Digestive system diseases exhibit high clinical incidence rates and complex pathogenesis, creating an urgent need for the development of safe and effective therapeutic agents. Hesperetin is a flavonoid compound widely presented in Rutaceae plants, and serves as the primary effective component of traditional Chinese medicines such as Chenpi (Citri Reticulatae Pericarpium), Zhiqiao (Aurantii Fructus) and Qingpi (Citri Reticulatae Pericarpium Viride) in preventing and treating digestive system dysfunction. Modern pharmacological studies have confirmed that hesperetin could exert potential therapeutic effects on digestive system diseases such as non-alcoholic fatty liver disease, liver fibrosis, ulcerative colitis, gastric cancer, and colorectal cancer through multiple pathways including inhibiting inflammatory responses, antioxidative stress, improving lipid metabolism, regulating intestinal flora, regulating mitochondrial autophagy, antitumor, etc. This article summarizes and analyzes the relevant research reports at home and abroad in the past 30 years, systematically reviews the pharmacological effects and molecular mechanisms of hesperetin in treatment of digestive system diseases, and provides theoretical basis and scientific reference for the development of new drugs and health products.

Objective Vitexin liposomes (Vit-Lips), chitosan modified Vit-Lips (CS-Vit-Lips) and deoxycholic acid and chitosan co-modified Vit-Lips (DA/CS-Vit-Lips) were prepared, and their oral pharmacokinetic behavior were compared. Methods Deoxycholic acid-chitosan complex (DA/CS) was synthesized. Film-ultrasonic method was used to prepare Vit-Lips. Encapsulation rate, drug loading and particle size were used as evaluation indexes, single factor investigation combined with Box-Behnken design-response surface methodology (BBD-RSM) were used to investigate the optimal prescriptions of Vit-Lips. CS-Vit-Lips and DA/CS-Vit-Lips were prepared by introducing chitosan and DA/CS, respectively. Transmission electron microscopy (TEM) was used to observe the microstructure of the three kinds of liposomes, and their particle stability in gastrointestinal fluids were compared. Using vitexin as reference, the drug release of Vit-Lips, CS-Vit-Lips and DA/CS-Vit-Lips in simulate gastrointestinal fluid were compared, and their drug release mechanism were also studied. The absorption of the vitexin and its three kinds of liposomes in different intestinal segments was investigated, and the intestinal absorption parameters were calculated. Blood samples were collected after gastric administration of vitexin and its three kinds of liposomes at a dose of 20 mg/kg (vitexin), respectively. Main pharmacokinetic parameters and relative oral bioavailability were also calculated. Results The optimal formulation of Vit-Lips: phospholipids to cholesterol dose ratio was 7.95:1, lipids to drug dose ratio was 9.63:1 and hydration time was 30.00 min. CS-Vit-Lips and DA/CS-Vit-Lips were prepared using chitosan and DA/CS solution with mass fraction of 0.8%, respectively. Entrapment efficiency of Vit-Lips, CS-Vit-Lips and DA/CS-Vit-Lips were (82.31 ± 0.96)%, (84.93 ± 1.17)%, (85.15 ± 1.38)%, drug loading were (7.33 ± 0.09)%, (6.24 ± 0.08)%, (6.30 ± 0.11)%, particles size were (197.70 ± 5.07), (233.06 ± 7.19), (237.93±6.96) nm, and ζ potential were (−28.81 ± 0.86), (27.75 ± 1.10), (26.14 ± 1.13) mV, respectively. The appearance of three kinds of liposomes was spherical vesicular, particle stability of CS-Vit-Lips and DA/CS-Vit-Lips was higher than that of Vit-Lips in simulated gastrointestinal fluid. Vit-Lips, CS-Vit-Lips and DA/CS-Vit-Lips enhanced cumulative release rate to 91.02%, 86.74% and 83.15%, respectively. The drug release process of the three kinds of liposomes conformed to Weibull model. The three kinds of liposomes effectively improved the absorption rate constant (K_a) and apparent absorption coefficient (P_app) of vitexin. Compared to vitexin, Vit-Lips, CS-Vit-Lips and DA/CS-Vit-Lips increased relative oral bioavailability to 1.10-fold, 3.08-fold and 4.70-fold, respectively. The half-life time (t_1/2) of CS-Vit-Lips and DA/CS-Vit-Lips had significantly prolonged (P < 0.01) and the peak concentration (C_max) was significantly increased (P < 0.01). Conclusion CS-Vit-Lips and DA/CS-Vit-Lips effectively promoted the oral absorption of vitexin, and the advantage of DA/CS-Vit-Lips were more obvious, laying an experimental foundation for further research.

