Soil salinity severely limits plant growth and productivity. Carex rigescens, a low-maintenance turfgrass species native to China, exhibits remarkable tolerance to abiotic stresses. Previous studies have highlighted the importance of the phenylalanine metabolic pathway in salt stress defense in C. rigescens; however, the specific mechanisms remain poorly understood. To elucidate the downstream metabolic and molecular components of this pathway, we analyzed flavonoid metabolism in two contrasting C. rigescens varieties-salt-sensitive 'Lvping No. 1' and salt-tolerant 'Lvping No. 2'-and functionally characterized the 4-hydroxyphenylpyruvate dioxygenase (CrHPPD) gene. Salt stress altered the abundance of several flavonoid metabolites, including 2'-hydroxygenistein, genistin, kaempferol, taxifolin, myricetin, and eriodictyol, identifying them as candidate salt-responsive metabolites. Naringenin and apigenin showed genotype- and tissue-dependent abundance patterns between the two varieties. We cloned CrHPPD, characterized its encoded protein, and found that CrHPPD-GFP displayed a cell periphery-associated fluorescence pattern in transient expression assays, although precise localization requires marker-based validation. CrHPPD expression was induced by NaCl and ABA, whereas PEG treatment elicited a weaker and more transient response. Furthermore, overexpression of CrHPPD in Arabidopsis thaliana enhanced germination rate, root length, catalase activity, and chlorophyll retention under salt stress, with the chlorophyll effect being most evident in OE6. Collectively, these findings reveal genotype- and tissue-specific flavonoid remodeling in C. rigescens under salt stress and demonstrate that CrHPPD positively contributes to salt tolerance when overexpressed in Arabidopsis. These results suggest that flavonoid metabolism and the HPPD-associated homogentisate/tocopherol antioxidant branch may represent two stress-responsive components of phenylalanine/tyrosine-derived metabolism, although their direct mechanistic connection requires further validation.

Background Pharmacological interventions of exercise adaptation offer a viable strategy to enhance athletic performance and improve the quality of life in both the elderly and manual workers. Gypenosides (Gyp), a group of bioactive dammarane-type triterpenoid saponins from genus Gynostemma, can increase exercise performance and muscle function in rodents and healthy males, whose mechanism remains unclear. Purpose We explored the role of the gut microbiota (GM) in exercise performance-potentiation by Gyp administration. Methods Multiple behavioral tests, immunofluorescence staining, 16s rRNA gene sequencing, and targeted metabolomics were assessed to demonstrate the altered endurance phenotype induced by Gyp. GM depletion and bile acid pool depletion models were introduced to evaluate the effects of these two factors on the performance-enhancing efficacy of Gyp. UPLC-Q/TOF-MS analysis and luciferase assays were performed to identify FXR antagonists in Gyp. In vitro and in vivo experiments were validated whether deoxycholic acid (DCA) can mimic the benefits of Gyp. Results Preventive Gyp treatment led to an improved endurance exercise phenotype and shifted oxidative metabolic pattern of myofibers from Type II to I in sedentary mice. Gyp reshaped the GM and enriched a group of secondary bile acids (BAs), which were the key signature of global metabolic remodeling. Mechanistically, Gypenosides including gypenoside V specifically inhibited the intestinal farnesoid X receptor (FXR) signaling and increased secondary BA levels such as DCA. DCA enhanced myogenic differentiation and myoblast fusion in C2C12 myotubes, and mimicked performance-enhancing effect of Gyp in vivo by activating TGR5-cAMP-PKA signaling pathway. Conclusion We identified gypenosides as a promising exercise mimetic and suggested the potential of the GM as a key target for physical performance enhancement. DCA-mediated TGR5 activation provides novel mechanistic insight into the pharmacological targeting of exercise adaptations.

