The inhibition of the proprotein convertase subtilisin/kexin type 9 (PCSK9) expression can reduce the circulating low-density lipoprotein cholesterol levels and the risk of atherosclerotic cardiovascular disease. As the production of PCSK9 inhibitors continues to advance, the development of quantitative and drug screening methodologies for PCSK9 becomes increasingly essential. Herein, a novel wash-free and sensitive chemiluminescence imaging sensing system was proposed for the user-friendly, affordable and reliable immunoassay of the PCSK9 protein. By integrating the imaging system with ultra-high performance liquid chromatography quadrupole-timeof-flight mass spectrometry, a high-throughput PCSK9 inhibitor screening platform was established to facilitate the rapid identification of active ingredients in lipid-lowering traditional Chinese medicines. The imaging system enabled simultaneous quantification of PCSK9 protein in up to 96 cell lysate samples within 24 min, and presented detection limit down to 0.24 pM, without the need for separation and rinsing, only requiring straightforward mixing and pipetting, thereby enhancing both screening efficiency and accuracy. Protogracillin, methylprotodioscin, and pseudoprotodioscin were effectively screened out, exhibiting significant PCSK9 inhibitory effects and safety profiles. These findings were further validated in the HepG2 cells and hyperlipidemic mice. This work opened a valuable and viable avenue for monitoring PCSK9 levels and discovering natural smallmolecule PCSK9 inhibitors, holding great potential in managing diseases associated with abnormal levels of PCSK9.

As a medicinal and edible herb, the quality grade of Codonopsis Radix (CR) is the key determinant of its market price. However, the current grading system remains significantly inadequate. Single-dimensional indices (including phenotypic traits, chemical components, or bioactivity) fail to accurately characterize the comprehensive quality of CR. Under this circumstance, this study established a multidimensional quality evaluation system integrating phenotypic traits, chemical components, and bioactivity. A multi-index weighted quality comprehensive evaluation index (QCEI) was constructed by combining the entropy weight method (EWM), criteria importance through inter-criteria correlation (CRITIC) method, technique for order preference by similarity to ideal solution (TOPSIS) method, and triangular area method. Hierarchical cluster analysis (HCA) based on QCEI classified 50 CR batches into 3 grades: Grade I (QCEI≥0.56, excellent quality), Grade II (0.38 ≤ QCEI<0.56, good quality), and Grade III (QCEI<0.38, general quality). In vivo validation showed a significant positive correlation between QCEI grades and immunomodulatory effects. Specifically, the CR of Grade I significantly enhanced the body weight growth rate, elevated thymus and spleen indices, and regulated serum levels of IL-2, IL-6, and TNF-α in immunosuppressed mice (p < 0.05-p < 0.0001). This study establishes a scientific and reliable quality grading standard for CR, providing a valuable reference for the comprehensive quality evaluation of other traditional Chinese medicinal materials.

As a classic typical of medicine and food homology, fennel essential oil (FEO) is at risk of fraud due to its high commercial value and soaring consumer demand. This study developed an integrated analysis strategy using electronic nose (E-nose), gas chromatography-mass spectrometry (GC-MS), and chemometrics to achieve rapid identification and quantitative analysis of FEO and its adulterants (anise essential oil [AEO], dill essential oil [DEO], caraway essential oil [CAEO], and cumin essential oil [CUEO]). E-nose combined with principal component analysis (PCA) successfully enabled rapid differentiation of FEO from four common adulterants. Using GC-MS-based untargeted metabolomics coupled with orthogonal partial least squares discriminant analysis (OPLS-DA), 17 differential metabolites were screened, of which 15 were further validated as key markers via gas chromatography-triple quadrupole mass spectrometry (GC-QQQ-MS)-targeted quantitative analysis. On this basis, partial least squares discriminant analysis (PLS-DA) and partial least squares regression (PLS-R) models were established to achieve adulterant type identification and adulteration ratio prediction. The results demonstrated that both established models exhibited good fitting and predictive capabilities. The PLS-DA model achieved 100% classification accuracy for samples with adulteration ratios exceeding 30%, whereas the prediction errors of the PLS-R models for all four adulteration systems were below 10%. This integrated strategy demonstrates the potential to detect FEO adulteration, thereby offering a valuable reference for the quality control of FEO in future studies.

