This study preliminarily explored the application of gastrodin (GAS) as a brain-targeting ligand for the construction of a novel glucose transporter (glucose transporter type 1, GLUT1) mediated brain-targeted lipid drug delivery system, aiming to achieve cascade-enhanced effects across the blood brain barrier and subsequent tumor targeting. Gastrodin-modified liposomes were successfully prepared using the thin-film hydration method, and their physicochemical properties, drug release characteristics, as well as in vitro and in vivo targeting abilities were preliminarily evaluated. Particular attention was paid to the in vitro and in vivo performance of gastrodin-modified liposomes prepared via a tri-branched conjugation strategy (GAS₃-Lip). The gastrodin-modified liposomes exhibited favorable targeting to bEnd.3 brain microvascular endothelial cells and GL261 glioma cells, as well as significant accumulation at the lesion site in vivo. In both in vitro and in vivo pharmacodynamic evaluations, the tri-branched gastrodin-modified doxorubicin-loaded liposomes (GAS₃-Lip/DOX) demonstrated stronger inhibitory effects against glioma compared with the classic glutathione (glutathione, GSH) modified brain-targeting liposomes, and the relevant animal experiments were approved by the Animal Experiment Ethics Committee of Southern Medical University (No: SMUL202412013). In conclusion, this strategy may provide a valuable approach for drug delivery research targeting glioma therapy.

This study developed a β-asarone (β-As)-modified nano delivery system loaded with tanshinone IIA (As-TSA-NPs) to enhance its therapeutic efficacy against cerebral ischemia-reperfusion injury (CIRI) via intranasal administration. As-TSA-NPs were prepared using the double emulsion-solvent evaporation method and characterized for morphology, particle size, zeta potential, drug loading capacity, and in vitro release behavior. Using coumarin 6 as a fluorescent probe, the 16HBE cell model was employed to investigate the effect of β-As modification on nanoparticle nasal mucosal permeability and inhibition of P-glycoprotein (P-gp) efflux. The therapeutic efficacy was evaluated in vivo using a rat middle cerebral artery occlusion (MCAO) model by assessing neurological function scores, cerebral infarction volume, and oxidative stress markers [superoxide dismutase (SOD), malondialdehyde (MDA)]. Results demonstrated that As-TSA-NPs were spherical, with an average size of approximately 180 nm, a zeta potential of about -33 mV, a drug loading capacity of approximately 3.5%, and exhibited favorable sustained release properties. β-As modification significantly enhanced cellular uptake (1.8-fold higher than unmodified NPs) and improved nasal mucosal permeability by inhibiting P-gp efflux. In vivo studies confirmed that the As-TSA-NPs group showed a 13.5% reduction in cerebral infarction volume, a 13.6% increase in SOD activity, a 21.5% decrease in MDA levels, and significantly improved neurological function scores compared to the model group. This study indicates that β-As-modified nanoparticles effectively enhance the nose-to-brain delivery efficiency of TSA, providing a novel targeted delivery strategy for CIRI treatment. All animal procedures and manipulations in this study were approved by the Animal Ethics Committee of Shanghai University of Traditional Chinese Medicine (PZSHUTCM210226002) and conducted in accordance with its guidelines.

Pulmonary hypertension (PH) is a fatal disease characterized by pulmonary vascular remodeling, in which pulmonary arterial hypertension (PAH) requires long-term combination drug therapy. In this study, we developed bosentan (BST) and sildenafil citrate (SIL) combination tablets to address the complexity of clinical combination drug regimens and poor patient compliance. An HPLC method for the determination of the content at pH 1.2, 4.5 and 6.8 was established through pre-prescription studies, which confirmed that the solubility of BST increased with the increase of pH, while the opposite was true for SIL. The prescription was optimized using wet granulation process with dissolution and content uniformity as the core indexes, and the optimal composition was determined as 47.8% BST, 31.2% SIL, 8%-18% maize starch-pre-gelatinized starch (2∶1), 2%-8% sodium carboxymethyl starch, 3% behenic acid glycerol ester and 1% magnesium stearate. Quality evaluation showed that the tablets complied with pharmacopoeial standards and had good stability. Pharmacokinetic studies in SD rats showed that the t_max of BST and SIL in the compounded tablets were 3.76 and 0.94 h, respectively, which were comparable to those of the commercially available preparations and the half-life was extended by 3-fold and 2-fold, respectively. This compound preparation significantly improves patient compliance by simplifying the dosing regimen (43 mg BST+28 mg SIL, 3 times daily). The two drugs can reach blood drug concentration at the same time, creating favorable pharmacokinetic conditions for achieving synergistic effects at the pharmacological level. This characteristic makes the compound preparation a potential new strategy for the treatment of PAH. The study was approved by the Animal Ethics Committee of China Pharmaceutical University, and all experiments complied with ethical regulations (Approval number: YSL-202505069).

