Multidrug-resistant (MDR) bacterial infections have emerged as a serious challenge of global public health crisis. The overuse and misuse of conventional antibiotics have dramatically accelerated the emergence, evolution and worldwide spread of drug-resistant bacterial strains, necessitating urgent exploration of novel antibacterial strategies. Bacteriophages serve as natural bacterial predators offering distinct advantages including high host specificity, autonomous self-replication capabilities and cost-effective large-scale production. However, wild-type phages present significant clinical limitations due to their narrow host ranges, susceptibility to rapid immune clearance and poor penetration of bacterial biofilms, which severely restrict their therapeutic applications. The convergence of synthetic biology, nanotechnology and advanced gene editing technologies has accelerated the development of engineered bacteriophage platforms, providing programmable, scalable and clinically translatable pathways to overcome these inherent biological constraints. Here, we systematically delineate four fundamental strategies for engineered bacteriophage development. Chemical modification utilizes reactive functional groups such as amino, carboxyl and thiol moieties on capsid proteins through esterification, amidation or click chemistry reactions to achieve precise drug conjugation and surface functionalization. In vivo editing encompasses ultraviolet or chemical mutagenesis for random mutation induction, homologous recombination for targeted genetic alterations, recombineering methodologies including electroporation-mediated bacteriophage recombination engineering, and CRISPR-Cas systems for precise genome editing to enable exact genetic reconstruction and host range reprogramming. In vitro synthesis leverages genome engineering platforms where intact phage genomes are transferred into yeast or host bacteria to facilitate highly efficient homologous recombination, enabling large DNA fragment assembly and cross-gene host range expansion without bacterial toxicity constraints. Directed evolution combines artificial selection through mutation library screening with rational design approaches involving chimeric receptor binding protein construction or site-specific mutagenesis, effectively balancing the discovery of unknown adaptive pathways with targeted host specificity modification. Moreover, we comprehensively discuss therapeutic applications across diverse clinical scenarios. Engineered bacteriophage effectively disrupt bacterial biofilms through sophisticated functionalized delivery platforms including nanozyme-conjugated phages, phage-liposome nanoconjugates and bio-responsive hydrogels, demonstrating significantly enhanced bactericidal efficiency compared to unmodified free phages. These bioengineered vectors attenuate bacterial virulence and resensitize pathogens to antibiotics by delivering CRISPR-Cas systems or base editors to disrupt critical virulence factors such as pili, capsule synthesis machineries and quorum sensing systems, or by inactivating antibiotic resistance determinants including beta-lactamase genes. As an intelligent nanomedicine delivery platform, engineered bacteriophage enable precise pathogen elimination an through photocatalytic reactive oxygen species generation, immunomodulatory interventions, or controlled release of antibacterial drugs. Furthermore, oral administration of engineered bacteriophage facilitates microbiota modulation, which selectively eliminate intestinal pathogens while preserve beneficial commensal microbiota, thereby restoring microbial community balance and preventing complications associated with dysbiosis. Finally, we critically analyze persistent challenges including host strain matching complexity, evolution of bacterial resistance mechanisms, pharmacokinetic optimization requirements, optimal administration route selection, large-scale production quality control standards and clinical dosing determination protocols. Through multidisciplinary integration of synthetic biology, infectious disease medicine and immunology, future translational medicine studies of bacteriophage should establish comprehensive technical platforms encompassing rapid phage screening, intelligent rational design, rigorous in vivo evaluation and standardized clinical validation processes, ultimately advancing engineered bacteriophage from laboratory innovations to clinically approved therapeutics for effectively combating MDR bacterial infections.

