Objective To elucidate the changes in the gut microbiota and molecular mechanism of huaier in enhancing the efficacy of oxaliplatin (OXA) chemotherapy in gastric cancer (GC). Methods The effects of huaier on the gut microbiota structure and intestinal barrier function in MKN45-bearing mice were analyzed by using 16S rRNA sequencing, RT-qPCR, and IHC. UPLC-MS and network pharmacology, as well as in vivo experimental approaches, were employed to investigate the key bioactive constituents of huaier systematically and elucidate the underlying mechanisms by which huaier exerts therapeutic effects against GC. The expression of the PI3K/AKT/SREBP pathway was detected in cancer cells and tissues via Western blot analysis. The safety profile of huaier combined with OXA was verified through serum biochemistry and HE-stained tissue section analyses. Results Huaier inhibited the PI3K/AKT/SREBP pathway by increasing the abundance of Akkermansia muciniphila, reduced lipid droplet deposition, and ultimately enhanced the sensitivity to OXA in the treatment of GC. Conclusion This study demonstrates the value of huaier in gastric cancer treatment by enhancing chemosensitivity and provides novel insights into its systemic therapeutic effects through molecular mechanisms and gut microbiota modulation.

Coagulation factor XIa (FXIa) plays a crucial role in thrombus formation; therefore, the development of potent and safe FXIa inhibitors is of great significance. In this study, compound F22, previously discovered by our group, was selected as the lead compound. Based on the principles of bioisosterism and fragment-based drug design, four series comprising 14 novel Asundexian derivatives not previously reported in the literature were designed and synthesized. The structures of the target compounds were confirmed by 1H NMR and HRMS, and their inhibitory activities against FXIa were evaluated using chromogenic substrate assay. Results showed that compound FD-1 exhibited the most potent activity, with an IC50 value of 2.8 nmol/L, which was superior to that of the lead compound F22 (IC50 = 4.5 nmol/L) and the reference drug Asundexian (IC50 = 5.0 nmol/L). Furthermore, in the activated partial thromboplastin time (aPTT) assay, compound FD-1 demonstrated excellent anticoagulant activity, outperforming Asundexian, showing no significant effect on prothrombin time (PT). These findings provide valuable insights for further structural optimization and rational design of small-molecule FXIa inhibitors.

Background: Influenza A virus (IAV) is a negative-sense RNA virus of the Orthomyxoviridae family and is the etiological agent of a highly contagious acute respiratory disease that can lead to acute lung injury. Objective: To elucidate the molecular mechanisms of IAV infection, an integrative research approach combining gene expression profiling, multinetwork analysis, and in vivo experimental validations was employed. Methods: First, a series of network-based analyses were performed, including protein-protein interaction network construction, weighted gene co-expression network analysis, and subsequent gene set enrichment analysis, to identify the major underlying mechanisms of IAV infection. Following gene expression analysis, core targets, both direct and indirect regulators, were screened. An IAV (H1N1) strain A/PR/8/34-induced acute lung injury mouse model was constructed for in vivo validations. Batch one included two groups to evaluate findings from the multi-network analysis: Mock (n = 10; 5 males and 5 females) and IAV (n = 10; 5 males and 5 females). Batch two included three groups to assess the role of toll-like receptor 3 (TLR3) in IAV infection: Mock (n = 6; 3 males and 3 females), IAV (n = 6; 3 males and 3 females), and TLR3 inhibitor (n = 6; 3 males and 3 females). Body weight was measured on days 0, 3, and 5 after infection. On day 5, lung tissues were collected to assess viral load and histopathological changes. Key targets were examined using enzyme-linked immunosorbent assay, Western blotting, and immunofluorescence staining, both in sera and lung tissues. Results: IAV infection was significantly associated with dysregulation of the immune-inflammation system, such as the LTR, nucle-otide-binding oligomerization domain-(NOD) like receptor, retinoic acid-inducible gene I-like receptor, and nuclear factor kappa-B signaling pathways. Gene set enrichment analysis further indicated that the TLR and necroptosis signaling pathways played crucial roles in the progression of IAV infection (TLR signaling pathway normalized enrichment score = 2.3941, P = 1.00 × 10 ⁻¹⁰; necroptosis normalized enrichment score = 1.9421, P = 6.21 × 10 ⁻⁷). Among the core targets, TLR3 and mixed lineage kinase domain-like protein (MLKL) may regulate gene expression at the transcriptional level (all P < 0.05). In vivo validation using an IAV (PR8) infected acute lung injury mouse model demonstrated increased viral load and lung index, alveolar structural damage, and inflammatory cell infiltration. Immunofluorescence staining exhibited large gaps in Lamin B1 staining and breaches in Emerin signals following IAV-PR8 infection. Expression levels of TLR3, p-receptor-interacting serine/threonine-protein kinase 3 (RIPK3)/RIPK3, and p-mixed lineage kinase domain-like protein (MLKL)/MLKL proteins in lung tissues, as well as proinflammatory factors and mediators in sera, were significantly elevated after IAV infection. Moreover, enhanced neutrophil infiltration (myeloperoxidase) and citrullinated histone H3 (a neutrophil extracellular trap-specific marker), both established indicators of neutrophil extracellular trap formation, were observed. Notably, treatment with a TLR3 inhibitor significantly ameliorated IAV-induced acute lung injury by regulating necroptosis-related targets. Conclusion: Our study provides network-based in vivo evidence that TLR3-receptor-interacting serine/threonine-protein kinase 3-MLKL-mediated necroptosis may underlie IAV-induced acute lung injury and could serve as a potential therapeutic target in severe influenza cases.

