Hair greying is a common aspect of the natural ageing process. Although it is generally not considered a medical problem, its high prevalence can substantially impact emotional state due to aesthetic concerns. A growing body of research has demonstrated that natural products and their derivatives derived from plants possess advantages and potential in the treatment of hair greying. To review the last research progress in the treatment of hair greying by natural products and their derivatives, focusing on the target and mechanism of action of natural products and their derivatives and providing a reference for future clinical use. We searched electronic databases (PubMed, Web of Science, ClinicalTrials.gov, and CNKI) for studies published between January 2005 and June 2025. The research focused on the pathogenesis of hair greying and the use of natural products and their derivatives to prevent and treat it, using the keywords: “hair greying”, “hair pigmentation”, “white hair”, “snow hair”, “melanocyte stem cells” and “melanin”. Account of natural products (e.g., Polygoni multiflori radix) and their derivatives (e.g., Epimedin B), are expected to treat hair greying due to their various qualities to regulate melanocyte stem cells, enhance melanin synthesis, or promote melanosome transport. Compared to oral administration, topical application represents a preferred approach for promoting hair pigmentation. We discussed and summarized the mechanism of natural products and derivatives in the treatment of hair greying, which provided a reference for future clinical use.

Hepatocellular carcinoma (HCC) remains challenging with limited immunotherapy response. Despite its clinical promise in advanced HCC, the mechanisms of icaritin, especially concerning ferroptosis induction and immune modulation, remain elusive. This study aims to determine if the antitumor effect of icaritin involves the induction of ferroptosis via NAD(P)H quinone oxidoreductase 1 (NQO1) and if it can augment the efficacy of programmed cell death 1 ligand 1 (PD-L1) therapy by potentiating natural killer (NK) cell activity. Using human HCC cell lines (Huh7, Hep3B, PLC/PRF/5, SNU-449, and MHCC97-H) and two synergistic mouse models (Hepa1-6 and SgPten/c-Met), we examined icaritin's inhibition of tumor growth and induction of ferroptosis via the NQO1 pathway, monitoring key markers (reactive oxygen species [ROS], glutathione peroxidase 4 [GPX4], ferritin heavy chain 1 [FTH1]). The NQO1 inhibitor dicoumarol was employed to validate the pathway. Tumor microenvironment (TME) remodeling was assessed through cancer-associated fibroblasts (CAFs) markers and immune cell profiling, focusing on NK cell infiltration. Combination therapy with anti-PD-L1 was tested in vivo. Icaritin significantly inhibited HCC growth in vitro and in vivo. Its antitumor effect was mediated by NQO1-mediated ferroptosis, via elevated ROS, diminished mitochondrial membrane potential, and downregulated GPX4 and FTH1. Analysis of The Cancer Genome Atlas (TCGA) data revealed that NQO1 is overexpressed in human HCC tissues. Icaritin enhanced NK cell infiltration while reducing CAF abundance and suppressing recombinant focal adhesion kinase (FAK) and discoidin domain receptor 1 (DDR1) signaling. Notably, icaritin synergized with anti-PD-L1 therapy to enhance tumor suppression without increasing toxicity, correlating with potentiated NK cell immunity. Our findings demonstrate that icaritin triggered NQO1-mediated ferroptosis and remodeled TME to enhance NK cell recruitment and PD-L1 therapy efficacy. This provides rationale for evaluating icaritin-based combination immunotherapy in HCC through dual action on ferroptosis and NK cell activation.

Aberrant activation of NLRP3 inflammasome has been associated with a variety of human inflammatory diseases, but no small molecule inhibitors of NLRP3 were applied in clinical practice. Our research group has previously shown that licochalcone B is an effective NLRP3 inflammasome inhibitor, however, its IC50 value is relatively high compared to compounds such as MCC950. Modifying bioactive natural products to find NLRP3 inflammasome inhibitors with stronger potency and higher specificity is a direction worthy of investigation. Our research aims to evaluate the impact of various licochalcone B derivatives on NLRP3 inflammasome, to screen for derivatives with better potency and elucidate the underlying mechanisms. We investigated the effects of licochalcone B and its derivatives on NLRP3 inflammasome by assessing the production of active caspase-1 and interleukin 1β (IL-1β). To elucidate the mechanism of CTG12, we employed co-immunoprecipitation. Furthermore, we evaluated CTG12 in LPS-induced acute systemic inflammation mouse models. The findings demonstrated that licochalcone B and its derivatives effectively inhibit NLRP3 inflammasome. CTG12 exhibits the most potent inhibitory activity, showing approximately tenfold increased effects compared to licochalcone B. Mechanistic investigations reveal that while CTG12 does not affect potassium (K⁺) efflux, calcium (Ca2⁺) influx, or mitochondrial reactive oxygen species (mtROS) production, it suppressed NLRP3 assembly by interfering with the ASC-NLRP3 interaction and the NLRP3-dependent ASC oligomerization process. In vivo, CTG12 provides significant therapeutic benefits in LPS-induced acute systemic inflammation models. Our results indicated that structurally modified licochalcone B derivative CTG12 inhibits NLRP3 assembly by interfering with the ASC-NLRP3 interaction, thereby inhibiting the NLRP3-dependent ASC oligomerization process in NLRP3 inflammasome activation. These studies would show that CTG12 is a valuable small molecule inhibitor that holds promise as a high-value drug candidate for the treatment of NLRP3-mediated inflammatory diseases.