Objective To screen characteristic genes driving gastric mucosal “inflammation-cancer transformation” based on multi-omics data and Mendelian randomization (MR) causal inference methods, and to explore potential traditional Chinese medicine (TCM) intervention strategies. Methods Transcriptomic data and large-scale genetic data were integrated. Differential analysis, MR analysis, and protein-protein interaction network construction were performed to screen characteristic genes causally associated with gastric cancer risk. Subsequently, survival analysis, immune infiltration analysis, and immune mediation mechanism analysis were conducted on the identified genes. Finally, molecular docking-based virtual screening was performed on a TCM small-molecule compound library using core characteristic genes as targets to predict potential active components. Results A total of 30 continuously up-regulated differentially expressed genes were identified during the “inflammation-cancer transformation” process. MR analysis identified five core characteristic genes (S100A8, CXCR1, S100A9, S100A12, ZBED2) showing positive causal associations with gastric cancer risk. Among them, high expression of S100A12 was significantly associated with poor prognosis in patients, positively correlated with neutrophil infiltration, and negatively correlated with B-cell and CD4⁺ T-cell infiltration. Mediation analysis revealed that S100A12 may promote carcinogenic effects by negatively regulating CD25⁺CD4⁺ T cells (primarily regulatory T cells). Phenome-wide MR analysis suggested good potential safety for targeting S100A12. Virtual screening identified several TCM small-molecule compounds with high binding affinity to the S100A12 protein (e.g., C-curarine, physalin D). Conclusion S100A12 is a key characteristic gene linking chronic inflammation to gastric cancer development, driving “inflammation-cancer transformation” by reshaping the immune microenvironment. The screening of TCM small molecules targeting this gene provides lead compound candidates for intervening in gastric precancerous lesions. The established research framework integrating multi-omics and reverse drug screening offers a new approach for the precise prevention and drug development of gastric cancer.

Ancient classical prescriptions, with their long clinical application experience and clear therapeutic effects, have become an important direction for the research and development of new traditional Chinese medicine drugs. With the continuous advancement of related research, it has not only provided a new path for explaining the scientific connotations of traditional prescriptions, but it has also significantly promoted the technological innovation of traditional Chinese medicine compound preparations. This has profound significance for the inheritance and innovative development of traditional Chinese medicine. In recent years, under the guidance of national policies, the development of classic prescription traditional Chinese medicine compound preparations has received much attention. Among them, solid preparations, due to their good stability, controllable dosage, and convenience in administration, have become the main dosage form in the development of classic prescription preparations and have gained wide application and market recognition. This article takes traditional Chinese medicine compound preparations as the entry point, systematically reviews the application status, problems and challenges, and solutions of solid preparation technology in the development of classic prescriptions, and discusses the prospects of solid preparation technology in the development of classic prescriptions, providing certain references for the future development of classic prescription traditional Chinese medicine compound preparations.

