by the coordinated restoration of SCFA and CDCA homeostasis. These findings offer a theoretical basis for leveraging LPTe as a functional dietary intervention for obesity management.

As a highly disabling chronic inflammatory disease, rheumatoid arthritis (RA) necessitates novel interventions. Liupao tea is a traditional Chinese dark tea known for its favorable anti-inflammatory properties. This study aims to elucidate the active ingredients and action mechanisms underlying the therapeutic effects of Liupao tea extract (LPTE) in RA. LPTE was preliminarily characterized by LC-MS technology. Network pharmacology and molecular docking predicted anti-RA compounds, targets, and pathways, with key compounds identified using chemical standards. The effect of LPTE on the collagen-induced arthritis mouse model was evaluated through serum biochemical analysis, micro-CT imaging, and histopathological analyses. Integrated serum metabolomics, 16S rRNA sequencing, MetOrigin analysis, SCFA metabolomics, and quantitative real-time PCR elucidated gut–joint axis mechanisms. LPTE effectively attenuated RA symptoms by reducing bone destruction and joint inflammation. Notably, LPTE reshaped gut microbiota by enriching key families such as Monoglobaceae, Eggerthellaceae, and Desulfovibrionaceae, thereby promoting SCFA production. Increased SCFA levels enhanced intestinal barrier integrity and exerted joint-protective and anti-inflammatory effects by upregulating tight junction proteins and activating SCFA receptors. LPTE also modulated arachidonic acid metabolism by affecting key genes such as Alox5, Ptgs2, and Cbr1. These effects collectively reduced the levels of pro-inflammatory cytokines and increased the expression of anti-inflammatory cytokines in joints. Additionally, quercetin, luteolin, ellagic acid, and kaempferol were identified as major anti-RA bioactive compounds in LPTE. Taken together, this study provides preliminary evidence that LPTE mitigates RA by regulating the gut–joint axis mediated via fatty acid metabolism.

Liupao tea (LPT) is a Chinese dark tea known to possess a unique flavour. Microbial fermentation plays a crucial role in flavour development and enrichment. Currently, the phytochemical profiles and bioactivities of LPT with and without fermentation are not fully known. In this study, we compared the chemical composition of raw tea (SF), stale-aroma (SA), and betelnut-aroma (BA) type LPT through the application of GC/LC-MS-based metabolomics, and experimentally investigated their bioactivities via antioxidant, anti-inflammatory, hypolipidemic, and hypoglycemic assays in vitro. The results indicated that fermentation enhanced the flavour of LPT as evidenced by the sweetness-producing substances, decreased bitterness and astringency-related compounds and enriched abundance of aroma-generating compounds. Two and four volatiles were detected to be major contributors to the aroma in SA and BA, respectively. Fatty acids and phosphatidylcholines were the primary lipids, among which the lysing diacylglycerol trimethyl homoserines were found to be a new class of lipids in LPT. Notably, the fermentation resulted in the degradation of compounds, particularly glycerophospholipids and saccharolipids. SF had the highest level of bioactivity, followed by BA and SA. These findings expand the present understanding regarding the development of flavour, nutrition, and medicinal value of LPT. Moreover, they provide a theoretical basis for the identification of BA and SA and serve as a reference value for consumers in their selection of LPT products.

Liupao tea is a post-fermented dark tea with bitterness and astringency as key sensory traits, though its chemical composition is not fully understood. Six Liupao tea samples with significant differences in bitterness and astringency were analyzed using non-targeted metabolomics and sensory evaluation. Thirty finished and five semi-finished Liupao tea samples were analyzed using UHPLC-MS-PRM for targeted quantification of bitter and astringent compounds. The results show that 477 non-volatile compounds were detected, including 18 potential bitter compounds and 22 potential astringent compounds. Six key bitter compounds (epigallocatechin gallate, catechin gallate, caffeine, quinic acid, neochlorogenic acid, and caffeic acid) and 11 key astringent compounds (e.g., epigallocatechin gallate, gallic acid, chlorogenic acid, ellagic acid) were identified. After fermentation, flavonoid glycosides and flavanols were reduced by 62.41 % to 97.46 %, while phenolic acids showed varied trends. Different rates of change in key compounds during fermentation resulted in variations in bitterness and astringency. This study offers insights for improving Liupao tea quality.