ObjectiveTo evaluate the Effect of Guishen Wan on learning and memory ability in D-galactose-induced aging model mice and to explore the application of the "Kidney-Brain Coordination" theory in traditional Chinese medicine （TCM）. MethodsAn subacute aging mouse model was established by subcutaneous injection of D-galactose （200 mg·kg^-1·d^-1） into the neck and back of KM mice. The mice were then administered GuiShen Wan at high， medium， and low doses （18， 9， 4.5 g·kg^-1·d^-1） or donepezil（2 mg·kg^-1·d^-1）， a standard clinical therapeutic agent for Alzheimer's disease， served as the positive control in this study，for 4 consecutive weeks. Learning and memory abilities were evaluated using the Morris water maze. The degree of renal fibrosis was observed via Masson staining. Hippocampal neuron injury was assessed by hematoxylin-eosin（HE） staining. Damage to Nissl bodies in the hippocampus was examined using Nissl staining. The fluorescence expression intensity of Klotho in the hippocampus was detected by immunofluorescence. Western blot was used to detect the protein expression of Klotho in the kidneys， as well as proteins related to the hippocampal inflammatory pathway and inflammatory factor-related signaling pathways. ResultsResults from the water maze test showed that compared with the blank group， the number of platform crossings and the time spent in the target quadrant were significantly decreased in the model group （P<0.01）. Compared with the model group， all dose groups of Guishen Wan significantly increased the number of platform crossings and the time spent in the target quadrant in model mice （P<0.01）.Masson staining results showed that compared with the blank group， the collagen volume fraction was significantly increased in the model group （P<0.01）. Compared with the model group， all dose groups of Guishen Wan significantly reduced the collagen volume fraction in the model group （P<0.01）. Kidney Western blot analysis showed that compared with the blank group， the Klotho protein was significantly decreased in the model group （P<0.01）. Compared with the model group， Guishen Wan elevated the klotho protein （P<0.01）.Neuronal cell count results showed that compared with the blank group， the number of neurons was significantly reduced in the model group （P<0.01）. Compared with the model group， Guishen Wan exhibited a dose-dependent trend in repairing neuronal damage and increasing the number of neuronal cells （P<0.01）.Immunofluorescence intensity results showed that compared with the blank group， the positive expression of Klotho was significantly decreased in the model group （P<0.01）. Compared with the model group， all dose groups of Guishen Wan enhanced Klotho fluorescence expression （P<0.01）.Hippocampal Western blot analysis showed that compared with the blank group， the ratios of interleukin（IL）-1β and IL-6 proteins were significantly increased in the model group （P<0.01）. Compared with the model group， all dose groups of Guishen Wan significantly reduced the ratios of IL-1β and IL-6 proteins（P<0.01）. Compared with the blank group， the ratio of phosphorylated-nuclear transcription factor-κB（p-NF-κB） protein was significantly increased （P<0.01）， while the klotho protein was significantly decreased （P<0.01） in the model group. Compared with the model group， all Guishen Wan groups significantly reduced the p-NF-κB protein （P<0.05， P<0.01） and significantly increased the Klotho protein（P<0.01）. ConclusionGuishen Wan effectively ameliorates D-galactose-induced aging in model mice. Its mechanism of action may be related to upregulating Klotho concentration and inhibiting inflammatory factors associated with the NF-κB signaling pathway.