Seventeen new dammarane-type saponins, gypenosides FJ1 - FJ17 (1-17), and five known compounds (18-22) including nine pairs of C-24 epimers were isolated from Gynostemma pentaphyllum. Their structures were determined through comprehensive 1D and 2D NMR spectroscopic analyses and HR-ESI-MS data. The absolute configurations of five pairs of C-24 epimer compounds (1-10) were assigned via electronic circular dichroism (ECD) analyses. Interestingly, a detailed spectroscopic analysis indicated that the C-24 configuration of epimers was associated with the chemical shift difference value of H-22 (ΔδH-22 > 0.5, 24R; ΔδH-22 < 0.5, 24S), leading to a simple and convenient method for the rapid assignment of C-24 configuration in C-24 epimers of dammarane-type saponins. This rule enabled rapid C-24 configuration assignment for compounds 11-22. Anti-hypoxic activity evaluation in PC12 cells under hypoxia demonstrated that compounds 8, 9, 11, 13 and 16-20 effectively protected PC12 cells from hypoxia-induced injury.

Paris polyphylla var. yunnanensis is a widely cultivated rhizomatous perennial woody plant known for producing highly valued steroidal saponins. Steroidal saponins serve as the primary active components and exhibit a variety of pharmacological effects. Their production is closely correlated with the growth environment and developmental stages of P. polyphylla. However, the medicinal value of P. polyphylla cultivated for varying durations remains unclear. In this study, we collected samples of P. polyphylla aged 3 to 8 years for metabolome and transcriptome sequencing analysis. The total saponin content was found to be significantly higher in 8-year-old P. polyphylla compared to younger plants. A total of 1,510 metabolites and 270.65 Gb of clean data were identified from these samples. This study provides insights into the medicinal values of P. polyphylla at different growth stages and elucidates the metabolic pathways for specific steroidal saponins.

Tea polyphenols, the primary bioactive constituents responsible for the various health benefits of tea, can be categorized into different subgroups according to their structural characteristics. However, the distinctions in antiviral activity among the diverse types of polyphenolic compounds remain unexplored. In the present study, fifty-eight tea polyphenols with varied structures, including eleven undescribed compounds, were isolated from Rougui Wuyi rock tea. Their molecular structures were elucidated using comprehensive analytical approaches of NMR, HRMS, CD spectroscopic data and acid hydrolysis. The isolated polyphenol analogs could be structurally classified into two main categories: flavan-3-ols, which include catechins, flavoalkaloids, procyanidins and theasinensins, and flavones, encompassing kaempferol, quercetin, myricetin, and their respective glycosides. The inhibitory activities of fifty-eight tea polyphenols against 3CLpro were assessed in vitro, and eighteen phenolic compounds exhibited inhibitory effects on 3CLpro, with IC50 values ranging from 9.8 μM to 61.1 μM. Among them, two types of tea polyphenols, catechin and flavoalkaloid derivatives, demonstrated superior inhibitory effects compared to other categories. The structure-activity relationship was further explored, and molecular docking analysis revealed that the differing inhibitory effects of catechin and flavoalkaloid derivatives were attributed to the variations in the number and positions of the hydrogen bond interactions with 3CLpro. This study provides a valuable understanding of tea polyphenols and supplies potential lead compounds for antiviral drugs.

This study identified 20 key odor-active compounds in cooked brown rice (CBR), cooked medium-milled rice (CMMR), and cooked well-milled rice (CWMR) of fragrant Simiao rice by gas chromatography-olfactometry-mass spectrometry and odor activity value (OAV). Based on the metabolomics analysis method, aroma fingerprints for CBR, CMMR, and CWMR were established, and 10 differential odor-active compounds contributing to their odor differences were identified, including hexanal, nonanal, heptanal, octanal, decanal, (E)-2-decenal, (E,E)-2,4-decadienal, tetradecanal, acetoin, and 2-pentylfuran. An analysis of the source, OAV, and correlation with lipid content of differential odor-active compounds confirmed that variations in raw rice lipid content led to odor differences of cooked rice with different degrees of milling. Upon the comprehensive evaluation of bran degree and ratio of germ-remained kernels of the samples, it was established that monitoring the ratio of germ-remained kernels during milling was crucial to ensure odor quality of CMMR. This research had significant implications for guiding the medium milling and advancing in the flavor and breeding of rice.

