Yan Liu

Gymnosperms represent an ancient lineage with diverse chemical profiles. Their terpenoids constitute a major class notable for substantial structural diversity. Terpene synthases (TPSs) are the driving force of terpene diversity. TPS-d is a gymnosperm-specific TPS subfamily, yet the evolution and functional diversity of TPS-d are still understudied. In this review, we analyze 134 functionally characterized gymnosperm TPSs to elucidate the evolutionary history and functional diversity of the predominant TPS-d subfamily. Phylogenetic analysis revealed substantial lineage-specific expansion and divergence within the TPS-d subfamily post-speciation. Crucially, we highlight the shifts in domain architecture during TPS-d evolution, providing structural evidence for the progressive attrition and eventual loss of the ancestral γ domain in TPS-d2 and TPS-d1 clades. Furthermore, we summarize the functional diversity of TPS-d enzymes in specialized metabolites, emphasizing their ecological roles in plant defense. Overall, this review provides a comprehensive overview of the evolutionary and functional landscape of the gymnosperm-specific TPS-d subfamily, serving as a valuable reference for future studies on TPSs in gymnosperms.

Rheumatoid arthritis (RA) is a chronic autoimmune disease with inflammation-mediated joint damage. Its pathogenesis involves synergistic impairment of multiple physiological balances, including immune cell subset dysregulation, abnormal inflammation, oxidative stress, pathological angiogenesis, bone homeostasis disorder and gut microbiota dysbiosis. Conventional therapies have high costs and adverse effects. Traditional Chinese medicine (TCM) follows a holistic balance-restoring concept and is a promising alternative, with Tripterygium wilfordii, Wutou Decoction and Toddalia asiatica extract as evidence-based representatives. This review summarizes 2020–2025 in vitro and in vivo studies on anti-RA TCM. Its core therapeutic mechanism is multi-target, network-based regulation. It rectifies various pathological imbalances simultaneously: modulating immune cell polarization, rebalancing inflammatory factors, scavenging ROS, inhibiting abnormal angiogenesis, regulating osteoblast-osteoclast crosstalk and reshaping gut microbiota. Single TCM agents act on multiple balance networks, showing complex pharmacological properties. TCM’s unique advantage for RA is reestablishing the body’s dynamic homeostasis by targeting interconnected pathological balances. Further research is needed to clarify TCM components’ synergistic regulatory mechanisms and promote their clinical translation.

To promote the development of high-quality Chinese patent medicine (CPM), we previously proposed a quality consistency evaluation (QCE) strategy based on intra- and inter-batch differences. In this study, Ultra-high performance liquid chromatography coupled with triple quadrupole mass spectrometry and gas chromatography coupled with triple quadrupole mass spectrometry (UPLC/GC-QQQ-MS) were separately developed and applied to determine non-volatile and volatile index components’ contents, representing the quality attributes of different raw medicines from Huoxiang Zhengqi Shui (HXZQS). Furthermore, the LPS-induced RAW264.7 cell model was applied to evaluate NO inhibitory activity, which ranged from 24.52% to 76.95%. Then the QCE strategy was applied to distinguish between superior and inferior levels from different manufacturer′s products based on the consistency of chemical contents and bioactivity. The results showed that intra-batch consistency (11.96% ‒ 47.93%) was better than inter-batch consistency (8.53% ‒ 131.25%), and inter-batch difference was the key factor affecting the quality consistency. The correlation coefficient (0.516) of chemical contents and NO inhibition rate suggested that their consistency basically followed a similar trend. The results were further graded by PCA analysis, where M7, M15, M6, M16, M3, and M5 were the best six manufacturers, and M7 had the highest content. Whereas M12 and M13 had the worst consistency and the lowest content. This study aimed to guide manufacturers to focus on process design and quality control, which not only improves the quality and safety of CPM but also promotes innovation and development of the pharmaceutical industry.

Bisbenzylisoquinoline and aporphine alkaloids are the two main pharmacological compounds in the ancient sacred lotus (Nelumbo nucifera). The biosynthesis of bisbenzylisoquinoline and aporphine alkaloids has attracted extensive attention because bisbenzylisoquinoline alkaloids have been reported as potential therapeutic agents for COVID-19. Our study showed that NnCYP80A can catalyze C-O coupling in both (R)-N-methylcoclaurine and (S)-N-methylcoclaurine to produce bisbenzylisoquinoline alkaloids with three different linkages. In addition, NnCYP80G catalyzed C-C coupling in aporphine alkaloids with extensive substrate selectivity, specifically using (R)-N-methylcoclaurine, (S)-N-methylcoclaurine, coclaurine and reticuline as substrates, but the synthesis of C-ring alkaloids without hydroxyl groups in the lotus remains to be elucidated. The key residues of NnCYP80G were also studied using the 3D structure of the protein predicted using Alphafold 2, and six key amino acids (G39, G69, A211, P288, R425 and C427) were identified. The R425A mutation significantly decreased the catalysis of (R)-N-methylcoclaurine and coclaurine inactivation, which might play important role in the biosynthesis of alkaloids with new configurations.

The edible rhizome variant of Nelumbo nucifera with various cultivars has a long history of use as a food in East Asia. In this study, 48 target metabolites were untargeted and identified in 212 rhizome cultivars (tropical and temperate types) using ultraperformance liquid chromatography-electrospray ionization quadrupole time-of-flight high-resolution mass spectrometry; among these, 32 compounds were newly reported in the rhizome. Combined with the browning phenotype of 212 lotus rhizomes, (epi) catechin, norarmepavine, and N-feruloyl-3-methoxytyramine were used as predominant chemical markers to separate different degrees of lotus rhizome browning. p-Coumaroyltyramine and N-trans-feruloyltyramine were selected as predominant chemical markers to investigate the differential expression between tropical and temperate lotus using principal component analysis and orthogonal partial least squares discriminant analysis. Shared and unique structure plots were used to compare the outcomes of the ecotype and browning OPLS model, showing that variation in tropical lotus rhizome browning is not obvious; this will be of great importance for genetic improvement by providing a hereditary basis.

Sacred lotus (Nelumbo nucifera) is a large economic crop, which is also cultivated as a horticultural crop. This study performed a systematic qualitative and quantitative determination of five anthocyanins and 18 non-anthocyanin flavonoids from the petals of 207 lotus cultivars. Among the compounds identified in this study, quercetin 3-O-pentose-glucuronide, quercetin 7-O-glucoside, laricitrin 3-O-hexose, and laricitrin 3-O-glucuronide were discovered for the first time in sacred lotus. The relationships between these pigments and petals colors were also evaluated. A decrease in the total content of anthocyanins and increase in the content of myricetin 3-O-glucuronide resulted in a lighter flower color. Furthermore, petals were yellow when the content of quercetin 3-O-neohesperidoside and myricetin 3-O-glucuronide were increased, whereas petals were red when the total anthocyanin content was high and the quercetin 3-O-sambubioside content was low. These investigations contribute to the understanding of mechanisms that underlie the development of flower color and provide a solid theoretical basis for the further study of sacred lotus.