1.Comparison of Wild and Cultivated Alpiniae Oxyphyllae Fructus Based on Traditional Quality Evaluation
Fengfan WANG ; Yajie XIANG ; Jian FENG ; Wencheng HOU ; Wenlan LI ; Yangyang LIU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(5):235-244
ObjectiveTo compare the differences between wild Alpiniae Oxyphyllae Fructus(WAOF) and cultivated Alpiniae Oxyphyllae Fructus(CAOF) through a traditional quality evaluation system for medicinal materials. MethodsA total of 10 batches of WAOF and 12 batches of CAOF samples were collected from various regions of Hainan province. Relevant analytical methods from the 2020 edition of the Pharmacopoeia of the People's Republic of China were employed to observe the characteristics of WAOF and CAOF, followed by microscopic identification, thin-layer chromatography(TLC) identification, moisture content(toluene method), total ash, acid-insoluble ash, water-soluble and alcohol-soluble extracts(hot dipping method), water-soluble protein, total polysaccharides and total flavonoids(ultraviolet spectrophotometry), and volatile oil content(method A under general rule 2204). The contents of five active components(protocatechuic acid, chrysin, kaempferol, tectochrysin and nootkatone) were quantified using ultra-performance liquid chromatography(UPLC), and the antioxidant activity was evaluated. Building upon traditional quality evaluation of AOF, quantitative measurements were conducted on its appearance traits including diameter, length, plumpness(diameter/length ratio), and color. Canonical correlation analysis was performed using SPSS 26.0 to explore relationships between appearance traits and intrinsic quality. ResultsNo significant differences were observed between WAOF and CAOF in microscopic observation, TLC identification, moisture content, protocatechuic acid content, kaempferol content, odor, or antioxidant activity measured by 2,2ʹ-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)(ABTS) method. WAOF exhibited significantly higher levels in water-soluble extracts, alcohol-soluble extracts, total polysaccharide content, water-soluble protein content, 100-grain weight, length, and total color difference(ΔE*ab) compared to CAOF(P<0.01). In contrast, CAOF showed significantly higher levels of total ash, acid-insoluble ash, content of total flavonoids, volatile oil content, chrysin content, tectochrysin content, nootkatone content, diameter, plumpness, lightness(L*), red-green chromaticity(a*), yellow-blue chromaticity(b*), and antioxidant activity measured by 1,1-diphenyl-2-picrylhydrazyl(DPPH) method compared to WAOF(P<0.01). Correlation analysis between 7 phenotypic traits and 8 quality traits revealed that among the phenotypic traits, plumpness, L*, a*, and b* exerted significant influence on intrinsic quality. Among the quality traits, total flavonoids, volatile oils, nootkatone, chrysin, and tectochrysin contributed substantially to intrinsic quality. ConclusionPlumpness, L*, a*, and b* of AOF significantly influence its intrinsic quality, and higher values of these parameters indicate relatively superior intrinsic quality. The comprehensive quality evaluation reveals that CAOF samples collected in this study are superior to their wild counterparts.
2.Exploring on Quality Evaluation Methods of Clinical Case Reports in Traditional Chinese Medicine Based on China Clinical Cases Library of Traditional Chinese Medicine
Kaige ZHANG ; Feng ZHANG ; Bo ZHOU ; Haimin CHEN ; Yong ZHU ; Changcheng HOU ; Liangzhen YOU ; Weijun HUANG ; Jie YANG ; Guoshuang ZHU ; Shukun GONG ; Jianwen HE ; Yang YE ; Yuqiu AN ; Chunquan SUN ; Qingjie YUAN ; Buman LI ; Xingzhong FENG ; Kegang CAO ; Hongcai SHANG ; Jihua GUO ; Xiaoxiao ZHANG ; Zhining TIAN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(1):271-276
As the core vehicle for preserving and transmitting traditional Chinese medicine(TCM) academic thought and clinical experience, the establishment of a robust quality evaluation system for TCM clinical case reports is a crucial component in the current standardization and modernization of TCM. Based on the practical experience of constructing the China Clinical Cases Library of Traditional Chinese Medicine by the China Association of Chinese Medicine, this study conducted a comprehensive analysis of critical challenges, including insufficient authenticity and unfocused evaluation criteria. It proposed a three-dimensional evaluation framework grounded in the structure-process-outcome logic, encompassing three dimensions of authenticity and standardization, characteristics and advantages, application and translational impact. This framework integrated 12 key evaluation indicators in a systematic manner. The model preserved the academic characteristics of TCM syndrome differentiation and treatment, while aligning with modern scientific research standards, achieving a balance between individualized TCM experience and standardized evaluation. Concurrently, this study provided theoretical foundations and methodological guidance for evaluating the quality of TCM clinical cases, contributing significantly to the inheritance of TCM knowledge, evidence-based practice, and the reform of talent evaluation mechanisms.
