Background Traditional Chinese Medicine (TCM) presents a unique therapeutic paradigm characterized by multi-compound, multi-target interventions, yet this complexity impedes mechanistic understanding and standardization. While artificial intelligence (AI) has been applied to isolated aspects of TCM research, a critical gap remains in integrating these applications across the inherent hierarchical structure of TCM—from the pharmacological effects of single compounds (SC) to the synergistic mechanisms of complex Chinese medicinal materials (CMM) and Chinese medicine formulae (CMF). Aim of review This review aims to introduce a novel, AI-driven framework that unifies the multi-scale target analysis continuum of TCM through a systematic, cross-scale data flow, positioning AI as the central catalyst for a holistic understanding across SC, CMM, and CMF levels. Key scientific concepts of review The proposed framework demonstrates how molecular targets predicted at the SC level serve as foundational inputs to decipher multi-SC synergistic networks within CMM. These modular networks are subsequently integrated to unravel the complex multi-target synergistic mechanisms of CMF, thereby paving the way for intelligent CMF recommendation (CMFR) and precise quantitative dosage prediction. Furthermore, the review critically addresses fundamental challenges such as the “semantic gap” between abstract TCM theories and molecular data, strongly advocating for a “computation-experiment” closed loop to validate in silico predictions. Finally, we propose transformative future directions, including the development of TCM-specific large language models (LLMs), to decode TCM’s pharmacological logic and chart a definitive path towards its scientific validation and global integration.

ETHNOPHARMACOLOGICAL RELEVANCE:Tremella fuciformis is an edible fungus with a cultivation and consumption history of over a thousand years in China. In traditional Chinese medicine, it has the effects of moistening the lungs, soothing the intestines, nourishing the skin, and prolonging life, and is often used as a tonic food or added to soups. Its core bioactive component is Tremella fuciformis polysaccharides, which are widely concerned for their potential in health maintenance and disease adjuvant treatment. AIM OF THE REVIEW:This review systematically summarizes the latest research progress in the extraction, structural characteristics, pharmacological activities, structure-activity relationships, quality control, and applications of TFPS. It deeply analyzes the controversies and limitations of existing studies, clarifies the correlation between structure and function, and provides a theoretical basis for the standardized production and high-value application of TFPS. MATERIALS AND METHODS:Databases like ScienceDirect, PubMed, CNKI, and field journals were searched for extraction, structural characterization, pharmacology, and applications of Tremella fuciformis polysaccharides, including recent English/Chinese literature. RESULTS:Tremella fuciformis polysaccharides have diverse extraction methods (hot water, ultrasonic/microwave-assisted, etc.), with composite tech boosting yield over 50%. Structurally, they have α-1,3-D-mannan backbone, varied monosaccharides, and conformations (random coil, triple-helical). Pharmacologically, they modulate immunity, reduce inflammation/tumors, resist oxidation/aging, protect nerves, regulate metabolism, and maintain intestinal homeostasis. Quality control uses HPLC/GC-MS fingerprints and HPSEC-MALLS for quantification. Applications span pharma (immunoadjuvants), food (additives), cosmetics (moisturizers), and new materials (drug carriers). CONCLUSION:Tremella fuciformis polysaccharides have great prospects but lack clarity on some mechanisms, higher-order structure research, and standardized quality criteria. With deeper study, they may bring new breakthroughs in industries of health maintenance, disease treatment, and cosmetics and skincare.

Ionization is usually difficult for underivatized dextran. After adding chemical additives, large polysaccharide ions can be investigated and their fragments are highly related with the structures.

Gymnodimine A (GYM-A) is a lipophilic cyclic imine toxin produced by marine dinoflagellates that accumulates in shellfish and poses potential risks to human health. However, regulatory limits have not been established worldwide, largely due to insufficient understanding of its metabolic pathways and toxicokinetic behavior. This study characterized the in vitro metabolism of GYM-A using human liver microsomes (HLM) and human liver S9 fractions (HS9) combined with liquid chromatography-high-resolution tandem mass spectrometry (LC-HRMS/MS). An integrated identification strategy incorporating segmented data-dependent acquisition (DDA), targeted/untargeted screening, MS/MS fingerprint comparison, density functional theory (DFT) calculation, and μElution solid phase extraction (SPE) enabled confident structural elucidation of metabolites derived from low-concentration substrates in complex matrices. Four phase I hydroxylated metabolites were identified: three mono-hydroxylated derivatives (M1-M3) and one di-hydroxylated derivative (M4), with 21-hydroxy-GYM-A (M1) as the highest relative abundance. No phase II glucuronide conjugates were detected under the conditions tested. Metabolic stability studies revealed high intrinsic clearance following first-order kinetics, with elimination primarily mediated by CYP450-dependent phase I oxidation. Enzyme phenotyping identified CYP3A5 as the principal isoform responsible for GYM-A hydroxylation. Detection of metabolite M2 in urine from dosed rats supported the in vitro findings. This work provides the first systematic metabolic profile of GYM-A and advances knowledge of its kinetic behavior, while the analytical workflow may serve as a useful tool for risk assessment and metabolism studies of related marine toxins.