Background: Type 2 diabetes (T2D) has become a serious global public health concern. Liubao tea (LBT) has demonstrated beneficial effects on gut microbiota and glucose-lipid metabolism, holding promising therapeutic potential for T2D; however, its underlying mechanisms remain unclear. This study aims to elucidate the potential mechanisms of Liubao tea extract (LBTE) against T2D. Methods: LC-MS technology was used to identify the chemical components of LBTE and combined with network pharmacology and molecular docking to screen its potential active ingredients and targets for improving T2D. Therapeutic efficacy was assessed in high-fat diet/streptozotocin (HFD/STZ)-induced diabetic mice via serum biochemical analyses and histopathological examinations. Serum metabolomics, 16S rRNA sequencing, quantification of short-chain fatty acids (SCFAs), quantitative real-time PCR (qPCR), and antibiotic-treated pseudo-germ-free models were employed to elucidate the underlying mechanisms. Results: LBTE effectively reduced blood glucose levels and improved lipid metabolism, primarily by promoting hepatic glycogen synthesis and suppressing glycerophospholipid synthesis. LBTE also alleviated hepatic inflammation by modulating inflammatory cytokine expression. Additionally, LBTE reshaped the gut microbiota profiles by decreasing harmful bacteria and increasing SCFA-producing bacteria, resulting in elevated fecal SCFAs. SCFAs contributed to improving hepatic metabolism and inflammation, enhancing intestinal barrier function. Notably, these effects were abolished by antibiotic-induced microbiota depletion, confirming the microbiota-dependent mechanism of LBTE. Quercetin, luteolin, genistein, and kaempferol were considered as potential active ingredients contributing to the antidiabetic effects of LBTE. Conclusions: These findings provide novel perspectives on the viability of LBTE as a complementary strategy for T2D prevention and management.

Background Understanding the metabolic changes in colorectal cancer (CRC) and exploring potential diagnostic biomarkers is crucial for elucidating its pathogenesis and reducing mortality. Cancer cells are typically derived from cancer tissues and can be easily obtained and cultured. Systematic studies on CRC cells at different stages are still lacking. Additionally, there is a need to validate our previous findings from human serum.Methods Ultrahigh-performance liquid chromatography tandem high-resolution mass spectrometry (UHPLC-HRMS)-based metabolomics and lipidomics were employed to comprehensively measure metabolites and lipids in CRC cells at four different stages and serum samples from normal control (NR) and CRC subjects. Univariate and multivariate statistical analyses were applied to select the differential metabolites and lipids between groups. Biomarkers with good diagnostic efficacy for CRC that existed in both cells and serum were screened by the receiver operating characteristic curve (ROC) analysis. Furthermore, potential biomarkers were validated using metabolite standards.Results Metabolite and lipid profiles differed significantly among CRC cells at stages A, B, C, and D. Dysregulation of glycerophospholipid (GPL), fatty acid (FA), and amino acid (AA) metabolism played a crucial role in the CRC progression, particularly GPL metabolism dominated by phosphatidylcholine (PC). A total of 46 differential metabolites and 29 differential lipids common to the four stages of CRC cells were discovered. Eight metabolites showed the same trends in CRC cells and serum from CRC patients compared to the control groups. Among them, palmitoylcarnitine and sphingosine could serve as potential biomarkers with the values of area under the curve (AUC) more than 0.80 in the serum and cells. Their panel exhibited excellent performance in discriminating CRC cells at different stages from normal cells (AUC = 1.00).Conclusions To our knowledge, this is the first research to attempt to validate the results of metabolism studies of serum from CRC patients using cell models. The metabolic disorders of PC, FA, and AA were closely related to the tumorigenesis of CRC, with PC being the more critical factor. The panel composed of palmitoylcarnitine and sphingosine may act as a potential biomarker for the diagnosis of CRC, aiding in its prevention.