Pure-silica DD3R zeolite membrane has been of significant interest in CO2 separation because of its appropriate pore size (0.36 nm x 0.44 nm) to distinguish CO2 from other gases. However, it is challenging to remove its organic template but simultaneously minimize defects by conventional calcination at high temperature. Herein, we proposed a non-thermal ultraviolet (UV) detemplation strategy to prepare high-quality DD3R zeolite membranes for CO2 separation. The 185/254 nm UV radiation not only excited the organic template, adamantanamine (ADA) filled in the membrane but also generated reactive oxygen species (ROSs), which caused decomposition of the template at near-ambient temperature. After being exposed in UV radiation for 4 days, the resultant membrane exhibited an ultrahigh CO2/CH4 selectivity of 876 as well as a good CO2 permeance of 1.37 x 10- 7 mol m- 2 s- 1 Pa- 1. The scalability of such strategy was demonstrated by a large-area hollow fiber membrane module (ca. 800 cm2), which performed a notably high CO2/CH4 selectivity of 170 at 4.1 MPa.

Oriented antibody immobilization has been widely employed in immunoassays and immunodiagnoses due to its efficacy in identifying target antigens. Herein, a heptapeptide ligand, HWRGWVC (HC7), was coupled to poly(glycidyl methacrylate) (PGMA) nanospheres (PGMA-HC7). The antibody immobilization behavior and antigen recognition performance were investigated and compared with those on PGMA nanospheres by nonspecific adsorption and covalent coupling via carbodiimide chemistry. The antibodies tested included bovine, rabbit, and human immunoglobulin G (IgG), while the antigens included horseradish peroxidase (HRP) and β-2-Microglobulin (β2-MG). The nanospheres were characterized using zeta potential and particle size analyzers, scanning electron microscopy, transmission electron microscopy, Fourier transform infrared spectroscopy, and reversed-phase chromatography, proving each synthesis step was succeeded. Isothermal titration calorimetry assay demonstrated the strong affinity interaction between IgG and PGMA-HC7. Notably, PGMA-HC7 achieved rapid and extremely high IgG adsorption capacity (~3 mg/mg) within 5 min via a specific recognition via HC7 without nonspecific interactions. Moreover, the activities of immobilized anti-HRP and anti-β2-MG antibodies obtained via affinity binding were 1.5-fold and 2-fold higher than those of their covalent coupling counterparts. Further, the oriented-immobilized anti-β2-MG antibody on PGMA-HC7 exhibited excellent performance in antigen recognition with a linear detection range of 0–5.3 μg/mL, proving its great potential in immunoassay applications.

Phytochemical investigation of Gynostemma pentaphyllum led to the purification of five novel dammarane-type triterpene isolates, gypenosides B1 - B5 (1-5). Their structures were determined through comprehensive 1D and 2D NMR spectroscopic analyses and HRESIMS data. Of note, 1-3 are inseparable mixtures of epimers due to their unstable nature, and a total of eight dammarane-type triterpene saponins were identified. Additionally, the protective activities of these new compounds against PC12 cell injury induced by hypoxia were evaluated.

Sixteen new dammarane-type triterpenoid saponins ( 1-16 ) featuring diverse structural variations in the side chain at C17, along with twenty-one known analogues ( 17-37 ), have been isolated from the rhizomes of Gynostemma longipes C. Y. Wu, a plant renowned for its medicinal and edible properties. The structural elucidation of these compounds was accomplished through comprehensive analyses of 1D and 2D NMR and HRMS spectroscopic data, supplemented by comparison with previously reported data. Subsequent assays on the isolates for their protective effects against hypoxia-induced damage in pheochromocytoma cells (PC12 cells) revealed that nine saponins exhibited significant anti-hypoxic activities. Further investigation into the anti-hypoxia mechanisms of the representative saponins demonstrated that compounds 22 and 36 markedly reduced the levels of hypoxia-induced apoptosis. Additionally, these compounds were found to decrease the release of lactate dehydrogenase (LDH) and malondialdehyde (MDA), while increasing the activity of superoxide dismutase (SOD), thereby indicating that the saponins could mitigate hypoxia-induced injuries by ameliorating apoptosis and oxidative stress. These findings offer substantial evidence for the future utilization and development of G. longipes , identifying dammarane-type triterpenoid saponins as its active anti-hypoxic constituents.

