Objective: To investigate the factors influencing intraoperative blood transfusion in patients with hip fractures and to develop a machine learning (ML) model for predicting this risk. Methods: A total of 424 patients with hip fractures who underwent surgical treatment between November 2022 and March 2025 in our hospital were selected. Key feature variables of intraoperative blood transfusion risk were identified using the Boruta algorithm. Four different ML algorithms—support vector machine (SVM), linear discriminant analysis (LDA), mixed discriminant analysis (MDA), and extreme gradient boosting (XGBoost)—were used to develop predictive models for intraoperative blood transfusion risk. The predictive performance of the four ML models were evaluated using accuracy, precision, receiver operating characteristic (ROC) curves, precision-recall curves (PRC), precision-recall gain curves (PRGC), and F1 scores. Shapley additive interpretation (SHAP) was used to interpret the final model. Results: Among the 424 patients, 77(18.2%) received intraoperative blood transfusion. The Boruta algorithm identified albumin (ALB), activated partial thromboplastin time (APTT), types of anesthesia, types of fracture, and hemoglobin (Hb) as key feature variables for predicting intraoperative blood transfusion risk. In model evaluation, the SVM model outperforms the other three models across multiple metrics, including the area under the receiver operating characteristic curve (AUC), recall, recall gain, accuracy, precision, F1 score, and the area under the precision-recall curve (PRC-AUC). The SVM model, interpreted and visualized based on SHAP values, effectively predicted intraoperative blood transfusion risk in patients with hip fracture. A visual online application was developed based on the SVM model (https://pbo-nomogram.shinyapps.io/blood/). Conclusion: Preoperative low ALB and Hb levels, prolonged APTT, general anesthesia, and intertrochanteric fractures are risk factors for intraoperative blood transfusion in hip fracture patients. The risk prediction model for intraoperative blood transfusion constructed based on the SVM algorithm has optimal performance, which provides new ideas and methods for the clinical early identification of hip fracture patients with high transfusion risk and the implementation of targeted interventions.

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 investigate the effect of mindfulness-based cognitive therapy on the sense of personal control and illness perception of young and middle-aged patients after percutaneous coronary intervention (PCI), so as to provide theoretical and practical guidance for the psychological care of patients after PCI.Methods:A randomized controlled trial was conducted. From January 2022 to December 2023, young and middle-aged patients after PCI admitted to the Department of Cardiology, Qinhuai Medical District, Eastern Theater Command General Hospital of the Chinese People′s Liberation Army were selected as the research subjects by convenient sampling method, patients were divided into the experimental group and the control group by the random number table method. The control group was given routine nursing, and the experimental group was given mindfulness-based cognitive therapy on the basis of routine nursing. The Personal Mastery Scale (PMS), Cardiac Self-Efficacy Questionnaire (CSEQ), and Brief Illness Perception Questionnaire (BIPQ) were used to evaluate personal sense of control, self-efficacy, and illness perception before and after the intervention respectively.Results:A total of 80 young and middle-aged patients after PCI were included. There were 40 cases in the control group, including 25 males and 15 females, aged (49.43 ± 5.55) years old. There were 40 cases in the experimental group, including 19 males and 21 females, aged (49.03 ± 4.19) years old. Before the intervention, there was no statistically significant difference in the PMS, CSEQ and BIPQ scores between the two groups of patients (all P>0.05). After the intervention, the PMS, CSEQ scores of the patients in the experimental group were (23.03 ± 2.54), (45.85 ± 8.16) points respectively, which were higher than (21.95 ± 2.28), (39.05 ± 8.78) points in the control group, the BIPQ score of the patients in the experimental group was(39.63 ± 8.12) points, which was lower the (45.45 ± 8.64) points in the control group, the differences were statistically significant ( t=-2.00, -3.59, 3.11, all P<0.05). Conclusions:Mindfulness-based cognitive therapy can effectively improve the sense of personal control and self-efficacy of young and middle-aged patients after PCI, reduce the sense of threat to the disease.