Tank fermentation is a novel fermentation technology; however, the role of microorganisms in Liupao tea quality is unknown. Using high-throughput sequencing, untargeted metabolomics, and targeted absolute quantification, the microbial community, chemical profile, metabolites, and metabolic pathway of Liupao tea were discovered. Sphingomonas, Aquabacterium, Wolbachia, Blastobotrys, Aspergillus, Rasamsonia, and Candida were found to be the most common microorganisms, and throughout fermentation, Blastobotrys was the dominant fungal genus. A total of 40 compounds were identified as differentially changed metabolites (p < 0.05, VIP >1) (VIP: variable important in projection) and significantly contributing to tank fermentation, flavonoids and amino acid derivatives decreased, while nucleotides significantly increased. Gallic acid, EGCG, CG, EGC, EC, etc. could be identified as markers to assess the degree or completion of Liupao tea fermentation. There were significant (p < 0.01) correlations between microbial populations and metabolites, with Sphingomonas being significantly (p < 0.01) associated with 27 metabolites and Blastobotrys being significantly (p < 0.01) associated with 12 metabolites. The metabolic pathways of catechins, flavonoids, and nucleotides elucidate the factors influencing tea quality formation. This research improved our understanding of how Liupao tea processing affects its chemical composition and quality formation.

Salicylic acid (SA) plays a crucial role in plant defense against biotrophic and semibiotrophic pathogens. In Arabidopsis (Arabidopsis thaliana), isochorismate synthase 1 (AtICS1) is a key enzyme for the pathogen-induced biosynthesis of SA via catalytic conversion of chorismate into isochorismate, an essential precursor for SA synthesis. Despite the extensive knowledge of ICS1-related menaquinone, siderophore, and tryptophan (MST) enzymes in bacteria, the structural mechanisms for substrate binding and catalysis in plant isochorismate synthase (ICS) enzymes are unknown. This study reveals that plant ICS enzymes catalyze the isomerization of chorismate through a magnesium-dependent mechanism, with AtICS1 exhibiting the most substantial catalytic activity. Additionally, we present high-resolution crystal structures of apo AtICS1 and its complex with chorismate, offering detailed insights into the mechanisms of substrate recognition and catalysis. Importantly, our investigation indicates the existence of a potential substrate entrance channel and a gating mechanism regulating substrate into the catalytic site. Structural comparisons of AtICS1 with MST enzymes suggest a shared structural framework with conserved gating and catalytic mechanisms. This work provides valuable insights into the structural and regulatory mechanisms governing substrate delivery and catalysis in AtICS1, as well as other plant ICS enzymes.

Buffalo colostrum is the initial mammary secretion after parturition, consisting of nutritional and bioactive components. In this study, we conducted a proteomic analysis of buffalo colostrum whey to identify bioactive proteins and peptides. A total of 107 differentially expressed proteins (DEPs) were identified in buffalo colostrum whey compared to those in mature milk. Gene Ontology analysis revealed that DEPs were primarily associated with immune response and tissue development. KEGG pathway enrichment suggested that colostrum actively enhances nascent immunity involved in interleukin and interferon signaling pathways. Furthermore, candidate antimicrobial peptides (AMPs) of whey protein hydrolysates from buffalo colostrum were characterized, which exhibits broad-spectrum activity against gram-positive and gram-negative pathogens. Overall, this study improves our understanding of protein variations in buffalo lactation, and contributes to the development of AMPs from buffalo colostrum.

Russula vinosa Lindbl (RVL) is a multifunctional edible fungus that has recently gained widespread attention due to its numerous nutritious, delicious, and medicinal properties. However, little attention has been directed toward analyzing comprehensive phytochemical profiling and bioactivity in RVL. This study aimed to evaluate the phytochemical and medicinal profiling of RVL systematically. Herein, we characterized global RVL phytochemical profiling using the LC/GC-MS-based multiomics strategy through initial methodological optimization. A total of 633 metabolites were identified using untargeted multiomics, with fatty acids, amino acids, polyphenols, terpenoids, organic acids, sugars, and phospholipids as the primary metabolites. Polyphenols served as the main active components by quantitative analysis and antioxidant evaluation, impling that they may be related to antitumor effects. Moreover, based on phytochemical profiling, the potential gene targets and mechanisms of anticancer properties of RVL were explored through network pharmacology. The significance of this study lies in expanding our understanding of the phytochemical and bioactive components of RVL and providing references for its further utilization and exploitation. Therefore, RVL could be an excellent raw material for the food, nutraceutical, and pharmaceutical industries due to its richness in nutrients and bioactive ingredients.