Objective In order to understand the current resistance status of Aedes albopictus(Ae. albopictus) adult population to commonly used insecticides and the resistance dynamics development in Zhejiang Province and provide a theoretical basis for the scientific and rational use of hygienic insecticides. Methods Resistance determination was carried out using the diagnostic dose method(GB/T 26347-2010)recommended by WHO in the adult mosquito contact cylinder method to monitor the resistance of Ae. albopictus field populations in 11 cities and districts in Zhejiang Province in 2024. Results The monitoring result showed that adult Ae. albopictus in 11 areas were resistant or suspected resistant populations to both cis-cypermethrin and permethrin; suspected and resistant populations to the organophosphate insecticide malathion were as high as 81.82%; while suspected resistance to the carbamate insecticide residual carbofuran accounted for 45.45%, and the rest of the monitoring sites were sensitive to it. Conclusions The result suggest that the generalized resistance of Ae. albopictus populations to pyrethroids and organophosphorus insecticides in the field in Zhejiang Province needs to be highly emphasized. In future mosquito vector and mosquito-borne disease control, insecticides should be selected based on the resistance monitoring information, while following scientific guidelines for insecticide use.

Objective To explore the value of support vector machine(SVM)model based on gray matter volume(GMV)for identifying amyotrophic lateral sclerosis(ALS),also to analyze the relevant brain regions.Methods MR 3D T1WI data of 60 ALS patients(ALS group)and 60 healthy volunteers(control group)were retrospectively analyzed.Taken GMV of each brain region obtained by voxel-based morphometry as the input features.F-score analysis was used to select feature with the highest classification accuracy to construct SVM model.Receiver operating characteristic curve was drawn to evaluate the efficacy of SVM model for identifying ALS,and top 10％was used as the weight threshold to obtain gray matter brain regions contributed the most to this model.Results SVM model constructed based on the top 40％GMV features had the highest classification accuracy(82.50％),with sensitivity,specificity and area under the curve(AUG)of 85.05％,80.40％and 0.890,respectively.The left precentral gyrus,left anterior cingulate gyrus and paracingulate gyrus,right middle temporal gyrus,opercular part of left inferior frontal gyrus,right dorsolateral superior frontal gyrus,left temporal pole:middle temporal gyrus,right superior occipital gyrus,orbital part of right middle frontal gyrus,right calcarine fissure and surrounding cortex,right fusiform gyrus were the top 1-10 gray matter brain regions contributed to this model.Conclusion ALS had specific GMV change pattern.SVM model based on GMV could be used to effectively identify ALS,while the left precentral gyrus was the most contributive brain region to this model.

Objective:To explore the coordinated development status of the medical service supply-economy-society trinity systems in Zhejiang Common Prosperity Demonstration Zone,and to offer references for formulating policies conducive to the efficient and coordinated development of these three systems.Methods:Based on the panel data from 2013 to 2022,this research was conducted on the 11 prefectural-level cities in Zhejiang Province.Firstly,an evaluation index system for the three systems was established,and the entropy method was employed to determine the weights of each index and calculate the comprehensive evaluation index.Secondly,a triangular model was introduced to delineate the relative relationships among the three systems,and the coupling coordination degree model was utilized to disclose the coordination degree of the three systems.Finally,the spatial autocorrelation analysis method was applied to investigate the spatial autocorrelation of the coupling coordination degree of the three systems.Results:(1)On the whole,the development status of the three systems has improved over time,yet the comprehensive development level remains to be improved.(2)Overall,each prefectural-level city has transited from a medical service supply-society-dominated development to a balanced and coordinated development of the medical service supply-economy-society trinity systems.The coupling coordination degree of the three systems has risen from 0.468(on the verge of imbalance)in 2013 to 0.609(primary coordination)in 2022,presenting an upward trend in general.However,the coordination level remains to be improved and there exist imbalances among regions.(3)The coupling coordination degree of the three systems in Zhejiang Province exhibits a significant positive spatial correlation,and the spatial distribution characteristics are relatively stable. Conclusion:The coupling coordination degree of the medical service supply-economy-society trinity systems in Zhejiang Province awaits further enhancement. At the level of Zhejiang Province,favorable policy support should be provided,the layout of medical resources should be rationally planned,and high-quality economic and social development should be promoted. Each prefectural-level city should formulate strategies for medical service supply,economic,and social development in accordance with its own development level and local conditions,strengthen inter-city linkage and cooperation,and thereby elevate the coordinated development level of the three systems.