Background Cholestatic liver disease (CLD) is a common disorder characterized by impaired bile flow that results in liver injury and fibrosis. However, current therapeutic options for CLD are limited and ineffective. Yu Dan Tong Formula (YDTF), a formulated Traditional Chinese Medicine (TCM), has been routinely used for over 50 years at Beijing Children's Hospital for the treatment of CLD in children. We previously found that YDTF can reduce jaundice and alleviate liver injury. However, the pharmacological mechanisms underlying the effects of YDTF on CLD remain unclear. Purpose This study aimed to investigate the pharmacological mechanisms of YDTF in the treatment of CLD, with a focus on NLRP3 inflammasome activation. Methods Macrophage inflammasomes were activated by classical and non-classical NLRP3 agonists, including taurochenodeoxycholic acid (TCDCA). The effects of YDTF on inflammasome activation and inflammatory responses were assessed in an lipopolysaccharide (LPS)-induced murine model of systemic inflammation in C57BL/6 mice, and the underlying mechanisms were evaluated. The effects of YDTF on NLRP3 activation and its therapeutic potential in CLD were explored using an α-naphthylisothiocyanate (ANIT)-induced CLD model in C57BL/6 mice Results YDTF inhibited NLRP3 activation in a dose-dependent manner with broad-spectrum inhibitory effects. Mechanistically, YDTF inhibited Ca²⁺ influx, thereby inhibiting apoptosis-associated speck-like protein (ASC) oligomerization, and subsequent inflammasome assembly and activation. In the ANIT-induced CLD mouse model, YDTF effectively suppressed NLRP3 inflammasome activation and reduced the magnitude of the inflammatory response. Furthermore, YDTF alleviated abnormal liver function and resolved liver injury. Conclusions YDTF effectively inhibited NLRP3 activation by inhibiting Ca²⁺ influx and significantly alleviated liver injury in the ANIT-induced CLD model. The inhibition of NLRP3 activation could represent a critical pharmacological strategy for the treatment of CLD. Additionally, our results suggest that YDTF may be beneficial in treating other NLRP3 inflammasome-driven diseases.

Emerging evidence underscores the pivotal role of Ten-eleven translocation 2 (Tet2), as an epigenetic regulator with neuroprotective functions, yet its temporal dynamics and pathogenic contributions to Alzheimer’s disease (AD) remain poorly understood. By integrating human longitudinal cohort data with experimental models, we demonstrated that Tet2 deficiency accelerated AD‑like neurodegeneration through oxidative stress-dependent cGAS-STING activation. Clinically, Tet2 loss-of-function carriers among Aβ-positive individuals exhibited significantly accelerated cognitive decline. In mice, Tet2 expression decreased with age and in late‑stage AD models, and constitutive Tet2‑mutant (Tet2mut) mice recapitulated AD-like behaviors and pathology by exacerbating neuroinflammation and impairing neurogenesis and synaptic plasticity. Crucially, Tet2 deficiency induced mitochondrial dysmorphology and heightened oxidative stress in the hippocampus, culminating in DNA damage and robust cGAS-STING activation. Mechanistically, integrative epigenomic analyses further revealed that Tet2 deficiency was associated with hypermethylation of antioxidant gene networks, thereby exacerbating oxidative stress and mitochondrial injury. Notably, antioxidant treatment with N-acetylcysteine (NAC) effectively alleviated oxidative damage, restored mitochondrial function and suppressed cGAS-STING signaling in Tet2-silenced microglia. In vivo, NAC administration in Tet2mut mice improved cognitive performance and synaptic plasticity while reducing neuroinflammation and neuronal loss through inhibition of cGAS-STING signaling. These findings delineate a previously unrecognized epigenetic-immune axis in AD, highlighting Tet2 enhancement and ROS modulation as promising therapeutic strategies.