Pinellia ternata, a widely used herbal medicine in the Western Pacific region, is frequently adulterated with morphologically similar species, posing challenges to quality control and safety. As part of the Forum for the Harmonization of Herbal Medicines (FHH) ATLAS project, this multi-national collaborative study aimed to evaluate and validate the applicability of practical and reliable methods to distinguish P. ternata from its adulterants, thereby ensuring standardized quality control across member countries. Multiple analytical approaches applicable to member countries were evaluated. Morphological analysis and high-performance thin-layer chromatography proved ineffective in distinguishing adulterants, whereas high-performance liquid chromatography (HPLC) and polymerase chain reaction (PCR) successfully identified them. HPLC, using triglochinic acid as a marker compound, differentiated P. ternata from P. pedatisecta but could not detect Typhonium flagelliforme. In contrast, PCR demonstrated higher specificity by amplifying distinct DNA fragments for P. ternata, P. pedatisecta, and P. tripartita, as well as a specific fragment for T. flagelliforme. Cross-validation among countries confirmed the reproducibility of these findings. By integrating chemical and molecular methods, this collaborative effort provides an effective framework for authenticating P. ternata, contributing to safer and more reliable herbal medicines in the Western Pacific and beyond.

BACKGROUND:Pogostemon cablin (P. cablin) is a valuable medicinal plant used in traditional medicine and the fragrance industry, but QC is challenging due to inconsistent stem-to-leaf ratios and frequent essential oil adulteration. OBJECTIVE:This study compares volatile components in different parts (aerial parts, stems, and leaves) and essential oil of P. cablin to support QC (not less than 20% leaf) and its rational use. METHODS:Volatile components in 21 batches of aerial parts, stems, and leaves, and 13 batches of essential oils, were analyzed using GC-MS. Multivariate curve resolution-alternating least-squares (MCR-ALS) was used for resolving co-eluted peaks, and chemical fingerprinting with chemometric techniques including hierarchical cluster analysis (HCA), principal component analysis (PCA), partial least-squares discriminant analysis (PLS-DA), and orthogonal partial least-squares discrimination analysis (OPLS-DA) were applied. RESULTS:Volatile profiling identified 56, 47, 28, and 45 components in the aerial parts, leaves, stems, and essential oil of P. cablin, respectively. MCR-ALS resolved 10 major volatile compounds to create chemical fingerprints for each analytical sample type. Quantitative analysis showed higher patchouli alcohol in leaves (12.47 mg/g) compared to stems (2.05 mg/g), while stems had more pogostone (2.71 mg/g versus 1.40 mg/g in leaves). Aerial parts and essential oil showed significant compositional differences. Based on the results of qualitative and quantitative analysis, chemometric methods, including HCA, PCA, PLS-DA, and OPLS-DA, clearly differentiated the four types of P. cablin analytical samples. CONCLUSION:Significant differences in volatile components across P. cablin parts and its essential oil support QC (not less than 20% leaf) and rational use. HIGHLIGHTS:This study is the first to use MCR-ALS and other chemometrics for qualitative and quantitative analysis of P. cablin parts and essential oils, aiding QC.

Glycans, also known as polysaccharides/saccharides/carbohydrates, commonly found in biological cell membrane/wall, and play important biological functions. Biomolecular recognition on the cell surface is governed by molecular interactions, making direct measurement essential for understanding biological processes. Atomic force microscopy (AFM) is the only microscopy technique that provides images in aqueous media with nanometer resolution, which greatly facilitates the specific recognition of macromolecules on the cell surface. Moreover, single-molecule force spectroscopy (SMFS) based on AFM has been commonly employed to investigate interactions between proteins and other molecules. However, there are fewer reviews on glycan molecular recognition. Therefore, this review focuses on the molecular recognition between glycans and cell membranes using AFM, covering glycans-receptor interactions and the important roles of glycans on the cell membrane. Finally, strategies for enhancing the study of molecular recognition between glycans and cell membranes through AFM are prospected and with the aims to address biomedical challenges and discover novel biomolecules.