Objective To investigate the molecular mechanism by which Linggui Zhugan Decoction (苓桂术甘汤, LGZGD) ameliorates heart failure (HF) after myocardial infarction (MI) via regulating mitophagy mediated by mitochondrial calcium uniporter (MCU)-mitochondrial calcium (mtCa²⁺) signaling pathway. Methods Rat models of post-MI HF and hydrogen peroxide (H₂O₂)-induced H9c2 cardiomyocyte injury models were established. For in vivo experiments, rats were assigned to sham group, model group, LGZGD group and captopril group. For in vitro experiments, H9c2 cells were divided into corresponding groups with additional interventions of MCU knockdown or overexpression. Echocardiography, Masson staining and transmission electron microscopy were performed to evaluate rat cardiac function, myocardial fibrosis and mitochondrial ultrastructure, respectively. The mRNA and protein expression levels of genes related to mitophagy and mitochondrial function were detected by qRT-PCR and Western blotting. The levels of reactive oxygen species (ROS) and mtCa²⁺ were measured, myocardial injury markers were quantitatively analyzed, and changes in cellular autophagic flux were observed using confocal laser scanning microscopy. Results Compared with sham group, rats with post-MI HF exhibited significantly impaired cardiac function, aggravated myocardial fibrosis and mitochondrial damage, dysregulated transcription of mitophagy-related genes, and markedly upregulated MCU transcription level (P < 0.001). In H₂O₂-stimulated H9c2 cells, ROS and mtCa²⁺ levels were significantly increased, autophagic flux was blocked, and myocardial injury marker levels were notably elevated (P < 0.01).Compared with model group, LGZGD treatment significantly improved cardiac function, alleviated myocardial fibrosis and mitochondrial damage, downregulated MCU expression (P < 0.01), regulated the mRNA expressions of autophagy and mitophagy-related genes in post-MI HF rats (P < 0.05, 0.01). At the cellular level, LGZGD reduced ROS and mtCa²⁺ levels in H₂O₂-injured H9c2 cells, restored mitochondrial morphology and autophagic flux, and decreased myocardial injury marker levels (P < 0.01). MCU knockdown enhanced the cardioprotective effect of LGZGD, whereas MCU overexpression exacerbated cellular injury. LGZGD effectively reversed MCU overexpression-induced cellular damage and ameliorated the dysregulation of oxidative stress-related enzymes (P < 0.05, 0.01). Conclusions LGZGD alleviates oxidative stress injury and improves post-MI heart failure by restoring MCU-mtCa²⁺ homeostasis and promoting mitochondrial autophagic flux.

Objective To clone the cycloartenol synthase (CAS) gene PcCAS from Polygonatum cyrtonema, perform bioinformatic and recombinant protein expression analyses, and determine its subcellular localization etc, thereby providing essential resources for further elucidating the pivotal role of PcCAS in saponin biosynthesis. Methods Based on the transcriptome data of P. cyrtonema, the PcCAS gene was screened and cloned, and bioinformatics approaches were employed to analyze the physicochemical properties, secondary structure, and tertiary structure of its encoded protein. qRT-PCR was used to analyze the tissue-specific expression of PcCAS. A prokaryotic expression vector was constructed and transferred into Escherichia coli for prokaryotic expression analysis. Yeast expression vectors were constructed and transformed into Pichia pastoris for subsequent yeast expression analysis. A subcellular-localization vector was built and infiltrated into Nicotiana benthamiana leaves for the analysis of heterologous expression in tobacco. Results The open reading frame (ORF) of PcCAS is 2 286 bp in length and encodes 759 amino acids. The protein is hydrophilic and lacks a signal peptide. The total saponin content and PcCAS expression level in the rhizome are significantly higher than those in the stem, leaf, and flower, indicating the expression level of PcCAS is positively related with the total saponin content. Yeast-expressed recombinant protein matched the expected molecular weight of 86 400. Subcellular-localization assays in tobacco leaves demonstrated that the PcCAS protein is predominantly localized to the endoplasmic reticulum. Conclusion PcCAS protein belongs to the OSC superfamily of enzymes and is localized in the cytoplasm, positively regulating the total saponin biosynthesis.

Objective To investigate the regulatory mechanism underlying the effect of low-temperature treatment on the accumulation of active components in Veronica polita and clarify its regulatory mechanism. Methods Plants subjected to low-temperature treatment were analyzed through phenotypic observation, physiological and biochemical indices, transcriptome and metabolome analyses. Results Prolonged low-temperature treatment significantly increased the plant height, fresh weight and dry weight, as well as the root length of V. polita. The activities of superoxide dismutase (SOD) and peroxidase (POD), as well as the contents of total flavonoids, total phenols and total sugars first rose and then decreased. The contents of malondialdehyde (MDA) and chlorophyll increased, whereas chlorophyll fluorescence parameters decreased. The contents of total flavonoids and total phenols also first increased and then decreased. Transcriptomic analysis showed that the differentially expressed genes (DEGs) exhibited a stage-specific expression patterns. Pathways such as secondary metabolite biosynthesis, phenylpropanoid biosynthesis, metabolic pathways, and plant hormone signal transduction were significantly enriched. Three flavonoid synthesis-related genes (GT5, JOX2, HO1) and five phenol synthesis and metabolism-related genes (LAC3, AOX4, LPR2, AS1, MT-CO2) were identified. Metabolomic analysis indicated that the contents of quercetin, myricetin, and benzoic acid showed a trend of first increasing and then decreasing with the extension of treatment time. Integrated multi-omics analysis demonstrated that the core mechanism by which V. polita responds to low-temperature stress involves the precise reprogramming of the phenylpropanoid/flavonoid metabolic pathway. Conclusion Appropriatelow-temperature treatment promotes the accumulation of secondary metabolites in V. polita, thereby providing a foundation for further research into its functional genes.

