Objective To investigate the mechanisms by which Compound Danshen Dropping Pills (复方丹参滴丸) ameliorate myocardial ischemia-reperfusion injury (MIRI) in mice using single-cell and spatial transcriptomics. Methods C57BL/6 mice were randomly divided into sham group, model group, Compound Danshen Dropping Pills low-, medium-, and high-dose (61.43, 122.85, 245.70 mg/kg) groups, with 15 mice in each group. Mice were continuously given drug intervention for 7 d, and a MIRI model was constructed using left anterior descending artery ligation reperfusion method 30 min after the last administration. Cardiac function was assessed by echocardiography 24 h after surgery, and myocardial histopathology was evaluated by hematoxylin-eosin (HE) and Masson staining. Hearts from sham group, model group, and Compound Danshen Dropping Pills medium-dose group were subjected to single-cell and spatial transcriptomic analyses. Single cell transcriptomics was used for cell type identification, cardiomyocyte subpopulation typing, differential gene expression analysis, pseudo temporal analysis, differential gene expression analysis and enrichment analysis. Spatial transcriptomics analysis was used to identify the spatial transcriptomic features of MIRI and the characteristic genes of injury boundary region. Western blotting was performed to detect the expression levels of nuclear receptor subfamily 3 group C member 1 (NR3C1), pyruvate dehydrogenase kinase 4 (PDK4), acyl-CoA synthetase long-chain family member 4 (ACSL4) and glutathione peroxidase 4 (GPX4) in myocardial tissue of mice. Results Compared with sham group, the left ventricular ejection fraction (LVEF) of mice in model group was decreased (P < 0.05), pathology showed significant necrosis of myocardial cells and increased collagen fibers. Compared with model group, Compound Danshen Dropping Pills significantly improved the cardiac function of MIRI mice (P < 0.05), reduced myocardial injury and decreased collagen fibers. The results of single-cell transcriptomics indicated that Compound Danshen Dropping Pills could increase the proportion of myocardial cells and reduce the proportion of fibroblasts and inflammatory cells. Spatial transcriptomics revealed key genes involved in the spatial transcriptomic characteristics and damage boundary regions of MIRI. Western blotting results showed that compared with sham group, the expression levels of NR3C1, PDK4 and ACSL4 proteins in myocardial tissue of mice in model group mice were significantly increased (P < 0.05), while the expression level of GPX4 protein was significantly decreased (P < 0.05). Compared with model group, the expression levels of NR3C1, PDK4 and ACSL4 proteins in myocardial tissue of mice in Compound Danshen Dropping Pills group were significantly reduced (P < 0.05), while the expression level of GPX4 protein was significantly increased (P < 0.05). Conclusion Compound Danshen Dropping Pills may alleviate MIRI in mice by modulating myocardial metabolism and attenuating ferroptosis.

Objective To study the chemical constituents from the ethyl acetate extract of Alhagi sparsifolia Shap. Methods The compounds were isolated and purified by silica gel, Sephadex LH-20, ODS column chromatography, and semi‑preparative HPLC, and their structures were identified by modern spectroscopic techniques. The anti‑cervical cancer activities of the compounds against human HeLa cells were evaluated by the MTT assay. Results Fifteen compounds were isolated from the ethyl acetate extract of A. sparsifolia and identified as 1-{1-[4-(4-hydroxyphenethoxy)phenethyl]-1H-pyrrol-2-l}ethan-1-one(1), 5,7-dihydroxy-6-methoxyflavone (2), callyspongidipeptide A (3), cyclo-(D-proline-D-valine) (4), 7-O-ethylguaiacyl glycerol (5), 7-O-ethylguaiacyl glycerol (6), isololiolide (7), N-phenethylacetamide (8), skullcapflavone II (9), 4-hydroxyphenethyl alcohol (10), N-phenethylformamide (11), 3′-methoxydaidzein (12), cyclo(alanine-proline) (13), N-(p-hydroxyphenethyl)formamide (14), and N-acetyltyramine (15). Conclusions Compound 1 is a new compound and named as alhagipyrrolizidine A. Compounds 1, 5 and 6 showed significant activity against human HeLa cells, with IC₅₀ values of 31.4, 26.6 and 37.3 μmol/L, respectively.

