The gut microbiota has emerged as a key modulator of host health, particularly because of its capacity to metabolize dietary glycosides. While C-glycosides demonstrate superior chemical stability and resistance to enzymatic cleavage compared to O-glycosides, specialized gut bacteria can selectively metabolize them, improving their bioavailability. Since the identification of microbial C-glycosidase enzymes, the precise reaction mechanisms remain incompletely understood, and standardized methodologies for studying these processes are still not well-established. To address this gap, we introduced an integrated multidisciplinary approach combining structural biology, enzymology, and analytical techniques, including liquid chromatography-tandem mass spectrometry (LC-MS/MS) and nuclear magnetic resonance (NMR), to systematically characterize these enzymatic activities. This comprehensive approach enables precise characterization of C-glycoside metabolism at molecular, enzymatic, and metabolic levels.

BACKGROUND:Changan Granule (CAG) is a drug product developed from a traditional Chinese medicine (TCM) empirical prescription for diarrhea-predominant irritable bowel syndrome (IBS-D). The action mechanism and effective compounds of CAG in the treatment of IBS-D are not well understood. PURPOSE:This study aimed to investigate the effectiveness, action mechanism and effective compounds of CAG for treating IBS-D. METHODS:Network pharmacology was used to screen the related pathways and active compounds of CAG in the treatment of IBS-D. Neonatal mother-infant separation, acetic acid enema and colorectal dilation were employed to construct IBS-D model for in vivo study. The effectiveness of CAG was evaluated in accordance with the results of body weight measurement, fecal water content determination, abdominal withdraw reflex test, open field test, sucrose preference test, forced swimming test and hematoxylin-eosin (HE) staining. The protein and mRNA levels of key molecules regulated by CAG were assessed through enzyme-linked immunosorbent assay (ELISA), western blotting, and reverse transcription quantitative polymerase chain reaction (RT-qPCR). The active compounds from CAG screened by network pharmacology were investigated with Caco-2 and RIN-14B cell models in vitro. RESULTS:Network pharmacological analysis showed that CAG regulated 5-hydroxytryptamine (5-HT) signaling pathway and tetrahydropalmatine, formononetin and corydaline might be the potential effective compounds. The validation experiments showed that CAG restored the decreased body weight, and alleviated intestinal sensitivity, low-grade inflammation, diarrhea, frequent defecation, anxiety and depression of IBS-D rats through regulating the expression levels of 5-HT, tryptophan hydroxylase (TPH)1/2, serotonin transporter (SERT), 5-hydroxytryptamine-3 and -4 receptors (5-HT3R and 5-HT4R) in brain-gut axis (BGA). Tetrahydropalmatine and formononetin were confirmed to be the potential effective compounds of CAG in regulating 5-HT signaling pathway. CONCLUSION:CAG exhibits therapeutic effect on IBS-D rats through regulating 5-HT signaling pathway in BGA. Tetrahydropalmatine and formononetin are major potential effective compounds. Our findings provide scientific basis for the clinical use and drug development of CAG for IBS-D.

Asarum sieboldii Miq. is an important medicinal plant valued for its diverse health benefits in the pharmaceutical industry. In the present study, we isolated and characterized isoeugenol synthase from A. sieboldii (AsIGS), an essential enzyme involved in the biosynthesis of volatile phenylpropenes. We hoped to elucidate the secondary metabolic network of eugenol in A. sieboldii plants, which constructed the prerequisite for quality improvement of the well-known TCM Asari Radix et Rhizoma. Bioinformatics analysis revealed high similarity between the DNA sequences of AsIGS and isoeugenol synthase genes from other plants, and that the association of the candidate protein AsIGS with the PIP reductase family. Moreover, the AsIGS protein displayed a molecular weight of about 34.96 kDa, with a theoretical isoelectric point of 6.01 and an average hydrophobicity of-0.092, indicating the protein's partial acidity, stability, and hydrophilic nature. Phylogenetic analysis showed that AsIGS had a close relationship with isoeugenol synthases and fewer eugenol synthases found in other species. Alphafold2 predicted the structure of the AsIGS protein, and CB-Dock2 predicted the binding sites of the ASIGSNADPH-coniferyl acetate ternary complex. In vitro enzymatic assay results demonstrated that the optimal temperature of the AsIGS-involved catalysis for coniferyl acetate was 30 degrees C, and several kinetics parameters were Km (12.21 mM), Vmax (27.9 U/mg), kcat (76.26 s-1), and kcat/Km (6.49 s-1 & sdot;mM-1). Furthermore, it was also determined that the AsIGS protein had varying performance at different pH levels. While the candidate protein converted coniferyl acetate into both isoeugenol and eugenol at pH 5.5, it just catalyzed the production of isoeugenol at pH 6.5. However, isoeugenol has never been detected in A. sieboldii. Altering AsIGS expression in transgenic plants impacted only eugenol contents. Compared with wild type, overexpression of AsIGS increased eugenol content by 23.3 %, while RNAi-induced down-regulation of AsIGS decreased it by 25.3 %. Taken together, these results confirmed that the AsIGS gene was involved in the biosynthesis of eugenol in A. sieboldii with a dual catalytic potential.