In this study, paeoniflorin (PF), the active constituent of Paeonia lactiflora, was encapsulated within Paeonia lactiflora derived extracellular vesicle-like particles (PL-EVLP) to augment anti-skin-aging efficacy. A hydrogel system (PF@PL-EVLP Gel) was subsequently engineered to further enhance the formulation's stability and transdermal delivery efficiency. PL-EVLP were isolated by differential centrifugation, and paeoniflorin-loaded PL-EVLP (PF@PL-EVLP) were prepared via ultrasound-assisted freeze-thaw cycling. The resulting PF@PL-EVLP exhibited cup-shaped, bilayer-membrane nanostructures with a mean diameter of approximately 148.07 nm, a high drug-loading capacity, and favorable stability. PF@PL-EVLP Gel was fabricated using Carbomer 934 as the gel matrix. Within the hydrogel system, extracellular vesicles maintained structural integrity and uniform dispersion, exhibiting excellent stability. In vitro release studies demonstrated that both PF@PL-EVLP and PF@PL-EVLP Gel exhibited sustained-release characteristics. In vitro pharmacodynamic studies revealed that PF@PL-EVLP exerts robust protection against oxidative damage, significantly elevating intracellular superoxide dismutase (SOD) and hydroxyproline (HYP) levels while markedly reducing malondialdehyde (MDA), matrix metalloproteinase-1 (MMP-1), and reactive oxygen species (ROS) contents. Compared with the solution group, PF@PL-EVLP Gel increased the cumulative transdermal permeation by approximately 10.82-fold, the local skin retention by 7.38-fold, and the cumulative permeation rate by 9.36-fold. In vivo pharmacodynamic evaluations demonstrated that PF@PL-EVLP Gel exerts pronounced anti-skin-aging effects, as evidenced by increased skin hydration, elevated dermal collagen-fiber density, and prevention of D-galactose-induced epidermal and dermal thinning. ELISA analyses revealed that the hydrogel significantly augmented cutaneous total antioxidant capacity (T-AOC), catalase (CAT), SOD, HYP, and tissue inhibitor of metalloproteinase-1 (TIMP-1) levels, concomitantly attenuating dermal MDA and MMP-1 contents; serum SOD was also upregulated and MDA downregulated. Western blot confirmed that the formulation mitigates keratinocyte apoptosis via BAX down-regulation. These findings provide a robust foundation for the topical application of Paeonia lactiflora extracellular vesicle-based hydrogel systems in transdermal delivery of active phytoconstituents for skin-aging retardation. The animal experiments were approved by the Animal Ethics Committee of Shanghai University of Traditional Chinese Medicine (approval numbers: PZSHUTCM2503120002, PZSHUTCM230907004).

Accurate authentication of components in traditional Chinese patent medicines (TCPMs) is fundamental to ensuring the safety and accuracy. Analysis of whole-GEnome (AGE) has been applied to species identification across plants, animals, and fungi. However, its applicability to the authentication of components in TCPMs remains to be evaluated. In this study, AGE was performed to identify Panax notoginseng in TCPMs. Bioinformatic analysis of nine genome assemblies from six Panax species yielded 4 289 410 specific target sequences of P. notoginseng, from which a species-specific target sequence database was constructed. Target sequences were selected and validated by Sanger sequencing. Four molecular techniques including PCR combined with agarose gel electrophoresis, PCR coupled with Sanger sequencing, TaqMan probe-based quantitative PCR, and high-resolution melting analysis were employed to detect the target sequence, thereby establishing identification methods for P. notoginseng. These assays were further applied to commercial TCPMs, all of which were successfully detected to contain P. notoginseng. Therefore, AGE enables accurate identification of components in TCPMs, offering new insights and technologies for quality control of TCPMs.