Creutzfeldt-Jakob disease(CJD)is a rapidly progressive and fatal neurodegenerative disorder caused by prions,with certain infectivity and iatrogenic transmission risks.With the rapid progress and application of new dia-gnostic biomarkers and detection methods,as well as the construction and improvement of surveillance and reporting systems,the detection of CJD in patients domestically and internationally has shown an increasing trend year by year.Due to its long incubation period and heterogeneity of early symptoms,early identification and diagnosis of the disease is difficult,increasing the risk of transmission within medical institutions.Currently,there is a lack of con-sensus on the infection prevention and control of CJD.In order to timely identify and diagnose CJD as well as effec-tively block its transmission in medical institutions,this consensus summarizes 15 clinical concerns and formulates 24 specific recommendations based on the latest domestic and international research findings and clinical evidence,as well as combines with clinical practice,aiming to standardize healthcare-associated infection prevention and control measures for CJD and reduce its transmission risk in medical institutions.

The scientific research and innovation capabilities of medical students are intrinsically linked to the sustained and high-quality development of national healthcare initiatives.Cultivating outstanding medi-cal students with independent scientific capabilities and innovative consciousness is a critical component in the education and training of high-level medical professionals.Our investigation revealed that within the imperfections of the cultivating model,some faculty and students at medical schools have an insufficient understanding of scientific research and innovation and lack motivation for engaging in such activities,which hinder the progression of scientific research activities.Consequently,we initiated a teaching practice and exploratory study on the"tutorial system"aimed at fostering medical students'scientific research and innovation abilities.Based on the principle of"research informing teaching,teaching and research advan-cing together,"this study implements a"tutorial system"coordinated by tutors,supplemented by graduate and undergraduate student mentors,to cultivate innovative thinking,stimulate interest in scientific re-search,and enhance practical and research skills among medical students.Through collaborative efforts within"scientific research innovation teams,"various educational methods—including preliminary re-search,in-class and extracurricular activities,intra-group and inter-group interactions,and theoretical and practical applications—are employed to improve and strengthen the cultivation of medical students'scientif-ic research and innovation abilities.This study aims to provide valuable references for optimizing medical education management systems and enhancing the quality of medical student training.

Creutzfeldt-Jakob disease(CJD)is a rapidly progressive and fatal neurodegenerative disorder caused by prions,with certain infectivity and iatrogenic transmission risks.With the rapid progress and application of new dia-gnostic biomarkers and detection methods,as well as the construction and improvement of surveillance and reporting systems,the detection of CJD in patients domestically and internationally has shown an increasing trend year by year.Due to its long incubation period and heterogeneity of early symptoms,early identification and diagnosis of the disease is difficult,increasing the risk of transmission within medical institutions.Currently,there is a lack of con-sensus on the infection prevention and control of CJD.In order to timely identify and diagnose CJD as well as effec-tively block its transmission in medical institutions,this consensus summarizes 15 clinical concerns and formulates 24 specific recommendations based on the latest domestic and international research findings and clinical evidence,as well as combines with clinical practice,aiming to standardize healthcare-associated infection prevention and control measures for CJD and reduce its transmission risk in medical institutions.

The scientific research and innovation capabilities of medical students are intrinsically linked to the sustained and high-quality development of national healthcare initiatives.Cultivating outstanding medi-cal students with independent scientific capabilities and innovative consciousness is a critical component in the education and training of high-level medical professionals.Our investigation revealed that within the imperfections of the cultivating model,some faculty and students at medical schools have an insufficient understanding of scientific research and innovation and lack motivation for engaging in such activities,which hinder the progression of scientific research activities.Consequently,we initiated a teaching practice and exploratory study on the"tutorial system"aimed at fostering medical students'scientific research and innovation abilities.Based on the principle of"research informing teaching,teaching and research advan-cing together,"this study implements a"tutorial system"coordinated by tutors,supplemented by graduate and undergraduate student mentors,to cultivate innovative thinking,stimulate interest in scientific re-search,and enhance practical and research skills among medical students.Through collaborative efforts within"scientific research innovation teams,"various educational methods—including preliminary re-search,in-class and extracurricular activities,intra-group and inter-group interactions,and theoretical and practical applications—are employed to improve and strengthen the cultivation of medical students'scientif-ic research and innovation abilities.This study aims to provide valuable references for optimizing medical education management systems and enhancing the quality of medical student training.