The therapeutic efficacy of traditional Chinese medicine has been widely acknowledged due to its extensive history of clinical effectiveness. However, the precise active components underlying each prescription remain incompletely understood. Polysaccharides, as a major constituent of water decoctions-the most common preparation method for Chinese medicinals-may provide a crucial avenue for deepening our understanding of the efficacy principles of Chinese medicine and establishing a framework for its modern development. The structural complexity and diversity of Chinese herbal polysaccharides present significant challenges in their separation and analysis compared to small molecules. This paper aims to explore the potential of Chinese herbal polysaccharides efficiently by briefly summarizing recent advancements in polysaccharide chemical research, focusing on methods of acquisition, structure elucidation, and quality control.

Immune-related skin diseases are a group of conditions directly driven by the abnormal activation or dysregulation of the immune system. Current clinical treatments for these diseases frequently exhibit limited target specificity, suboptimal sustained efficacy, and potential adverse effects. As an emerging immunomodulatory "organ", gut microbiota has been confirmed by clinical data as a feasible method for the therapy of immune-related skin disorders. Therefore, it is imperative to explore the specific mechanisms by which gut microbiota affects immune-related skin diseases. As potent immunomodulatory agents, accumulating evidence has confirmed that polysaccharides are closely related to gut microbiota. At the same time, researches on polysaccharides targeting gut microbiota for the management of immune-related skin disorders are also increasing annually. This work endeavored to provide a comprehensive summary and exploration of the mechanisms by which gut microbiota affected immune-related skin diseases, and the effect of polysaccharides on gut microbiota. To elucidate the mechanisms by which polysaccharides modulated the gut-skin axis in treating immune-related skin diseases, thereby offering novel therapeutic insights and advancing clinical research and applications.

The interpretation of the chemical quality differences during Dendrobium huoshanense (D. huoshanense) traditional processing into 'Fengdou' is critical for scientifically evaluating the medicinal value, optimizing the processing methods, and ensuring the stability and predictability of its efficacy. In this study, the quality differences of polysaccharides during the processing of D. huoshanense were conducted utilizing high performance size exclusion chromatography coupled with multi-angle laser light scattering and refractive index detector (HPSEC-MALLS-RID), nuclear magnetic resonance (NMR) and saccharide mapping based on polysaccharides analysis by using carbohydrate gel electrophoresis (PACE) and high-performance anion exchange chromatography-pulsed amperometric detection (HPAEC-PAD). The results showed that the concentration of non-starch D. huoshanense polysaccharides (NDHP) rose and exhibited greater stability during the processing. The molecular weight (Mw), molecular weight distribution, cumulative molar mass, radius of gyration (Rz), and monosaccharide composition molar ratio of NDHP did not have significant changes during the processing, and these parameters would be affected by the presence of starch. The NMR investigation indicated that the primary glycosidic linkages of β-1,4-Manp and β-1,4-Glcp, together with the acetyl group substitutions at O-2 and O-3 of 1,4-linked Manp, were unaltered. Additionally, the PACE and HPAEC-PAD profiles of acidic and enzymatic hydrolysates of NDHP were similar. These findings contribute to a deeper understanding of the changes in bioactive polysaccharides during the processing of D. huoshanense into 'Fengdou', providing theoretical support for improving its processing techniques.

Introduction As a key technique in saccharide mapping, polysaccharides analysis using carbohydrate gel electrophoresis (PACE) analysis platform has always been self-assembled and built in various laboratories independently, but there is no unified commercial analysis instrument platform so far. Objective This research aimed to establish an integrated commercially viable PACE analysis platform to standardize the application of PACE in the field of saccharides analysis and the quality control of polysaccharides. Methods We established an online detection platform of saccharide-mapping apparatus (SMA) and took advantage of it to characterize the oligosaccharides for quality control of polysaccharides in herbal medicine. The polysaccharides from Gastrodia elata Bl. were finally selected to verify the potential of SMA including precision, repeatability, stability, and sensitivity. Results The online detection platform of SMA was more integrated and commercially viable, which can investigate oligosaccharides from polysaccharides after hydrolysis more quickly and accurately for quality control. Consequently, it could enable real-time monitoring of oligosaccharide migration, facilitate the determination of the optimal endpoint of polyacrylamide gel electrophoresis (PAGE), and improve the sensitivity of PACE. In addition, the refrigeration device configured in SMA can solve the problem of temperature fluctuation caused by joule heat during gel electrophoresis, which greatly enhances the separation resolution. Finally, the proposed platform was applied to real samples of polysaccharides from Gastrodia elata Bl., and the results successfully distinguished the polysaccharides from Gastrodia elata Bl. with different cultivars. Conclusion The developed online detection platform of SMA offers a rapid and convenient approach for quality control of polysaccharides in Chinese medicines and can be applied for future advancements toward commercialization.