C-glycosides are commonly found in medicinal plants and exhibit extensive structural diversity along with various bioactivities, including antibacterial, anti-inflammatory, antiviral, antioxidant, and antineoplastic activities. In C-glycosides, the anomeric carbon of the sugar moiety is directly connected to an aglycone through carbon-carbon bonding. Compared with O-glycosides, C-glycosides are structurally stable and resistant to acids and enzymes. Consequently, they are typically unbreakable, resulting in poor absorbability and low bioavailability. Interestingly, some intestinal bacteria can cleave C-C glycosidic bonds, providing a specific and environmentally friendly biological approach to degrade C-glycosides. In this study, a set of standard operating procedures (SOPs) was developed for screening intestinal bacteria capable of cleaving C-C glycosidic bonds based on the biotransformation model of natural compounds. The SOPs include the preparation and enrichment of intestinal bacteria, activity-oriented screening, and activity validation in a low-carbon source medium. This methodology provides a foundational reference for researchers aiming to isolate and study these specialized functional bacteria.

Thymus quinquecostatus Celak., a traditional aromatic edible plant from Lamiaceae, is widely used as food additive, food condiment, spice, and herbal teas. Polyphenol-rich fraction of T. quinquecostatus (PRF) has been proven to be effective protective effect for cerebral ischemia reperfusion injury (CIRI) in our previous study. In this study, we developed a novel "Gut flora-Compound-Target-Pathway" (GCTP) network based on network pharmacology coupled with gastrointestinal metabolism for screening bio-active components, key targets and gut floras through the classical technique for order preference by similarity to ideal solution (TOPSIS). This compensates for the lack of gut floras and gastrointestinal metabolism in network pharmacology. Firstly, four incubation models covering simulated gastric juice, simulated intestinal juice, gut floras of normal and transient middle cerebral artery occlusion (tMCAO) rat in vitro were applied to PRF. The 109 proto-components and 64 metabolites were elucidated by ultra-high performance liquid chromatography-Q exactive orbitrap-mass spectrometry (UPLC-QE-Orbitrap-MS). Then, the key targets of matrix metalloproteinase 9 (MMP9), prostaglandin-endoperoxide synthase 2 (PTGS2), tyrosine-protein kinase fyn (FYN), estrogen receptor 1 (ESR1), amyloid precursor protein (APP), and epidermal growth factor receptor (EGFR), and gut floras of Enterococcus avium LY1 were selected. Moreover, the selected key proto-components were rosmarinic acid, daidzein, quercetin, luteolin, apigenin, methyl rosmarinate, kaempferol, luteoloside, and caffeic acid, and the key metabolites were isokaempferide, isorhamnetin, isoquercetin, and mangiferin. Binding of compounds to the key proteins was analyzed by molecular docking, and also verified though an 2,2'-azobis(2-amidinopropane)dihydrochloride (AAPH) induced oxidative stress zebrafish model and real-time quantitative polymerase chain reaction (RT-qPCR) assays. This study provides a new idea and a better understanding of PRF for its protective effects on CIRI and its underlying mechanisms.