Objective To investigate the mechanism by which sodium new houttuyfonate (SNH) inhibits hepatocellular carcinoma (HCC) progression through autophagy regulation via ubiquitin-specific peptidase 22 (USP22)-mediated silent mating type information regulation 2 homolog 1 (SIRT1) deubiquitination. Methods Human liver cancer HepG2 cells were treated with SNH or sorafenib for 24 h, cell viability was detected by CCK-8 assay, cell apoptosis was detected by flow cytometry, and cell invasion ability was detected by Transwell assay. After plasmid transfection induced overexpression of USP22, the USP22-SIRT1 interaction was verified by co-immunoprecipitation (Co-IP). USP22 and SIRT1 protein expressions, as well as SIRT1 ubiquitination were detected by Western blotting. After treatment with SIRT1 activator, autophagy activity was evaluated through mRFP-GFP-LC3 dual fluorescence labeling, adenosine triphosphate (ATP) level measurement, lactate production assay, and analysis of USP22, SIRT1, microtubule-associated protein light chain 3 (LC3)-II/I and p62 protein expressions. For in vivo experiments, 40 BALB/c-nu nude mice were eutectopically transplanted with HepG2 cells and intervened with SNH or sorafenib for 12 d. Tumor volume and weight were monitored every 3 d. Hematoxylin-eosin (HE) staining and TUNEL staining were performed on tumor tissues, and immunohistochemistry was used to detect the protein expressions of nuclear proliferation antigens Ki67, USP22 and SIRT1. Results SNH dose-dependently suppressed HepG2 cell viability and invasion (P < 0.01, 0.001), induced cell apoptosis (P < 0.001). The effect of high-dose SNH was comparable to sorafenib. Co-IP confirmed USP22-SIRT1 protein interaction, which was significantly reduced by high-dose SNH (P < 0.01). High-dose SNH significantly down-regulated USP22 and SIRT1 protein expressions (P < 0.01, 0.001), increased SIRT1 ubiquitination (P < 0.001). These effects were reversed by USP22 overexpression (P < 0.05, 0.001). SNH inhibited autophagy in HepG2 cells, manifested by enhanced GFP/mRFP fluorescence signal (P < 0.01), decreased ATP level (P < 0.001), increased lactate level (P < 0.001), increased p62 expression and decreased expression levels of USP22, SIRT1 and LC3-II/I proteins (P < 0.05, 0.001). SIRT1 activation partially counteracted the autophagic inhibition of SNH (P < 0.05, 0.01, 0.001). In vivo, high-dose SNH significantly reduced tumor volume, weight and malignancy degree (P < 0.001), induced tumor cell apoptosis and decreased Ki67, USP22, and SIRT1 expressions (P < 0.001). Conclusion SNH inhibits HCC progression by suppressing autophagy through USP22-mediated regulation of SIRT1 deubiquitination, providing a potential therapeutic strategy for HCC.

Objective High-throughput sequencing of Rehmannia chrysantha L.was performed to elucidate the structure of its chloroplast genome and its phylogenetic relationship with related species. Methods Codon preference analysis of chloroplasts of R. chrysantha was performed using CodonW v1.3 software. The codon usage bias was analyzed by using CodonW v1.3 software. Then, the obtained data were imported into IRscope and mVISTA to clarify sequence variations between R. chrysantha and related species. This study constructed a phylogenetic tree of the genus Rehmannia and its related genera on MEGA11 software. Results The chloroplast genome of R. chrysantha was a typical circular double-stranded molecule with a quadripartite structure and a total length of 153 789 bp. It has 28 high-frequency codons. Neutral plot analysis, ENC-plot analysis, and PR2-plot analysis all indicated that natural selection was the main cause of codon usage bias in R. chrysantha. It was also found that there were varying degrees of variation in the inverted repeat (IR) boundaries of the Rehmannia genus, and these changes were mainly concentrated in JLA and JLB. The genes rpl2, psbI, petN, psbZ, ycf1, and ndhB were identified as potential DNA barcodes for distinguishing closely related genus Rehmannia. In addition, phylogenetic analysis revealed a close evolutionary relationship between R. chrysantha and R. glutinosa L. Conclusion Protein-coding genes in the chloroplast genomes of genus Rehmannia exhibited a preferential use of codons ending in A/U, with natural selection being the primary factor influencing this codon usage bias. These findings will provide a foundation for future studies on molecular evolution, phylogeny, and chloroplast genetic engineering within this genus.

