Green prescription is a written prescription aimed at improving health by promoting physical activity and improving diet， with advantages such as high cost-effectiveness， strong feasibility， and minimal harm to patients. The theory of traditional Chinese medicine （TCM） green prescription integrates the health philosophy of "following rule of yin and yang， and adjusting ways to cultivating health"， the exercise philosophy of balancing yin-yang and the five elements， and the dietary philosophy of moderation and balance， which embody core TCM concepts such as treating disease before its onset and harmony between humans and nature. It has also developed traditional exercise practices like Tai Chi， Baduanjin， Wuqinxi， Yi-Gin-Ching， and Qigong， as well as dietary adjustments like medicated diet and herbal wines. However， it is believed that the TCM green prescription currently suffers from insufficient evidence-based research， low patient awareness and acceptance， and weak basic research. Based on this， it is proposed that large-sample clinical trials should be conducted in the future to improve the quality of evidence-based medicine， basic research can be carried out with the help of artificial intelligence and other methods in research design， the hospital information system （HIS） can be used for control at the implementation level， and publicity and patient education can be strengthened through the new media， so as to promote the development and application of the TCM green prescriptions in the field of global health treatment.

Objective To investigate the acupoint selection law in acupuncture therapy for post-stroke respiratory dysfunction through data mining techniques with R language;To provide references for acupoint selection of clinical acupuncture practice for this disease.Methods Clinical research literature about acupuncture therapy for post-stroke respiratory dysfunction was retrieved from CNKI,Wanfang Data,VIP,CBM,PubMed,Cochrane Library,Web of Science and Embase from the establishment of the databases to February 20,2025.An acupuncture prescription database was established using Excel 2021.R 4.4.3 was used to analyze acupoint application frequency,meridian tropism,regional distribution and specific acupoints,and clustering analysis and association rule analysis were performed.Results Totally 123 articles were obtained,including 123 acupuncture prescriptions,involving 110 acupoints with a total application frequency of 723 times.High-frequency acupoints included Feishu(BL13),Hegu(LI4),Zusanli(ST36)and Fengchi(GB20),etc.The most frequently used meridians were the Bladder Meridian,Ren Meridian,Lung Meridian and Stomach Meridian.Acupoints were predominantly distributed in the head,neck and back regions,with crossing acupoints being the most commonly employed specific acupoints.Clustering analysis identified five meaningful acupoint combinations.The acupoint pair"Feishu(BL13)-Pishu(BL20)"demonstrated the strongest association.Conclusion Acupuncture treatment for post-stroke respiratory dysfunction follows the principle of strengthening the spleen to resolve phlegm and ventilating lung qi.The core acupoint combination is Feishu(BL13)-Pishu(BL20),with additional a acupoints adjusted based on syndrome differentiation to optimize therapeutic efficacy.

Osteoporotic fractures are characterized by reduced bone mass and microstructural abnormalities, leading to differences in the healing process compared to traumatic fractures. The fracture healing process is generally divided into the inflammatory phase, the repair phase and the remodeling phase. In patients with osteoporotic fractures, due to factors such as decreased bone density, reduced bone quality and aging, there are partial alterations in inflammatory cells and osteoimmunity associated with those three healing phases, ultimately affecting fracture healing. This article focuses on the differences in fracture healing between non-osteoporotic fractures and osteoporotic fractures, reviewing relevant research literature and consensus. It analyzes and discusses changes in bone marrow mesenchymal stem cells and bone metabolism during osteoporosis, the effects of inflammatory aging, underlying diseases, and anti-osteoporosis medications on fracture healing, as well as the changes observed after pharmacological intervention. The aim is to emphasize personalized treatment approaches that account for individual and bone-specific factors in managing osteoporotic fractures, striving to promote better fracture healing outcomes and improving patients' quality of life.