Ethnopharmacological relevance: In traditional Chinese medicine (TCM), Yigong San (YGS) is mainly used to treat dyspepsia caused by deficiency of spleen and stomach qi. Although the chemical composition and bioactivity of YGS has been well studied, the main in vivo compounds and their distribution in tissues still need to be made clearer. Aim of the study: To elucidate the pharmacokinetic profiles and tissue distribution of eight main compounds of YGS in rats, and provide a reference for clinical application and new drug development. Materials and methods: UPLC-Q-Exactive-Orbitrap-MS was used to qualitatively characterize the parent compounds and their metabolites in the plasma of rats after oral administration of YGS. A sensitive, reliable, and accurate ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method using UPLC-AB Sciex QTRAP 5500 MS was established to quantitatively determine eight main compounds of YGS in rat plasma and tissues, including liquiritin, isoliquiritin, hesperidin, ginsenosides Rb1, Re and Rg1, atractylenolides I and II. Results: The mean area under the concentration-time curve (AUC) values of ginsenoside Rb1, hesperidin, and liquiritin at low, medium, and high doses were greater than 150 ng h/mL. The elimination half-life (t1/2) values of ginsenoside Rb1, atractylenolides I and II (low and medium doses) were longer than 10 h. Peak time (Tmax) values of all compounds were shorter than 10 h. Except for atractylenolides, the maximum concentration (Cmax) values of the compounds were greater than 10 ng/mL. The eight compounds were detected in the heart, brain, liver, spleen and kidney at 0.25 h after oral administration. Liquiritin and isoliquiritin had higher exposure in the liver and heart. Hesperidin and ginsenosides Rb1, Re, and Rg1 are mainly distributed in the spleen and kidney. Atractylenolides I and II are mainly distributed in spleen, liver and kidney. Conclusions: All main compounds of YGS, i.e., liquiritin, isoliquiritin, hesperidin, ginsenosides Rb1, Re, and Rg1, and atractylenolides I and II are absorbed into plasma and widely distributed in various tissues. Among them, hesperidin, ginsenoside Rb1, and atractylenolide I are main in vivo compounds. They are mainly distributed in spleen, liver and kidney. The results of this study provide a basis for further in-depth development and application of YGS.

Yigong San (YGS) is a traditional Chinese medicine formula used for pediatric anorexia, chronic atrophic gastritis, and irritable bowel syndrome. In this study, the excretion of eight main compounds, including liquiritin; isoliquiritin; hesperidin; ginsenosides Rb1, Re, and Rg1; and atractylenolides I and II, in rat urine, feces, and bile, was investigated by ultra-high performance liquid chromatography-tandem mass spectrometry. The results showed that the cumulative excretion rates of the compounds in rat urine, feces, and bile were 0.018-1.15%, 0.024-19.89%, and 0.0025-0.72%, respectively. Among the eight compounds detected, liquiritin was the richest in urine, and ginsenosides Re and Rg1 and atractylenolide I were mainly found in feces and bile. In summary, the main components of YGS are excreted via multiple approaches. Liquiritin is mainly through urine, whereas isoliquiritin; hesperidin; ginsenosides Rb1, Re, and Rg1; and atractylenolides I and II are mainly through feces. The excretion of these compounds in bile is usually positively correlated with that in feces. This study lays a foundation for further pharmacological research and application of YGS.

A bacteriophage BD49 specific for Citrobacter braakii was screened out and purified by double-layer plate method. It consists of a polyhedral head of 93.1 ± 1.2 nm long and 72.9 ± 4.2 nm wide, tail fibers, collar, sheath and baseplate. The bacteriophage was identified by morphology observed with transmission electron microscope (TEM), whole genome sequencing carried out by Illumina next generation sequencing (NGS) technique, and gene annotation based on Clusters of Orthologous Groups of proteins (COG) database. It was identified primarily as a member of Caudovirales by morphology and further determined as Caudovirales, Myoviridae, and Citrobacter bacteriophage by alignment of its whole genome sequence with the NCBI database and establishment of phylogenetic tree. The bacteriophage showed good environmental suitability with optimal multiplicity of infection (MOI) of 0.01, proliferation time of 80 min, optimum living temperature of 30–40 °C, and living pH of 5–10. In addition, it exhibited synergistic effect with ciprofloxacin against C. braakii in antibacterial tests.