Objective To screen the optimal purification and drug-loading processes of exosomes (Exo), construct an in vitro blood-brain barrier (BBB) cell model and an Alzheimer’s disease (AD) mouse model, and investigate the efficiency of human embryonic kidney 293T (HEK 293T) Exo-mediated ginsenoside Rg₁ (Rg₁) across the BBB, so as to provide a basis for the treatment of brain diseases. Methods Exosomes were purified using ultracentrifugation, tangential flow filtration, and dual-coupled harmonic oscillation technology. The optimal purification method was selected based on concentration, particle size, and polydispersity index (PDI). On the basis of single-factor screening of voltage, pulse duration, pulse number, and Rg₁ concentration, the electroporation process for constructing Exo-Rg₁ complexes was optimized using orthogonal design with drug encapsulation efficiency as the evaluation index. An in vitro BBB model was established using brain endothelial cell line 3 (bEnd.3). Model integrity was verified by measuring transendothelial electrical resistance (TEER), liquid level permeability, and sodium fluorescein permeability. The Exo-Rg₁ complex group (experimental group), free Rg₁ group (control group), exosome alone group, and empty exosome + free Rg₁ group were set up. The concentration of Rg₁ penetrating the in vitro BBB model at different time points was detected by HPLC, and the permeability efficiency was calculated. In vivo therapeutic effects were evaluated through behavioral tests, histological observation, and detection of protein and inflammatory factor levels in AD model mice. Results The optimal exosome purification process was dual-coupled harmonic oscillation technology, yielding an exosome concentration of 2.9 × 10¹¹ particles/mL, a particle size of (91.8 ± 1.5) nm, and a PDI of 0.21 ± 0.01. The optimized electroporation conditions were voltage of 100 V, pulse duration of 15 ms, pulse number of four times, and Rg₁ concentration of 0.4 mg/mL. Under these conditions, the encapsulation efficiency of the complexes was 61.58%, which was superior to that of the traditional ultrasonic method. The in vitro BBB model using bEnd.3 cells was successfully established, with TEER values stably above 300 Ω·cm². A significant liquid level difference was maintained in the in vitro BBB model group after 4 h. The sodium fluorescein permeability in the model group was significantly lower than that in the blank control group [(41.34 ± 1.83)% vs (98.87 ± 2.09)%, P < 0.01], meeting the requirements for barrier function. In vitro penetration experiments showed that the penetration efficiency of Rg₁ in the experimental group (15.3%) was significantly higher than that in the control group (3.3%) at 8 h (P < 0.05). Animal experiments demonstrated that the complex significantly improved learning and memory abilities in model mice, protected hippocampal neurons, upregulated the expression of choline acetyltransferase (ChAT) and α7 nicotinic acetylcholine receptor (α7 nAChR) proteins, and reduced the levels of interleukin-1β (IL-1β), IL-6, and tumor necrosis factor-α (TNF-α), with effects superior to those of the free Rg₁ group and the exosome-only group. Conclusion Exosomes can effectively enhance the efficiency of Rg₁ in crossing the BBB, providing an experimental basis for improving the therapeutic effect of brain diseases.