3.Differentiation and Treatment of Small Cell Lung Cancer from the Perspective of "Internal Wind in Hidden Circulation"
Shengjuan HU ; Li HOU ; Tao SUN ; Li FENG
Journal of Traditional Chinese Medicine 2026;67(9):1003-1007
Small cell lung cancer (SCLC) is characterized by rapid onset, high invasiveness, and a strong tendency for recurrence and metastasis, which aligns with the pathogenic characteristics of wind pathogen in traditional Chinese medicine (TCM). This paper explores the pathological mechanism and dynamic pattern identification and treatment of SCLC from the perspective of "internal wind in hidden circulation". It is proposed that the core pathogenesis of SCLC is rooted in depletion of healthy qi, with binding of phlegm, stasis, and toxin. When pathogenic factors become excessive, the ascending and descending of yang qi becomes disordered, transforming into wind. This leads to internal wind in hidden circulation, which moves erratically and damages healthy qi. In the limited stage, cancer toxin accumulates and internal wind arises covertly, treatment for which should focus on regulating qi and resolving toxin, defending against wind and resisting pathogen with modified Bufei Decoction (补肺汤) and Shengjiang Powder (升降散). In the early extensive stage, phlegm and stasis generate wind, and internal wind spreads through collate-rals; treatment should resolve phlegm and dispel stasis, extinguish wind and resolve toxin, with modified Lingjiao Gouteng Decoction (羚角钩藤汤) combined with Tianma Gouteng Beverage (天麻钩藤饮). During the treatment stage, there is qi and yin depletion, and deficient wind harassing the interior, for which it is recommended to boost qi and nourish yin, soften the liver and extinguish wind, with modified Zhengan Xifeng Decoction (镇肝熄风汤) combined with Qingzao Jiufei Decoction (清燥救肺汤). In the progression stage, internal wind stirs again and cancer toxin scurries; treatment should focus on strengthening the healthy qi and replenishing essence, restraining wind and penetrating toxin, with modified Sanjia Fumai Decoction (三甲复脉汤). In the terminal stage, yin and yang are on the verge of dissociation and depleted yang floats upward; treatment should constrain and astringe to prevent collapse, rescue yang and contain yin, with modified Dihuang Drink (地黄饮子) combined with Laifu Decoction (来复汤).
4.Differentiation and Treatment of Small Cell Lung Cancer from the Perspective of "Internal Wind in Hidden Circulation"
Shengjuan HU ; Li HOU ; Tao SUN ; Li FENG
Journal of Traditional Chinese Medicine 2026;67(9):1003-1007
Small cell lung cancer (SCLC) is characterized by rapid onset, high invasiveness, and a strong tendency for recurrence and metastasis, which aligns with the pathogenic characteristics of wind pathogen in traditional Chinese medicine (TCM). This paper explores the pathological mechanism and dynamic pattern identification and treatment of SCLC from the perspective of "internal wind in hidden circulation". It is proposed that the core pathogenesis of SCLC is rooted in depletion of healthy qi, with binding of phlegm, stasis, and toxin. When pathogenic factors become excessive, the ascending and descending of yang qi becomes disordered, transforming into wind. This leads to internal wind in hidden circulation, which moves erratically and damages healthy qi. In the limited stage, cancer toxin accumulates and internal wind arises covertly, treatment for which should focus on regulating qi and resolving toxin, defending against wind and resisting pathogen with modified Bufei Decoction (补肺汤) and Shengjiang Powder (升降散). In the early extensive stage, phlegm and stasis generate wind, and internal wind spreads through collate-rals; treatment should resolve phlegm and dispel stasis, extinguish wind and resolve toxin, with modified Lingjiao Gouteng Decoction (羚角钩藤汤) combined with Tianma Gouteng Beverage (天麻钩藤饮). During the treatment stage, there is qi and yin depletion, and deficient wind harassing the interior, for which it is recommended to boost qi and nourish yin, soften the liver and extinguish wind, with modified Zhengan Xifeng Decoction (镇肝熄风汤) combined with Qingzao Jiufei Decoction (清燥救肺汤). In the progression stage, internal wind stirs again and cancer toxin scurries; treatment should focus on strengthening the healthy qi and replenishing essence, restraining wind and penetrating toxin, with modified Sanjia Fumai Decoction (三甲复脉汤). In the terminal stage, yin and yang are on the verge of dissociation and depleted yang floats upward; treatment should constrain and astringe to prevent collapse, rescue yang and contain yin, with modified Dihuang Drink (地黄饮子) combined with Laifu Decoction (来复汤).