Ethnopharmacological relevance: The flowers of Trollius chinensis Bunge (Ranunculaceae) is a traditional Chinese medicine used to treat various inflammatory diseases, including upper respiratory infections, chronic tonsillitis, and pharyngitis. Recently, there has been growing research on the antiviral role of the flowers of T. chinensis Bunge. However, little is known about its anti-influenza virus effects and the underlying mechanisms. Aim of the study: This study aims to evaluate the therapeutic effects of the crude extract from the flowers of T. chinensis Bunge (CEFTC) on mice infected with influenza virus. We further explored its mechanism by detecting the expression of vital proteins (TLR3, TBK1, TAK1, IKK alpha, IRF3, and IFN-beta) related to TLR3 signaling pathway. Materials and methods: Mice were infected with influenza A virus (H1N1) through the nasal cavity and were intragastrically administered CEFTC at the dose of 0.2 mg/g once daily. The therapeutic effects of CEFTC were evaluated by blood cell count, lung index, spleen index, alveolar lavage fluid testing, and HE staining. Network pharmacology analysis predicted the potential signaling pathway between the flowers of T. chinensis Bunge and pneumonia. The expression of TLR3, TBK1, TAK1, IKK alpha, IRF3, and IFN-beta in lung tissues were examined by Western blot assay. In addition, the immunofluorescence assay was applied to assess the effect of CEFTC on the distribution of IRF3 and IFN-beta between nuclei and cytoplasm. Results: Compared with the infected group, the lung index was markedly reduced, and the pathological damage of the lungs was also attenuated in the CEFTC treatment group. The network pharmacology analysis indicated that the NF-kappa B pathway was a potential signaling pathway in the flowers of T. chinensis Bunge for the treatment of pneumonia, TLR3, IRF3, and TBK1 were crucial targets associated with pneumonia. Western blot assay demonstrated that in the high-dose virus infected group, CEFTC reduced the expression of TLR3, TAK1, TBK1, and IRF3. Furthermore, CEFTC could increase the nuclear distribution of IRF3 in alveolar epithelial cells after virus infection. Conclusions: These results suggested that different doses of influenza virus could cause varying infection symptoms in mice. Moreover, CEFTC could exert anti-influenza virus effects by regulating the expression of TLR3, IRF3, IFN-beta, TAK1, and TBK1 in the TLR3 signaling pathway.

目的 考察金莲花汤中葛根素、牡荆素、迷迭香酸和咖啡酸的肠吸收特性,确定金莲花汤中活性成分.方法 采用HPLC法测定肠吸收样品中咖啡酸、葛根素、牡荆素、迷迭香酸,以大鼠肠外翻吸收实验和Caco-2细胞模型转运实验计算各成分的累积吸收量、吸收速率常数和表观渗透系数.结果 随着金莲花汤的质量浓度增加,咖啡酸、葛根素、牡荆素、迷迭香酸的吸收速率常数呈线性关系增加,符合零级吸收速率,且吸收率均小于1.4％,在肠道中吸收效果普遍较差,其中咖啡酸的吸收较为明显.各成分的表观渗透系数值大小为迷迭香酸>咖啡酸>葛根素>牡荆素.结论 金莲花汤中葛根素、牡荆素、迷迭香酸和咖啡酸均可在肠道中被吸收,吸收方式为被动扩散,咖啡酸和迷迭香酸可能是金莲花汤中发挥药效的活性物质,葛根素和牡荆素可能是前药.

