Chinese Traditional and Herbal Drugs

Preparation and in vitro anti-tumor evaluation of targeted nanoparticles co-loaded with homohardenine/triptolide

Chinese Traditional and Herbal Drugs. 2026, 57(18): 7281-7294.

Objective To prepare prostate specific membrane antigen (PSMA)-targeting peptide-modified lipid-polymer nanoparticles co-loaded with homoharringtonine and triptolide (HHT/TPL-PSMA-Lips), optimize its preparation process, and evaluate its in vitro anti-prostate cancer activity. Methods The solubility and oil-water partition coefficient (lgP) of HHT and TPL were determined, and PSMA-targeting peptide-modified PEGylated phospholipids were synthesized. HHT/TPL-PSMA-Lips was prepared by the emulsification-solvent evaporation method, and the formulation was optimized using central composite design-response surface methodology (CCD-RSM). Indicators including particle size, ζ potential, and encapsulation efficiency were investigated. The in vitro release behavior of the formulation in different media was evaluated. MTT assay and flow cytometry were used to assess the in vitro inhibitory effect and apoptosis-inducing activity of the nanoparticles on prostate cancer cells, while cell uptake assay was performed to verify the tumor cell targeting efficiency of the nanoparticles. Results HHT and TPL showed low solubility in various media and strong lipophilicity. The molecular weight of the synthesized PSMA-targeting peptide-modified PEGylated phospholipids was 4 643.8. HHT/TPL-PSMA-Lips prepared with the optimal formulation exhibited uniform particle size, with an average particle size of (209.12 ± 3.24) nm, a ζ potential of (-34.4 ± 2.5) mV, and a comprehensive encapsulation efficiency of (81.20 ± 0.40)% [HHT (80.73 ± 0.86)%, TPL (82.00 ± 0.40)%]. In vitro release experiments demonstrated that HHT/TPL-PSMA-Lips could effectively improve the solubility of HHT and TPL, and exhibite a certain pH-responsive release property. Cell experiments showed that the HHT/TPL combined administration group displayed stronger inhibitory effect on prostate cancer cells and better apoptosis-inducing activity than the single drug groups. HHT/TPL-Lips enhanced the anti-tumor activity, and the targeted modified nanoparticles (HHT/TPL-PSMA-Lips) showed the strongest inhibitory effect on prostate cancer cells. Cell uptake assay indicated that modification with PSMA-targeting peptide could effectively enhance the cellular uptake of the nanoformulation in tumor cells. Conclusion The prepared HHT/TPL-PSMA-Lips not only solves the solubility problem of the drugs but also significantly enhances tumor targeting and anti-tumor efficacy, providing a new approach for the development of new traditional Chinese medicine formulations for the treatment of prostate cancer.

Analysis of key components in anti-browning process of medicine-food homology formula Wuzhiyin and its gastroprotective function

Chinese Traditional and Herbal Drugs. 2026, 57(18): 7295-7308.

Objective Given that the liquid formulation of the medicine-food homology formula Wuzhiyin is susceptible to non-enzymatic browning during storage, this study systematically investigates the underlying browning mechanism and evaluates the gastroprotective effect of Wuzhiyin prepared using an anti-browning processing technology against alcoholic gastric injury. Methods The differential metabolites and browning intermediates before and after browning of Wuzhiyin were analyzed by non-targeted ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) and Spe-Pak microcolumn separation technology. The degree of browning was characterized by color changes, browning degree, and alterations in nutritional components. An anhydrous ethanol-induced acute gastric injury mouse model was established to evaluate the gastroprotective function of anti-browning prepared Wuzhiyin. Results Total sugars, total polyphenols, and total amino acids were significantly decreased by browning (P < 0.05). Nineteen key differential compounds were screened, among which fructose, sucrose, valine, isoleucine, tyrosine, and p-coumaric acid were identified as major driving factors. 2-Hydroxy-3-methyl-2-cyclopenten-1-one was identified as the core browning intermediate, and the browning mechanism was elucidated as a synergistic process involving the Maillard reaction, acid-promoted sugar degradation, and oxidative cross-linking of phenolic compounds. High-dose anti-browning prepared Wuzhiyin (43.72 g/kg) significantly reduced the gastric ulcer index (inhibition rate 72.42%) and alleviated pathological damage to the gastric mucosa. The underlying mechanisms include inhibiting nuclear factor-κB (NF-κB) pathway, reducing inflammatory cytokines [interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), IL-6], regulating oxidative stress [elevation of superoxide dismutase (SOD)/malondialdehyde ratio], and protecting mucosal barrier [modulation of NO, prostaglandin E₂ (PGE₂), trefoil factor 2 (TFF2)]. Conclusion Fructose, sucrose, valine, isoleucine, tyrosine, and p-coumaric acid are the key drivers of browning in Wuzhiyin; the Maillard reaction, acid-catalyzed sugar degradation, and phenolic oxidative cross-linking constitute the major browning pathways; and Wuzhiyin prepared via the anti-browning processing technology exhibits a significant gastroprotective effect against alcoholic gastric injury.