Oligosaccharides constitute fundamental components in numerous traditional Chinese medicines (TCMs). Conventional chromatographic methods for natural product analysis are not suitable for oligosaccharides due to their large polarity and structural similarity. Herein, an ultra-high performance liquid chromatography with charged aerosol detector (UHPLC-CAD) method was developed for the profiling of oligosaccharides using 9 neutral (DP3-DP11) reference oligosaccharides. Various factors, including columns, mobile phase, elution conditions, flow rate, and column temperature were systematically examined. Optimal separation was achieved using an Amide column with gradient elution within 18min, at 0.5mL/min flow rate and 30℃ column temperature. Moreover, an ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF/MS) method was also optimized to provide structural information. The developed method was applied to detect oligosaccharides in several TCMs, including Morindae Officinalis Radix (MOR), Ziziphi Spinosae Semen (ZSS), Menthae Haplocalycis Herba (MHH) and Chrysanthemi Indici Flos (CIF), revealing 9 and 16 oligosaccharides being uncovered from MHH and CIF respectively for the first time. This study presents a versatile UHPLC-CAD and UHPLC-Q-TOF/MS method with the potential for advancing oligosaccharides discovery and contributing to the quality analysis of TCMs.

Jiaogulan herbal tea, known for its pharmacological benefits, is derived from various species within the genus Gynostemma, particular for G. pentaphyllum (GP) and G. longipes (GL). Herein, a global approach integrating chemometrics with qualitative and quantitative methods was utilized to investigate the chemical profiles and variations in GP and GL from different geographic origins. A total of 227 triterpenoid saponins were identified in two Gynostemma species, among which 23 differential markers ware screened out by multivariate statistical analysis. Furthermore, the contents of significant markers were determined after the demalonylation process. The chemical composition of GP samples sourced from diverse regions were firstly summarized into three chemotypes (I-III), along with an additional chemotype (IV) for GL. This study contributes to a deeper understanding of Gynostemma plants and provides valuable theoretical basis for the quality control in the production of Jiaogulan herbal tea.

Ethnopharmacological relevance: Epimedii Folium (Yin-yang-huo in Chinese), a traditional and commonly used herbal medicine (HM), is a representative of multi-plant sources. To date, little is known about the reasons for similar therapeutic effects of this HM from multi-plant sources.Aim of the study: To investigate the underlying reasons for the similar pharmacological effects of Epimedii Folium from two botanical sources (Epimedium koreanum Nakai and Epimedium wushanense T. S. Ying).Materials and methods: Firstly, the phytochemicals of the extracts of E. koreanum and E. wushanense were systematically analyzed. Meanwhile, their pharmacological effects on kidney-yang deficiency (KYD) syndrome were evaluated in rats induced by hydrocortisone. Subsequently, we proposed a combined effect index (CEI) to assess the effects of two plants on the secretion of testosterone by combing the system exposure of twelve active components in vivo with their regulation activities of testosterone production in vitro. Moreover, the label-free proteomics and Western blot analysis were conducted to evaluate the possible mechanism of Epimedii Folium from two botanical sources.Results: E. koreanum and E. wushanense exhibited similar pharmacological effects on KYD syndrome with promoting the mating behaviors and testosterone levels of rats, although there is a certain difference in the main components between two plants. The CEI analysis showed that there was no difference (P > 0.05) in the sum of CEIs of two Epimedium, indicating that their similar therapeutic effects are attributed to bioactive metabolites in vivo. Furthermore, Epimedii Folium can regulate testosterone production in rat Leydig cell via reversing expressions of key steroidogenic enzymes, such as steroidogenic acute regulatory protein (StAR) and 38-hydroxysteroid dehydrogenase (38-HSD).Conclusion: Our results supply critical evidence for the similar pharmacological effects of two Epimedium species, acting by consistent bioactive components directly exposing in vivo, not chemical compositions presenting in herbs. It provides a reasonable scientific basis for understanding of the HMs originated from multi-plant sources for the same clinical application.

The husks of Xanthoceras sorbifolia Bunge have gradually attracted widespread attention in recent years due to the abundant resources and ideal pharmacological activities, with barrigenol-like triterpenoid saponins being its biological constituents. In this study, a feature-based molecular networking (FBMN) was utilized to perform the targeted isolation of triterpenoids. As a result, six undescribed barrigenol-type saponins (1-6) along with fourteen known analogues (7-22) were isolated from the extract of X. sorbifolia husk. Their structures were determined through a comprehensive analysis of NMR and HRMS spectroscopic data. Among them, compounds 1-3 are a specific type of saponin featuring a fucose moiety attached at C-21. The antitumor activities of isolated compounds were evaluated and compounds 7, 9 and 10 showed significant inhibitory activities against A549 and HepG2 cell lines in a dose-dependent manner.