Background: Xiaoer Chaige Tuire Oral Liquid (XCT) is a compound preparation composed of 7 traditional Chinese medicines including Bupleuri Radix, Puerariae Lobatae Radix, Scutellariae Radix, Gypsum Fibrosum, Artemisiae Annuae Herba, Paeoniae Radix Alba and Zhigancao in proportion. It is mainly aimed at treating upper respiratory tract infection in children caused by exogenous wind-heat. Modern pharmacological studies have indicated that XCT has a variety of activities such as antibacterial, anti-inflammatory, antiviral and immune regulation.Purpose: To analyze the oral administration metabolism process of XCT by means of sequential metabolism in vitro and verification experiment in vivo, and to screen potential quality control components of XCT.Methods: The main components of prepared XCT were analyzed by high performance liquid chromatography with diode-array detection (HPLC-DAD). The employed models in vitro of XCT sequential metabolism were artificial gastric fluid, artificial intestinal fluid, intestinal bacteria, Caco-2 cells and liver S9 in order. The quantitative analysis of sequential metabolism samples were also analyzed by HPLC-DAD. Ultra performance liquid chromatography QExactive Orbitrap tandem mass spectrometry (LC-QExactive-HF-x-Orbitrap-MS) was used to determine the components in medicated plasma and metabolites in liver S9 of rats after applying XCT orally.Results: A total of 25 components were identified from XCT, of which 19 with high content. Sequential metabolism studies in vitro indicated that baicalin, baicalein, genistin and saikosaponin A were digested in artificial gastric fluid. Albiflorin, glycyrrhizic acid, gallic acid and baicalein were unstable in artificial intestinal fluid. Daidzin, liquiritin and genistin were severally hydrolyzed into daidzein, liquiritigenin and genistein by intestinal bacteria. In addition, there were 7 well absorbed components were detected in the samples of Caco-2 cell model. After incubated by liver S9 system, 7 metabolites which were mainly the products of hydroxylation and glycosylation were identified. A total of 10 components were determined in the drug-containing plasma of rats, among which daidzein, puerarin and benzoic acid were the common components both in absorption model samples and the plasma samples.Conclusions: A sequential metabolism methodology of XCT in vitro and in vivo was established. Puerarin, daidzein, benzoic acid, baicalin, baicalein, wogonin, wogonoside, oroxylin A, 3′-methoxypuerarin, paeoniflorin, scopoletin and liquiritigenin were screened to be potential quality control components of XCT, which provides a material basis for quality control standard and redevelopment of XCT.

Background: Xiaoer Chaige Tuire Oral Liquid (XCT) is a preparation composed of 7 traditional Chinese medicines including Bupleuri Radix, Puerariae Lobatae Radix, Scutellariae Radix, Gypsum Fibrosum, Artemisiae Annuae Herba, Paeoniae Radix Alba and Glycyrrhizae Radix Et Rhizoma Praeparata Cum Melle in proportion. According to traditional Chinese medicine theory, it has the function of dispelling wind evil and relieving exterior syndrome, clearing summer heat and dampness, and reducing internal heat. So, it is indicated for pediatric upper respiratory tract infection caused by exogenous wind-heat. Modern pharmacological studies have indicated that XCT has a variety of activities such as anti-inflammation and antivirus. Purpose: To screen potential quality markers (Q-markers) of XCT by tracking in vivo bioactive compounds concomitantly using in vitro sequential metabolism and in vivo biopharmaceutical analysis. Methods: In vitro metabolic models including artificial gastric juice, intestinal juice, intestinal microbiota, Caco-2 cell monolayer and liver S9 were employed to simulate metabolism of main compounds of XCT in the body. High performance liquid chromatography with diode-array detection (HPLC-DAD) was used to quantitatively determine main components of XCT preparation and its sequential metabolism samples. Ultra performance liquid chromatography with QExactive Orbitrap tandem mass spectrometry (UPLC-QExactive-HF-x-Orbitrap-MS) was used to qualitatively determine in vivo components of XCT preparation in rat plasma and metabolites obtained with liver S9 fraction of rats. Results: Twenty-five compounds were identified from the preparation of XCT. Sequential in vitro metabolism studies indicated that most of these compounds except baicalin and baicalein were stable in artificial gastric juice, albiflorin, glycyrrhizic acid, gallic acid and baicalein were unstable in artificial intestinal juice, daidzin, liquiritin and genistin were hydrolyzed into their aglycones daidzein, liquiritigenin and genistein by intestinal microbiota, and 7 compounds thereout including benzoic acid, puerarin, 3'-methoxypuerarin, paeoniflorin, scopoletin, daidzein and liquiritigenin were shown to be well absorbed with Caco-2 cell monolayer model. These 7 compounds were demonstrated to be metabolized via hydroxylation and glycosylation by liver S9 system. Ten components of XCT preparation including puerarin, baicalin, wogonoside, benzoic acid, daidzein, baicalein, wogonin, oroxylin A, isoscopoletin and isoliquiritigenin were identified from rat plasma by in vivo biopharmaceutical analysis. Most of the compounds screened with both in vitro and in vivo metabolic studies were shown to be active against inflammation and influenza virus. Conclusions: A screening strategy for potential quality markers (Q-markers) of XCT preparation based on tracking in vivo bioactive compounds using the combination of in vitro sequential metabolism and in vivo biopharmaceutical analysis was established. With this strategy, a total of 12 compounds including puerarin, daidzein, benzoic acid, baicalin, baicalein, wogonoside, wogonin, oroxylin A, 3'-methoxypuerarin, paeoniflorin, scopoletin and liquiritigenin were screened to be potential Q-markers of XCT, which provides a material basis for quality control and development of XCT.

