(+)-Borneol, the main component of "Natural Borneol" in the Chinese Pharmacopoeia, is a high-end spice and precious medicine. Plant extraction cannot meet the increasing demand for (+)-borneol, while microbial biosynthesis offers a sustainable supply route. However, its production was extremely low compared with other monoterpenes, even with extensively optimizing the mevalonate pathway. We found that the key challenge is the complex and unusual dephosphorylation reaction of bornyl diphosphate (BPP), which suffers the side-reaction and the competition from the cellular dephosphorylation process, especially lipid metabolism, thus limiting (+)-borneol synthesis. Here, we systematically optimized the dephosphorylation process by identifying, characterizing phosphatases, and balancing cellular dephosphorylation metabolism. For the first time, we identified two endogenous phosphatases and seven heterologous phosphatases, which significantly increased (+)-borneol production by up to 152%. By engineering BPP dephosphorylation and optimizing the MVA pathway, the production of (+)-borneol was increased by 33.8-fold, which enabled the production of 753 mg/L under fed-batch fermentation in shake flasks, so far the highest reported in the literature. This study showed that rewiring dephosphorylation metabolism was essential for high-level production of (+)-borneol in Saccharomyces cerevisiae, and balancing cellular dephosphorylation is also helpful for efficient biosynthesis of other terpenoids since all whose biosynthesis involves the dephosphorylation procedure.

Ligusticum sinense cv.Fuxiong derived from"Xiongqiong",and in the Song Dynasty's Tai Ping Hui Min He Ji Ju Fang,it was already distinguished from Chuanxiong Rhizoma in usage.Doctors in the Ming Dynasty further clarified the differences in efficacy between the two.However,with the widespread circulation of Chuanxiong Rhizoma,Ligusticum sinense cv.Fuxiong gradually became marginalized,serving as a substitute,and is now only cultivated and used in small quantities within Jiangxi Province.This article reviewed the prescriptions and the ancient Chinese medical books that have records of Ligusticum sinense cv.Fuxiong,analyzed its specific applications alongside Chuanxiong Rhizoma in prescriptions and case studies to elucidate their differences in efficacy:Ligusticum sinense cv.Fuxiong is pungent and has strong dispersing power and is good at unblocking meridians and promoting qi circulation,which is suitable for excessive syndromes;Chuanxiong Rhizoma is good at promoting blood circulation and relieving pain,and is good at regulating and nourishing,which is suitable for patients with deficiency syndromes,aiming to provide insights and recommendations for the further development and rational clinical medication of Ligusticum sinense cv.Fuxiong.

In recent years， hyperuricemia （HUA） has shown a rapidly increasing incidence and tends to occur in increasingly young people， with a wide range of cardiac， renal， joint， and cancerous hazards and all-cause mortality associations. Western medicine treatment has limitations such as large liver and kidney damage， medication restriction， and easy recurrence. The intestine is the major extra-renal excretion pathway for uric acid （UA）， and the intestinal microecology can be regulated to promote UA degradation. It offers great potential to develop UA-lowering strategies that target the intestinal microecology， which are promising to provide safer and more effective therapeutic approaches. Traditional Chinese medicine （TCM） can treat HUA via multiple targets and multiple pathways from a holistic view， with low toxicity and side effects. Studies have shown that intestinal microecology is a crucial target for TCM in the treatment of HUA. However， its specific mechanism of action has not been fully elucidated. Focusing on the key role of intestinal microecology in HUA， this review explores the relationship between intestinal microecology and HUA in terms of intestinal flora， intestinal metabolites， intestinal UA transporters， and intestinal barriers. Furthermore， we summarize the research progress in TCM treatment of HUA by targeting the intestinal microecology， with the aim of providing references for the development of TCM intervention strategies for HUA and the direction of future research.

Ligusticum sinense cv.Fuxiong derived from"Xiongqiong",and in the Song Dynasty's Tai Ping Hui Min He Ji Ju Fang,it was already distinguished from Chuanxiong Rhizoma in usage.Doctors in the Ming Dynasty further clarified the differences in efficacy between the two.However,with the widespread circulation of Chuanxiong Rhizoma,Ligusticum sinense cv.Fuxiong gradually became marginalized,serving as a substitute,and is now only cultivated and used in small quantities within Jiangxi Province.This article reviewed the prescriptions and the ancient Chinese medical books that have records of Ligusticum sinense cv.Fuxiong,analyzed its specific applications alongside Chuanxiong Rhizoma in prescriptions and case studies to elucidate their differences in efficacy:Ligusticum sinense cv.Fuxiong is pungent and has strong dispersing power and is good at unblocking meridians and promoting qi circulation,which is suitable for excessive syndromes;Chuanxiong Rhizoma is good at promoting blood circulation and relieving pain,and is good at regulating and nourishing,which is suitable for patients with deficiency syndromes,aiming to provide insights and recommendations for the further development and rational clinical medication of Ligusticum sinense cv.Fuxiong.