Objective: This study aimed to investigate the effects of serum levels of apelin and CD40L on major adverse cardiovascular events (MACEs) after percutaneous coronary intervention (PCI). Methods: A case–control study was conducted to select patients undergoing PCI in our hospital from June 2020 to June 2022. Patients were divided into the occurrence group and the non-occurrence group according to whether MACEs occurred during the 12-month follow-up after surgery. Enzyme-linked immunosorbent assay was used to detect the expression levels of serum apelin and CD40L in the two groups, and the correlation between the expression of apelin and CD40L and prognosis was analyzed. Logistic regression analysis was performed on the indicators with differences to analyze the influencing factors of the prognosis of PCI. Results: Compared with the non-occurrence group, the occurrence group had a significantly lower level of apelin and a significantly higher level of CD40L (p < 0.001). Apelin was negatively correlated with the occurrence of MACEs after PCI (r = –0.583, p < 0.001), and CD40L was positively correlated with the occurrence of MACEs after PCI (r = 0.569, p < 0.001). Logistic regression analysis showed that apelin was a protective factor for MACEs after PCI (odds ratio (OR) = 0.248, p < 0.001); CD40L, age, hypertension, and the number of diseased vessels were risk factors for MACEs after PCI (OR = 8.684, 0.018, 0.003, 0.020, p < 0.05). The area under curve (AUC) of apelin combined with CD40L was large, and the predictive value was higher than that of apelin and CD40L alone (AUC values were 0.956, 0.857, 0.905, p < 0.001; p < 0.001; p < 0.001). Conclusions: This study showed that the levels of apelin and CD40L were correlated with MACEs after PCI. Clinicians should pay close attention to the levels of apelin and CD40L in patients after PCI and be alert to the occurrence of MACEs.

RATIONALE Shuang-Huang-Lian powder injection (SHLPI) is the famous modern traditional Chinese medicine formula preparations (TCMFs) widely applied to treat acute upper respiratory infections. However, SHLPI is an injection extracted from pure Chinese medicine, and many adverse reactions have been reported clinically. Therefore, it is necessary to deeply characterize the chemical composition of SHLPI and quantitatively analyze its potential allergenic components. METHODS In this study, the samples were analyzed by ion mobility quadrupole time-of-flight mass spectrometer coupled to ultra-high performance liquid chromatography (UHPLC-QTOF-MS) combined with self-built database. Furthermore, the parallel reaction monitoring (PRM) model of ultra-performance liquid chromatography-quadrupole-Orbitrap mass spectrometry (UHPLC-Q-Orbitrap-MS) was used to successfully quantify 10 representative bioactive components. RESULTS Through this strategy, a total of 90 compounds identified and the fragmentation pathways of five representative compounds in the five main components of SHLPI were summarized and 10 components (neochlorogenic acid, chlorogenic acid, sweroside, forsythiaside A, luteoloside, isochlorogenic acid B, isochlorogenic acid C, baicalin, phillyrin, baicalein) as the quality markers of SHLPI based on UPLC-Q-Orbitrap-MS. CONCLUSIONS This work comprehensively characterized the material basis of SHLPI, summarized the cracking laws of representative substances, and quantitatively analyzed 10 potential allergenic components. Therefore, this study could provide a basis for the quality control of SHLPI and the clinical rational use of drugs to reduce its adverse reactions.