Objective:To compare postoperative anal function recovery between laparoscopic subtotal colectomy with antiperistaltic cecorectal anastomosis and laparoscopic total colectomy with ileorectal anastomosis for slow transit constipation.Methods:This multicenter retrospective cohort study enrolled patients meeting the following criteria: (1) severe constipation symptoms (<2 bowel movements/week), absent or insignificant defecation urge, abdominal distension, requiring laxatives to maintain bowel movements or laxatives being ineffective; (2) constipation symptoms for over 5 years, ineffective after >2 years of medical treatment, with strong desire for surgery; (3) significantly prolonged colon transit time (>72 hours) without significant gastric or small intestinal transit dysfunction; (4) no organic colonic lesions confirmed by colonoscopy and abdominal CT. Exclusion criteria: (1) patients undergoing open surgery; (2) exclusion of outlet obstruction constipation (e.g., rectocele, rectal prolapse, puborectalis spasm) by functional defecation MRI; (3) comorbid psychiatric disorders; (4) missing clinical data or loss to follow-up (postoperative follow-up <24 months). Based on these criteria, clinical and follow-up data were collected from 220 patients who underwent either laparoscopic subtotal colectomy with antiperistaltic cecorectal anastomosis (LSC group, n = 115) or laparoscopic total colectomy with ileorectal anastomosis (LTC group, n = 105) for slow transit constipation between January 2013 and December 2022. Subjective anal function (Constipation Severity Score and Wexner Fecal Incontinence Score) and objective anal function (positive rate of rectoanal inhibitory reflex [RAIR] and anorectal manometry) were observed preoperatively and at 6, 12, and 24 months postoperatively. Results:No significant differences were found in baseline characteristics between the two groups (all P >0.05). All surgeries were completed successfully without major significant complications. Subjective anal function assessment: At 24 months postoperatively, Constipation Severity Scores decreased significantly compared to preoperative scores in both groups [LSC group: (25.2±2.8) vs. (2.9±1.8), P <0.001; LTC group: (25.8±2.9) vs. (2.8±1.9), P<0.001]. No significant differences were found between the groups at 6, 12, and 24 months postoperatively (all P>0.05). Wexner Fecal Incontinence Scores at 24 months were significantly lower than those at 6 months in both groups [LSC group: (12.9±1.8) vs. (3.9±2.5), P<0.001; LTC group: (12.6±1.8) vs. (5.4±2.4), P<0.001]. Although no significant difference was found at 6 months ( P = 0.190), the LSC group had significantly lower Wexner scores than the LTC group at 12 and 24 months postoperatively (both P < 0.001). Objective anal function assessment: (1) Positive RAIR rate: Preoperative positive RAIR rates were 33.0% (38/115) in the LSC group and 25.7% (27/105) in the LTC group ( P > 0.05). At 24 months, positive rates increased significantly in both groups [LSC: 66.1% (76/115); LTC: 63.8% (67/105)] compared to preoperative rates (both P<0.001), but no significant differences were found between groups at 6, 12, and 24 months (all P>0.05). (2) Resting pressure (RP) and squeeze pressure (SP): No significant differences were found in preoperative RP and SP between groups (all P>0.05). The LSC group had significantly higher RP and SP than the LTC group at 6 and 12 months postoperatively (all P<0.05), but no significant differences were found at 24 months ( P>0.05). Conclusion:Both laparoscopic subtotal colectomy with antiperistaltic cecorectal anastomosis and laparoscopic total colectomy with ileorectal anastomosis are safe for patients with slow transit constipation. However, laparoscopic subtotal colectomy with antiperistaltic cecorectal anastomosis offers superior postoperative anal function recovery.