Objective To address the difficulty of early sex identification in Cannabis sativa and the instability of existing molecular markers, this study developed stable and accurate sex identification molecular markers based on the structural characteristics of sex chromosomes of C. sativa. This approach aims to provide a reliable tool for early sex determination at the seedling stage. Methods By comprehensively utilizing multiple chromosome-level haplotype genomic data of C. sativa, the male-specific segments in the male-specific of Y (MSY) region were systematically compared and screened through bioinformatics methods. Specific molecular markers were then designed from large MSY fragments and tested for specificity and stability using 36 C. sativa samples with known sex (15 males and 21 females) through PCR amplification and agarose gel electrophoresis, and the newly developed markers were compared with previously reported ones (MADC5 and MADC6) for validation. Results A total of 15 648 Y chromosome-specific sequences were identified, from which 12 SCAR markers were developed. Among them, five markers (MSY99M-3, MSY99M-4, MSY99M-5, MSY100M-1, and MSY100M-7) consistently produced distinct male-specific bands in all male plants, without amplification in female samples, achieving 100% identification accuracy, which was superior to that of previously reported markers. Conclusion The SCAR markers developed from Y chromosome-specific regions demonstrated high specificity, stability, and relia, enabling rapid and precise early sex identification in C. sativa. These markers offer significant potential to enhance the production efficiency of medicinal cannabis and related products.

Wound healing is a complex biological process regulated by multiple cells, factors, and signaling pathways. Delayed healing, non-healing wounds, and excessive scar formation seriously affect patients’ quality of life. Therefore, accelerating the healing speed and improving aesthetic outcomes have become major clinical concerns. Zicao (Arnebiae Radix) is a traditional Chinese medicine with the effects of “cooling the blood, promoting circulation, detoxifying, and generating flesh,” and has been widely used since antiquity to treat ulcers, burns, and other wounds. Modern studies have revealed that its main active components include naphthoquinones, polysaccharides, and phenolic acids, which exert multiple pharmacological effects such as anti-inflammatory, antibacterial, antioxidant, immunomodulatory, pro-angiogenic, and re-epithelialization-promoting activities. During wound healing, Arnebiae Radix can act on multiple targets and at multiple stages to regulate inflammation, promote granulation tissue maturation, accelerate epithelial regeneration, and improve scar structure, thereby comprehensively facilitating wound healing. This review systematically summarizes the mechanisms of Arnebiae Radix and its topical preparations in different phases of wound healing, and provides an overview of both traditional and modern dosage forms, aiming to offer a reference for further research and clinical application of Arnebiae Radix in wound healing.

Breast cancer bone metastasis (BCBM) is a common complication of breast cancer, which often induces bone pain, pathologic fractures, hypercalcemia, etc. The osteoprotegerin (OPG)/receptor activator of nuclear factor-κB (RANK)/RANK ligand (RANKL) signaling pathway plays a crucial role in development and progression of BCBM. Results has shown that this signaling pathway is closely related to osteoclast activation. The clinical treatment strategies for BCBM include bone-modifying drugs, molecularly targeted therapies, and immunotherapies. Both traditional Chinese medicine (TCM) monomers and compound preparations are able to upregulate OPG expression to inhibit osteoclast activation and block the RANKL-mediated nuclear factor-κB signaling pathway activation, thereby suppressing the pathological progression of BCBM. However, the clinical application of TCM is still limited by factors such as uncertain pharmacokinetic characteristics, incomplete quality standards, and variations in therapeutic efficacy. Elucidating the regulatory mechanisms of the OPG/RANK/RANKL signaling pathway in BCBM may therefore provide a theoretical foundation for developing TCM-based treatment strategies.