Objective To investigate the effect of electroacupuncture on nutritional obese mice through TLR5-gut microbio-ta pathway,and the underlying mechanism.Methods A total of 60 SPF C57BL/6 mice were selected.Among them,10 blank mice were assigned to normal group,and the nutritive obesity model was established in the other 50 mice,which was induced by high-fat diet.Twenty nutritive obesity model mice were screened and randomly divided into model group and electroacupuncture group,with 10 mice in each group.The electroacupuncture group was needled at Zhongwan,Tianshu,Guanyuan and Fenglong points.And after obtaining Qi,electroacupuncture treatment was conducted on those mice for 21 days continuously.The change in differences of general indexes(body weight,Lee's index),tissue indexes(perirenal adipose-tissue mass),serum indexes[cho-lesterol(CHO),triglyceride(TG),high-density lipoprotein(HDL-C),low-density lipoprotein(LDL-C),blood glucose),molecular indexes[leptin(LEP),adiponectin(ADPN),interleukin-6(IL-6),interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α),lipopo-lysaccharide-binding protein(LBP)and cholecystokinin(CCK)]were observed.TLR5 protein distribution was detected by immu-nohistochemical reaction,and TLR5 gene expression was detected by RT-PCR.Structural differences of intestinal flora was ana-lyzed by second generation 16S rDNA gene sequencing technology.Results The body weight,Lee's index,fat mass around kid-ney,serum contents of CHO,TG,HDL-C,the contents of LEP and FFA in fat tissue,and contents of IL-6,IL-1β,TNF-α,LBP in intestinal mucosa tissue of model group were significantly higher than that of the normal group,and the content of adipocyto-kine ADP was significantly lower.After electroacupuncture treatment,those indicators were significantly improved.The protein distribution and gene expression of TLR5 in the intestinal mucosa of the model group were significantly higher than those of the normal group,and were significantly down-regulated after electroacupuncture intervention.The number of intestinal flora spe-cies in the model group was significantly lower than those in the normal group,and were significantly increased after electroacu-puncture intervention.Proportions of Bacteroidia,Gammaproteobacteria,Eggerthellacrae and Sutterellaceae were significantly increased,while proportions of Bacilli,Clostridia,Clostridiaceae and Actinobacteria were significantly decreased.Changes in gut flora were significantly reversed after electroacupuncture intervention.Conclusion Electroacupuncture has a satisfying effect in treatment of nutritional obesity.It can regulate the distribution and expression of TLR5 protein in intestinal mucosa by balancing the structure of intestinal flora,reduce the chronic inflammatory reaction of intestinal mucosa,promote lipid metabo-lism of the body,and achieve the purpose of weight loss and lipid reduction.

Steatotic liver diseases (SLD) are the principal worldwide cause of cirrhosis and end-stage liver cancer, affecting nearly a quarter of the global population. SLD includes metabolic dysfunction-associated alcoholic liver disease (MetALD) and metabolic dysfunction-associated steatotic liver disease (MASLD), resulting in asymptomatic liver steatosis, fibrosis, cirrhosis and associated complications. The immune processes include gut dysbiosis, adiposeliver organ crosstalk, hepatocyte death and immune cell-mediated inflammatory processes. Notably, various immune cells such as B cells, plasma cells, dendritic cells, conventional CD4+ and CD8+ T cells, innate-like T cells, platelets, neutrophils and macrophages play vital roles in the development of MetALD and MASLD. Immunological modulations targeting hepatocyte death, inflammatory reactions and gut microbiome include N-acetylcysteine, selonsertib, F-652, prednisone, pentoxifylline, anakinra, JKB-121, HA35, obeticholic acid, probiotics, prebiotics, antibiotics and fecal microbiota transplantation. Understanding the immunological mechanisms underlying SLD is crucial for advancing clinical therapeutic strategies.