Background Estrogen-deficient osteoporosis demands safe and effective therapies. Psoralea corylifolia fruit (PCF), a phytoestrogenic herbal medicine for osteoporosis, lacks clarified key active components and synergistic combinations, which limits its application. Purpose This study established an AI-driven stepwise strategy to identify anti-osteoporotic synergistic combination from PCF, quantify their synergism, and validate pharmacodynamics, thus providing a reference for complex disease treatment. Methods An AI-driven stepwise integrated strategy was proposed: Ⅰ) Artificial intelligence-assisted preliminary screening of PCF active components via dual estrogenic/anti-osteoporotic models; Ⅱ) molecular simulation (docking/dynamics) for secondary screening of candidate components; Ⅲ) multi-dimensional evaluation for precision targeting of lead components; Ⅳ) Chou-Talalay method used to determine synergism (combination index, CI<0.9), additivity (0.9≤CI≤1.1), or antagonism (CI>1.1). Ultimately, the pharmacodynamic activity of the screened synergistic combination was validated through systematic in vitro and in vivo pharmacodynamic evaluations, combined bioequivalence analysis based on 90% confidence interval. Results Machine learning (ML) screening identified 20 active components, narrowed to 6 via molecular simulations. Three leads viz neobavaisoflavone (NBIF), bavachin (BA) and isobavachalcone (IB) were finalized. The NBIF+BA synergistic combination with significant synergism (CI<0.9) showed superior efficacy to individual components, and equivalence to PCF extract (90% confidence interval: 78.12%-132.15%). Conclusion This AI-driven strategy successfully identified a synergistic combination of NBIF+BA with potent anti-osteoporotic effects from PCF, presenting a promising approach for the treatment of complex diseases.

Paris species, known as “Chonglou” in traditional Chinese medicine, have been used clinically for millennia. Nevertheless, the variation patterns of their chemical constituents and biological activities among different species are still poorly understood and urgently need to be elucidated. Current research has largely focused on a few pharmacopoeial species, while many with ethnomedicinal value remain scientifically unexplored. This study presents the first integrated metabolomic and pharmacological comparison of two pharmacopoeial species, P. polyphylla Smith var. chinensis (PPC) and P. polyphylla Smith var. yunnanensis (PPY), with two ethnomedicinal species, P. thibetica France (PT) and P. fargesii France (PF). A comprehensive phytochemical profile was established using UPLC-Q-TOF-MS/MS, identifying 119 compounds, 85.7% of which were steroidal saponins. Significant inter-species variation in total saponin content was observed, with PF showing the highest average. Multivariate statistical analysis successfully identified species-specific markers and revealed unexpected chemotaxonomic affinities. Pharmacologically, all four species exhibited significant dual bioactivities. They demonstrated potent hemostatic effects, significantly reducing tail bleeding time and directly inducing platelet aggregation. Concurrently, the extracts showed considerable anti-hepatocarcinoma activity. Mechanistically, this anti-cancer effect involved the induction of S-phase cell cycle arrest and apoptosis, which were associated with down-regulation of Bcl-2 and suppression of c-Myc signaling. The above-mentioned findings confirmed PF and PT as promising complementary resources to the official pharmacopoeial species. This work provides a critical chemical and pharmacological foundation for their quality standardization and potential therapeutic application in hemostasis and liver cancer management.