ObjectiveTo investigate the effect and mechanism of Banxia Shumitang plus Zhizichi Tang （BZ） on PTEN-induced kinase 1 （PINK1）/Parkin signaling pathway in regulating mitochondrial autophagy in preventing insomnia with depression. MethodsSeventy-two 6-week-old male Kunming mice were randomized into a normal group， a model group， low-dose， medium-dose， and high-dose BZ groups （BZ-L， BZ-M， BZ-H， 2.92， 5.85， 11.7 g·kg^-1）， and an estazolam （0.21 mg·kg^-1） group. Insomnia model was established on days 1 to 3. p-Chlorophenylalanine （PCPA） was injected intraperitoneally at a dose of 400 mg·kg^-1 into mice of the model group and administration groups for 3 consecutive days. Then， the classic depression model was established with chronic unpredictable mild stress （CUMS） for 28 consecutive days. Administration and modeling were performed simultaneously. After gavage， 24 h spontaneous activity test was performed to test sleep state， and sugar preference test and open field test were performed to evaluate anxiety-like state. The morphological changes of the hypothalamus were observed by hematoxylin-eosin （HE） staining， and the number of Nissl bodies was observed by Nissl staining. The ultrastructure of mitochondria was observed by transmission electron microscopy. The mitochondrial membrane potential assay kit （JC-1） was used to examine the mitochondrial function， and a biochemical assay kit was used to measure the ATP content in the hypothalamus. The level of reactive oxygen species （ROS） was determined by the immunofluorescence assay， and the serum levels of 5-hydroxytryptamine （5-HT）， noradrenalin （NE）， and cortisol （CORT） were determined by enzyme-linked immunosorbent assay （ELISA）. Western blot and Real-time quantitative polymerase chain reaction （Real-time PCR） were employed to determine the expression of PINK1， Parkin， microtubule-associated protein 1 light chain 3 （LC3）， and isolator protein 1 （p62） at protein and mRNA levels， respectively. ResultsThe 24 h spontaneous activity test showed that compared with the normal group， the model group did not exhibit circadian rhythm characteristics in their spontaneous activity time， while the circadian rhythm characteristics were restored in each treatment group （P<0.05）. The sucrose preference test indicated that compared with that in the normal group， the sucrose preference rate in the model group reduced （P<0.01）， and compared with that in the model group， the sucrose preference rate in each treatment group increased （P<0.01）. The open field test revealed that compared with the normal group， the model group exhibited decreased movement distance， average speed， frequency of visits to the central area， and time spent in the central area （P<0.01）， and compared with those in the model group， these parameters in each treatment group increased （P<0.05， P<0.01）. HE staining showed that compared with the normal group， the model group exhibited disarrangement of neurons in the hypothalamus， enlarged intercellular space， and severe nuclear condensation， while the degree of hypothalamic neuronal damage in each treatment group was alleviated. Nissl staining indicated that compared with the normal group， the model group demonstrated aggravated neuronal damage and increased damaged cells （P<0.01）. Compared with that in the model group， the number of damaged cells in each treatment group decreased （P<0.01）. Transmission electron microscopy showed that compared with the normal group， the model group exhibited mitochondrial swelling and rupture in the hypothalamus， with visible autophagic lysosomes in the cytoplasm. Compared with those in the model group， only a few autophagic lysosomes were observed in the BZ-L group， and the degree of mitochondrial swelling and rupture was alleviated in the other treatment groups. ROS immunofluorescence assay showed that compared with the normal group， the model group displayed enhanced red fluorescence intensity of mitochondrial ROS （P<0.01）， which was attenuated in each treatment group （P<0.01）. Mitochondrial membrane potential assay showed that compared with the normal group， the model group had declined mitochondrial membrane potential （P<0.01）， which was recovered in each treatment group （P<0.05， P<0.01）. The biochemical assay of ATP levels indicated that compared with the normal group， the model group exhibited decreased ATP content （P<0.01）， which rebound in each treatment group （P<0.05， P<0.01）. ELISA showed that compared with the normal group， the model group exhibited decreased 5-HT content and increased NE and CORT content （P<0.01）. Compared with the model group， each treatment group showed increased 5-HT content （P<0.01） and reduced NE content （P<0.01） and CORT content （P<0.05， P<0.01）. Western blot of the hypothalamus showed that compared with the normal group， the model group exhibited downregulated protein levels of PINK1， Parkin， and LC3 （P<0.01） and upregulated protein level of p62 （P<0.01）. Compared with the model group， each treatment group showed upregulated protein levels of PINK1， Parkin， and LC3 （P<0.05， P<0.01） and downregulated protein level of p62 （P<0.05， P<0.01）. Real-time PCR showed that compared with the normal group， the model group exhibited downregulated mRNA levels of PINK1， Parkin， and LC3 （P<0.01） and upregulated mRNA level of p62 （P<0.01）. Compared with the model group， each treatment group showed upregulated mRNA levels of PINK1， Parkin， and LC3 （P<0.01） and downregulated mRNA level of p62 （P<0.01）. ConclusionBZ can ameliorate the insomnia and anxiety state of mice by regulating PINK1/Parkin signaling pathway to improve the mitochondrial function， enhance mitochondrial autophagy， protect neurons， and repair damaged nerve cells.