Idiosyncratic Drug-Induced Liver Injury (IDILI) challenges drug development and clinical use. Its unpredictable nature complicates mechanistic research. Polygonum multiflorum Thunb (PM), a traditional tonic herb, has been used for centuries in China and East Asia. Now, PM-induced IDILI is a focus of traditional Chinese medicine safety research. Nevertheless, the underlying mechanisms of PM-triggered IDILI require further investigation. Previous clinical studies identified elevated Tumor Necrosis Factor-alpha(TNF-α) levels in patients susceptible to PM-induced liver injury. This finding prompted us to employ integrated network pharmacology, targeted lipidomics, and transcriptomics to investigate the combined effect and mechanism of PM and the susceptibility factor TNF-α in C57 mice. Histochemical staining, biochemical assays, and inflammatory cytokine analysis revealed significant hepatic inflammatory infiltration in the TNF-α+PM co-exposure group versus controls. This group exhibited markedly elevated levels of liver injury markers (ALT, AST, DBIL) and inflammatory mediators (IL-6, TNF-α) (P < 0.05, P < 0.01). Subsequent administration of the TNF-α antagonist etanercept restored hepatic architecture and significantly reduced cytokine levels (P < 0.01), approaching baseline values. Notably, neither TNF-α nor PM monotherapy induced liver injury in mice. Network pharmacology analysis identified 33 bioactive PM components and 11 core targets. Targeted lipidomics revealed 32 differential lipid metabolites (DLMs), primarily glycerolipids (GL) and glycerophospholipids (GP). Transcriptomics demonstrated cooperative regulation of 199 differentially expressed genes(DEGs) by TNF-α and PM. Network enrichment analysis further uncovered lipid dysregulation mediated by Pparg, Cyp4a12a, Cyp4a12b, and Cyp4a31 genes, with RT-qPCR validation confirming these findings. In summary, TNF-α plays a central driving role in PM-induced liver injury. When co-administered with PM, it activates the PPAR signal ing pathway via downregulation of Cyp4a12a, Cyp4a12b, and Cyp4a31, thereby dysregulating lipid metabolism, promoting inflammatory mediator production, and ultimately causing liver injury.

ETHNOPHARMACOLOGICAL RELEVANCE:Hepatic fibrosis (HF) is a prevalent consequence of chronic liver injury, resulting from the persistent interplay of inflammation, oxidative stress, dysregulated cell death, metabolic reprogramming, and remodeling of the immune microenvironment, which collectively establish a self-amplifying profibrotic network. In traditional medical systems, HF corresponds to syndromes such as hypochondriac pain, mass formation, blood stasis, and jaundice, indicating a chronic and systemically imbalanced disease trajectory. Guided by holistic and multi-target principles, traditional Chinese medicine (TCM) has amassed considerable ethnopharmacological knowledge in managing chronic liver diseases and fibrosis. AIM OF THE STUDY:This review seeks to transcend single-mechanism explanations by integrating contemporary evidence regarding the antifibrotic effects of TCM from a systems-pathology perspective. We aim to elucidate how TCM interventions coordinately modulate the interactions among inflammation, oxidative stress, programmed cell death, metabolic dysregulation, and remodeling of the immune microenvironment, thereby disrupting the self-sustaining fibrotic network. MATERIALS AND METHODS:Bibliometric analyses were performed to characterize research trends and thematic evolution in TCM-related antifibrotic studies. Concurrently, a systematic literature search was executed in PubMed, Web of Science, and CNKI for experimental and clinical studies published between 2000 and 2025. The Latin binomials of all medicinal plants discussed were standardized and verified using the Medicinal Plant Names Services (MPNS) database to ensure taxonomic accuracy and nomenclatural consistency. RESULTS:Current evidence suggests that TCM does not exert antifibrotic effects by targeting isolated pathological nodes; instead, it attenuates key profibrotic feedback loops through multi-level coordinated regulation. At the inflammatory level, TCM suppresses amplification axes centered on NF-κB and NLRP3, thereby diminishing the cumulative injury burden imposed by persistent immune activation on both parenchymal and non-parenchymal liver cells. At the levels of oxidative stress and metabolism, TCM modulates Nrf2-associated antioxidant networks in conjunction with energy and lipid metabolic pathways, disrupting the reciprocal reinforcement between oxidative stress and metabolic reprogramming. Regarding programmed cell death, TCM influences the dynamic balance among apoptosis, pyroptosis, necroptosis, and ferroptosis, thereby limiting the continuous pro-inflammatory stimulation of the fibrotic microenvironment. In terms of immune microenvironment remodeling, TCM regulates macrophage polarization, lymphocyte function, and sinusoidal endothelial phenotypes, facilitating a transition from a profibrotic state to reparative immune homeostasis. These convergent effects ultimately integrate at critical signaling nodes, including TGF-β/Smad, PI3K/Akt/mTOR, and Wnt/β-catenin pathways, thereby attenuating extracellular matrix accumulation and its associated mechanotransductive feedback that sustains fibrosis. CONCLUSION:Collectively, TCM demonstrates a systems-level antifibrotic profile by modulating the interactions among multiple pathological processes, rather than relying solely on single-target inhibition. This mode of action is consistent with the complex pathophysiology of HF. However, current evidence primarily relies on associative mechanisms and phenotypic improvements, while offering limited insight into multicellular causal interactions, critical regulatory nodes, and the pharmacodynamic foundations of TCM interventions. Consequently, future research should conceptualize HF as a disorder characterized by disrupted inter-organ regulation, integrate the gut-liver axis and immune involvement, employ single-cell and multi-omics approaches to elucidate causal pathways, and validate system-level effects through rigorously designed long-term clinical studies.