5.Guidelines for standardized implementation of pharmacist-managed clinics (2026 edition)
Pengxiang ZHOU ; Maobai LIU ; Xiaoli DU ; Xiaoyang LU ; Mei DONG ; Rong DUAN ; Ruigang HOU ; Xiaoyu LI ; Qi CHEN ; Yanxiao XIANG ; Weiyi FENG ; Rong CHEN ; Deshi DONG ; Yong YANG ; Li LI ; Xiaocong ZUO ; Jinfang HU ; Hongliang ZHANG ; Qingchun ZHAO ; Qi LIN ; Yang HU ; Jiaying WU ; Rongsheng ZHAO
China Pharmacy 2026;37(9):1105-1112
OBJECTIVE To formulate Guidelines for the standardized implementation of pharmacist-managed clinics ( 2026 edition ) in response to the challenges faced by such clinics in China, including uneven development, large discrepancies in service specifications, insufficient patient awareness, and limited medical insurance coverage. METHODS Led by the Pharmaceutical Affairs Professional Committee of the Chinese Hospital Association, the Evidence-based Pharmacy Professional Committee of the Chinese Pharmaceutical Association, and the Hospital Pharmacy Professional Committee of the Cross-strait Medical and Health Exchange Association, a total of 19 domestic hospital pharmacy experts were organized. Through a systematic review of national policies and literature research, current practical experience was summarized. Consensus on the contents of the guidelines was reached after in-depth discussions. RESULTS &CONCLUSIONS The guidelines covered five sections: definition and connotation of pharmacist-managed clinics, establishment requirements, implementation and management, post competency, and practical research. Firstly, the definition and connotation included three operational forms of pharmacist-managed clinics (independent mode, physician-pharmacist joint mode, and online pharmacist-managed clinic mode) and classified service modes (specialty-specific, drug-specific, and disease-specific pharmacist-managed clinics). The establishment requirements were further refined, covering system construction (pharmaceutical service management system, quality control and assessment mechanism), personnel qualifications (professional credentials, continuing education and professional training, etc), service recipients, as well as service venues and facilities. Subsequently, the implementation and management of pharmacist-managed clinics were proposed, involving service procedures, intervention measures, documentation and records, patient education and follow-up, humanistic care, as well as risk management and quality control. Finally, post competency encompassed the competency requirements for pharmacists providing services in pharmacist-managed clinics, as well as the suggestions on teaching methods; practical research encouraged the conduct of high-quality pharmaceutical practice in the setting of pharmacist-managed clinics. The guidelines provide valuable guidance for the standardized implementation of pharmacist-managed clinics in China in terms of establishment, management, teaching, and research, fill the guideline gap in this field, and can promote the high-quality development of pharmacist-managed clinics.
6.Construction of a residual astigmatism prediction model after Toric IOL implantation based on preoperative parameters
Yafei WANG ; Mingjia HOU ; Xiaoyan WANG ; Du FENG
International Eye Science 2026;26(7):1287-1292
AIM: To explore preoperative risk factors affecting residual astigmatism after Toric intraocular lens(IOL)implantation, and construct and verify a clinically applicable prediction model.METHODS: This retrospective study included patients who underwent Toric IOL implantation at Puyang Oilfield General Hospital and the Second People's Hospital of Puyang City from October 2023 to August 2025. Patients were categorized into two groups according to residual astigmatism at 3 mo postoperatively: >0.25 D group and ≤0.25 D group. Preoperative clinical data and ocular parameters of the two groups were collected and compared. Multivariate Logistic regression analysis was used to screen risk factors, and a nomogram prediction model was established accordingly. Receiver operating characteristic(ROC)curve, calibration curve, and Hosmer-Lemeshow test were adopted to evaluate discrimination, calibration, and clinical applicability of the model. Bootstrap method was applied for internal validation.RESULTS: Totally 103 patients(103 eyes)were included, with 72 patients(72 eyes)in the group with residual astigmatism ≤0.25 D, including 38 males and 34 females, with an average age of 67.35±6.12 y; There were 31 cases(31 eyes)in group with residual astigmatism >0.25 D, including 19 males and 12 females, with an average age of 69.87±6.45 y. The proportion of patients with a history of diabetes, as well as the values for axial length, horizontal corneal diameter, flat corneal curvature(K1), steep corneal curvature(K2), astigmatic axis and proportion of lens nuclear hardness grade III-IV were significantly higher in the residual astigmatism group >0.25 D than those in the residual astigmatism ≤0.25 D group(all P<0.05). Multivariate analysis identified increased horizontal corneal diameter, higher K1 value, higher K2 value, and higher lens nuclear hardness(grade III-IV)as independent risk factors for postoperative residual astigmatism >0.25 D(all P<0.05). The nomogram constructed from these factors demonstrated that the area under the ROC curve(AUC)was 0.794, with a sensitivity of 80.31% and a specificity of 75.20%. The concordance index(C-index)of internal validation was 0.865. Calibration curve and Hosmer-Lemeshow test(χ2=5.231, P=0.734)indicated good consistency between predicted and actual values of the model.CONCLUSION: The nomogram model established based on horizontal corneal diameter, K1, K2 and lens nucleus hardness exhibits favorable predictive performance. It can effectively predict the risk of residual astigmatism prior to Toric IOL surgery, providing reference for individualized clinical surgical planning.