Relationship between two phase states and component changes during decoction process of Baihe Dihuang Decoction

Chinese Traditional and Herbal Drugs. 2026, 57(18): 7309-7320.

Objective To investigate the phase changes and component distribution patterns during the decoction process by performing phase separation and characterization of Baihe Dihuang Decoction (BDD, 百合地黄汤) at different decoction times, and provide a reference for elucidating the material basis of synergistic effects of components in traditional Chinese medicine formulas. Methods BDD was decocted for 20, 40, 60, 80, 100, and 120 min to obtain the decoctions. The sediment phase and nanophase solutions were separated using the “high-speed centrifugation-dialysis” method. The two phases were characterized by scanning electron microscope, laser particle size analyzer, ultraviolet-visible spectroscopy and Fourier transform infrared spectroscopy. The fingerprint spectra of BDD and its different phases were constructed using HPLC method, and chemometric methods were applied to screen key differential components among different phases, followed by content determination of the main differential components. Results The nanophase of BDD exhibited the smallest particle size, while the original decoction showed the largest particle size. The particle size of the nanophase initially increased and then decreased with prolonged decoction time, eventually stabilizing, and the stability of nanophase gradually enhanced. The contents of polysaccharides and proteins increased progressively with prolonged decoction time, and both the nano-sized fraction and the precipitated fraction were found to be rich in polysaccharides. Spectral analysis revealed that the chemical structures of components in the original decoction and each phase at different decoction times were generally similar, but differences in functional group contents were observed. Fingerprint analysis combined with hierarchical cluster analysis (HCA) and partial least squares-discriminant analysis (PLS-DA) showed that the types of chemical components in different phases of BDD were similar, but their contents differed significantly. The effect of decoction time on components was smaller than that of phase state. A total of 12 key differential components among different phases were screened. The average contents of five indicator components catalpol, regaloside C, regaloside A, regaloside B, isoverbascoside in the nanophase were higher than those in the sediment phase and the original decoction. Conclusion During the decoction process, BDD interacts with each other, resulting in significant differences in the content of active ingredients among various phases. The nanophase achieves enrichment of active components, suggesting that it may be the main effective phase of BDD.

Research on quality control of soft material fine powder of Guizhi Fuling Capsules based on cascade fault-tolerant model

ZHONG Xiaohui, TONG Feng, HOU Huarui, ZHANG Yongchao, WANG Tuanjie, ZHANG Chenfeng, XU Fangfang, LIU Hengxu

Chinese Traditional and Herbal Drugs. 2026, 57(18): 7321-7333.

Objective To address the potential uneven distribution of active ingredients indicated by uneven color within batches of soft material fine powder during the production process of Guizhi Fuling Capsules (GFC, 桂枝茯苓胶囊), a “cascade fault-tolerant model” was constructed based on color difference characteristics to explore a new low-cost approach for intermediate quality control. Methods A total of 177 samples of soft material fine powder from five batches were collected. The paeoniflorin content, determined by HPLC, was divided into low, medium, and high grades at a ratio of 2:6:2. Based on a traditional three-class model, a cascade fault-tolerant quality control model was constructed by comparing 126 algorithm combinations, which involved a primary algorithm for early warning and a secondary physical direction for correction. External validation was introduced to assess the generalization ability, and quality control experiments were conducted based on the model results. Finally, near-infrared spectroscopy (NIRS) technology was used for benchmarking against the upper limit of efficacy. Results Correlation analysis established paeoniflorin as the core grading indicator, with its content showing a significant positive correlation with red-green chroma a^* (r = 0.64, P < 0.001). The analysis indicated that the accuracy of the benchmark model was only 70.34% (RSD was 12.70%), which was difficult to meet quality control requirements. However, the cascade fault-tolerant model achieved an overall industrial accuracy of 78.62%, with significantly enhanced robustness (RSD was 6.79%), and its external validation accuracy was 80.56%. Quality control experiments showed that the RSD of paeoniflorin content in mixed samples decreased from 7.74% in random mixing to 2.34%, indicating a significant increase in material uniformity, and a simultaneous reduction in total mixing process time and load. Benchmarking results with NIRS (accuracy of 90.68%, RSD was 6.22%) indicated that the colorimeter had a high cost-performance ratio and could be used for batch screening; NIRS was more accurate and suitable for high-precision grading. Conclusion The cascade fault-tolerant model constructed in this study achieved the synergistic empowerment of algorithm early warning and physical correction, effectively enhancing the discriminative robustness of low-cost optical sensors. This provides a new strategy for quality control of active ingredients in batches of intermediates during the production process of traditional Chinese medicine in the future.