Aim To investigate the effects of the fangchinoline derivative LYY-32 on the biological prop-erties of the BLM642-1290 DNA helicase,in order to lay a foundation for further research on its antitumor activity.Methods Fluorescence polarization assay,malachite green-phosphate and ammonium molybdate colorime-try,and fluorescein-labeled DNA gel electrophoresis experiments were conducted to study the effects of fangchinoline derivative LYY-32 on the DNA binding activity,ATPase activity,and DNA unwinding activity of BLM642-1290 DNA helicase.The effects of LYY-32 on the DNA unwinding activity of DNA helicase in cells were studied using fluorescent techniques and time-lapse microscopy.Ultraviolet spectral scanning was used to investigate the effects of LYY-32 on the confor-mation of the BLM642-1290 DNA helicase.Results At a concentration of 10 μmol·L-1,the inhibition rate of LYY-32 on BLM642-1290 DNA helicase binding to dsDNA was 53.17％.At a concentration of 5 μmol·L-1,the inhibition rate of LYY-32 on BLM642-1290 DNA helicase binding to ssDNA was 88.49％.The inhibition rate of LYY-32 on the ATPase activity of BLM642-1290 DNA he-licase was 89.3％at a concentration of 50 μmol·L-1.When the concentration of LYY-32 exceeded 5μmol·L-1,its inhibition rate on the DNA unwinding activity of BLM642-1290 DNA helicase was 100％.LYY-32 also significantly inhibited the DNA unwinding ac-tivity of DNA helicase in cells.However,LYY-32 had no effect on the conformation of BLM642-1290 DNA heli-case.Conclusion The DNA binding activity,AT-Pase activity,and DNA unwinding activity of BLM642-1290 DNA helicase could be significantly inhibi-ted by the fangchinoline derivative LYY-32.

AIM To investigate the effects of volatile oil from Acorus tatarinowii Schott(A.tatarinowii)on neuroinflammation in a rat model of tic disorders.METHODS The SD rats were randomly divided into the blank group(8 rats)and the model group(40 rats).The rat models of tic disorders established successfully by intraperitoneal injection of iminodiapropionitrile(IDPN)were further divided into the model group,the tiapride group and the high-dose,moderate-dose and low-dose A.tatarinowii volatile oil groups,with 8 rats in each group.The 4-week intragastric treatment of respective drug was initiated the next day after the completion of modeling,and normal saline was dosed upon the blank group and the model group,during which the rats' behavioral changes were assessed by stereotyped behavior and motor behavior score every week.After the administration,the rats had their morphological changes of striatal neurons observed by Nissl staining;their levels of TGF-β,IL-10,TNF-αand IL-1β in serum and striatum detected by ELISA;their striatal protein expressions of CX3CL1 and CX3CR1 detected by Western blot and immunohistochemistry;and their striatal expressions of M1,M2 microglia marker proteins CD86,CD206,SYN and PSD-95 detected by immunofluorescence co-staining.RESULTS Compared with the model group,the A.tatarinowii volatile oil groups demonstrated improved twitch-like behavior;decreased scores of motor behavior and rigid behavior(P＜0.01);alleviated damage of Nissl bodies in neurons;increased serum and striatum levels of TGF-β and IL-10(P＜0.05,P＜0.01);decreased levels of TNF-α and IL-1β(P＜0.01);decreased striatal protein expressions of CX3CL1 and CX3CR1(P＜0.01);increased protein expressions of PSD95 and SYN(P＜0.05,P＜0.01);and decreased CD86/Iba1(P＜0.01)and increased CD206/Iba1(P＜0.01)in terms of the fluorescence intensity.CONCLUSION A.tatarinowii volatile oil contributes an anti-tic effect and improves the neuroinflammation in the brain of the rat model of tic disorders by promoting the transformation of microglia into M2 type via CX3CL1/CX3CR1 signal axis.