The profound phenotypic heterogeneity of circulating tumor cells (CTCs) presents a major analytical challenge, demanding technologies capable of high-plex, quantitative single-cell profiling. Here, we introduce PRISM (Phenotypic Resolution via Immuno-SERS Mapping), a biosensing platform engineered to meet this challenge. The platform integrates a high-efficiency dual-antibody (anti-EpCAM/anti-CSV) capture substrate with a suite of seven spectrally orthogonal SERS nanoprobes, enabling crosstalk-free, multiplexed quantification of epithelial, mesenchymal, and stem-like (E-M-S) markers. We demonstrate the platform's robust analytical performance, including high capture efficiency for heterogeneous cell lines and excellent linearity. Its superior phenotypic resolving power was validated by quantitatively distinguishing canonical cell line archetypes and tracking dynamic protein expression shifts during induced epithelial-mesenchymal transition. Applying PRISM to CTCs from pancreatic cancer patients, we introduce a novel data analysis framework, including a 'Metastasis Potential Score' (MPS), to translate high-dimensional spectral data into a clinically relevant metric for risk stratification. Furthermore, longitudinal analysis of patient samples demonstrates the platform's utility as a dynamic monitoring tool, capable of tracking therapy-induced phenotypic shifts. PRISM establishes a powerful analytical methodology for high-dimensional single-cell analysis, providing a robust tool for both fundamental cancer biology research and translational clinical applications.

Puerariae Lobatae Radix (PLR) is a commonly used herbal medicine in traditional Chinese medicine, whereas Puerariae Lobatae Caulis (PLC), its aerial part, is frequently utilized as an adulterant due to similar morphological features. Conventional identification methods and pharmacopoeial criteria fail to effectively distinguish these two homologous herbs, creating a demand for a precise identification method. Therefore, a Digital ID method was established for their identification. UPLC-QE-Orbitrap-MS coupled with Progenesis QI software was applied to analyze multi-batch samples of PLR and PLC. Common ion information was extracted, and a differential ion matrix was constructed after eliminating blank ions and cross-shared components. High-intensity characteristic ions were defined as exclusive Digital ID for authenticity identification. The Matching Confidence (MC) values of both herbs matching their own Digital IDs were >85%, while cross-matching MC values were <4%. Unlike subjective macroscopic authentication, compendial TLC and single-marker quantification that fail to discriminate the two materials, and spectroscopic methods incompatible with powdered samples, this digital ID framework enables automated objective authentication via quantitative Matching Confidence without manual spectral comparison. This technique achieves accurate and efficient digital identification of the two herbs, facilitating the quality control of PLR and offering a new strategy for the quality assessment of traditional Chinese medicine (TCM).

The leaves of Cyclocarya paliurus (Batal) Iljinsk., a plant native to China that has long been used in traditional Chinese medicine to treat diabetes. It remains to be determined what chemical constituents are responsible for this effect. The aim of this study was to identify the antidiabetic components of C. paliurus using the spectrum-effect relationship and confirmed using molecular docking. The components of C. paliurus were detected using ultra-high-performance liquid chromatography mass spectrometry/mass spectrometry (UHPLC-MS/MS) and ultra-high-performance liquid chromatography-diode array detector (UHPLC-DAD). C. paliurus could significantly reduce glycosylated haemoglobin, blood glucose levels and blood lipid profile. Esculetin, shikimic acid, isoquercitrin, quercitrin, kaempferol-3-O-rhamnoside, tricin, and pterocaryoside A were determined to be the main antidiabetic components of C. paliurus. Results revealed that the antidiabetic effect of C. paliurus was due to the combination of multiple chemical components. The method developed is valuable for identifying the active ingredients of this plant species.

INTRODUCTION:Multiorigin phenomenon is quite common in Chinese herbal medicine, whereas quality consistency evaluation among the involved plant species is seldom considered. OBJECTIVES:The present study, taking Taraxaci herba as a typical case, aimed to develop a strategy based on a combination of chemical similarity and in vitro bioequivalence to evaluate the quality consistency of three Taraxacum species. MATERIALS AND METHODS:The chromatographic fingerprints were established through two approaches, namely, high-performance liquid chromatography coupled with photodiode array detector and quadrupole time-of-flight mass spectrometry (HPLC-PDA/QTOF-MS) and gas chromatography coupled with quadrupole time-of-flight mass spectrometry (GC-QTOF-MS). The common characteristic peaks from chromatographic fingerprints were quantified. The bioactivities were tested by antioxidant and anti-inflammatory activity assays. Quality consistency of the three Taraxacum species was globally evaluated by using a combination of chemical similarity and in vitro bioequivalence through univariate and multivariate statistical analysis. RESULTS:The statistical results showed no classification of the three species based on contents of quantified analytes and in vitro bioactivities, respectively. CONCLUSION:This study not only demonstrated the medicinal interchangeability of three species of Taraxaci herba but also proposed a comprehensive strategy to evaluate quality consistency of multiorigin Chinese herbal medicines by integrating chemical similarity and in vitro bioequivalence.