Preparation, pharmacokinetics and protective effect on hepatic ischemia reperfusion injury of galangin-whey protein-nanoparticles

Chinese Traditional and Herbal Drugs. 2026, 57(18): 7334-7346.

Objective To prepare galangin-whey protein-nanoparticles (Gal-WP-NPs), and study its oral pharmacokinetic behavior and protective effect on the hepatic ischemia reperfusion injury in vivo. Methods Single-factor experiments were conducted to investigate the effects of various formulation processes on Gal-WP-NPs. Box-Behnken design-response surface methodology (BBD-RSM) was used to obtain its optimal prescriptions. Entrapment efficiency, drug loading, particle size and ζ potential were determined. Gal-WP-NPs powder was prepared by freeze-drying method. Transmission electron microscopy (TEM) was used to observe the microstructure of Gal-WP-NPs. Galangin crystal in Gal-WP-NPs powder was analyzed by X-ray powder diffraction (XRPD). Solubility of galangin and Gal-WP-NPs powder were compared, and drug release behavior of Gal-WP-NPs in gastrointestinal fluid was determined. SD rats were administered galangin and Gal-WP-NPs via ig at a dose of 150 mg/kg (calculated as galangin), the main pharmacokinetic parameters and relative bioavailability of Gal-WP-NPs were calculated. The model of hepatic ischemia reperfusion injury was established, and then the protective effects of galangin and Gal-WP-NPs were compare on the hepatic ischemia reperfusion injury. Results Optimal formulation processes of Gal-WP-NPs: the volume ratio of the anti-solvent phase to the solvent phase was 15.40:1, the amounts ratio of whey protein to galangin was 15.05:1, and the ultrasonic power was 82.00 W. Envelopment efficiency, drug loading, particle size and ζ potential were (94.07 ± 0.98)%, (5.76 ± 0.09)%, (197.39 ± 4.46) nm and (-23.79 ± 0.75) mV, respectively. Gal-WP-NPs are spherical nanoparticles. Galangin existed as an amorphous state in Gal-WP-NPs powder. Solubility of galangin was significantly increased (P < 0.01), and the release process of Gal-WP-NPs in vitro conformed to the Weibull model. The peak time (t_max) of Gal-WP-NPs was delayed to (2.17 ± 0.64) h, half-life period (t_1/2) was increased to (5.11 ± 0.95) h, and oral relative bioavailability was increased to 6.58 times. Gal-WP-NPs significantly enhanced the protective effect of galangin on the hepatic ischemia reperfusion injury. Conclusion Gal-WP-NPs significantly increased the oral relative bioavailability of galangin and enhanced the protective effect on the hepatic ischemia reperfusion injury.

Dynamic spatiotemporal response mechanisms of multi-component and multi-target effects of Buyang Huanwu Decoction based on BiGRU-integrated temporal attention network

Chinese Traditional and Herbal Drugs. 2026, 57(18): 7347-7358.

Objective To establish an interpretable deep learning model based on a bidirectional gated recurrent unit (BiGRU) and a temporal attention mechanism to elucidate the dynamic spatiotemporal response mechanisms of the multi-component–multi-target interactions underlying the effects of Buyang Huanwu Decoction (BYHWD) against ischemic stroke (IS). Methods A rat model of middle cerebral artery occlusion (MCAO) was established. Plasma and brain tissue samples were collected at multiple time points after administration. Plasma concentrations of 14 active components of BYHWD were quantified by UPLC–MS/MS, and the expression levels of 15 key targets in brain tissue were determined by ELISA to construct a multi-component–multi-target time series dataset. A BiGRU-based temporal attention network was established using a sliding window strategy, and leave-one-batch-out cross-validation (LOBO-CV) combined with bootstrap resampling was employed to evaluate model performance and stability. The model was further interpreted to identify highly contributive time points, potential key components, and responsive target features. Results The model demonstrated good predictive performance in LOBO-CV, with an R² value greater than 0.80 and a mean squared error (MSE) below 0.20 across all test sets. Temporal attention analysis identified 10 h post-administration as a time point receiving high model attention, with an attention peak value of 0.602 6. Gradient attribution analysis identified paeonol, calycosin, and formononetin as the major driving components, while brain derived neurotrophic factor (BDNF), protein kinase G type II (PKG2), and calmodulin-dependent protein kinase II (CAMK2) were identified as the key responsive targets. Bootstrap resampling showed that the 10 h critical time window was identified with a frequency of 90%, indicating good model stability. Conclusion This study established a BiGRU and temporal attention-based approach for dynamic time series analysis of the multi-component–multi-target actions of BYHWD. The proposed method provides new temporal evidence for elucidating the mechanisms underlying the anti-IS effects of BYHWD and offers a reference for investigating the dynamic pharmacological mechanisms of traditional Chinese medicine formulas.