Ethnopharmacological relevance: Changan Granule (CAG) is a Chinese patent drug developed based on an empirical prescription in accordance with the formulation theory of Traditional Chinese Medicine. The pre-scription is composed of eight herbal drugs which have been traditionally used by Chinese people for a long history. It has effects of invigorating spleen and supplementing qi, as well as regulating liver and ceasing diar-rhea, and is indicated for the treatment of irritable bowel syndrome (IBS).Aim of the study: This study was aimed to investigate the interaction between CAG and its main components and cytochrome P450 (CYP450) enzymes so as to characterize the major metabolites and metabolic enzymes and evaluate the safety concerns to its clinical use.Materials and methods: Both in vivo and in vitro experiments using such as diarrhea-predominant IBS (IBS-D) rat model, HepG2 cells, and human liver microsomes (HLM) were carried out to investigate the interaction between CAG and its main components and CYP450 enzymes. Real-time quantitative PCR (qPCR), ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS), and cocktail probes were employed to qualitatively or quantitatively measure the metabolites and metabolic enzymes.Results: CAG inhibited the enzyme activities of CYP1A2, CYP2E1, CYP2D6, CYP2C9, and CYP3A4 and the mRNA expressions of CYP2E1, CYP2C9, CYP3A4, and CYP2D6 in vitro. CAG down-regulated the increased expression of CYP1A2 and up-regulated the decreased expression of CYP3A1 in vivo. Twenty-two metabolites were charac-terized from the main components of CAG after incubation with HLM in vitro. CYP2D6, CYP2E1, CYP3A4 and CYP2C9 were identified as the characteristic metabolic enzymes.Conclusions: This study provides a reference for clinical application of CAG in safety. CAG and CYP450 enzymes are interacted. CAG is mainly metabolized by CYP2E1 and CYP2D6. The expression of CYP2E1 and CYP2D6 are more susceptible to be influenced by CAG in comparison with that of CYP3A4, CYP2C9 and CYP1A2. It implies the potential risk of interaction when CAG is taken together with the drugs metabolized by CYP2E1 and CYP2D6.

The C-glycosidic bond that connects the sugar moiety with aglycone is difficult to be broken or made due to its inert nature. The knowledge of C-glycoside breakdown and synthesis is very limited. Recently, the enzyme DgpA/B/C cascade from a human intestinal bacterium PUE was identified to specifically cleave the C-glycosidic bond of puerarin (daidzein-8-C-glucoside). Here we investigated how puerarin is recognized and oxidized by DgpA based on crystal structures of DgpA with or without substrate and biochemical characterization. More strikingly, we found that apart from being a C-glycoside cleaving enzyme, DgpA/B/C is capable of efficiently converting O- to C-glycoside showing the activity as a structure isomerase. A possible mechanistic model was proposed dependently of the simulated complex structure of DgpB/C with 3″-oxo-daidzin and structure-based mutagenesis. Our findings not only shed light on understanding the enzyme-mediated C-glycosidic bond breakage and formation, but also may help to facilitate stereospecific C-glycoside synthesis in pharmaceutical industry.

Bacterial drug-resistance is a serious problem afflicting pharmacologists all over the world. Many strategies have been developed and practiced to overcome it, but almost no one is satisfactory due to the continual change of bacteria.