For wild natural medicine, unanticipated biodiversity as species or varieties with similar morphological characteristics and sympatric distribution may co-exist in a single batch of medical materials, which affects the efficacy and safety of clinical medication. DNA barcoding as an effective species identification tool is limited by its low sample throughput nature. In this study, combining DNA mini-barcode, DNA metabarcoding and species delimitation method, a novel biological sources consistency evaluation strategy was proposed, and high level of interspecific and intraspecific variations were observed and validated among 5376 Amynthas samples from 19 sampling points regarded as "Guang Dilong" and 25 batches of proprietary Chinese medicines. Besides Amynthas aspergillum as the authentic source, 8 other Molecular Operational Taxonomic Units (MOTUs) were elucidated. Significantly, even the subgroups within A. aspergillum revealed here differ significantly on chemical compositions and biological activity. Fortunately, this biodiversity could be controlled when the collection was limited to designated areas, as proved by 2796 "decoction pieces" samples. This batch biological identification method should be introduced as a novel concept regarding natural medicine quality control, and to offer guidelines for in-situ conservation and breeding bases construction of wild natural medicine.

Tripterygium wilfordii is a valuable medicinal plant rich in biologically active diterpenoids, but there are few studies on the origins of these diterpenoids in its secondary metabolism. Here, we identified three regions containing tandemly duplicated diterpene synthase genes on chromosomes (Chr) 17 and 21 of T. wilfordii and obtained 11 diterpene synthases with different functions. We further revealed that these diterpene synthases underwent duplication and rearrangement at approximately 2.3-23.7 million years ago (MYA) by whole-genome triplication (WGT), transposon mediation, and tandem duplication, followed by functional divergence. We first demonstrated that four key amino acids in the sequences of TwCPS3, TwCPS5, and TwCPS6 were altered during evolution, leading to their functional divergence and the formation of diterpene secondary metabolites. Then, we demonstrated that the functional divergence of three TwKSLs was driven by mutations in two key amino acids. Finally, we discovered the mechanisms of evolution and pseudogenization of miltiradiene synthases in T. wilfordii and elucidated that the new function in TwMS1/2 from the terpene synthase (TPS)-b subfamily was caused by progressive changes in multiple amino acids after the WGT event. Our results provide key evidence for the formation of diverse diterpenoids during the evolution of secondary metabolites in T. wilfordii.

Sterol C24-methyltransferase (SMT) plays multiple important roles in plant growth and development. SMT1, which belongs to the family of transferases and transforms cycloartenol into 24-methylene cycloartenol, is involved in the biosynthesis of 24-methyl sterols. Here, we report the cloning and characterization of a cDNA encoding a sterol C24-methyltransferase from().(GenBank access number KU885950) is a 1530 bp cDNA with a 1041 bp open reading frame predicted to encode a 346-amino acid, 38.62 kDa protein. The polypeptide encoded by thecDNA was expressed and purified as a recombinant protein from() and showed SMT activity. The expression ofwas highly up-regulated incell suspension cultures treated with methyl jasmonate (MeJA). Tissue expression pattern analysis showed higher expression in the phellem layer compared to the other four organs (leaf, stem, xylem and phloem), which is about ten times that of the lowest expression in leaf. The results are meaningful for the study of sterol biosynthesis ofand will further lay the foundations for the research in regulating both the content of other main compounds and growth and development of

The 4-hydroxy-3-methylbut-2-enyl diphosphate reductase (HDR) is the last step key enzyme of the methylerythritol phosphate (MEP) pathway, synthesizing isopentenyl diphosphate and its allyl isomer dimethylallyl diphosphate, which is important for regulation of isoprenoid biosynthesis. Here the full-length cDNA of, designated(GenBank Accession No. KJ933412.1), was isolated fromfor the first time. TwHDR has an open reading frame (ORF) of 1386 bp encoding 461 amino acids. TwHDR exhibits high homology with HDRs of other plants, with an N-terminal conserved domain and three conserved cysteine residues.cDNA was cloned into an expression vector and transformed into anmutant. Since loss-of-functionmutant is lethal, the result showed that transformation ofcDNA rescued themutant. This complementation assay suggests that thecDNA encodes a functional HDR enzyme. The expression ofwas induced by methyl-jasmonate (MJ) insuspension cells. The expression ofreached the highest level after 1 h of MJ treatment. These results indicate that we have identified a functional TwHDR enzyme, which may play a pivotal role in the biosynthesis of diterpenoid triptolide in.