Objective: To develop a convenient, direct, and highly sensitive method for screening trace chemical additives in complex Chinese patent medicines, thereby addressing core technological bottlenecks in pharmaceutical analysis and quality control. Methods: A brush ionization probe device was independently designed and constructed, and an efficient detection method was established through systematic optimization of key parameters. Twenty-three Chinese patent medicine samples, representing 6 dosage forms (capsules, tablets, pills, granules, powders, and liquid preparations), were analyzed using 10 common chemical additives as target analytes. Results: All samples were successfully analyzed without complex pretreatment, and 5 chemical additives were detected in 7 Chinese patent medicines. The brush ionization probe device exhibited cost-effectiveness (~0.2 USD per probe), operational simplicity, rapid analysis (~10s per sample), high efficiency, and minimal reagent consumption (~10 μL per sample). Conclusion: This advancement is expected to provide an innovative scientific tool for improving the generality and convenience of on-site quality control, while promoting technological progress in disciplines such as pharmacology and traditional Chinese medicine.

OBJECTIVE To investigate the apoptosis-inducing effect of esculetin （Esc） on acute myeloid leukemia （AML） HL-60 cells by regulating the protein kinase B （AKT）/S-phase kinase-associated protein 2 （SKP2）/MutT homolog 1 （MTH1） pathway. METHODS AML HL-60 cells were randomly divided into control group （routine culture）， Esc low-concentration group （L-Esc group， 25 μmol/L Esc）， Esc medium-concentration group （M-Esc group， 50 μmol/L Esc）， Esc high-concentration group （H-Esc group， 100 μmol/L Esc）， and high-concentration of Esc+ SC79 （AKT agonist） group （100 μmol/L Esc+5 μmol/L SC79）. Cell proliferation in each group was detected by MTT assay and colony formation assay. The level of reactive oxygen species （ROS） in cells was measured by using the CM-H2DCFDA fluorescent probe. Cell apoptosis was analyzed by flow cytometry. Western blot assay was performed to detect the expression levels of apoptosis-related proteins [B-cell lymphoma 2 （Bcl-2）， Bcl-2-associated X protein （Bax）， cleaved caspase-3]， AKT/SKP2/MTH1 pathway-related proteins （p-AKT， AKT， SKP2， MTH1）， along with the upstream and downstream proteins of AKT phosphatidylinositol 3-kinase （PI3K）， cyclin-dependent kinase inhibitor 1 （P21） and cyclin-dependent kinase inhibitor 1B （P27）. RESULTS Compared with control group， the cell viability， colony number， and the phosphorylation levels of AKT and PI3K proteins as well as protein expressions of SKP2， MTH1 and Bcl-2 were significantly decreased （P＜0.05）， while ROS level， apoptosis rate， and the expression levels of Bax， cleaved caspase-3， P21 and P27 proteins were significantly increased （P＜0.05）. Moreover， the effects of Esc exhibited concentration-dependence （P＜0.05）. Compared with H-Esc group， above indexes of high-concentration of Esc+ SC79 group were reversed significantly （P＜0.05）. CONCLUSIONS Esc may promote massive ROS production and induce activation of apoptosis in HL-60 cells by inhibiting the AKT/SKP2/MTH1 pathway， thus inhibiting the proliferation of HL-60 cells.