Objective:To study the intestinal absorption and dynamic characteristics of the main components of Yigongsan and determine the main absorbable components.Methods:The intestinal absorption experiment was carried out by using the rat model of everted intestinal sac in vitro,and the absorption parameters of each component were calculated.Results:The apparent permeability coefficient (P app ) values of liquiritin,hesperidin,glycyrrhizic acid,narirutin,isoliquiritin apioside,and ginsenoside Re were 1×10 –6 -1×10 –5 cm·s –1 .Except those of isoliquiritin apioside and glycyrrhizic acid,the absorptionrateconstant （K a ） values of the other four compounds existed statistical significant differences among groups with different concentrations （P＜0.05）,and increased linearly with concentration （r＞0.97）.Conclusion:The six components are well absorbed mainly through passive transportation in the intestine.The intestinal absorption of ginsenoside Re is the best,followed by liquiritin,glycyrrhizic acid,narirutin,isoliquiritin apioside,and hesperidin.The results of this study can provide references for determining the effective substances of Yigongsan.

目的 研究以山楂核干馏液为主要成分的天然保鲜除味剂对饭菜抑菌保鲜效果和主要菌群结构的影响.方法 采用该天然保鲜除味剂对饭菜进行保鲜处理,观察其对饭菜样品中菌落总数、亚硝酸盐含量、菌群结构变化的影响,并测定其最小抑菌浓度(MIC).结果 该天然保鲜除味剂能明显抑制饭菜样品中的菌落数增长并调节菌群结构,对大肠埃希菌、铜绿假单胞菌和肺炎克雷伯菌均有良好的抑制作用,MIC为母液浓度的1/8.结论 以山楂核干馏液为主要成分的天然保鲜除味剂可延长饭菜等食品的保鲜时间,其作用机制与抑制菌群增殖和调节菌群结构有关.

The quality of Chinese herbal medicine (CHM) raw materials is essential, and mass spectrometry (MS)-based technologies have been playing key roles in the quality control of CHMs. However, the use of miniature mass spectrometry (mini-MS) for quality control of CHMs has rarely been reported. In this work, we developed a rapid analytical method for the quality evaluation of CHMs based on paper spray ionization (PSI)-mini-MS/MS. The quality evaluation of Lygodium japonicum (Thunb.) Sw. was used as an example. Following a "multi-component" quality evaluation strategy, nine active constituents of L. japonicum were selected to be used as analytes for quality control. We confirmed that the precursor-product ion information in the MS/MS spectra of each analyte in the herbal extracts was consistent with the standards. Also, we developed a mini-MS-based quantitative method for each analyte using its quantification ion. The quantitative methodology was rigorously validated using quality control samples. Finally, the quality evaluation of L. japonicum was carried out using the established MS/MS method combined with statistical analysis. A wide range of common quality issues with L. japonicum can be effectively determined, including whether it is adulterated with sand and distinguishing among different parts and species. This study demonstrates that mini-MS for quality evaluation of CHMs is feasible. Mini-MS for quality evaluation of herbal medicines will potentially have a good prospect due to its many advantages such as low cost, low power consumption, and portability in the future.

课题组前期实验发现黄芪注射液可通过调控TLR3 信号通路保护流感病毒感染的 Raw264. 7 细胞[1].本次研究选择了不同黄芪剂型,即黄芪水煎剂(Astragalus decoction, AD)进行后续实验,重点探讨 AD 对流感病毒的作用及机制,为其临床应用提供实验依据.

Lonicera japonica Flos (LJF) is taken orally as a health food and medicinal plant in China for a long time. The gastrointestinal metabolism of LJF was investigated in vitro by three independent models (gastric juice, intestinal juice, and human intestinal bacteria), qualitative analyzed by UPLC-LTQ-Orbitrap-MSn and quantified by HPLC-DAD. 72 prototype compounds were detected in LJF water extraction (LJF-WE), including 14 organic acids, 43 iridoids, 14 flavonoids and one other compound. The prototype and metabolic components of LJF-WE bio-transformed by simulated gastric fluid (70 and 12), intestinal fluid (69 and 12) and human fecal bacteria (29 and 70) were characterized, respectively. The metabolites were formed through desaccharization, isomerization, hydrogenation, methylation, dehydration, and then cyclization, glucuronization and dimethylation followed. 8 bioactive compounds including neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, sweroside, secoxyloganin, isochlorogenic acid B, isochlorogenic acid A and isochlorogenic acid C were much stable in simulated gastric fluid and intestinal fluid, compared with human fecal bacteria. Especially, sweroside and secoxyloganin with glucoside bonds degradated extraordinarily fast, because of the abundant β-glucosidases in human fecal bacteria.