Objective:To explore the distribution of pathogenic bacteria causing postoperative infections in chil-dren undergoing surgery for Hirschsprung's disease(HD),and the clinical significance of serum triggering receptor ex-pressed on myeloid cells-1(sTREM-1)and high mobility group box 1(HMGB1).Methods:98 children with HD admit-ted from January 2021 to January 2025 were divided into infection group(n=42)and non-infection group(n=56)accord-ing to whether the children were complicated with infection after operation.Automatic bacterial identification system was used to detect the distribution of pathogenic bacteria in secretion samples from infected sites of HD children after operation.ELISA was used to measure serum sTREM-1 and HMGB1.ROC was used to explore the diagnostic value of serum sTREM-1 and HMGB1 for severe infection in children with HD.Results:A total of 49 strains of pathogenic bac-teria were isolated from 42 children with HD in infection group,of which 63.27%(31/49)were gram-negative bacteria,accounting for the highest proportion,followed by Gram positive bacteria at 30.61%(15/49)and fungi at 6.12%(3/49).Among them,Pseudomonas aeruginosa had the highest proportion of 24.49%(12/49)among Gram negative bacteria.there were 15 cases of mild infection(15 strains of pathogens),16 cases of moderate infection(17 strains of pathogens)and 11 cases of severe infection(17 strains of pathogens).With the aggravation of the infection degree of HD children,the distribution pattern of pathogenic bacteria changed,and gradually changed to mainly gram-negative bacteria infec-tion.Compared with non infected group,the levels of serum sTREM-1 and HMGB1 in infected group were higher(P<0.05).The levels of serum sTREM-1 and HMGB1 in children with severe HD infection are higher than those in children with mild and moderate HD infection(P<0.05).The AUC of the combined diagnosis of serum sTREM-1 and HMGB1 for severe infection in HD children was the highest,at 0.985,which was superior to the separate diagnosis of serum sTREM-1 and HMGB1(Zcombination-sTREM-1=1.988,P=0.047;Zcombination-HMGB1=2.126,P=0.034).Conclusion:Serum sTREM-1 and HMGB1 are elevated in HD children with postoperative infection,and the pathogenic bacteria are mainly gram-negative bacteria.In addition,the combined detection of the two provides a reference for the clinical diagnosis of severe infection in HD children.

Obesity is a common chronic metabolic disease mainly characterized by excessive accumulation of adipose tissue.Recently,the global prevalence of obesity-related metabolic diseases has increased significantly,seriously affecting the physical and mental health of patients.Adipokines are pleiotropic bioactive substances secreted by adipose tissue,which have physiological functions such as regulating energy metabolism,inflammatory response and insulin sensitivity.Abnormal hyperplasia of adipose tissue can induce chronic inflammatory responses in the body,stimulate the production or secretion disorders of adipokines,and alter glucose and lipid homeostasis,thereby leading to the occurrence and development of obesity-related metabolic diseases.However,the specific mechanism remains unclear.Exercise prescription is a planned exercise guidance program based on the results of the patient's physical fitness test to achieve the expected goal by adopting the prescribed exercise methods.In recent years,previous studies have found that exercise prescriptions can regulate adipokines,thereby preventing and treating obesity-related metabolic disorders,which may become a potential treatment for obesity-related metabolic diseases in clinical practice.This article reviews the mechanism and clinical effect of targeted regulation of adipokines by exercise prescriptions in the treatment of obesity-related metabolic disorders,in order to provide some new ideas and directions for finding new therapies for obesity-related metabolic diseases.

OBJECTIVE To investigate the impact of inflammatory cytokines related to different endotypes of chronic rhinosinusitis on the transcriptome of nasal epithelial cells.METHODS Human primary nasal epithelial cells were cultured in vitro,and treated with type 1,type 2,and type 3 inflammatory cytokines IFN-γ(10 ng/ml),IL-4(50 ng/ml),and IL-17A(50 ng/ml),respectively.After 48 hours,cells were collected for RNA extraction and transcriptome analysis.Differentially expressed genes were identified,followed by functional annotation and pathway enrichment analysis.Additionally,common regulated genes among different cytokines were analyzed.RESULTS Treatment of primary nasal epithelial cells with IFN-γ,IL-4,and IL-17A induced significant transcriptomic alterations,with 650,540,and 192 differentially expressed genes,respectively,compared to the control group.Functional annotation and pathway enrichment analysis showed that IFN-γ was mainly associated with defense response to virus,antigen processing and presentation,leukocyte and lymphocyte mediated immunity,and NOD-like receptor signaling pathway;IL-4 was mainly related to the regulation of leukocyte chemotaxis,cell-cell adhesion,ECM-receptor interaction,and MAPK signaling pathway;IL-17A was involved in the regulation of neutrophil chemotaxis,response to molecule of bacterial origin,chemokine signaling pathway,and NF-κB signaling pathway.Genes commonly regulated by IFN-γ,IL-4,and IL-17A were identified,with DUOX2 being upregulated by all the three cytokines.CONCLUSION IFN-γ,IL-4,and IL-17A reshapes the transcriptomic profile of nasal epithelial cells,which mainly regulates signaling pathways related to defense response to virus,leukocyte chemotaxis,and response to bacteria,respectively.