7.The mechanism of magnoflorine in inhibiting colon cancer based on network pharmacology and in vitro experiment
Lulu TAN ; Lina ZHU ; Shujin ZHANG ; Yuxuan WANG ; Huimei LI ; Yuke WANG ; Jiayi HOU ; Qilong FENG ; Jianyun SHI
Acta Universitatis Medicinalis Anhui 2026;61(6):1021-1031
ObjectiveTo explore the function and related molecular mechanisms of magnoflorine against colon cancer via network pharmacology, molecular docking, and in vitro cell experiments. MethodsIn this study, the canonical SMILES of magnoflorine was obtained from the PubChem database, and the potential targets of magnoflorine were predicted by the Swiss Target Prediction database, while the disease targets of colon cancer were obtained from the DisGeNET, GeneCards and OMIM databases. The intersecting targets between magnoflorine's predicted targets and colon cancer disease targets were taken, and a protein-protein interaction (PPI) network was constructed and analyzed. The DAVID online database was employed to conduct Gene Ontology (GO) and KEGG pathway enrichment analyses on core targets. The top five key targets screened were docked with magnoflorine using AutoDock software. Finally, cellular experiments including CCK-8 assays, EdU experiments, cell scratch assays, and Transwell assays were conducted to validate the results from network pharmacology and molecular docking. Results44 key targets of magnoflorine in resisting colon cancer were acquired. The molecular docking results showed that magnoflorine had a strong binding activity with the core target signal transducer and activator of transcription 3 (STAT3) in the top five of the PPI network. Cellular experiments confirmed that magnoflorine could inhibit the proliferation and migration of colon cancer cells by suppressing the JAK/STAT3 signaling pathway. ConclusionMagnoflorine may inhibit the proliferation and migration of colon cancer cells by regulating the JAK/STAT3 signaling pathway.
8.α7 Nicotinic Acetylcholine Receptor: From Structural Mechanisms to Therapeutic Translation
Yu-Han LIU ; Jiao-Feng CHEN ; Hong-Juan WANG ; Huan CHEN ; Hong-Wei HOU
Progress in Biochemistry and Biophysics 2026;53(9):2378-2391
The α7 nicotinic acetylcholine receptor (α7 nAChR) is a ligand-gated ion channel characterized by high Ca2+ permeability, rapid activation, and rapid desensitization. It is widely expressed in the nervous system and various non-neuronal cell types. α7 nAChR is involved in regulating neuronal excitability, synaptic transmission and plasticity, cognitive function, and inflammatory responses. These properties have made α7 nAChR a potential therapeutic target for neuropsychiatric and inflammatory diseases. In recent years, advances in cryo-electron microscopy have provided structural insights into α7 nAChR gating, ligand recognition, allosteric modulation, and allosteric activation. The receptor mainly undergoes transitions among resting, open, and desensitized states. Recent structural studies have further revealed asymmetric conformational changes and multiple intermediate states during receptor activation and recovery from desensitization. Conformational rearrangements in the extracellular domain-transmembrane domain coupling region and the pore-forming M2 helices are closely associated with channel opening and desensitization. However, structural characterization of the α7 nAChR gating cycle still largely depends on pharmacological stabilization. Open-state structures have been resolved in the presence of PNU-120596 or GAT107, whereas a high-resolution structure of the open state induced by acetylcholine alone remains to be determined. These physiological roles underpin the involvement of α7 nAChR in multiple disease processes. In the hippocampus, α7 nAChR regulates local neuronal circuits, theta oscillations, synaptic transmission, and synaptic plasticity, thereby contributing to learning, memory, attention, and other cognitive processes. In macrophages, microglia, and other immune-related cells, α7 nAChR activation regulates inflammatory responses and oxidative stress. These effects involve several signaling pathways, including Janus kinase 2 (JAK2)/STAT3, phosphoinositide 3-kinase (PI3K)/AKT, and NF-κB signaling, as well as regulation of the NLRP3 inflammasome and IL-1β production. Such anti-inflammatory effects may also contribute to α7 nAChR-mediated