Solid tumors frequently develop drug resistance by therapy‐induced collagen‐dense barriers that impede drug penetration and immune cell infiltration. Here, a traditional Chinese medicine (TCM)‐inspired carbon dots (NRG‐CDs) system is reported that reprograms cancer cell death from apoptosis to necroptosis, effectively dismantling the collagen barrier typically caused by drug‐induced collagen up‐regulation while activating immunogenic cell death. By integrating naringenin (NRG) with ε‐poly(L‐lysine) through dimethyl sulfoxide‐assisted solvothermal processing, the resulting NRG‐CDs achieve effective cancer cell killing capacity with minimal cytotoxicity toward normal cells. Compared to NRG, the NRG‐CDs shift cell death modality from apoptosis to necroptosis, reducing collagen secretion while releasing damage‐associated molecular molecules for antigen‐presenting cell maturation. Further incorporating oxidized dextran creates an injectable pH‐responsive hydrogel (CO‐Gel) that can locoregionally release NRG‐CDs in the acidic tumor microenvironment. In the aggressive murine 4T1 model, peritumoral CO‐Gel administration achieves dual therapeutic outcomes: primary tumor suppression and pulmonary metastasis inhibition. The strategy is further validated in orthotopic hepatic tumors in a rat model, demonstrating broad applicability. This work provides a paradigm shift from conventional cytotoxic drug design to death modality‐engineered nanotherapeutics.

The polysaccharides from L. barbarum L. fruits (LBPs) are a kind of complex polysaccharides, which mainly contribute to the bioactivity of L. barbarum L. fruits and riches in acidic polysaccharides fractions. And the pharmacological research showed that the immunomodulatory effect of LBPs was closely related to alpha-1,4-Dgalactosiduronic linkages. Therefore, as one of the main active polysaccharide fractions in LBPs, acidic polysaccharides fraction should also be used for the quality control of L. barbarum L. fruits. In this study, a novel quantitative evaluation strategy of polysaccharides from L. barbarum L. fruits based on simultaneous quantification of total polysaccharides and polysaccharides with active structure characteristics was proposed to improve their performance in the medicinal and functional food area more comprehensively and effectively, and beneficial to improve the quality control of L. barbarum L. fruits. The content of total polysaccharides in L. barbarum L. fruits was determined by HPSEC-RID method combined with universal dn/dc (0.15 mL.g(-1)). Additionally, HPAEC-PAD method combined with pectinase hydrolysis and m-hydroxybiphenyl colorimetric method were developed for the determination of acidic polysaccharide components related to active structural characteristics in L. barbarum fruits. The results revealed that the trends of total polysaccharides and acidic polysaccharides in L. barbarum L. fruits did not entirely coincide, further emphasizing the necessity of simultaneously quantifying both fractions for quality control. Furthermore, the m-hydroxybiphenyl method can be used as an alternative method to determine the content of acidic polysaccharides by HPAEC-PAD combined with pectinase hydrolysis.

Background/Objectives: Allergic diseases (e.g., asthma, chronic urticaria) are increasing globally, but current anti-allergic drugs exhibit limitations in efficacy and safety. Traditional Chinese Medicine (TCM) emphasizes constitutional regulation for allergic diseases management. The allergic constitution prescription (ACP), a TCM formulation, lacks clear mechanistic insights. Methods: This study employs a novel network pharmacology approach integrating ultra-high performance liquid chromatography quadrupole time-of-flight tandem mass spectrometry (UHPLC-Q-TOF-MS/MS) to identify ACP’s chemical components and compare its mechanisms with anti-allergic drugs. Chemical components of ACP were analyzed via UHPLC-Q-TOF-MS/MS, and allergic disease-related targets were collected from public databases. Anti-allergic drug targets were intersected with ACP-disease targets to identify unique and common pathways. Molecular docking and dynamics simulations assessed binding affinity between key compounds and core targets. Results: We identified 126 compounds in ACP. Compared to anti-allergic drugs, ACP targeted 10 unique and five common key pathways (e.g., MAPK signaling), 10 unique and nine common core targets (e.g., Tumor Necrosis Factor (TNF), IL-6), and 14 unique and 15 common key compounds. Simulations confirmed high binding affinity of ACP compounds to core targets. Conclusions: These findings highlight ACP’s potential multi-target mechanisms for allergic diseases treatment, identifying unique and shared pathways, targets, and compounds compared to anti-allergic drugs, offering new insights for further mechanistic studies. However, it is crucial to note that these mechanistic predictions and compound-target interactions are primarily derived from computational analyses, and experimental validation (e.g., in vitro or in vivo assays) is essential to confirm these computational findings.