Objective To establish an oxidative stress mouse model of atopic dermatitis (AD) by applying 2,4-dinitrofluorobenzene (DNFB) to the back and post-auricular skin of KM mice, and to evaluate the regulatory role of the RAGE-NLRP3 axis (receptor for advanced glycation end products-NOD-like receptor family, pyrin domain containing 3 axis) in AD-related oxidative stress, thereby providing a potential therapeutic target for AD treatment. Methods Twenty SPF-grade female KM mice were randomly divided into a control group (Control group) and an experimental group (DNFB group), with 10 mice in each group. Mice in the Control group were treated with an acetone-olive oil vehicle (acetone: olive oil = 3:1) on their back and post-auricular skin. Mice in the DNFB group were treated with 0.5% DNFB (prepared by adding 0.5 g DNFB per 100 mL of acetone-olive oil vehicle) on the same areas, once daily for 14 consecutive days. The severity of skin lesions was scored on days 2, 4, 6, 9, 12, and 14 of treatment. On day 14, scratching behavior and ear thickness were evaluated. Ear swelling was evaluated on the final day by measuring bilateral ear thickness three times with a vernier caliper; the three measurements were averaged. HE staining was used to observe morphological and structural changes of cells in the back skin tissues. The mRNA and protein expression levels of RAGE (receptor for advanced glycation end products) in skin tissues were detected by quantitative real-time PCR, Western blot, and immunohistochemical staining. The mRNA expression levels of oxidative stress-related molecules, including NLRP3 (NOD-like receptor family, pyrin domain containing 3), caspase-1 (cysteine-dependent aspartate-specific protease 1), and IL-1β (Interleukin-1β), were detected by quantitative real-time PCR. Results On day 14, the back skin lesion scores of the Control group and DNFB group were (0.20±0.42) and (9.93±1.30) (P<0.000 1), respectively. Scratching behavior scores were (5.00±2.05) and (49.26±8.49) episodes, respectively (P<0.000 1), and ear thicknesses were (213.00±11.87) μm and (765.93±140.47) μm (P<0.000 1), respectively. The DNFB group exhibited marked skin dryness, desquamation, and thickening. HE staining results showed that skin inflammation was obvious in the DNFB group, consistent with the pathological features of AD. Quantitative real-time PCR and Western blot results showed that compared with the Control group, the mRNA expression level of RAGE in skin tissues of the DNFB group was significantly increased (P<0.05), and the protein expression level of RAGE was also significantly increased (P<0.01). Immunohistochemical staining results showed that compared with the Control group, skin tissue sections of the DNFB group exhibited thickened stratum corneum and fibrotic proliferation of fibroblasts in the interstitium under microscopic observation, with a significant increase in RAGE protein expression in the skin tissues (P<0.01). Quantitative real-time PCR results showed that the mRNA expression levels of NLRP3, caspase-1, and IL-1β in skin tissues of the DNFB group were all significantly increased (P<0.01). Conclusion The AD mouse oxidative stress model has been successfully established by topical DNFB application. RAGE may promote the development of AD by regulating the NLRP3 inflammasome and IL-1β release, forming an oxidative-inflammatory cascade, suggesting that it could be a potential therapeutic target for AD.

Inteins are unique protein insertion sequences capable of self-excision, enabling the covalent ligation of flanking extein peptides via amide bond formation. This process proceeds spontaneously without requiring external enzymes, cofactors, or chemical reagents, granting inteins exceptional biocompatibility and traceless performance in protein engineering applications. Split inteins represent a specialized and versatile subclass whose splicing domains are encoded by two separate gene fragments rather than a single continuous open reading frame. These fragments, known as the N-terminal (IntN) and C-terminal (IntC) split inteins, associate through non-covalent interactions including hydrophobic forces, hydrogen bonds, and van der Waals forces to assemble into an active three-dimensional structure, which then drives efficient extein ligation and enables protein trans-splicing. Protein trans-splicing mediated by split inteins has become a cornerstone for traceless protein ligation owing to its high specificity and irreversibility, fundamentally reshaping strategies for protein modification, assembly, and functional regulation. Compared with traditional chemical ligation methods, split intein systems require no complex chemical derivatization of peptide fragments and can operate efficiently at micromolar concentrations under physiological conditions, thus avoiding structural and functional damage caused by organic reagents. In contrast to enzymatic ligation tools such as sortase, split inteins eliminate the need for additional enzymes or cofactors, simplifying reaction systems, reducing costs, and minimizing non-specific side products. These distinctive advantages render split inteins highly promising for applications in chemical biology, synthetic biology, and biopharmaceutical development. In recent years, deepened mechanistic understanding has established structure-guided rational design as the primary approach to overcoming key limitations of split inteins, including intrinsic aggregation propensity, strict extein sequence dependence, and limited splicing efficiency. Bioinformatic tools have been used to identify aggregation-prone regions in the IntN fragment, and site-directed mutagenesis of hydrophobic residues, relocation of split sites, or removal of misfolding-prone sequences has substantially reduced in vitro aggregation and improved soluble expression and assembly activity. Rational engineering of catalytic residues and adjacent flexible loops has relaxed strict amino acid preferences at extein junctions, enhancing sequence tolerance and reducing the risk of functional impairment in target proteins. Consensus design based on multiple sequence alignments has yielded ultra-fast splicing variants such as Cfa DnaE and Cat-TerL, which exhibit significantly accelerated kinetics and improved tolerance to denaturing conditions. Meanwhile, advances in structural biology have further clarified the conformational dynamics and catalytic mechanisms of splicing, supporting the precise design of high-performance intein modules. On this basis, electrostatic interaction tuning and metagenomic screening have yielded multiple mutually orthogonal split intein pairs, enabling selective multi-fragment protein ligation and providing new routes for the efficient synthesis of large multi-domain functional proteins. With these engineered split inteins offering continuously improved performance and expanded applicability, protein trans-splicing has been widely applied in numerous cutting-edge areas of protein research and biomedicine. In gene delivery, split intein-based systems overcome the packaging limit of adeno-associated viral vectors, enabling the accurate reconstitution of large therapeutic proteins and base editors in target cells, thereby enhancing the efficacy and scope of gene therapy for genetic diseases. In internal protein sequence editing, split inteins mediate precise sequence replacement and modification in flexible regions or loops of target proteins, without the need for complex multi-step ligation and protein refolding involved in traditional protein semisynthesis. In protein-protein interaction studies, intein-mediated splicing covalently captures transient and weak intracellular complexes, enabling sensitive, high-throughput interaction detection and drug screening. In synthetic biology, conditionally controllable splicing systems support the construction of diverse intracellular and cell-surface biological logic gates for the precise regulation of cellular behavior. In mechanistic biochemical research, split inteins enable photocatalytic proximity labeling and site-specific tagging, allowing the preparation of homogeneous protein samples carrying precise post-translational modifications such as ubiquitination and polyglutamylation for chromatin interactome analysis and epigenetic studies. Moreover, covalent trapping strategies using split inteins stabilize transient enzymatic intermediates, providing unprecedented insights into molecular mechanisms such as nucleosome ubiquitination that are difficult to elucidate using conventional methods. This review systematically summarizes key technological advances in split inteins over the past decade, highlighting engineering strategies, mechanistic insights, and the development of orthogonal components. It comprehensively surveys emerging applications at the frontiers of protein research, analyzes current core challenges, and proposes future directions, particularly emphasizing artificial intelligence-driven de novo design and novel splicing pathways to break existing technical bottlenecks. By enabling traceless, efficient, and versatile protein manipulation, split inteins continue to serve as indispensable tools that drive innovation in protein engineering and fundamental life science research.