Objective To evaluate the quality of Lycium barbarum leaves from various production areas in Ningxia, an HPLC fingerprinting technique combined with chemometrics and quantitative determination of multiple alkaloids constituents with LC-MS/MS was used to construct the alkaloidal fingerprints of L. barbarum leaves (S1—S50) from five bases in three production areas in Ningxia. Methods The HPLC method and Similarity Evaluation System for Chromatographic Fingerprinting in Chinese Medicine (version 2012) were used to generate fingerprints, and similarity evaluation and common peak confirmation were carried out. Meanwhile, LC-MS/MS was used to establish a method for the determination of the contents of seven alkaloids constituents (choline, betaine, trigonelline, lyciumin A, lyciumin B, N-trans-feruloyltyramine, N-feruloyl-3-methoxytyramine) to evaluate the leaves of L. barbarum from different origins in Ningxia. Results The obtained fingerprints had a total of 18 common peaks. The similarity between the alkaloid fingerprints of 50 batches of L. barbarum leaves. The control fingerprints ranged from 0.692 to 0.981. Zhongning and Guyuan origin L. barbarum leaves fingerprints have higher similarity to the control fingerprints, while Yinchuan origin L. barbarum leaves fingerprints have lower similarity to the control fingerprints. Hierarchical clustering analysis (HCA) clustered the 50 batches of samples into four categories; principal component analysis (PCA) screened out four principal components, with a cumulative variance of 89.014%; orthogonal partial least squares-discriminant analysis (OPLS-DA) screened out the components with larger contributions to inter-group variance, which were, in the order of the size of the VIP, peak 5 > peak 3 > peak 10 > peak 11 > peak 13 > peak 2 > peak 4 > peak 9; content determination showed that seven alkaloids from Guyuan origin had the highest content. The results of content determination showed that the seven alkaloids of Guyuan origin had the highest content. Conclusion A stable HPLC fingerprint of L. barbarum leaves of different origins and a quantitative LC-MS/MS method for the determination of seven alkaloids were constructed, which can be used for the analysis of the difference in the origins of L. barbarum leaves and quality evaluation by combining with chemometrics.

Objective Novel self-assembled nanoparticles (RMN-SAN) of Dahuang (Rhei Radix et Rhizoma) and Mudanpi (Moutan Cortex) were prepared by the micro-precipitation method, and their characterization, formation mechanism and anti-colorectal cancer (CRC) efficacy were investigated. Methods The process parameters were optimized through single-factor experiments, and characterization methods such as dynamic light scattering, ζ potential and transmission electron microscopy were performed. The core active components were identified by UPLC-Q-TOF-MS, and the self-assembly mechanism of RMN-SAN was revealed by FTIR and UV spectroscopy. The in vitro CT26 cell experiments and in vivo tumor-bearing mouse models were used to verify its anti-CRC efficacy. Results The optimized preparation method for RMN-SAN involves taking 10 g each of Rhei Radix et Rhizoma and Moutan Cortex, adding 10 times their volume of water, soaking for 30 min, and refluxing for 0.5 h, followed by filtration. The residue is then refluxed with 8 times its volume of 70% ethanol for 0.5 h and filtered again. The two extracts are combined, magnetically stirred at 600 r/min and 25 ℃ for 30 min, and the alcohol is removed by rotary evaporation at 60 ℃, concentrating to a final concentration of 0.2 g/mL. The solution is subjected to gradient centrifugation (4 ℃, 6 000, 8 000, and 10 000 r/min, each for 10 min), followed by dialysis (cut-off molecular weight 3 500) for 12 h, yielding the final product. The results showed that the average particle size of the optimized RMN-SAN was (109.30 ± 4.20) nm, the PDI was 0.353 ± 0.009, and the ζ potential was (-24.5 ± 1.3) mV, presenting a uniform spherical structure. Five core components such as rhein, aloe-emodin, sennoside B, paeonol, and paeoniflorin were identified, and their self-assembly was driven by hydrogen bonds and π-π stacking interactions, which was verified by molecular docking. The in vitro experiments confirmed that RMN-SAN could inhibit the proliferation of CT26 cells in a concentration-dependent manner, induce apoptosis and block the cell cycle at the G₁ phase. In the in vivo experiments, the tumor suppression effect of the high-dose RMN-SAN group (6.0 g/kg) was significantly higher than that of the traditional decoction group, and it could regulate the levels of immune and inflammatory factors more efficiently. Conclusion This study enabled the utilization of the therapeutic advantages of traditional Chinese medicine through modern processing, providing new ideas for the optimization of the preparation of self-assembled nanoparticles of traditional Chinese medicine, and also providing an efficient and stable new formulation for the treatment of CRC with traditional Chinese medicine.