Objective:To investigate the current status of referral for critically ill pregnant women in Beijing and Chongqing,provide improvement strategies for the management of referral for critically ill pregnant women.Meth-ods:From May 2022 to June 2024,a self-made questionnaire was used to survey medical staff at different levels of midwifery institutions in Beijing and Chongqing through the internet.The survey targets 18 midwifery institutions in Haidian District and Changping District of Beijing,as well as 50 midwifery institutions in 25 districts and counties of Chongqing.Collect and analyze basic information of the surveyed subjects,the operation of the referral process for critically ill pregnant women(including familiarity and rationality of the referral process),referral indications,dif-ficulties and problems encountered during the referral process,etc.Results:①In this study,a total of 761 valid questionnaires were collected,of which 539 were from Beijing and 222 were from Chongqing.Among 761 surveyed individuals,physicians accounted for 51.4％and nurses(including midwives)accounted for 48.6％;39.7％of the cases came from third level midwifery institutions,while 60.3％came from second level or lower midwifery institu-tions.②93.1％of the surveyed medical staff were familiar with the referral process,with 92.8％in Beijing and 94.2％in Chongqing.There was no statistically significant difference between the two regions(x2=2.843,P=0.241);98.3％of the surveyed medical staff believed that the referral process in their unit was reasonable,with 98.9％in Beijing and 96.8％in Chongqing.There was no statistically significant difference between the two re-gions(x2=4.121,P=0.127).③ The main problems encountered by referral institutions were delayed patient ref-errals(51.0％)and unclear case reporting(42.9％),while the most common difficulties encountered by referral institutions were poor referral transportation(29.5％),inability to respond to referral requests in a timely manner(17.9％),and difficulties in inter hospital communication(17.7％).Conclusions:Medical staff in Beijing and Chongqing are familiar with the referral system for critically ill pregnant women and have achieved good operation-al results.In response to the main problems and difficulties in the current referral process,it is recommended to combine online and on-set consultations to promote continuous communication,standardize referral case records,establish a two-way referral model and regular feedback mechanism,and strengthen targeted personnel commu-nication.

Objective:To explore the genotype and the clinical phenotype of SCN2A-related developmental delay in children. Methods:A case series study was adopted. Collect clinical data from 10 cases of children with SCN2A gene variants diagnosed with global developmental delay/intellectual disability who were admitted to the Children′s Hospital between July 2019 and March 2023. Summarize the clinical phenotype and genotype based on clinical data such as general information, clinical manifestations, imaging examinations, laboratory tests, genetic testing results, and comprehensive pediatric neuropsychological development assessment. Results:A total of 10 patients were recruited, including 7 males and 3 females, with an age range of 27 days to 5 years and 9 months. 9 patients underwent children′s neuropsychological and behavioral assessments, and the results were consistent with global developmental delay, including 2 mild cases, 4 moderate cases, and 3 severe cases. 3 cases had autism spectrum disorder, and 2 cases had epilepsy. 6 patients underwent complete head MRI examination, and 4 of them showed abnormalities, including delayed myelination, widening of the local extra brain space in the frontal lobe, and abnormal frontal lobe morphology. All 10 cases had point variants. Among them, 9 cases are de novo and 1 case is maternal inheritance. Out of 10 cases, there were 5 cases with copy number variations, but all of them were of unknown significance. Among the 10 variants, 8 have been reported and 2 have not been reported, namely c.4145A>T(p.N1382I) and c.4937T>A(p.I1646N). In this study, 4 out of 10 patients with SCN2A variants had variation sites located in the S4 segment of domain which constitute Nav1.2, the sodium ion channel encoded by SCN2A. The developmental quotient level was lower when the variation sites were located in the S4 segment of domain, and the difference was statistically significant ( t=-3.101, P=0.017), indicating that the severity of developmental delay may be related to the localization of amino acids corresponding to variant sites within the protein domain. Conclusion:SCN2A mutations are strongly associated with diverse neurodevelopmental disorders. In this study, the phenotypic spectrum of SCN2A variants encompassed epilepsy, global developmental delay, and autism spectrum disorder. Affected individuals exhibited early-onset developmental delays, predominantly moderate to severe in severity. Voltage-sensing domain dysfunction in sodium channels may constitute a critical pathomechanism underlying neurodevelopmental impairments. Further electrophysiological characterization and molecular mechanistic studies are warranted todelineate the genotype-phenotype correlations between specific variant loci and clinical severity.

Traditional Chinese Medicine(TCM)emphasizes syndrome differentiation and treatment,characterized by"maintaining the prescription if effective"and"changing the prescription if ineffective".Traditional randomized controlled trials(RCTs)are inadequate for evaluating the efficacy of dynamic treatment adjustments.The Sequential Multiple Assignment Randomized Trial(SMART)is an emerging adaptive research design that incorporates randomization at multiple stages,allowing for adjustments in subsequent interventions based on treatment responses.This approach is suitable for evaluating dynamic treatment regimens while retaining the low bias risk of traditional RCTs,making it highly promising for clinical research in TCM.This paper summarizes recent methodological advancements in SMART design,including different sample size estimation and statistical analysis methods for primary effect objectives,embedded adaptive intervention objectives,and optimization objectives,along with providing corresponding operational software.Additionally,it offers considerations for applying SMART design in TCM research,such as the selection of disease types,interventions,decision points,tailoring variables,sample size calculation,statistical methods,the importance of pilot trials,ethical considerations,and limitations.The aim is to promote the exploration and practice of this method in the field of TCM,thereby contributing to the generation of high-quality evidence-based evidence for TCM.