OBJECTIVE To formulate the Expert Consensus on the Management of Radiopharmaceuticals in Medical Institutions （hereinafter referred to as “the Consensus”）， focusing on key aspects such as the procurement of radiopharmaceuticals， the management of self-made radiopharmaceutical preparations， standardized clinical application， and patient medication safety management， so as to provide a reference for improving the whole-process management of radiopharmaceuticals in medical institutions.METHODS The Consensus was initiated by the Chinese Society of Nuclear Medicine， the Clinical Pharmacy Professional Committee of the China Anti-Cancer Association， and the Committee of Integrative Oncology Pharmacy of the China Anti-Cancer Association. Organized by the consensus drafting group more than 90 experts from the fields of nuclear medicine， nuclear pharmacy， and pharmacy science across China were involved. The Consensus was developed through systematic review of relevant laws， regulations， policy documents， and existing experience in radiopharmaceutical management， followed by extensive discussion and deliberation to finalize the consensus. RESULTS &CONCLUSIONS The Consensus establishes 10 key points for whole-process management of radiopharmaceuticals in medical institutions， covering organizational management and personnel requirements， procurement， preparation， acceptance， storage， risk prevention and control during use， radiation protection， and waste disposal. The Consensus provides a reference for improving radiopharmaceutical management in medical institutions and for ensuring their efficacy and safety in clinical application.

Objective:To analyze the willingness and related factors of pregnant women in Shanghai City to receive influenza vaccines during pregnancy.Methods:An online questionnaire survey was conducted among pregnant women who registered in maternal and child health hospitals in 16 districts of Shanghai. The basic information, health status during pregnancy, attitude towards influenza vaccination, and knowledge of influenza and influenza vaccine were collected, and χ2 test and multivariate logistic regression analysis were used to analyze the willingness and related factors of pregnant women to receive influenza vaccines during pregnancy. Results:A total of 3 219 valid questionnaires were received, with 10.90% of pregnant women willing to receive influenza vaccination during pregnancy, 57.50% unwilling to receive vaccination and 31.59% holding an uncertain attitude. Among the pregnant women who were willing to receive the vaccine, 82.91% would take the initiative to know the relevant information about the influenza vaccine, and 87.18% understood the transmission routes of the flu. 32.09% believed that pregnant women could receive the influenza vaccination at any stage of pregnancy. Among those who were unwilling and held an uncertain attitude, the proportions of the above three questions were 47.06%, 83.95% and 17.80%, respectively. Among pregnant women who were unwilling to receive vaccination and held an uncertain attitude, the main reason for their vaccine hesitation was the concern about the safety of the vaccine for pregnant women and fetuses during pregnancy. Multivariate logistic regression analysis showed that pregnant women with higher education ( OR=3.278, 95% CI: 1.678-6.406, P=0.001), second trimester ( OR=1.542, 95% CI: 1.159-2.054, P=0.003), third pregnancy ( OR=2.318, 95% CI: 1.008-5.331, P=0.048), and previous influenza vaccination ( OR=1.552, 95% CI: 1.119-2.152, P=0.008) had a higher willingness to receive influenza vaccines during pregnancy. Conclusion:Pregnant women in Shanghai have a relatively low willingness to receive influenza vaccines during pregnancy, mainly due to concerns about vaccine safety.