neuroprotection. Although several compounds targeting α7 nAChR have entered clinical studies, no α7 nAChR-selective drug has been approved for clinical use. Limited clinical translation may be related to rapid receptor desensitization, inconsistent efficacy across clinical studies, insufficient or variable target engagement, patient heterogeneity, and clinical endpoint selection. TC-5619 showed promising effects on cognitive impairment and negative symptoms in an early exploratory study of schizophrenia, but these effects were not confirmed in a subsequent, larger phase II trial. Encenicline (EVP-6124) also showed cognitive improvement in early studies, whereas two global phase III trials in schizophrenia failed to meet their co-primary cognitive and functional endpoints. Pharmacological strategies targeting α7 nAChR have expanded from orthosteric agonists to positive allosteric modulators (PAMs) and allosteric agonist-positive allosteric modulators (ago-PAMs). Orthosteric agonists directly activate the receptor by binding to the orthosteric site, but their effects are limited by rapid receptor desensitization. Type I PAMs mainly enhance agonist-evoked responses with relatively small effects on desensitization, whereas type II PAMs markedly potentiate receptor responses and reduce or delay desensitization. Ago-PAMs such as GAT107 can directly activate α7 nAChR through an allosteric mechanism while also potentiating responses to orthosteric agonists. Together, these findings have broadened the pharmacological approaches to α7 nAChR regulation. In summary, future drug development should focus on precise regulation of α7 nAChR according to receptor structure and disease mechanisms, rather than simply enhancing receptor activation. Further studies are needed to clarify how candidate ligands affect receptor gating, conformational transitions, recovery from desensitization, and downstream signaling. Pharmacological differences between neuronal and non-neuronal α7 nAChRs, between homomeric α7 and heteromeric α7β2 receptors, and between α7 nAChR and other nAChR subtypes also require further investigation. Integrating structure prediction, artificial intelligence-assisted virtual screening, molecular docking, molecular dynamics simulations, and electrophysiological validation may facilitate the discovery and optimization of new α7 nAChR modulators.
9.From Blood-brain Barrier Penetration to Barrier Functional Remodeling: New Intervention Strategies via Nanodelivery Systems for Alzheimer’s Disease
Xu-Ran WANG ; Chang-Feng YIN ; Huan CHEN ; Hong-Wei HOU ; Yi-Kun WANG
Progress in Biochemistry and Biophysics 2026;53(9):2414-2429
Alzheimer’s disease (AD) is pathologically characterized by cerebral amyloid β‑protein (Aβ) aggregation, neurofibrillary tangles and progressive cognitive deterioration. There is an urgent clinical demand for targeted therapeutic agents against AD, whereas the blood-brain barrier (BBB) acts as a critical physical barrier that blocks over 98% small-molecule drugs and nearly all biomacromolecules from entering brain parenchyma. Nanomedicine-based drug delivery systems (NDDSs) with tunable physicochemical properties can cross the BBB via multiple transcytosis pathways including adsorptive-mediated, receptor-mediated and transporter-mediated routes, opening a promising avenue for targeted AD treatment. A core academic viewpoint proposed herein is that robust in vitro endothelial penetration of nanocarriers cannot guarantee effective accumulation in brain target cells. Comprehensive evaluation of BBB-crossing delivery efficiency should not merely rely on in vitro permeability tests, but cover the full multi-step transcytosis cascade, cellular tropism in brain tissues and in vivo therapeutic outcomes. This review systematically sorts out diverse nanoplatforms applicable to BBB penetration for AD intervention. Inorganic nanomaterials such as gold and ceria nanoparticles possess large specific surface areas and intrinsic antioxidant capacity, which eliminate reactive oxygen species and hinder Aβ fibrillization. Liposomal formulations and solid lipid nanoparticles exhibit superior biocompatibility with biomimetic phospholipid bilayer architectures, capable of co-loading hydrophilic nucleic acids for Tau regulation and lipophilic Aβ inhibitors; relying on receptor-mediated