ETHNOPHARMACOLOGICAL RELEVANCE:Celosia Semen (CS) serves as a traditional Chinese medicine (TCM) for promoting liver health and enhancing vision, with extensive clinical applications. Triterpenoid saponins represent the primary active components of CS, with the highest concentration of Celosin I (CI) detected. The urgent need for effective NAFLD treatments motivated us assess the beneficial effects of total saponins from CS (TSCS) and CI. AIMS OF THE STUDY:To investigate the therapeutic effects of TSCS and CI on NAFLD and its underlying molecular mechanisms. MATERIALS AND METHODS:The impact of TSCS and CI on NAFLD was evaluated through in vitro and in vivo methodologies, utilizing high-fat diet (HFD) and palmitic acid/oleic acid modeling on C57BL/6J mice and AML12 cells, respectively. Biochemical analysis, H&E and Oil red O staining were used to characterize the lipid-lowering and hepatoprotective activities of TSCS and CI. Lipidomics discerned the impact of TSCS and CI interventions on liver lipid composition, distribution and alteration in NFALD mice. RT-qPCR and western blotting detected the influence of TSCS and CI on genes linked to de novo lipogenesis, fat calculation uptake, oxidation and esterification. The docking analysis anticipated the interaction of six major triterpenoid saponins within TSCS with SREBP1. RESULTS:TSCS and CI markedly diminished lipid accumulation induced by high fat both in vivo and in vitro. TSCS and CI also mitigated hepatic steatosis and liver injury induced by HFD through the reduction of TC, TG, FAs, ALT, and AST, even at minimal dose of 25 mg/kg. Lipidomics indicated that TSCS and CI had the potential to modulate the lipid metabolism network, rectify lipid metabolic dysregulation induced by NAFLD, decrease the levels of harmful lipids, and elevate the levels of advantageous lipids. Furthermore, TSCS and CI exhibited a strong affinity to SREBP1, thereby might directly influence the expression of SREBP1 and a cascade of essential enzymes involved in de novo lipogenesis, and finally resulting in a diminished synthesis of novel lipids. CONCLUSION:TSCS and CI were confirmed firstly as key active components of CS in amending hepatic steatosis and mitigate liver damage in NAFLD, outlining the preliminary mechanism. They warrant further exploration as drug candidates for NAFLD treatment, especially in light of the current shortage of medications and limited therapeutic options.

BACKGROUND:Triptolide (TP), a bioactive compound derived from Tripterygium wilfordii, exhibits potent immunosuppressive activity but is limited in clinical application due to hepatotoxicity effects. The direct molecular targets mediating this toxicity remain unclear. PURPOSE:This research was conducted to pinpoint the principal hepatic molecular target underlying TP-induced liver injury and to elucidate the associated toxicological mechanisms, providing insights for potential hepatoprotective interventions. METHODS:Chemical proteomics and metabolomic profiling were integrated to identify direct TP targets. The interaction between TP and adenosylhomocysteinase (AHCY) was evaluated using molecular dynamics (MD) simulations and surface plasmon resonance (SPR) analysis. The protective potential of AHCY overexpression against TP-induced liver injury was investigated in experimental models. RESULTS:Proteomics identified 29 candidate TP-binding proteins associated with liver injury. Metabolomics profiling indicated disturbances in amino acid metabolism, highlighting AHCY as a central mediator. TP exhibited high-affinity binding to AHCY (KD = 3.179 × 10-11 M), resulting in S-adenosylhomocysteine (SAH) accumulation, DNA hypomethylation, metabolic dysfunction, and oxidative stress. Notably, AHCY overexpression attenuated these TP-induced hepatotoxicity. CONCLUSION:Inhibition of AHCY plays a pivotal role in TP-induced hepatotoxicity. Modulating AHCY activity may offer a promising strategy to enhance the hepatic safety profile of TP in therapeutic applications.