ObjectiveTo investigate the effect and mechanism of Banxia Shumitang plus Zhizichi Tang （BZ） on PTEN-induced kinase 1 （PINK1）/Parkin signaling pathway in regulating mitochondrial autophagy in preventing insomnia with depression. MethodsSeventy-two 6-week-old male Kunming mice were randomized into a normal group， a model group， low-dose， medium-dose， and high-dose BZ groups （BZ-L， BZ-M， BZ-H， 2.92， 5.85， 11.7 g·kg^-1）， and an estazolam （0.21 mg·kg^-1） group. Insomnia model was established on days 1 to 3. p-Chlorophenylalanine （PCPA） was injected intraperitoneally at a dose of 400 mg·kg^-1 into mice of the model group and administration groups for 3 consecutive days. Then， the classic depression model was established with chronic unpredictable mild stress （CUMS） for 28 consecutive days. Administration and modeling were performed simultaneously. After gavage， 24 h spontaneous activity test was performed to test sleep state， and sugar preference test and open field test were performed to evaluate anxiety-like state. The morphological changes of the hypothalamus were observed by hematoxylin-eosin （HE） staining， and the number of Nissl bodies was observed by Nissl staining. The ultrastructure of mitochondria was observed by transmission electron microscopy. The mitochondrial membrane potential assay kit （JC-1） was used to examine the mitochondrial function， and a biochemical assay kit was used to measure the ATP content in the hypothalamus. The level of reactive oxygen species （ROS） was determined by the immunofluorescence assay， and the serum levels of 5-hydroxytryptamine （5-HT）， noradrenalin （NE）， and cortisol （CORT） were determined by enzyme-linked immunosorbent assay （ELISA）. Western blot and Real-time quantitative polymerase chain reaction （Real-time PCR） were employed to determine the expression of PINK1， Parkin， microtubule-associated protein 1 light chain 3 （LC3）， and isolator protein 1 （p62） at protein and mRNA levels， respectively. ResultsThe 24 h spontaneous activity test showed that compared with the normal group， the model group did not exhibit circadian rhythm characteristics in their spontaneous activity time， while the circadian rhythm characteristics were restored in each treatment group （P<0.05）. The sucrose preference test indicated that compared with that in the normal group， the sucrose preference rate in the model group reduced （P<0.01）， and compared with that in the model group， the sucrose preference rate in each treatment group increased （P<0.01）. The open field test revealed that compared with the normal group， the model group exhibited decreased movement distance， average speed， frequency of visits to the central area， and time spent in the central area （P<0.01）， and compared with those in the model group， these parameters in each treatment group increased （P<0.05， P<0.01）. HE staining showed that compared with the normal group， the model group exhibited disarrangement of neurons in the hypothalamus， enlarged intercellular space， and severe nuclear condensation， while the degree of hypothalamic neuronal damage in each treatment group was alleviated. Nissl staining indicated that compared with the normal group， the model group demonstrated aggravated neuronal damage and increased damaged cells （P<0.01）. Compared with that in the model group， the number of damaged cells in each treatment group decreased （P<0.01）. Transmission electron microscopy showed that compared with the normal group， the model group exhibited mitochondrial swelling and rupture in the hypothalamus， with visible autophagic lysosomes in the cytoplasm. Compared with those in the model group， only a few autophagic lysosomes were observed in the BZ-L group， and the degree of mitochondrial swelling and rupture was alleviated in the other treatment groups. ROS immunofluorescence assay showed that compared with the normal group， the model group displayed enhanced red fluorescence intensity of mitochondrial ROS （P<0.01）， which was attenuated in each treatment group （P<0.01）. Mitochondrial membrane potential assay showed that compared with the normal group， the model group had declined mitochondrial membrane potential （P<0.01）， which was recovered in each treatment group （P<0.05， P<0.01）. The biochemical assay of ATP levels indicated that compared with the normal group， the model group exhibited decreased ATP content （P<0.01）， which rebound in each treatment group （P<0.05， P<0.01）. ELISA showed that compared with the normal group， the model group exhibited decreased 5-HT content and increased NE and CORT content （P<0.01）. Compared with the model group， each treatment group showed increased 5-HT content （P<0.01） and reduced NE content （P<0.01） and CORT content （P<0.05， P<0.01）. Western blot of the hypothalamus showed that compared with the normal group， the model