Clinical efficacy and potential mechanisms of kidney-tonifying versus blood-activating traditional Chinese medicines in treatment of distal radius fractures: An integrated study based on Meta-analysis and network pharmacology

Chinese Traditional and Herbal Drugs. 2026, 57(18): 7359-7375.

Objective To systematically evaluate the clinical efficacy of kidney-tonifying and blood-activating traditional Chinese medicines as adjunctive therapies for distal radius fractures using a Meta-analysis, and to preliminarily explore their mechanisms of action through network pharmacology. Methods PubMed, Web of Science, Cochrane Library, Embase, CBM, CNKI, Wanfang, and VIP databases were searched from inception to June 2026 to identify clinical controlled studies comparing traditional Chinese medicine decoctions combined with conventional Western medicine versus conventional Western medicine alone for distal radius fractures. RevMan 5.4 software was used for Meta-analysis, and network pharmacology was employed to screen core active components, predict targets, and conduct pathway enrichment analysis. Results A total of 23 studies involving 3 377 patients were included. The Meta-analysis results showed that both kidney-tonifying and blood-activating traditional Chinese medicines significantly improved the clinical effective rate, shortened fracture healing time, reduced visual analog scale (VAS) pain scores, increased osteocalcin levels, and improved wrist joint function, with no statistically significant difference in overall efficacy between the two categories (all P > 0.05). Blood-activating traditional Chinese medicines showed superior improvements in wrist dorsiflexion range of motion, while kidney-tonifying traditional Chinese medicines exhibited more stable efficacy with lower heterogeneity. Network pharmacology identified core targets such as tumor protein p53 (TP53), proto-oncogene tyrosine-protein kinase Src (SRC), signal transducer and activator of transcription 3 (STAT3), primarily regulating pathways including phosphatidylinositol-3-hydroxykinase (PI3K)-protein kinase B (Akt), mitogen-activated protein kinase (MAPK), nuclear factor-κB (NF-κB). Blood-activating traditional Chinese medicines exerted anti-inflammatory and microcirculation-improving effects mainly through vascular endothelial growth factor (VEGF), tumor necrosis factor (TNF), and Janus kinase (JAK)-STAT pathways, whereas kidney-tonifying traditional Chinese medicines promoted osteogenic differentiation and bone metabolism via the Wnt/β-catenin pathway and cell cycle pathways. Conclusion Kidney-tonifying and blood-activating traditional Chinese medicines are effective and safe as adjunctive therapies for distal radius fractures, both promoting fracture healing through multi-component, multi-target, and multi-pathway mechanisms. Blood-activating traditional Chinese medicines are more suitable for relieving swelling and pain in the early stage, while kidney-tonifying traditional Chinese medicines are better for strengthening bone and promoting healing in the middle and late stages.

Visualization analysis of application of mass spectrometry imaging technology in field of traditional Chinese medicine research based on CiteSpace knowledge graph

Chinese Traditional and Herbal Drugs. 2026, 57(18): 7376-7387.