Objective:By sequencing and analyzing the biliary flora by 16S rDNA high-throughput sequencing technology,to identify the main flora associated with gallbladder stone formation and the main flora regulated by the Dahuang lingxian(DHLX),pre-liminary investigation the effect of DHLX on the biliary flora.Methods:The 50 male guinea pigs were randomly divided into Normal group,Model group,DHLX group.There were 15 guinea pigs in the normal group and 20 guinea pigs in the DHLX group,and 20 guin-ea pigs in the model group:the guinea pig model of gallstone was replicated with high-fat lithogenic diet,which was simultaneously ad-ministrated by gavage.After continuous intervention for 8 weeks,bile and gallbladder tissue samples were collected,and the gallstone formation rate of guinea pigs in each group was calculated,the pathological morphological changes of gallbladder tissue were detected by HE,T-CHO,TBA,LPS,IL-6,TNF-α were detected by ELISA,and the changes of biliary flora were detected by 16S rDNA;the correlation between biliary flora and bile index was detected by Pearson statistical method.Results:The stone formation rate of guinea pigs in the normal group was 8.3%,the rate of model stone composition was 90.8%,and the stone composition rate of DHLX group was 36.4%,and the HE staining results showed that compared with the normal group,the mucous membrane of the guinea pig in the model group was thickened,the columnar epithelial cells were arranged in a large number of inflammatory cells,and the columnar epi-thelial cells of the gallbladder mucosa in the chinese medicine group were arranged neatly compared with the model group,the thick-ness of the mucosa was reduced compared with the model group,and some inflammatory cells were infiltrated;ELISA results showed that compared with the normal group,the expressions of T-CHO,LPS,IL-6 and TNF-α in the bile of guinea pigs in the model group were significantly increased(P<0.01),while the content of TBA was significantly reduced(P<0.01);compared with the model group,the expression of LPS and IL-6 in the bile of the DHLX group were significantly reduced(P<0.01).The results of 16S rDNA showed that compared with the normal group the Ace index and Chao1 index of the model group were significantly reduced(P<0.01),and the Chao1 index of the DHLX group was significantly higher than that of the model group(P<0.05);biliary flora at the genera level was mainly composed of Burkholderia,Sphingomycetes,Breghamus,Delfortella,Pseudomonas;correlation analysis showed that total cholesterol was negatively correlated with the abundance of Methyloversatilis(P<0.05),and total bile acids were positively corre-lated with the abundance of Burkholderia(P<0.05),and Pseudomonas erythrocytes,Rhizobia,the abundance of Phreatobacter was negatively correlated(P<0.01)and LPS was positively correlated with the abundance of Pseudomonas erythrocytes(P<0.01).Conclu-sion:Biliary microflora disorder exists in the formation of biliary stones,and biliary microflora may participate in the formation of stones by regulating cholesterol,bile acids and LPS.DHLX can regulate the changes in the abundance of the microflora,make the structure of the microflora become normal,reduce the inflammation of the gallbladder,and prevent the formation of gallstones.