ETHNOPHARMACOLOGICAL RELEVANCE:Hyperlipidemia as a major health issue has attracted much public attention. As a geographical indication product of China, Liupao tea (LPT) is a typical representative of traditional Chinese dark tea that has shown good potential in regulating glucose and lipid metabolism. LPT has important medicinal value in hyperlipidemia prevention. However, the active ingredients and metabolic mechanisms by which LPT alleviates hyperlipidemia remain unclear. AIM OF THE STUDY:This study aimed to systematically investigate the metabolic mechanisms and active ingredients of LPT extract in alleviating hyperlipidemia. MATERIALS AND METHODS:Firstly, we developed a mouse model of hyperlipidemia to study the pharmacodynamics of LPT. Subsequently, network pharmacology and molecular docking were performed to predict the potential key active ingredients and core targets of LPT against hyperlipidemia. LC-MS/MS was used to validate the identity of key active ingredients in LPT with chemical standards. Finally, the effect and metabolic mechanisms of LPT extract in alleviating hyperlipidemia were investigated by integrating metabolomic, lipidomic, and gut microbiome analyses. RESULTS:Results showed that LPT extract effectively improved hyperlipidemia by suppressing weight gain, remedying dysregulation of glucose and lipid metabolism, and reducing hepatic damage. Network pharmacology analysis and molecular docking suggested that four potential active ingredients and seven potential core targets were closely associated with roles for hyperlipidemia treatment. Ellagic acid, catechin, and naringenin were considered to be the key active ingredients of LPT alleviating hyperlipidemia. Additionally, LPT extract modulated the mRNA expression levels of Fxr, Cyp7a1, Cyp8b1, and Cyp27a1 associated with bile acid (BA) metabolism, mitigated the disturbances of BA and glycerophospholipid (GP) metabolism in hyperlipidemia mice. Combining fecal microbiota transplantation and correlation analysis, LPT extract effectively improved species diversity and abundance of gut microbiota, particularly the BA and GP metabolism-related gut microbiota, in the hyperlipidemia mice. CONCLUSIONS:LPT extract ameliorated hyperlipidemia by modulating GP and BA metabolism by regulating Lactobacillus and Dubosiella, thereby alleviating hyperlipidemia. Three active ingredients of LPT served as the key factors in exerting an improvement on hyperlipidemia. These findings provide new insights into the active ingredients and metabolic mechanisms of LPT in improving hyperlipidemia, suggesting that LPT can be used to prevent and therapeutic hyperlipidemia.

BACKGROUNDPile fermentation is one of the key steps in developing the Liupao tea (LBT) quality and unique characteristics. The complex biochemical profile of LBT results from microorganisms present during the pile-fermentation process. However, the critical underlying microorganisms and the marker compounds still need to be determined.RESULTSStaphylococcus, Brevibacterium, Kocuria, Aspergillus, and Blastobotrys were the common dominant microorganisms at the end of the pile fermentation of LBT. Staphylococcus, Aspergillus, Blastobotrys, and nine other genera carried by raw tea are the core microorganisms in the LBT during pile fermentation. A total of 29 critical compounds contributed to the metabolic changes caused by the processing of LBT. Of these, gallic acid, adenine, hypoxanthine, uridine, betaine, 3,4-dihydroxybenzaldehyde, and alpha-linolenic acid could be characterized as potential marker compounds. Correlation analysis showed that the core microorganisms, including Sphingomonas, Staphylococcus, Kocuria, Aureobasidium, Blastobotrys, Debaryomyce, and Trichomonascus, were closely related to major chemical components and differential compounds. Moreover, the mutually promoting Staphylococcus, Kocuria, Blastobotrys, and Trichomonascus were correlated with the enrichment of marker compounds. Integrated molecular networking and metabolic pathways revealed relevant compounds and enzymes that possibly affect the enrichment of marker compounds.CONCLUSIONThis study analyzed the LBT fermentation samples by omics analysis to reveal the stable microbial community structure, critical microorganisms, and markers compounds affecting the quality of LBT, which contributes to a better understanding of pile fermentation of LBT and the fermentation theory of dark tea. (c) 2023 Society of Chemical Industry.