BackgroundHepatocellular carcinoma (HCC) poses ongoing difficulties for public health systems due to its high incidence and poor prognosis. Huqi formula (HQF), a well-known prescription in traditional Chinese medicine, has demonstrated notable clinical effectiveness in the treatment of HCC. However, the mechanisms underlying its therapeutic effects have yet to be completely elucidated.PurposeThis study aimed to investigate the anti-HCC effects of HQF and its underlying mechanisms.MethodsChemical profiling and quantification of HQF were conducted by LC-MS and HPLC. Orthotopic and subcutaneous tumor models were established through hydrodynamic injection of Akt/Nras plasmids and subcutaneous injection of c-Met/sgPten cells, respectively, to evaluate the therapeutic effects of HQF on HCC. Network pharmacology, RNA-Seq, molecular docking, Western blot, and flow cytometry were employed to assess the anti-HCC mechanisms.ResultsLC-MS analysis identified 41 components, with HPLC quantification showing salvianolic acid B as the most abundant compound (0.303%). In Akt/Nras and c-Met/sgPten-induced HCC models, HQF significantly reduced tissue damage, improved liver function, and inhibited HCC progression. Mechanistic studies revealed that HQF induced apoptosis in HCC cells by downregulating p-PI3K, p-AKT, and p-mTOR expression, with molecular docking indicating the strongest binding affinity between salvianolic acid B and PI3K. HQF further enhanced CD4+ and CD8+ T cell infiltration within the tumor microenvironment. When combined with PD-1 therapy, HQF improved therapeutic efficacy against HCC. Finally, toxicity assays confirmed the safety profile of HQF.ConclusionHQF demonstrated significant anti-HCC effects and a synergistic effect with PD-1, could be used as an alternative therapeutic agent for HCC.

The multi-modal lateral flow immunosensor (LFIS) with integrated advantages and mutual built-in calibration ability could avoid the limitations of traditional single-signal output mode to meet the growing demands for trace contaminants. In this study, an innovative "three-in-one" LFIS platform was developed for colorimetric, grayscale, and fluorescent detection of aflatoxin B1 (AFB1). It integrated polydopamine (PDA)-coated ferric metal-organic framework (MOF) (MIL@PDA) and the functionalized zirconium MOF of a UiO linker enriched with abundant AIEgens (UiOL@AIEgens) as signal probes. The synthesized MIL and UiOL with large surface area and high porosity could enrich numerous PDA and AIEgens probes, respectively, for signal amplification, not only significantly enhancing the colorimetric signal outputs, and grayscale and fluorescence responses, but also largely improving the detection sensitivity and analytical accuracy. The MIL@PDA-monoclonal antibodies (mAbs) probes could specifically identify AFB1 antigens on the test (T) line to produce a visible dark-grey band for qualitative colorimetric detection, and the grayscale intensities were monitored for the quantitation of AFB1 via a portable device. The fluorescence mode was realized through the MIL@PDA-mAbs probes quenching the fluorescence of UiOL@AIEgens on the T line, and the fluorescence intensities were recorded by using a smartphone for accurate quantitation. Under optimal conditions, this MIL@PDA-UiOL@AIEgens driven three-modal LFIS platform allowed for AFB1 detection in a wide range of 0.01-5 ng/mL with a low limit of detection of 0.01 ng/mL that was more than 59-fold lower than the traditional AuNPs-based LFIS. The feasibility and practicability of this LFIS platform was verified for AFB1 detection in lotus seeds. This study opened up a new avenue for developing high-performance LFIS platforms for the trace detection more harmful analytes in complex matrices.