ETHNOPHARMACOLOGICAL RELEVANCE:Paridis Rhizoma, derived from two pharmacopoeial Paris species viz. Paris polyphylla Smith var. yunnanensis (Franch.) Hand. - Mazz. (PPY) and P. polyphylla Smith var. chinensis (Franch.) Hara (PPC), has the traditional efficacy of heat-clearing and detoxifying, and is recognized for its anti-inflammatory and hemostatic effects. Despite significant chemical differences, these two species are clinically prescribed without discrepancy, potentially limiting optimal therapeutic outcomes. AIM OF THE STUDY:The aim was to compare the anti-inflammatory and hemostatic bioactive ingredients of two pharmacopoeial Paris species, providing a foundation for more precise medicinal application. MATERIALS AND METHODS:Chemical profiling of PPY and PPC was conducted using UHPLC-QTOF-MS. Anti-inflammatory and hemostatic activities were assessed in zebrafish models. Active compounds were identified through partial least squares-regression (orthogonal partial least squares-regression) and Spearman correlation analyses, and their structure-activity relationships were subsequently elucidated. Network pharmacology, RT-qPCR and Western blot were employed to explore potential targets and pathways. HPLC-UV quantitative method was established to compare active component contents between two species. RESULTS:Spectrum-effect analysis identified 5 anti-inflammatory and 3 hemostatic steroidal saponins from two species. Structural features such as the C17-OH group, C3 oligosaccharide chain and F-ring were critical to bioactivity. Paris saponin II and polyphyllin I, enriched in PPY, demonstrate stronger anti-inflammatory effects via the NF-κB signaling pathway. Chonglouoside H and polyphyllin VI, abundant in PPC, exhibited better hemostatic activity, potentially related to tnfa downregulation. CONCLUSIONS:This study elucidated distinct bioactivity bias of PPY and PPC due to their difference in chemical compositions, suggesting the necessity of precise medication.

INTRODUCTION:The quality of traditional Chinese medicine (TCM) is a prerequisite for clinical efficacy. However, the existing quality evaluation methods are not strongly correlated with efficacy, and they are unable to adequately reflect the quality grade of Changii Radix (CR). OBJECTIVES:In this study, a biology-related chemical indicator quality grading prediction model was developed to predict the quality grade of CR. MATERIALS AND METHODS:Firstly, the quality grade of CR was pre-classified based on immunological activity. Subsequently, one-way analysis of variance, gray correlation analysis, and Pearson correlation analysis were employed to identify the chemical indicators associated with immunological activity. Finally, separately using chemical indicators as independent variables and quality grades as dependent variables, the logistic regression model and a multi-index weighted quality comprehensive evaluation index (QCEI) were constructed to predict the quality grade of CR. RESULTS:The results indicated that 27 batches of CR samples could be divided into three grades of I, II, and III. The gray correlation degrees and Pearson correlation coefficients between water-soluble extractives, polysaccharide, amino acid, and immunological activity all exceeded 0.8 and 0.4 (p < 0.05), respectively. Additionally, both the logistic regression model and QCEI could effectively predict the quality grade of CR, with the logistic regression model showing superior performance. CONCLUSION:This study is the first to establish a chemistry-biology integrated strategy for evaluating the quality grade of CR, providing a novel insight into the assessment of TCM quality grade.