Steroidal saponins are major active constituents of Maidong (Ophiopogon japonicus (L. f.) Ker Gawl.), and their structures and pharmacological activities are strongly influenced by glycosylation, whereas glycosyltransferases from Maidong remain poorly studied. In this study, we performed de novo transcriptome sequencing, assembly and annotation of roots, stems and leaves of O. japonicus, and combined domain-based screening to identify and conduct phylogenetic analysis of the UDP-dependent glycosyltransferase (UGT) gene family. In total, we obtained 109 UGT candidate coding sequences (CDSs) and identified 34 full-length UGT CDSs (OjUGT1-OjUGT34). Phylogenetic analysis grouped OjUGTs together with Arabidopsis UGTs into 15 subfamilies, among which clade D (UGT73 subfamily) and clade G (UGT85 subfamily) contained relatively more members. Conserved motif analysis and multiple sequence alignment showed that 31 OjUGT proteins possess the canonical plant secondary product glycosyltransferase (PSPG) motif. Expression profiling revealed pronounced tissue-specific differences in OjUGT transcripts, with OjUGT32 (UGT73 subfamily) highly expressed in tuberous roots and OjUGT34 (UGT73 subfamily) highly expressed in leaves. Comparative analyses across species suggested lineage-specific expansion of UGT73 and UGT85 genes within Asparagaceae, which may be associated with steroidal saponin biosynthesis. By integrating phylogenetic relationships, PSPG motif conservation, three-dimensional structure prediction and tissue-specific expression patterns, we propose that OjUGT32 and OjUGT34 are likely involved in the glycosylation of steroidal saponins in O. japonicus. Overall, this study establishes the expression profile of the UGT gene family in Maidong and predicts the physicochemical properties and sequence features of OjUGTs, providing candidate genes for elucidating the glycosylation steps in steroidal saponin biosynthesis.

(-)-Germacrene D is a monocyclic sesquiterpenoid with significant biological activity, and serves as a key precursor in the synthesis of other bicyclic sesquiterpene skeletons. Although plant-derived germacrene D synthases have been widely reported, fungal germacrene D synthases, particularly from genus Penicillium, remain rarely documented. In this study, a sesquiterpene synthase gene, PhGDS, was cloned from the endophytic fungus Penicillium herquei of Huperzia serrata. The encoded protein contains the canonical DDXXD and NSE/DTE motifs of type Ⅰ terpene synthases. To elucidate its function, PhGDS was heterologously expressed in Saccharomyces cerevisiae and Escherichia coli, the resulting products were identified by gas chromatography-mass spectrometry (GC-MS) and nuclear magnetic resonance spectroscopy. Two compounds were isolated from the fermentation products of S. cerevisiae, which were identified as (-)-germacrene D (66.01 mg·L^-1) and germacra-1(10),5-dien-4α-ol (0.54 mg·L^-1). Soluble expression of PhGDS was successfully achieved in E. coli, and in vitro enzymatic assays demonstrated that it catalyzes farnesyl diphosphate (FPP) to produce (-)-germacrene D and germacra-1(10),5-dien-4α-ol. This study reports the discovery and characterization of a germacrene D synthase from genus Penicillium, providing a valuable genetic resource for the scalable production of (-)-germacrene D and its derived sesquiterpenoids via synthetic biology.