ETHNOPHARMACOLOGICAL RELEVANCE:Xihuang Pill (XHP) is a traditional Chinese medicine formula that was originally used to treat malignant ulcers. Recent studies revealed its therapeutic effects on various malignant tumors. However, its potential efficacy and mechanisms in primary liver cancer (PLC) were not thoroughly investigated. AIM OF THE STUDY:This study aimed to elucidate the efficacy and potential mechanisms of XHP in the treatment of PLC. METHODS:An orthotopic PLC mice model was established adopting hydrodynamic tail vein injection method. Human liver cancer cell lines and organoids were utilized to assess the effect of XHP in vitro. The expressions of alpha-fetoprotein (AFP) and Yes-associated protein (YAP) were evaluated with western blotting. The mRNA expressions of YAP downstream targets were detected with qRT-PCR. Data from Liver Hepatocellular Carcinoma Collection of the Cancer Genome Atlas (TCGA-LIHC) were extracted to identify the potential targets of HCC. The major active components of XHP methanol extract and XHP medicated serum were detected by UHPLC-MS/MS. Human liver cancer cell lines were used to assess the efficacy and potential mechanisms of these active components in XHP in vitro. Finally, molecular docking was conducted to predict the binding affinities of XHP's active components with AFP and YAP. RESULTS:XHP inhibited PLC tumor growth in the mice model with decreased AFP and Ki-67 index. In vitro, XHP suppressed the proliferation and migration of liver cancer cell lines in a time- and dose-dependent manner. Furthermore, even with a low concentration (5 mg/mL), XHP paralyzed the growth of PLC organoids derived from patients. Mechanistically, XHP downregulated the expression of AFP and YAP signaling in vitro and in vivo. UHPLC-MS/MS analysis identified 25 active components in XHP medicated serum. Among them, certain active compounds suppressed PLC cell proliferation and downregulated AFP and YAP signaling, suggesting their therapeutic potentials in PLC. Molecular docking indicated that several components in XHP exhibited strong binding affinities with both AFP and YAP. CONCLUSION:XHP inhibited PLC growth by suppressing AFP and YAP signaling. This study provides an experimental basis for XHP application in PLC treatment.

Objective: Stress is the main factor that leads to hair loss, and no effective therapeutics have been developed for stress alopecia. Polygonum multiflorum Thunb. (PM) is a famous traditional Chinese medicine for tonifying the kidney and nourishing the liver, which is used to treat hair loss. This study aimed to demonstrate that PM is a promising agent for the treatment of chronic stress-induced hair loss. Materials and Methods: C57BL/6J male mice were subjected to chronic restraint stress to inhibit hair follicle growth. Subsequently, the effect of PM was examined by oral administration of the PM water extract or cortisone, followed by histological analysis, immunofluorescence of follicle samples, serum metabolomics, and adrenal proteomics. Results: Hair regeneration in a mouse model was severely inhibited by chronic restraint stress. After PM treatment, hair regeneration in model mice was promoted, including an increased length of hair shafts in skin sections, an increased number of 5-Ethynyl-2’- deoxyuridine-positive cells, and a decreased number of apoptotic cells around the hair follicles. The various test indicators of these PM-treated mice almost recovered to levels comparable to those of the control mice. Through metabolomics and proteomics analyses, we identified 181 endogenous differential metabolites and 75 differential proteins, which may be the potential targets for PM. Pathway enrichment analysis showed that steroid biosynthesis (the main pathway through which stress leads to an imbalance in glucocorticoid secretion by the adrenal gland) may play an important role in the therapeutic effects of PM. In addition, omics analysis revealed that glycolysis/gluconeogenesis, pyruvate metabolism, lipid metabolism, and other glucocorticoid-related metabolic pathways differed significantly after PM treatment. Conclusions: PM is an important natural drug that regulates hair regeneration in mice under stress, and its underlying mechanism is closely related to the regulation of adrenal glucocorticoids and their metabolism.