Fucosylated chondroitin sulfate from Pearsonothuria graeffei (FCS-Pg), a natural macromolecular polysaccharide, has been proven to prevent obesity, but its underlying molecular mechanism is still unclear. C57BL/6 J mice fed on high fat diet (HFD) were administered FCS-Pg lasting for ten weeks. The results demonstrated that FCS-Pg supplementation reduced body weight with dosage manner compared with HFD group. The expressions of intestinal lipid synthesis related proteins such as cluster of differentiation 36 (CD36) in FCS-Pg group were lower than those in the HFD group. Compared with the normal group, HFD caused gut microbiota disorder in the colon. FCS-Pg supplementation at high dosage restored the gut microbiota composition with higher abundance of Alistipes and Bacteroidetes and lower abundance of Colidextribacter, Bilophila and Firmicutes compared with HFD group. Moreover, FCS-Pg decreased the expression of proteins involved in triglyceride synthesis such as glycerol 3-phosphate dehydrogenase 1 (GPD1) and increased the expression of proteins involved in lipolysis and thermogenesis such as adipose triglyceride lipase (ATGL) and uncoupling protein 1 (UCP1) in the white adipose tissue (WAT) compared with HFD group. In conclusion, our study suggested that FCS-Pg significantly prevented obesity and improved WAT function in HFD-fed mice by regulating intestinal lipid metabolism and microflora composition.

Chemical-induced cholestasis (CIC) has become a concern in chemical safety risk assessment in pharmaceutical, food, cosmetic, and industrial manufacturing. Currently, known animal and in vitro liver models are unsuitable as high-throughput screening tools due to their high cost, time-consuming, or poor screening accuracy. Herein, a cohort of chemicals validated as cholestatic hepatotoxic in humans, rodents, and in vitro liver models was established for testing. The accuracy and reliability of the detection of CIC in zebrafish larvae were assessed by liver phenotype, bile flow inhibition rate, bile acid distribution, biochemical indices, and RT-qPCR. In addition, the nomogram prediction model was constructed using binomial logistic regression analysis. The model was constructed with three variables: aspartate aminotransferase (AST.FC) level, total bile acid (TBA.FC) level, and fold change in the number of bile acid nodes per unit of bile ducts in the zebrafish liver (NPL.FC), which showed high predictive power (areas under the ROC curve: 0.983). Furthermore, this study demonstrated that zebrafish larvae have some model specificity for CIC risk assessment of estrogen endocrine disruptors and that testing after 10 dpf provides more scientific results. Overall, combining zebrafish larval phenotyping and nomograms is an efficient and powerful tool for CIC risk monitoring of chemicals.

BACKGROUND:YY1 plays a crucial part in the onset and progression of numerous liver diseases, yet the significant contribution of YY1 to drug-induced liver injury (DILI) appears to have been underestimated by researchers. PURPOSE:To reveal the underlying role of YY1 in DILI. METHOD:The compounds that interact with YY1 were queried in the Comparative Toxicogenomics Database (CTD), with the majority found to be hepatotoxic, which includes certain widely used drugs. Molecular docking and SPR characterized the robust binding of hepatotoxic compounds to YY1. The duty of YY1 in DILI was investigated in Diosbulbin B (DIOB), a recently identified hepatotoxic compound that tightly associates with YY1, and further validated on ANIT, LCA, APAP, and CDDP. Transcriptomic analysis disclosed the underlying mechanisms involved in DIOB-induced liver injury. RT-qPCR, immunohistochemistry, immunofluorescence, western blotting, and cellular transfection techniques were employed to validate the specific mechanism. RESULTS:Among the 94 compounds affecting YY1 expression in the CTD, 59 compounds exhibited hepatotoxicity, showing close interactions with YY1 and almost consistent binding sites by molecular docking. The SPR validated the tough binding of several hepatotoxic compounds to YY1, including five FDA-approved hepatotoxic drugs. Mechanistically, the involvement of YY1 in DILI was uncovered through the cholestasis lens, mice hepatic YY1 was up-regulated by hepatotoxic DIOB and transcriptionally inhibited FXR and its downstream BSEP and MRP2 expression, initiating early in cholestatic liver injury and persisting to drive the progression of cholestasis. ANIT and LCA-induced model of cholestasis provided evidence for the hypothesis that YY1 frequently mediates drug induced cholestasis (DIC). APAP and CDDP indicated that YY1 may also be involved in hepatocellular and mixed type DILI. CONCLUSION:YY1 widely mediated the development of DIC and also might be engaged in other types of DILI. YY1 presented a common target for hepatotoxic medications and the targeting of liver YY1 for drug development may offer a novel approach for managing DILI.