transcytosis, they achieve sustained drug retention in the brain. Polymeric nanocarriers including nanogels and polyamidoamine dendrimers enable multi-target combinatorial therapy, and can be engineered to release cargo in response to inflammatory microenvironments, thereby suppressing excessive microglial activation and protecting neuronal mitochondria. Beyond conventional nanocarriers, this work elaborates two cutting-edge BBB-crossing delivery platforms: biomimetic nanosystems and metal-organic frameworks (MOFs). Biomimetic nanoparticles camouflaged with erythrocyte, platelet or macrophage membranes, as well as natural exosomes, evade immune clearance, prolong systemic circulation and inherently home to inflammatory lesions. Serving as “nano-decoys”, they neutralize Aβ neurotoxins and remodel cerebral inflammatory microenvironments simultaneously. MOFs feature high porosity and customizable pore channels for co-delivery of multiple therapeutics, and can be integrated with near-infrared photothermal and photooxidation modalities to facilitate focal brain lesion therapy. This review highlights a transformative paradigm shift in the field of BBB-targeted AD therapy: research focus has shifted from simply maximizing cerebral drug penetration toward active modulation and functional restoration of the BBB. Impaired BBB transporters intrinsically impede endogenous Aβ clearance. Accordingly, BBB-regulating nanocarriers are designed to remodel the low-density lipoprotein receptor-related protein 1 (LRP1) trafficking cascade, redirecting endocytic vesicles from lysosomal degradation to non-degradable transcytosis and restoring the intrinsic Aβ efflux capacity of the BBB. Distinct from conventional strategies that only exert local lesion inhibition, this systemic clearance strategy eliminates cerebral Aβ deposits by accelerating peripheral excretion.
10.Cloning and Functional Characterization of UDP-rhamnose Synthases from Ornithogalum saundersiae
Panpan ZHANG ; Kangxin HOU ; Feng ZHANG ; Yue ZHANG ; Jinlong LIU ; Caixia WANG ; Xuemi HAO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(23):196-203
ObjectiveTo screen and identify uridine diphosphate (UDP)-rhamnose synthase (RHM) genes from Ornithogalum saundersiae and verify their UDP-rhamnose (UDP-Rha) biosynthetic functions, thereby providing reliable candidate genes for the efficient biosynthesis of neohesperidin and other rhamnoside natural products. MethodsBased on previously reported plant RHM sequences, two candidate RHM genes, OsRHM1 and OsRHM2, were identified from O. saundersiae through transcriptome screening and sequence alignment. The two genes were cloned and subjected to bioinformatics analysis and multiple sequence alignment. An in vitro RHM-Cm1,2RhaT cascade reaction was established, and the ability of OsRHM1 and OsRHM2 to catalyze the conversion of UDP-glucose (UDP-Glc) to UDP-Rha was evaluated based on neohesperidin production. Real-time fluorescence quantitative polymerase chain reaction PCR (Real-time PCR) was performed to analyze the relative expression levels of the two genes in roots, bulbs, and leaves. ResultsThe open reading frame (ORF) of OsRHM1 was 2 076 bp and encoded 692 amino acids, with a predicted protein molecular mass of 77.97 kDa and a theoretical isoelectric point of 7.25. The ORF of OsRHM2 was 2 019 bp and encoded 673 amino acids, with a predicted protein molecular mass of 75.82 kDa and a theoretical isoelectric point of 7.24. Multiple sequence alignment showed that OsRHM1 and OsRHM2 contained the typical conserved motifs GxxGxxG/A and YxxxK of the plant RHM family and shared high amino acid sequence identity with previously reported RHMs. The in vitro cascade reaction demonstrated that both OsRHM1 and OsRHM2 were capable of catalyzing the conversion of UDP-Glc to UDP-Rha. The neohesperidin production in the OsRHM2 cascade reaction system was approximately 2.58-fold that in the OsRHM1 system. Real-time PCR analysis showed that the relative expression levels of OsRHM1 and OsRHM2 were higher in roots than in bulbs and leaves. ConclusionOsRHM2 exhibited a relatively high capacity for UDP-Rha supply and may serve as a candidate enzyme resource for the subsequent biosynthesis of rhamnoside natural products and the construction of UDP-Rha supply systems.

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