Despite having a generally conserved structure, chloroplast genome data have been helpful for plant population genetics and evolution research. To mine Pueraria montana chloroplast genome variation architecture and phylogeny, we investigated the chloroplast variation architecture of 104 P. montana accessions from across China. P. montana’s chloroplast genome showed high diversity levels, with 1674 variations, including 1118 single nucleotide polymorphisms and 556 indels. The intergenic spacers, psbZ-trnS and ccsA-ndhD, are the two mutation hotspot regions in the P. montana chloroplast genome. Phylogenetic analysis based on the chloroplast genome dataset supported four P. montana clades. P. montana variations were conserved among and within clades, which showed high gene flow levels. Most P. montana clades were estimated to have diverged at 3.82–5.17 million years ago. Moreover, the East Asian summer monsoon and South Asian summer monsoon may have accelerated population divergence. Our results show that chloroplast genome sequences were highly variable and can be used as molecular markers to assess genetic variation and relationships in P. montana.

Objective: To address the critical gap that existing medical large language model evaluation systems are predominantly based on western medical paradigms and lack specialized assessment standards for the field of traditional Chinese medicine (TCM), this study constructs a comprehensive evaluation framework specifically designed for TCM clinical large language models, providing scientific assessment tools for their development, validation, and clinical application. Methods: This framework addresses the critical issue of missing TCM evaluation standards by incorporating TCM-specific elements such as pattern differentiation and treatment determination, as well as formula and herb recommendations, while emphasizing patient safety and data privacy. Results: The framework encompasses 3 core dimensions: (1) clinical scenario coverage and authenticity, including renowned physician case mining, pattern identification assistance, and therapy recommendations; (2) core diagnostic and therapeutic capability support, including symptom terminology recognition, pattern identification reasoning, and medical record generation; and (3) clinical application maturity and safety assurance, including data diversity, output reliability, and privacy protection. Conclusions: This evaluation system provides essential tools for the development and clinical deployment of TCM large language models, facilitating the responsible integration of AI technology into traditional medicine practice while preserving the theoretical integrity of TCM.

Leonurus japonicus is a vital herb known for its blood-activating and menstruation-regulating properties, with stachydrine being a crucial secondary metabolite contributing to these effects. Enhancing the accumulation of active ingredients is a key concern in cultivation. This study utilized a hydroponic experiment with varying inorganic nitrogen (N) concentrations (supplied as Ca(NO₃)₂·4H₂O and KNO₃) as nutrient substrates, demonstrating that exogenous nitrogen input significantly increased biomass and promoted stachyridine synthesis in L. japonicus over an extended incubation period. Analysis of key intermediates in the stachydrine synthesis pathway suggests that exogenous nitrogen enhanced the amino acid synthesis pathway in primary metabolism, promoting glycolysis and accumulating tricarboxylic acid (TCA) cycle substances, namely, pyruvate and α-ketoglutarate. This, in turn, enhanced the production of stachydrine precursors—glutamic acid and ornithine. Nitrogen’s involvement redirected the amino acid synthesis pathway more towards the alkaloid synthesis pathway. The study confirmed the essential role of nitrogen in promoting the quality formation of L. japonicus and provided initial evidence for the mechanism behind stachydrine formation, offering theoretical and practical insights for improving L. japonicus cultivation technology.