INTRODUCTION:Determining the hepatotoxic potential of Euodiae Fructus (EF) and exploring the methods and mechanisms of detoxification after processing and compatibility are critical for its rational use. The changes in components and endogenous metabolites after administration might provide a pathway to resolve the above issues. OBJECTIVE:This study aims to investigate whether boiling water washing (BWW), drying after BWW, or compatibility in Wuzhuyu decoction (WZYD) can mitigate the hepatotoxicity of EF, and to explore the underlying mechanisms through chemical composition and metabolomics analysis. METHODS:The hepatotoxicity of EF, processed EF, and WZYD were evaluated in normal mice, then the hepatotoxicity of WZYD was evaluated in migraine model mice. General physical signs (e.g., weight loss, reduced activity, and dull fur), biochemical markers (e.g., ALT, AST, TBIL, and ALP levels), and histopathological examination were observed. Ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was used to quantify the change of 14 specific ingredients in EF after processing and compatibility, including alkaloids (e.g., evodiamine, rutaecarpine), phenolic acids (e.g., chlorogenic acid), and flavonoids. Metabolomics based on ultra-high-performance liquid chromatography quadrupole time-of-flight tandem mass spectrometry (UHPLC-Q-TOF-MS) was employed to identify key pathways, and protein expression in these pathways was assessed to confirm detoxification mechanisms. RESULTS:The decocted extract of raw EF (SEF) induced liver injury in normal mice at a human equivalent dose, while the decocted extracts of boiling water-washed EF (TEF) and dried boiling water-washed EF (GEF) reduced this injury. WZYD did not induce liver injury in normal and migraine model mice, further supporting its safety profile. Significant differences in chemical composition were observed among SEF, TEF, GEF, and WZYD, including a reduction in phenolic acids (e.g., chlorogenic acid and caffeic acid) and an increase in alkaloids (e.g., evodiamine and rutaecarpine). Metabolomics analysis revealed that both boiling water washing (BWW) processing and WZYD compatibility influenced linoleic acid metabolism, oxidative stress, and inflammation. Specifically, CYP2E1 expression was significantly reduced in the TEF and WZYD groups, accompanied by decreased oxidative markers (MDA) and inflammatory cytokines (TNF-α and IL-6). CONCLUSION:In summary, BWW processing and WZYD compatibility reduce EF-induced hepatotoxicity by modulating linoleic acid metabolism, CYP2E1 activity, oxidative stress, and inflammation. These processes are interconnected and play a central role in the detoxification of EF. By altering the chemical composition of EF, these processes reduce the levels of potential hepatotoxic components such as phenolic acids, while increasing the concentration of hepatoprotective alkaloids like evodiamine and rutaecarpine. These changes, combined with the modulation of key metabolic pathways, provide a scientific basis for the safer clinical use of EF.

Non-clinical pharmacology of traditional Chinese medicine (TCM) formulae plays a crucial role in elucidating their therapeutic mechanisms. With the integration of cutting-edge disciplines such as systems biology, big data, and artificial intelligence, the research depth and breadth in this field continue to expand. However, technical standards for non-clinical pharmacology research on TCM formulae have not yet been established. To address this, technical guidelines for non-clinical pharmacology research of TCM formulae were formed, aiming to scientifically standardize research content, elevate research quality, and facilitate the mutual translation between clinical and basic research. Guided by integrative pharmacology of TCM and driven by clinical value, these guidelines employ multi-dimensional qualitative pharmacokinetics (PK) and pharmacodynamics (PD) studies alongside quantitative "PK-PD" studies of key components. This approach enables the transition from panoramic description to precise mechanistic research, establishing a complete evidence chain system for the effectiveness of TCM formulae and demonstrating their holistic action characteristics of "multi-component, multi-pathway, multi-target". Meanwhile, the guidelines emphasize that animal models for TCM research should ensure stability and reproducibility. Additionally, pharmacodynamic evaluation is characterized by its measurability and quantifiability. The mechanisms of action at the molecular level should be in-depth and interpretable. These aspects collectively provide guidance for basic non-clinical pharmacology research on TCM formulae and the development of new TCM drugs. The formulation of these guidelines leveraged the research strengths of the drafting units' teams, incorporated literature review findings, and achieved consensus through extensive discussions among multidisciplinary experts. Broad expert opinions from universities, research institutes, and enterprises engaged in new drug development were solicited, ensuring the guidelines' practicality, standardization, scientific rigor, and feasibility.

Continuous improvement in food safety standards poses new challenges to the detection of pesticide residues. Recently, time-resolved fluoroimmunoassay (TRFIA) has been widely concerned with rapid development in the detection of diverse pesticides in foods due to its simple pretreatment, convenient operation, low cost, high sensitivity, and fast analysis speed. The relevant publications have increased, attracting growing research interest. Thus, in this review, the general situation and main types of TRFIA were first introduced, then, the real applications of different materials assisted TRFIA including lanthanide elements (europium, terbium, and samarium), lanthanide element-doped fluorescent microspheres and nano-beads, in combination with biotin-avidin system amplification strategies in the detection of pesticide residues in food samples, as well as their advantages, faced challenges and future prospects were discussed. It is hoped that this review will provide leading insights into the future development and wide application of TRFIA for researchers in different fields to ensure the quality and safety of foods, as well as peopleu2019s health.