Authentication and adulteration detection of closely related herbal medicines is a thorny issue in the quality control and market standardization of traditional Chinese medicine. Taking Fritillariae Bulbus (FB) as a case study, we herein proposed a three-step strategy that integrates mass spectrometry-based metabolomics and multivariate statistical analysis to identify specific markers, thereby accurately identifying FBs and determining the adulteration level. First, an ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry-based untargeted metabolomics method was employed to profile steroid alkaloids in five sorts of FB and screen potential differential markers. Then, the reliability of the screened markers was further verified by the distribution in different FB groups acquired from ultra-high performance liquid chromatography triple quadrupole mass spectrometry-based pseudotargeted metabolomics analysis. As a result, a total of 16 compounds were screened out to be the specific markers, which were successfully applied to distinguish five FBs by using discriminant analysis model. Besides, partial least squares regression models based on specific markers allowed accurate prediction of three sets of adulterated FBs. All the models afforded good linearity and good predictive ability with regression coefficient of prediction (R2p) > 0.9 and root mean square error of prediction (RMSEP) < 0.1. The reliable results of discriminant and quantitative analysis revealed that this proposed strategy could be potentially used to identify specific markers, which contributes to rapid chemical discrimination and adulteration detection of herbal medicines with close genetic relationship.

Quality consistency evaluation of traditional Chinese medicines (TCMs) is a crucial factor that determines the safe and effective application in clinical settings. However, TCMs exhibit diverse, heterogeneous, complex, and flexible chemical compositions, as well as variability in preparation processes. These characteristics pose greater challenges in researching the consistency of TCMs compared to chemically synthesized and biological drugs. Therefore, it is paramount to develop effective strategies for evaluating the quality consistency of TCMs. From the starting point of quality properties, this review explores the strategy used to evaluate quality consistency in terms of chemistry-based strategy (chemical consistency) and the biology-based strategy (bioequivalence). Among them, the chemistry-based strategy is the mainstream, and biology-based strategy complements the chemistry-based strategy each other. Furthermore, the emerging chemistry-biology strategies (overall evaluation) is discussed, including individually combining strategy and integration strategy. Finally, this review provides insights into the challenges and future perspectives in this field. By highlighting current status and trends in TCMs quality consistency, this review aims to contribute to establishment of generally applicable chemistry-biology integrated evaluation strategy for TCMs. This will facilitate the advancement toward a higher stage of overall quality evaluation.

The saponin-enriched extract from Celosiae Semen is a promising resource owing to its lipid-lowering activity. However, triterpenoid saponins are difficult to extract owing to their high molecular weight and strong water solubility. The aim of this paper was to explore an eco-friendly and effective technology of extraction and enrichment of total triterpenoid saponins to obtain high lipid-lowering fractions. Initially, Box-Behnken design experiments were employed to optimize the heat reflux extraction process on the basic of mono-factor experiments. Afterwards, the crude extract was further purified using D-101 resin, and the purification parameters were investigated based on adsorption/desorption experiments and biological activity assay. Under optimal conditions, the purity of the finally obtained total triterpenoid saponins was increased by 7.28-fold. The lipid-lowering activities of the six main triterpenoid saponins were evaluated in HepG2 cells induced by palmitic acid. The results of Oil Red O staining showed that the compounds all exhibited potential lipid-lowering activity. The structure-activity relationship analysis suggested that the oligosaccharide chain at C-28 played an essential role in their lipid-lowering activity and the substituent group at C-23 site also showed important effects. The optimal extraction and purification methods may facilitate the utilization of Celosiae Semen for the industrial production as a functional food and drug.