Objective To analyze the law of acupoint selection and compatibility of acupuncture and moxibustion and the application law of acupuncture and moxibustion in the treatment of lactational mastitis using complex network technology.Methods The clinical research literature about acupuncture and moxibustion treatment of lactational mastitis was retrieved from CNKI,CBM,Wanfang Data,VIP and PubMed from the establishment of the databases to March 15,2025.The literature was screened according to the standards to build a prescription database of acupuncture and moxibustion treatment of lactational mastitis.SPSS Modeler 18.0 software was used to analyze association rules,and Gephi 0.9 software was used for complex network analysis.Results A total of 108 articles were included.141 acupuncture and moxibustion prescriptions were extracted,involving 74 acupoints,with a total use frequency of 677 times.The high-frequency acupoints were Jianjing,Danzhong,Rugen,Zusanli,Neiguan,etc.Specific acupoints were mainly composed of Wushu acupoints(169 times,21.10%);the acupoints were mainly distributed in the limbs(36);the most frequently used meridian was stomach meridian(168 times,24.82%).The combination of acupoints with the highest correlation was Jianjing-Neiguan-Rugen.Complex network analysis identified 22 core acupoints,with the most commonly used acupuncture method being filiform needle acupuncture(79 times).Conclusion Acupuncture and moxibustion treatment of lactational mastitis pays special attention to the selection of stomach meridian.The compatibility mode is mainly from top to bottom,and the corresponding acupoints are selected according to syndrome differentiation.

Accurate characterization of the chemical composition of complex traditional Chinese medicine(TCM)is an essential foundation for the modern scientific interpretation of TCM principles.Mass spectrometry is the most dominant technique in current research on the material basis of TCM,offering the highest sensitivity and the richest information provision.Establishing mass spectrometry databases represents the most effective approach to facilitating the structural analysis of TCM chemical components.This paper systematically searches and reviews literature published from January 2005 to January 2025 through online databases such as China National Knowledge Infrastructure,PubMed,and Web of Science,using"mass spectrometry database"and"traditional Chinese medicine"as keywords.It reviews the current status of seven TCM chemical component mass spectrometry databases and seven natural product mass spectrometry databases.The key advancements of these mass spectrometry databases for natural products are summarized,detailing their characteristics,search methodologies,included information,and data sources.Additionally,challenges related to data quality,standardization,timely updates,database interaction,retrieval functionality,and data sharing and security are discussed in depth.Furthermore,the paper explores prospective development directions for TCM mass spectrometry databases,emphasizing the importance of open data sharing,technological innovation,and data security.Through this analysis,the paper aims to offer theoretical guidance and practical recommendations for the precise identification of TCM components,as well as for the construction and application of these databases.

[Purpose]To analyze the incidence and mortality of thyroid cancer in Zhejiang cancer registration areas in 2021 and trends from 2000 to 2021.[Methods]Based on cancer data from 22 registries across Zhejiang Province between 2000 and 2021,the crude incidence/mortality rates,age-standardized incidence/mortality rates of thyroid cancer by Chinese and world standard population(ASIRC/ASMRC,ASIRW/ASMRW),cumulative rate(0～74 years old)and truncated rate(35～64 years old)were analyzed.Joinpoint regression model was employed to calculate the annual percentage change(APC)and average annual percentage change(AAPC)with 95％confidence in-terval(CI)for analyzing trends of thyroid cancer incidence and mortality rates from 2000 to 2021.[Results]The crude incidence rate of thyroid cancer increased from 3.62/105 in 2000 to 79.66/105 in 2021,and ASIRC rose from 3.11/105 to 69.49/105(AAPC=15.55％,95％CI:14.65％～16.61％).The AAPC for ASIRC was slightly higher in female(15.54％,95％CI:14.71％～16.49％)than that in male(15.02％,95％CI:13.53％～16.82％).The increase was significantly more pronounced in rural areas(AAPC=23.34％,95％CI:21.48％～25.33％)compared to urban areas(AAPC=14.12％,95％CI:13.15％～15.43％).Among age groups,the age group of 15～44 years old showed the fastest increase in crude incidence rate(AAPC=20.37％,95％CI:18.46％～22.31％),followed by the age group of 45～64 years old(AAPC=18.41％,95％CI:16.65％～20.19％).Between 2000 and 2021,the crude mortality rate of thyroid cancer rose from 0.34/105 to 0.56/105(AAPC=5.52％,95％CI:3.45％～7.70％),the trend of ASMRC was relatively stable.[Conclusion]From 2000 to 2021,the incidence of thyroid cancer in Zhejiang Province increased markedly,especially among females and younger to middle-aged adults,while the age-standardized mortality rate remained stable.