BACKGROUND:Non-alcoholic fatty liver disease (NAFLD) poses a significant global public health concern. Liupao tea (LPT) is a Chinese national geographical indication product renowned for its lipid-lowering properties. However, the precise mechanisms and active constituents contributing to the efficacy of LPT against NAFLD remain unclear. PURPOSE:This study aims to comprehensively explore the therapeutic potential of Liupao tea extract (LPTE) in alleviating NAFLD through an integrated strategy. METHODS:Initially, network pharmacology analysis was conducted based on LPTE chemical ingredient analysis, identifying core targets and key components. Potential active ingredients were validated through chemical standards based on LC-MS/MS. To confirm the pharmacological efficacy of LPTE in NAFLD, NAFLD mice models were employed. Alterations in hepatic lipid metabolism were comprehensively elucidated through integration of metabolomics, lipidomics, network pharmacology analysis, and real-time PCR analysis. To further explore the binding interactions between key components and core targets, molecular docking and microscale thermophoresis (MST) analysis were employed. Furthermore, to investigate LPTE administration effectiveness on gut microbiota in NAFLD mice, a comprehensive approach was employed. This included Metorigin analysis, 16S rRNA sequencing, molecular docking, and fecal microbiome transplantation (FMT). RESULTS:Study identified naringenin, quercetin, luteolin, and kaempferol as the potential active ingredients of LPTE. These compounds exhibited therapeutic potential for NAFLD by targeting key proteins such as PTGS2, CYP3A4, and ACHE, which are involved in the metabolic pathways of hepatic linoleic acid (LA) and glycerophospholipid (GP) metabolism. The therapeutic effectiveness of LPTE was observed to be comparable to that of simvastatin. Furthermore, LPTE exhibited notable efficacy in alleviating NAFLD by influencing alterations in gut microbiota composition (Proteobacteria phylum, Lactobacillus and Dubosiella genus) that perhaps impact LA and GP metabolic pathways. CONCLUSION:LPTE could be effective in preventing high-fat diet (HFD)-induced NAFLD by modulating hepatic lipid metabolism and gut microbiota. This study firstly integrated bioinformatics and multi-omics technologies to identify the potential active components and key microbiota associated with LPTE's effects, while also primally elucidating the action mechanisms of LPTE in alleviating NAFLD. The findings offer a conceptual basis for LPTE's potential transformation into an innovative pharmaceutical agent for NAFLD prevention.

Context There are still comparatively few methods for the detection of drug residues in buffalo milk. Aims This study aims to develop a method for the detection of drug residues in buffalo milk. Methods In this study, we developed an analytical method for veterinary drug residues in buffalo milk using liquid chromatography–tandem mass spectrometry. The multi-residue method was established for the simultaneous identification and quantitation of eight common veterinary drugs, including metronidazole, salbutamol, atropine, trimethoprim, hydrocortisone, kitasamycin, roxithromycin and tylosin. Key results The sample processing method for buffalo milk was compared and optimised. The precision, recovery and matrix effects of the method were validated. The precision and accuracy of all analytes ranged from 1.81% to 12.35% and 1.25% to 14.57%, respectively. The average recovery percentages varied between 93.59% and 114.57%, and the average matrix effect ranged from 87.12% to 103.76%. All eight analytes in buffalo milk exhibited stability under different treatment conditions. Conclusions The developed method was successfully applied to laboratory analysis and routine sample analysis. The method was demonstrated to be rapid, sensitive and reliable for the rapid monitoring of veterinary drug residues in buffalo milk. Implications The findings of this study contribute to the risk assessment of veterinary drug residues for preventing the human consumption of contaminated buffalo milk and its derivatives.