In the wake of the continuous advancement of social technology and the economy, the pressure on people in work, study, and life has escalated. The chronic stress state persisting for an extended period has become the norm and may give rise to diverse health problems. Hence, choosing appropriate approaches to alleviate stress is of paramount importance. Common methods for stress relief encompass exercise, moderate alcohol consumption, stress preadaptation, and so forth. For individuals with more severe symptoms, medications such as paroxetine might be requisite. Although these methods can to some extent ameliorate or mitigate stress, there is currently a dearth of systematic comparative research on the efficacy of different intervention measures. In this paper, the chronic restraint stress model was used to induce anxiety behavior in mice to simulate the state of stressed people. The effects of five intervention methods, namely paroxetine, TSG, exercise, moderate drinking and stress preadaptation, on relieving stress and regulating stress hormones were comprehensively analyzed. The results showed that when the mice were in normal physiological state, none of the five intervention methods had any effect on the behavior of the mice. Except for TSG, which could significantly reduce the content of serum stress hormones (P<0.01), the other four intervention methods had no significant effect on stress hormones. When the mice were in a state of anxiety, TSG, paroxetine and stress preadaptation could improve the behavioral state of anxiety in mice, such as the number of entering the center (P<0.05), the central activity time (P<0.05), and the number of standing (P<0.05), and the serum levels of stress hormones were significantly reduced in these groups (P<0.05). Exercise and alcohol improved only some of the measures in anxious mice. The overall improvement effect of each index was TSG > paroxetine > stress preadaptation >exercise > alcohol. In conclusion, some pharmacological and non-pharmacological interventions can improve the anxiety-prone caused by stress to varying degrees, among which TSG has the best effect.

ETHNOPHARMACOLOGICAL RELEVANCE:Biejia Ruangan Tablet (BJRG) is a patented traditional Chinese medicine formula with demonstrated efficacy against liver fibrosis. However, its underlying molecular mechanisms remain unclear. AIM:This study aimed to investigate the anti-fibrotic effects of BJRG and elucidate its molecular mechanisms. METHODS:Liver fibrosis was induced in murine models using carbon tetrachloride (CCl4) or bile duct ligation (BDL) to evaluate the efficacy of BJRG. Network pharmacology was employed to predict the mechanisms of BJRG. The expression of PDGF-AA was assessed in both murine and human samples. The interaction between BJRG and PDGF-AA was investigated in vitro using primary mouse bone marrow-derived macrophages (BMDMs) and LX-2 cells. Ultra-performance liquid chromatography/mass spectrometry (UPLC/MS) was used to identify liver-entry active components of BJRG. Molecular docking was performed to predict their binding affinity with PDGF-AA. RESULTS:BJRG significantly alleviated liver fibrosis in both CCl4-and BDL-induced murine models. Network pharmacology suggested that BJRG targeted the PDGF/PI3K/AKT signaling pathway. Then we revealed that BJRG specifically downregulated PDGF-AA expression and PI3K/AKT signaling in M2-like macrophages, thereby attenuating hepatic stellate cell (HSC) activation. Clinical data demonstrated that PDGF-AA was associated with liver cirrhosis, and BJRG suppressed PDGF-AA levels in patients with lower Ishak inflammation scores. Molecular docking indicated that the liver-entry components of BJRG exhibited strong binding affinity with PDGF-AA. CONCLUSION:BJRG ameliorated liver fibrosis by disrupting the crosstalk between M2-like macrophages and HSCs through downregulation of the PDGF-AA/PI3K/AKT pathway.

Alcoholic liver disease (ALD) is a leading cause of premature death globally yet remains under-controlled. In this study, we investigated the protective effects of tetramethylpyrazine (TMP), an aromatic compound found in fermented alcohol, against ALD in a National Institute on Alcohol Abuse and Alcoholism (NIAAA) mice model. Our results demonstrated that TMP significantly reduced alcohol-induced liver injury, steatosis, oxidative stress, and mitochondrial damage, while restoring NAD+ levels and the NAD+/NADH ratio, increasing ATP production, regulating energy metabolism disorders, and restoring metabolic balance (P < 0.05). Liver transcriptomic analysis identified 906 ALD-associated genes enriched in energy and lipid metabolism pathways, with a molecular signature of NAD-dependent oxidoreductase activity. Protein interaction analysis predicted Nicotinamide Phosphoribosyltransferase (NAMPT) as a key rate-limiting enzyme in NAD metabolism. Cellular Thermal Shift Assay (CETSA) experiments and molecular docking studies further confirmed that TMP can restore the level of NAD+ by stabilizing the NAMPT protein. TMP is present in various foods, including Semen Sojae Preparatum, a TMP-rich fermented food commonly used in Traditional Chinese Medicine for ALD treatment. This food exhibited significant protective effects against ALD. In conclusion, TMP, an aromatic compound in fermented alcohol, could protect the liver from alcohol-induced damage. Enhancing TMP content in fermented alcohol holds significant promise for mitigating the adverse effects of alcohol consumption on the liver.