As a commonly used bulk Chinese medicinal herb, Baizhu (Atractylodis Macrocephalae Rhizoma) has a long history of medicinal use and wide application, possessing effects such as strengthening the spleen and boosting qi, drying dampness, and promoting diuresis. Its processing began during the Northern and Southern Dynasties, evolving over generations into more than ten methods primarily involving slicing and stir-frying. Among these, Atractylodis Macrocephalae Rhizoma and stir-frying Atractylodis Macrocephalae Rhizoma with honey bran remain in use today. Current quality control for Atractylodis Macrocephalae Rhizoma primarily relies on pharmacopoeia and local standards, employing multi-indicator monitoring. However, these standards predominantly focus on chemical constituents, often detaching from clinical practice. Furthermore, standards for processed slices highly overlap with raw material standards, lacking distinct characteristics for processed forms. This review synthesizes ancient herbal texts, processing treatises, successive editions of pharmacopoeias, regional processing regulations, and modern literature to trace the historical evolution of Atractylodis Macrocephalae Rhizoma processing, quality control standards and technical methodologies of Atractylodis Macrocephalae Rhizoma. It aims to provide theoretical foundations for refining Atractylodis Macrocephalae Rhizoma quality standards and advancing the production of high-quality, efficacious Atractylodis Macrocephalae Rhizoma.

Inflammatory bowel disease (IBD) is a chronic, recurrent, non-specific gastrointestinal disease with an unclear pathogenesis, and there is no curative treatment at present. As a multi-component natural traditional Chinese medicine, Coptidis Rhizoma shows the advantages of multi-target and multi-pathway regulation in the treatment of IBD with its rich chemical composition and unique pharmacological activity. Its core active ingredients are isoquinoline alkaloids, including berberine, coptisine, palmatine, berberrubine, etc., supplemented by flavonoids, lignans, organic acids, polysaccharides, terpenoids and other components, which have the effect of clearing heat and drying dampness,purgingfire and removing toxin. Modern pharmacological studies have confirmed that Coptidis Rhizoma and its active components exert therapeutic effects on IBD through multiple mechanisms, including antioxidative stress, anti-inflammatory, immunomodulatory, intestinal mucosal barrier protection, regulation of intestinal flora, glucose metabolism modulation, and cholinergic system regulation. This review summarizes the recent advances in the mechanisms and applications of Coptidis Rhizoma and its active ingredients in the treatment of IBD, providing a reference for the development of standardized clinical preparations of Coptidis Rhizoma.

At present, there are certain technical review standards and practical cases for the technical review of traditional Chinese medicine preparation registration in medical institutions in Jiangxi Province. However, with the continuous development of traditional Chinese medicine and the increasingly updated clinical needs, it is particularly important to conduct more in-depth discussion and improvement of technical review requirements. The purpose of this article is to deeply explore the relevant requirements for the technical review of the filing of traditional Chinese medicine preparations in medical institutions, with a view to providing a scientific and reasonable reference basis for effectively standardizing the technical review of the filing of traditional Chinese medicine preparations in medical institutions. Through the sorting and analysis of 127 registration and filing applications in the past five years, including the clinical needs of traditional Chinese medicine preparations prepared by traditional techniques, the challenges faced by medical institutions in the development of preparations, and the guiding role of technical review in the development of preparations, the focus is on the key points of technical review (i.e. prescription composition, preparation process, quality standards, etc.) were analyzed in depth, with a view to making the key points of drug technical review play a greater role in refining the content of application materials, standardizing the integrity of registration application materials, unifying review and approval standards, and improving registration (filing) technical review capabilities. Therefore, reasonable registration (filing) technical review points are crucial to improving the registration (filing) management of medical institutions’ preparations. They can guide medical institutions to improve the research and development level of traditional Chinese medicine preparations, standardize registration application requirements, ensure the safety and effectiveness of preparations, and promote the healthy development of traditional Chinese medicine preparations in medical institutions.