group exhibited downregulated protein levels of PINK1， Parkin， and LC3 （P<0.01） and upregulated protein level of p62 （P<0.01）. Compared with the model group， each treatment group showed upregulated protein levels of PINK1， Parkin， and LC3 （P<0.05， P<0.01） and downregulated protein level of p62 （P<0.05， P<0.01）. Real-time PCR showed that compared with the normal group， the model group exhibited downregulated mRNA levels of PINK1， Parkin， and LC3 （P<0.01） and upregulated mRNA level of p62 （P<0.01）. Compared with the model group， each treatment group showed upregulated mRNA levels of PINK1， Parkin， and LC3 （P<0.01） and downregulated mRNA level of p62 （P<0.01）. ConclusionBZ can ameliorate the insomnia and anxiety state of mice by regulating PINK1/Parkin signaling pathway to improve the mitochondrial function， enhance mitochondrial autophagy， protect neurons， and repair damaged nerve cells.

Adipose tissue is a critical energy reservoir in animals and humans, with multifaceted roles in endocrine regulation, immune response, and providing mechanical protection. Based on anatomical location and functional characteristics, adipose tissue can be categorized into distinct types, including white adipose tissue (WAT), brown adipose tissue (BAT), beige adipose tissue, and pink adipose tissue. Traditionally, adipose tissue research has centered on its morphological and functional properties as a whole. However, with the advent of single-cell transcriptomics, a new level of complexity in adipose tissue has been unveiled, showing that even under identical conditions, cells of the same type may exhibit significant variation in morphology, structure, function, and gene expression——phenomena collectively referred to as cellular heterogeneity. Single-cell transcriptomics, including techniques like single-cell RNA sequencing (scRNA-seq) and single-nucleus RNA sequencing (snRNA-seq), enables in-depth analysis of the diversity and heterogeneity of adipocytes at the single-cell level. This high-resolution approach has not only deepened our understanding of adipocyte functionality but also facilitated the discovery of previously unidentified cell types and gene expression patterns that may play key roles in adipose tissue function. This review delves into the latest advances in the application of single-cell transcriptomics in elucidating the heterogeneity and diversity within adipose tissue, highlighting how these findings have redefined the understanding of cell subpopulations within different adipose depots. Moreover, the review explores how single-cell transcriptomic technologies have enabled the study of cellular communication pathways and differentiation trajectories among adipose cell subgroups. By mapping these interactions and differentiation processes, researchers gain insights into how distinct cellular subpopulations coordinate within adipose tissues, which is crucial for maintaining tissue homeostasis and function. Understanding these mechanisms is essential, as dysregulation in adipose cell interactions and differentiation underlies a range of metabolic disorders, including obesity and diabetes mellitus type 2. Furthermore, single-cell transcriptomics holds promising implications for identifying therapeutic targets; by pinpointing specific cell types and gene pathways involved in adipose tissue dysfunction, these technologies pave the way for developing targeted interventions aimed at modulating specific adipose subpopulations. In summary, this review provides a comprehensive analysis of the role of single-cell transcriptomic technologies in uncovering the heterogeneity and functional diversity of adipose tissues.

The traditional Chinese medicine(TCM) industry is a crucial part of China's pharmaceutical sector and plays a strategic role in ensuring public health and promoting economic and social development. In response to the practical demand for high-quality development of the TCM industry, this paper focused on the bottlenecks encountered during the digital and intelligent transformation of TCM production systems. Specifically, it explored technical strategies and methodologies for constructing the best TCM production mode. An innovative artificial intelligence(AI)-centered technical architecture for TCM production was proposed, focusing on key aspects of production management including process modeling, state evaluation, and decision optimization. Furthermore, a series of critical technologies were developed to realize the best TCM production mode. Finally, a novel AI-driven TCM production mode characterized by a closed-loop system of "measurement-modeling-decision-execution" was presented through engineering case studies. This study is expected to provide a technological pathway for developing new quality productive forces within the TCM industry.