Objective To analyze the current application status and hotspots of mass spectrometry imaging (MSI) in traditional Chinese medicine (TCM) research, and use knowledge graphs to display the research process in this field, providing reference for in-depth research and application. Methods Relevant Chinese and English literature from China National Knowledge Infrastructure (CNKI), Wanfang Data Knowledge Service Platform (Wanfang), VIP Data Knowledge Service Platform (VIP) and Web of Science (WOS) databases were retrieved. Literature was screened by NoteExpress literature management software. The CiteSpace software was used to draw a knowledge graph of publishing volume, publishing institutions, authors, keywords, etc. Results A total of 124 Chinese literature and 306 English literature were included. The quantities of related publications significantly increased since the 2021. The Chinese Traditional and Herbal Drugs and Chinese Journal of Chinese Materia Medica are the journals with the highest quantities of publications in Chinese. Journal of Ethnopharmacology is the journal with the highest quantities of publications in English. Sun Bo’s team and Cai Zongwei’s team were the teams with the highest quantities of publications in Chinese and English, respectively. The China Academy of Chinese Medical Sciences and Chinese Academy of Science were the research institutions with the highest quantities of publications in Chinese and English, respectively. Research in Chinese-language literature focused primarily on metabolomics, chemical composition, and tissue distribution, whereas English literature research emphasized component identification, metabolites, and biosynthesis. The main research focus of Chinese and English literature research is the same, both involving the characterization and spatial distribution of chemical components, biosynthesis, and metabolic pathways. Conclusion At present, MSI has been widely used in TCM research, primarily covering the characterization and spatial localization of chemical component, quality evaluation of TCM, biosynthesis of active components, in vivo metabolism and distribution, and elucidation of the TCM mechanism of action. Future research should focus on the following aspects: improvement of MSI-related databases, quantitative MSI technology optimization, the integration of MSI with other analytical technologies, clinical research and application promotion of TCM, the development and application of three-dimensional and in vivo MSI.

Systematic screening of reference genes and expression analysis of genes involved in gastrodin biosynthesis pathway in Gastrodia elata

Chinese Traditional and Herbal Drugs. 2026, 57(18): 7388-7396.

Objective To screen stable reference genes in Gastrodia elata and analyze the expression patterns of key enzyme genes in the gastrodin biosynthesis pathway, providing reliable normalization tools for molecular studies of G. elata. Methods Two transcriptome datasets, encompassing various tissues (tubers, stems, and flowers) at the jian-ma stage and four developmental stages post-inoculation, were integrated. Expression matrices were constructed using Transcripts Per Million (TPM) and normalized via the TMM method. Candidate reference genes were screened based on a coefficient of variation (CV) threshold of < 0.05, and specific primers were designed using the PrimerServer2 package. The expression levels of these candidates across different tissues and developmental stages were validated by qRT-PCR. Stability was comprehensively evaluated using delta-Ct, geNorm, NormFinder, BestKeeper, and RefFinder. Furthermore, the identified optimal reference gene was used to normalize the expression of six key enzyme genes in the gastrodin biosynthesis pathway. Results Gelref1 exhibited the highest stability across all samples, serving as a universal reference gene for various tissues and developmental stages of G. elata. Expression analysis revealed distinct patterns for the six key enzyme genes, with most showing high expression levels during the mid-to-late stages of tuber development. Conclusion This study establishes a systematic workflow for reference gene screening in G. elata based on RNA-Seq data, providing a reliable foundation for further research into the molecular mechanisms and functional validation of genes in gastrodin biosynthesis.

Cloning and functional characterization of chalcone synthase gene SsCHS1 from Spatholobus suberectus

Chinese Traditional and Herbal Drugs. 2026, 57(18): 7397-7406.

Objective To clone the chalcone synthase gene SsCHS1 from Spatholobus suberectus and conduct bioinformatics and functional analyses, in order to elucidate its role in flavonoid biosynthesis. Methods Using cDNA from S. suberectus as a template, SsCHS1 was cloned by RT-PCR. Systematic bioinformatics analyses were performed on the sequence characteristics, protein structure, and function of the gene. An SsCHS1 overexpression vector was constructed and genetically transformed into Nicotiana benthamiana. The total flavonoid content and the expression levels of key enzyme genes in the flavonoid biosynthesis pathway in transgenic plants were determined. Results The full-length SsCHS1 gene was 1 176 bp, encoding a protein consisting of 391 amino acids, with a theoretical relative molecular mass of 42 825.35, an isoelectric point of 6.10, and was characterized as a hydrophilic protein. It showed high homology with CHS proteins from leguminous plants such as Glycyrrhiza uralensis and Glycine max. Promoter analysis revealed that SsCHS1 contains multiple cis-acting elements associated with hormone response, light response, stress response, and transcription factor binding sites. Functional analysis demonstrated that overexpression of SsCHS1 significantly promoted flavonoid accumulation in transgenic tobacco and upregulated the expression levels of key enzyme genes in the flavonoid biosynthesis pathway. Conclusion The SsCHS1 gene from S. suberectus promotes flavonoid biosynthesis. The identification of this gene provides an important theoretical basis and a candidate gene for further elucidation of the biosynthetic mechanisms of active components in S. suberectus.