The MYC transcription factor is a core regulator in the jasmonic acid signaling pathway, widely involved in plant growth and development, secondary metabolism, stress response, and hormone signal transduction. Based on the genomic data of Lonicera japonica, this study identified members of the LjMYC gene family through bioinformatics methods, and analyzed their physicochemical properties, evolutionary relationships, conserved domains and gene structures, as well as promoter cis-acting elements. The expression patterns of this gene family in different tissues, different flowering stages, and under methyl jasmonate (MeJA) treatment were also investigated. The results showed that a total of 25 LjMYC genes were identified in Lonicera japonica, distributed across eight chromosomes. Subcellular localization prediction indicated that 22 LjMYCs proteins were localized in the nucleus, LjMYC13 and LjMYC14 in chloroplasts, and LjMYC4 in the cytoplasm. Cis-acting element analysis revealed that the promoter regions of the LjMYCs gene contain elements responsive to light, growth and development, hormones, and stress. RT-qPCR analysis demonstrated that eight genes, including LjMYC8, exhibited tissue-specific expression in floral organs. The expression trends of ten genes, including LjMYC1, were generally consistent during the flowering process (S1-S6): Expression was slightly lower at S2 than at S1, increased at S3, slightly decreased at S4, and remained high at S5 and S6. Exogenous MeJA treatment promoted flowering, whereas delayed flowering treated with sodium diethyldithiocarbamate trihydrate (DIECA). Except for LjMYC20, MeJA treatment induced the expression of nine members, including LjMYC1, while DIECA treatment suppressed their expression. This study systematically reveals the fundamental characteristics of the LjMYC gene family and its expression patterns in different tissues, flowering stages, and in response to jasmonic acid signaling, providing key insights for exploring the functions of this gene family in jasmonic acid signal transduction in Lonicera japonica.

Drought is a critical stress factor severely restricting the growth and development of Thesium chinense Turcz., while the WOX (WUSCHEL-related homeobox) transcription factor family plays an important role in plant growth, development, and stress response. To explore the drought resistance mechanisms of T. chinense and identify relevant WOX genes, this study simulated drought stress using 20% PEG6000, combined with physiological and transcriptomic analyses to reveal its response strategies. The results indicated that drought stress significantly inhibited plant growth and photosynthesis but activated the antioxidant enzyme system (with CAT, POD, SOD, and APX activities reaching 4.09, 1.57, 3.05, and 58.44 times those of the control, respectively). Additionally, it promoted the accumulation of secondary metabolites such as total polysaccharides, total flavonoids, and thesiumine Ⅰ. Through transcriptomic analysis, the candidate gene TcWOX8 was identified. TcWOX8 was successfully cloned from T. chinense, with a full length of 897 bp encoding 298 amino acids, a protein molecular mass of 33.6 kD, and containing a conserved Homeodomain. Phylogenetic analysis revealed its closest relationship to WOX8 from Vitis vinifera and Hordeum vulgare, suggesting its potential involvement in plant abiotic stress responses. Further experiments confirmed that the TcWOX8 protein can be solubly expressed, possesses transcriptional activation activity, and localizes to the nucleus. The expression of this gene was not only induced by drought stress but also significantly responded to salt, low-temperature, and high-temperature stresses. These results provide a solid molecular foundation for elucidating the drought resistance mechanisms of T. chinense and revealing the key regulatory role of TcWOX8 in broad-spectrum stress defense in plants.

Lysine-specific demethylase 1 (LSD1) is a member of the flavin adenine dinucleotide (FAD)-dependent monoamine oxidase family and is highly expressed in various tumor cells. LSD1 plays a crucial role in tumor progression by participating in multiple processes such as tumor cell proliferation, invasion, differentiation, and immune cell killing. Studies have shown that LSD1 exerts its oncogenic functions not only by demethylating histone and non-histone substrates but also through demethylase activity-independent protein-protein interactions, thereby diversifying its mechanisms of action. With the increasing depth of research into the tumor-promoting mechanisms of LSD1, targeting LSD1 for cancer therapy has become a major focus in the field. This review summarizes the oncogenic functions of LSD1 through its interactions with different proteins and provides an overview of LSD1 inhibitors that have entered clinical stages. A comprehensive understanding of the roles of LSD1 in tumors will offer theoretical support for developing more effective and specific LSD1 inhibitors, thereby guiding future directions for enhanced anti-tumor therapies targeting LSD1.