Artificial Intelligence ; Drugs, Chinese Herbal ; Medicine, Chinese Traditional/methods* ; Humans

OBJECTIVE: To investigate the midterm clinical efficacy of medial patellofemoral complex (MPFC) reconstruction for recurrent patellar dislocation with high-grade trochlear dysplasia. METHODS: A retrospective analysis was carried out among adult patients who underwent arthroscopically assisted MPFC reconstruction between January 2014 and December 2020. Dejour classification was evaluated to grade trochlear dysplasia; tibial tubercle-trochlear groove (TT-TG) distance and Insall-Salvati index were measured. Preoperative and postoperative patient-reported outcome measures (PROMs) were compared, including International Knee Documentation Committee (IKDC) score, Kujala score, Lysholm score and Tegner score. Information regarding returning-to-sport rate, re-instability events and complications was collected. Patellar tilt (PT), lateral patellar displacement (LPD) and bisect offset (BSO) ratio were measured based on axial computed tomography before and after surgery to assess the patellofemoral congruence. RESULTS: A total of 46 MPFC reconstructions in 43 patients were enrolled, including 16 male and 27 female. Mean age at surgery was (22.2±7.6) years (range: 14-44 years). Mean follow-up was (49.9±22.6) months (range: 18-102 months). The percentages of Dejour B, C and D dysplasia were 37.0% (17/46), 43.5% (20/46), and 19.6% (9/46), respectively. Mean Insall-Salvati index was 1.2±0.2 (range: 0.85-1.44), and mean TT-TG distance was (19.6±3.5) mm (range: 10.6-28.7 mm). At latest follow-up, there were significant improvements in all PROMs (P < 0.001): IKDC score, from 56.3±15.1 to 86.2±8.1; Kujala score, from 58.9±15.6 to 92.6±5.4; Lysholm score, from 63.7±15.0 to 94.0±5.7; Tegner score, from 3.1±1.4 to 4.7±1.4, and there were no significant differences in the improvements of the scores between the patients with Dejour B, C and D dysplasia. Overall, ninety percent of the patients returned to their preoperative sports level. One patient reported a postoperative subluxation, while no cases of infection, limited range of motion or patella fracture were observed. PT, LPD and BSO ratio were all significant altered (P < 0.001) after MPFC reconstruction. CONCLUSION Arthroscopically assisted MPFC reconstruction yielded satisfactory midterm clinical results for recurrent patellar dislocation with high-grade trochlear dysplasia. No significant differences of improvements in knee function were observed among the three types of high-grade trochlear dysplasia.
Humans ; Patellar Dislocation/surgery* ; Male ; Female ; Adult ; Arthroscopy/methods* ; Retrospective Studies ; Adolescent ; Young Adult ; Patellofemoral Joint/surgery* ; Recurrence ; Plastic Surgery Procedures/methods* ; Patella/surgery* ; Treatment Outcome

Ergopeptines or their derivatives are widely used for treating neurodegenerative and cerebrovascular diseases. The nonribosomal peptide synthetase-d-lysergyl peptide synthetase A (LPSA) determines ergopeptine formation but the detailed mechanism remains to be elucidated. Here, we characterized two LPSAs from Claviceps purpurea Cp-1 strain through heterologous expression in Aspergillus nidulans feeding with d-lysergic acid. We proved that Cp-LPSA1 catalyzed the formation of ergocornine, α-ergocryptine, and β-ergocryptine, precisely controlled by the substrate specificity of its three modules. Cp-LPSA2 was initially inactive but could be restored to catalyze α-ergosine formation. Using this platform, we validated that P1-LPSA1 and P1-LPSA2 from the reported C. purpurea P1 strain catalyzed ergotamine and α-ergocryptine formation, respectively. Typically, the non-ribosomal peptide codes implicated in every module of the LPSAs were defined and elucidated, in which certain key residues could play a switched role for substrate specificity and product interconversion. By constructing chimeric LPSAs through module assembly, the production of the desired ergopeptines was achieved. Notably, 1.46 mg/L of α-ergocryptine and 1.09 mg/L of ergotamine were produced respectively by mixed-culture of C. paspali No. 24 (fermentation supernatant) and the recombinants of A. nidulans. Our findings provide insights into the biosynthetic mechanism of ergopeptines and lay a foundation for directed ergopeptine biosynthesis.