RATIONALE:Shuang-Huang-Lian powder injection (SHLPI) is a well-known modern traditional Chinese medicine formula preparation (TCMFP) widely used to treat acute upper respiratory infections. However, SHLPI is extracted from pure Chinese medicine and administered through an injection, and many adverse reactions have been reported clinically. Therefore, it is necessary to characterize in depth the chemical composition of SHLPI and quantitatively analyze its potential allergenic components.METHODS:In this study, the samples were analyzed using ion mobility ultra-high-performance liquid chromatography-quadrupole time-of-flight-mass spectrometry (UHPLC-QTOF-MS) combined with a self-built database. Furthermore, the parallel reaction monitoring (PRM) model of ultra-high-performance liquid chromatography-quadrupole-Orbitrap-mass spectrometry (UHPLC-Q-Orbitrap-MS) was used to successfully quantify 10 representative bioactive components.RESULTS:Using this strategy 90 compounds were identified, the fragmentation pathways of five representative compounds in the five main components of SHLPI were summarized, and 10 components (neochlorogenic acid, chlorogenic acid, sweroside, forsythiaside A, luteoloside, isochlorogenic acid B, isochlorogenic acid C, baicalin, phillyrin, and baicalein) were determine as the quality markers of SHLPI based on UPLC-Q-Orbitrap-MS.CONCLUSIONS:This work comprehensively characterized the material basis of SHLPI, summarized the cracking laws of representative substances, and quantitatively analyzed 10 potential allergenic components. Therefore, this study could provide a basis for the quality control of SHLPI and the clinical rational use of drugs to reduce its adverse reactions.

Gypenosides (Gps) are the major bioactive components in Gynostemma species. They include neutral Gps and acidic malonylgypenosides (MGps). MGps are abundant in Gynostemma species and can be transformed into corresponding Gps via extraction, concentration, and drying. If only the Gps were quantified and MGps were ignored, the quality of Gynostemma species would be underestimated. This study aimed to develop a sample preparation method involving demalonylation and ultrahigh-performance liquid chromatography-charged aerosol detector (UHPLC-CAD) analysis to determine the contents of gypenoside XLIX (Gp XLIX) and gypenoside A (Gp A). First, the optimized ultrasonic extraction method was established to extract G. longipes powder ul-trasonically. Then, the extracted solution was put into a closed container (centrifuge tube) and heated in a water bath at 95 degrees C. Then, MGps were converted into corresponding Gps. The proposed preparation method was compared with the other three methods, including water bath reflux heating, alkali hydrolysis, and extraction of heated powder, and was shown to exhibit higher conversion and better convenience. Subsequently, an UHPLC-CAD method was established and validated. Gp XLIX and Gp A showed excellent linear correlations between 15.55 and 248.8 mu g/mL and 24.10-385.5 mu g/mL, respectively (R2 > 0.999). The limit of detection was 1.40 ng (Gp XLIX) and 2.41 ng (Gp A), and the limit of quantification was 7.77 ng and 14.46 ng, respectively. The relative standard deviation for precision, stability, and repeatability was 0.63-3.15%. The average recovery of Gp XLIX and Gp A was 98.97% and 98.23%, respectively. The established method was applied for determining Gp XLIX and Gp A contents in wild or cultivated G. longipes samples collected from the Qinba Mountains area. The contents of Gp XLIX and Gp A were 5.16-23.02 mg/g and 15.78-54.55 mg/g, respectively. Conclusively, the proposed sample preparation and analysis method could be used for the quality control and evaluation of G. longipes.

为了对绞股蓝皂苷A结构进行确证,对绞股蓝总甙、绞股蓝总甙片等质量标准、档案及相关文献进行了查阅和研究,结果发现绞股蓝总甙片和绞股蓝总甙(苷)胶囊质量标准中使用的绞股蓝皂苷A对照品的结构存在差异.经过调研,发现使用绞股蓝皂苷A名称的化合物存在4种可能的结构(长梗绞股蓝皂苷A、绞股蓝皂苷ⅩⅥ、绞股蓝皂苷XLⅢ和(23S)-19-羰基-21,23-环氧-3β,20,21-三羟基达玛-24-烯-3-O-[α-L-吡喃鼠李糖基-(1→2)][β-D-吡喃木糖基(1→3)]-α-L-吡喃阿拉伯糖苷).结合绞股蓝及其制剂的研究现状,认为绞股蓝皂苷A的结构应为(23S)-19-羰基-21,23-环氧-3β,20,21-三羟基达玛-24-烯-3-O-[α-L-吡喃鼠李糖基-(1→2)][β-D-吡喃木糖基(1→3)]-α-L-吡喃阿拉伯糖苷,CAS号为157752-01-7.目前,绞股蓝总甙、绞股蓝总甙片、绞股蓝总甙(苷)胶囊等5个药品标准所涉及的绞股蓝皂苷A对照品实际都不是绞股蓝皂苷A化合物本身,建议对相关的质量标准进行修订.