Cuproptosis, a recently identified form of regulated cell death triggered by excess intracellular copper, has emerged as a promising cytotoxic strategy for cancer therapy. However, the therapeutic efficacy of copper ionophores such as elesclomol (ES) is often hindered by cellular copper homeostasis mechanisms that limit copper influx and cuproptosis induction. To address this challenge, we developed a nanoagent utilizing outer membrane vesicle (OMV) derived from Akkermansia muciniphila (Akk) for co-delivery of antioxidant 1 copper chaperone (Atox1)-targeting siRNA and ES (siAtox1/ES@OMV) to tumors. In vitro, we demonstrated that Atox1 knockdown via siRNA significantly disrupted copper export mechanisms, resulting in elevated intracellular copper levels. Simultaneously, ES facilitated efficient copper influx and mitochondrial transport, leading to Fe-S cluster depletion, increased proteotoxic stress, and robust cuproptosis. In vivo, siAtox1/ES@OMV achieved targeted tumor delivery and induced pronounced cuproptosis. Furthermore, leveraging the immunomodulatory properties of OMVs, siAtox1/ES@OMV promoted T-cell infiltration and the activation of tumor-reactive cytotoxic T cells, enhancing tumor immune responses. The combination of siAtox1/ES-induced cuproptosis and immunogenic cell death synergistically suppressed tumor growth in both subcutaneous breast cancer and orthotopic rectal cancer mouse models. This study highlights the potential of integrating copper homeostasis disruption with a copper ionophore using an immunomodulatory OMV-based vector, offering a promising combinatorial strategy for cancer therapy.

Kirsten rat sarcoma viral oncogene homolog (KRAS) protein inhibitors are a promising class of therapeutics, but research on molecules that effectively penetrate the blood-brain barrier (BBB) remains limited, which is crucial for treating central nervous system (CNS) malignancies. Although molecular generation models have recently advanced drug discovery, they often overlook the complexity of biological and chemical factors, leaving room for improvement. In this study, we present a structure-constrained molecular generation workflow designed to optimize lead compounds for both drug efficacy and drug absorption properties. Our approach utilizes a variational autoencoder (VAE) generative model integrated with reinforcement learning for multi-objective optimization. This method specifically aims to enhance BBB permeability (BBBp) while maintaining high-affinity substructures of KRAS inhibitors. To support this, we incorporate a specialized KRAS BBB predictor based on active learning and an affinity predictor employing comparative learning models. Additionally, we introduce two novel metrics, the knowledge-integrated reproduction score (KIRS) and the composite diversity score (CDS), to assess structural performance and biological relevance. Retrospective validation with KRAS inhibitors, AMG510 and MRTX849, demonstrates the framework's effectiveness in optimizing BBBp and highlights its potential for real-world drug development applications. This study provides a robust framework for accelerating the structural enhancement of lead compounds, advancing the drug development process across diverse targets.

Phages, as obligate bacterial and archaeal parasites, constitute a virus group of paramount ecological significance due to their exceptional abundance and genetic diversity. These biological entities serve as critical regulators in Earth's ecosystems, driving biogeochemical cycles, energy fluxes, and ecosystem services across terrestrial and marine environments. Within soil microbiomes, phages function as microbial "dark matter," maintaining the soil-plant system balance through precise modulation of the microbial community structure and functional dynamics. Despite the growing research interests in soil phages in recent years, the proportion of such studies in environmental virology remains disproportionately low, which is primarily attributed to researchers' limited familiarity with the research methodologies for phage microecology, incomplete technical frameworks, and inherent challenges posed by soil environmental complexity. To address these challenges, this review synthesizes cutting-edge methodologies for soil phage investigation from four aspects: (1) tangential flow filtration (TFF)-based phage enrichment strategies; (2) integrated quantification approaches combining double-layer agar plating, epifluorescence microscopy, and flow cytometry; (3) multi-omics analytical pipelines leveraging metagenomics and viromics datasets; and (4) computational frameworks merging machine learning algorithms with eco-evolutionary theory for deciphering phage-host interaction networks. Through comparative analysis of methodological principles, technical merits, and application scopes, we establish a comprehensive workflow for soil phage research. Future research in this field should prioritize: (1) construction of soil phage resource libraries, (2) exploration of RNA phages based on transcriptomes, (3) functional characterization of unknown genes, and (4) deep integration and interaction validation of multi-omics data. This systematic methodological synthesis provides critical technical references for addressing fundamental challenges in characterizing soil phages regarding the community structure, functional potential, and interaction mechanisms with hosts.
Bacteriophages/physiology* ; Soil Microbiology ; Ecosystem ; Microbiota ; Metagenomics/methods*