Most organelle-targeting probes require the removal of excess dye to enhance the signal-to-noise ratio before microscopic imaging experiments. However, this washing step may cause cellular damage and interfere with the continuous observation of cellular activities. Here, we report a series of wash-free probes based on small molecule phenothiazine/phenoxazine. Simple modification of phenothiazine/phenoxazine by nitro group significantly enhances the polarity-sensitive characteristic, which are well-suited for wash-free cellular imaging. These probes feature low molecular weight, excellent photostability, and large Stokes shifts (up to 191 nm). By conjugation with targeting groups, a series of probes are developed for specific imaging in different organelles such as lysosomes, mitochondria, the endoplasmic reticulum, lipid droplets, and the plasma membrane, without any washing step. Moreover, by simply replacing S atom in the central core with O atom, the emission of probes shifts from red to green-yellow. Using these two probes, high-contrast, dual-color wash-free imaging can be achieved. Among them, the PXZ-Lipid probe was successfully applied for real-time monitoring of dynamic changes in lipid droplets within living cells. This work establishes a general strategy for designing small-molecule, wash-free fluorescent probes based on phenothiazine/phenoxazine scaffolds, enabling real-time, multi-organelle imaging with minimal cellular disturbance.

Active ingredients discovery has always been a hot topic for the research of Chinese medicines (CMs). The component-effect relationship is one of the widely used strategies for finding herbal active ingredients. This article systematically reviewed component-effect relationship analysis in the application of active components discovery in CMs, including sampling and sample preparation, chemical fingerprint establishment, pharmacological effect evaluation, and component-effect relationship analysis. The challenges to this method, such as the effects of dose–response relationship, data normalization, and the relationship analysis methods, were also discussed, which is helpful to well understand the method and improve the research quality.

Skull defect repair is a complex and critical medical challenge, and there is an urgent need to develop multifunctional tissue engineering scaffolds for skull regeneration. The success of bone tissue engineering depends on the construction of scaffolds that can regulate the immune microenvironment of bone regeneration and mimic the liquid crystal and viscoelastic properties of natural bone extracellular matrix. Hence, a smart hydrogel (PEGDA5/AM15/CLC-BMP-4@MBG) with good biocompatibility and the ability to modulate the wound immune microenvironment has been developed for the repair of skull defects. The hydrogel consists of chitin liquid crystal hydrogel (PEGDA5/AM15/CLC) and mesoporous bioactive glasses (MBGs) loaded with bone morphogenetic protein-4 (BMP-4). The liquid crystal hydrogel not only offers the necessary biological support and mechanical properties but also maintains the stability of the liquid crystal state, facilitating adhesion and regeneration of surrounding bone tissue. In addition, BMP-4@MBG intelligently regulates the release rate of BMP-4 in response to changes in wound microenvironment, thus effectively promoting the transformation of macrophages from M1 to M2 macrophages. At the same time, Ca2+ and Si4+ released by MBG degradation and BMP-4 synergically promote bone repair process. The PEGDA5/AM15/CLC-BMP-4@MBG hydrogel shows excellent immunomodulatory and osteogenic properties of bone microenvironment and is a promising scaffold material for bone tissue engineering.

The P. heterophylla and its adulterants were identified by HPLC-CAD fingerprint of sucrose and oligosaccharides in P. heterophylla. The improved quantitative analysis of multi-components with a single marker (iQAMS) was further established for simultaneous determinations of sucrose and oligosaccharides in P. heterophylla. The HPLC-CAD fingerprint and similarity coefficients between P. heterophylla and its adulterants showed significant differences. The relative errors (REs) between iQAMS method and external standard method (ESM) were below 3.00%, but significant difference was shown between iQAMS (different marker for whole program with gradient elution) and QAMS (one marker for whole program with gradient elution), indicating that QAMS method should be improved, especially for gradient elution which influence the response of analytes. The accuracy, precision, reproducibility, and stability of this method were validated which exhibited satisfactory results, indicating that iQAMS method could be used for quantitative analysis of sucrose and oligosaccharides in P. heterophylla instead of ESM. The iQAMS combined with HPLC-CAD fingerprint could be used to determine the content of each oligosaccharide, and it can be used for quality control of P. heterophylla.