ETHNOPHARMACOLOGICAL RELEVANCE:Dysregulation of the upper respiratory tract microbiota, host metabolite profiles, and immune lymphocyte subpopulations is critical in the pathogenesis of respiratory tract infectious diseases (RTIs). Sang Zhu Yang Zheng herbal tea (SZYZ) has emerged as a promising intervention for RTIs, yet its underlying mechanism remains unclear. AIM OF THE STUDY:This study aims to investigate the immunomodulatory mechanisms of SZYZ underlying its role in the prevention and treatment of RTIs. MATERIALS AND METHODS:Human studies were conducted to investigate SZYZ's immunomodulatory effects in preventing RTIs in healthy individuals. In healthy human subjects, SZYZ administration was evaluated for its impact on immune cell counts, airway microbiota composition, and metabolic profiles using advanced sequencing and metabonomic techniques. Murine models were utilized to explore SZYZ's preventive and therapeutic effects, as well as its underlying mechanisms against RTIs, with a focus on influenza A virus (IAV) infection. Murine models were employed with H1N1 PR8 virus infection, and study the therapeutic efficacy and mechanisms of SZYZ alone or in combination with oseltamivir. RESULTS:In healthy humans, SZYZ profoundly increased the total T cells, B cells, CD4+ T cells, and CD8+ T cells. 16S rRNA sequencing showed decreased airway microbes' diversity, enrichment of the genera Bacteroides and Bifidobacterium, and lowered phyla Actinobacteria, Synergistetes, and several opportunistic pathogens, including Actinomyces, Fretibacterium, Mobiluncus, and Cloacibacillus, and inhibited microbe functions of human disease and bacterial infectious pathways, after administration. Serum and fecal metabonomic identified changes in 138 and 197 metabolites, respectively. Airway Actinobacteria were negatively corelated to total T cells and CD8+T cells count. Serum methyl hexadecanoic acid levels were negatively correlated with T cell counts and positively with Actinobacteria abundance. In IAV-infected mice, SZYZ, administered either as a prophylactic intervention or in combination with oseltamivir, significantly reduced mortality and pathological damage, which were related to immunomodulatory mechanisms of oxidative stress, inflammatory cytokines, and lymphocyte ratio. CONCLUSION:We comprehensively elucidate the immunomodulatory effects of SZYZ's on microbiota, metabolism, and lymphocyte subpopulations, which provides a new therapeutic approach for the prevention and treatment of RTIs.

BACKGROUND:Intrahepatic cholangiocarcinoma (ICC) ranks second among primary liver cancers in terms of prevalence, characterized by poor prognosis and scarce therapeutic interventions. Fufang-Biejia-Ruangan tablet (BJRG), a traditional Chinese herbal remedy, has been widely used for liver diseases. However, its therapeutic efficacy and underlying mechanisms in ICC remain poorly understood. AIM OF THE STUDY:This study aims to systematically investigate the anti-ICC effects of BJRG, focusing on tumor progression and microenvironment modulation, through experimental and transcriptomic analyses. METHODS:The chemical composition of BJRG was analyzed employing ultra-performance liquid chromatography-mass spectrometry (UPLC-MS). In vitro assays were performed with QBC939 and LX-2 cell lines. Two primary ICC models (AKT/YAP and sgP53/KRAS) were established via hydrodynamic tail-vein injection of corresponding plasmids. A co-culture system for subcutaneous tumor formation was developed using cancer-associated fibroblasts (CAFs) derived from the AKT/YAP model and primary tumor cells derived from the sgP53/KRAS model. RESULTS:UPLC-MS analysis identified 1091 chemical components, primarily terpenoids, sugars, glycosides, and phenylpropanoids. The therapeutic efficacy of BJRG was evaluated for the treatment of ICC. BJRG treatment slowed down the growth of both human ICC cell lines and AKT/YAP ICC mouse model. Mechanistically, BJRG inhibited HIPPO-PI3K/AKT signaling pathway in ICC tumor cells. Importantly, BJRG significantly inhibited the growth of CAFs via HIPPO-PI3K/AKT cascades. Of note, co-culture CAFs with ICC cell lines substantially sensitized the efficacy of BJRG in sgP53/KRAS syngeneic tumor model. Furthermore, BJRG therapy not only affected CAFs but also induced alterations in vascular structures and hypoxic conditions within lesions in the AKT/YAP model. This intervention promoted the infiltration of T lymphocytes and macrophages into the tumor microenvironment, which may further augment the anti-proliferative effects of BJRG by enhancing the immune response within ICC tumor tissues. CONCLUSION:Our research demonstrates BJRG's anti-ICC efficacy via diverse pathways, including the suppression of tumor cell proliferation, regulation of CAFs activity, and promotion of immune cell infiltration. These findings underscore BJRG as a promising therapeutic candidate for ICC, offering novel mechanistic insights and highlighting its potential for clinical translation.