Objective To observe the clinical effect of doing Huatuo five-animal play treating blood-stasis-type silent myocardial ischemia(SMI)and its influence on coronary hemodynamics.Methods Ninety patients diagnosed as silent myocardial ischemia were randomly divided into a five-animal play group(n=45,age:51.56±11.00 years,24 males and 21 females)and a regular exercise group(n=45,age:52.44±7.19 years old,24 males and 21 females)according to a random number table.The five-animal play group practised the traditional five-animal play,while the regular exercise group con-ducted daily 30-minute moderate-intensity treadmill aerobic exercise,five times a week,for 4 weeks.The changes in the number of abnormal ST-T segment occurrences,myocardial oxygen consumption(CMO),coronary ischemia threshold(CIT),and blood smoothness index(BSD)within 24-hour ambu-latory electrocardiogram before and after the intervention were observed in both groups,with the thera-peutic effect and hemodynamic characteristics of both groups evaluated.Results The average number of ST-T segment abnormalities decreased after intervention in both groups(P<0.001),with significantly greater improvement in the five-animal play group than the regular exercise group(P<0.001).More-over,the average CMO decreased significantly,while the average CMR and BSD increased significant-ly in both groups after intervention(P<0.05),with significantly greater improvement in the five-ani-mal play group than the other group.Meanwhile,the total effective rate in the five-animal play group was significantly higher than the regular exercise group(P<0.01).Conclusion Undergoing the five-ani-mal play and moderate intensity treadmill aerobic exercise both are effective in treating silent myocardi-al ischemia.However,the former therapy is superior to the latter in bettering CMO,CIT and BSD.

OBJECTIVE To observe the effect of Shixiang plaster on promoting the wound healing of diabetic foot ulcer.METHODS Totally 50 male SPF grade SD rats were prepared to establish the diabetic models by feeding with high glucose and high fat forage and intraperitoneal injection of streptozotocin,the rats models that were es-tablished successfully were randomly divided into the model group,the Shixiang plaster group and the Kangfuxin solution group.The models of diabetic ulcers were established.The Shixiang plaster group was treated with exter-nal Shixiang plaster,the Kangfuxin solution group was given external Kangfuxin solution,and the model group was treated with coverage with sterile gauze.The wound healing status of the rats was observed,the wound tis-sues were collected for bacterial culture and hematoxylin-eosin(HE)staining after drug administration for 14 and 28 days,respectively.The expression levels of nuclear factor-red cell-2-related factor 2(Nrf-2),heme oxygenase-1(HO-1),tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)and vascular endothelial growth factor(VEGF)were detected by immunohistochemistry(IHC),the expression levels of Nrf-2 and HO-1 were detected with the use of Western Blot,and the levels of serum malondialdehyde(MDA),superoxide dismutase(SOD)and reactive oxygen species(ROS)were detected by enzyme-linked immunosorbent assay(ELISA).RESULTS After the drug administration for 14 and 28 days,the wound healing rates of the Shixiang plaster group were(62.15±3.82)％and(81.68±3.83)％,respectively,higher than(47.14±2.80)％and(69.96±6.49)％of the Kangfuxin solu-tion group and(29.14±9.52)％and(57.91±6.63)％of the model group,and there were significant differences(F=21.716,12.626,P=0.002,0.007).The bacterial colony counts of the Shixiang plaster group were less than those of the Kangfuxin solution group and the model group after the drug administration for 14 and 28 days(P＜0.05).The result of HE staining showed that the Shixiang plaster group had a better wound healing.The result of IHC indicated that the expression levels of Nrf-2,HO-1 and VEGF of the Kangfuxin solution group and the Shix-iang plaster group were up-regulated after the drug administration for 28 days,while the expression levels of TNF-α and IL-6 were down-regulated.The expression levels of Nrf-2 and HO-1 proteins of the Shixiang plaster group were higher than those of the Kangfuxin solution group and the model group after the drug administration for 14 and 28 days,the levels of serum MDA and ROS of the Shixiang plaster group were lower than those of the Kangfuxin solution group and the model group,and the serum SOD level of the Shixiang plaster group was higher than that of the model group(P＜0.05).CONCLUSIONS Shixiang plaster can effectively promote the wound heal-ing of the rats with diabetic foot ulcers and reduce the bacterial colony counts of wound surfaces.The mechanism may be associated with the alleviation of oxidative stress injury by mediating the Nrf-2/HO-1 signaling pathways,promotion of angiogenesis and inhibition of excessive inflammatory reactions.