Background: and Purpose Vertebrobasilar fusiform aneurysms (VBFAs) carry substantial morbidity and mortality, but optimal management for unruptured VBFAs remains unclear. We compared the safety and efficacy of conservative management (CM), stent-assisted coiling (SAC), and flow diverters (FDs) in patients with unruptured VBFAs, focusing on long-term prognosis. Methods: This study included data from a nationwide Chinese cohort of patients with vertebrobasilar dissecting aneurysms. Inverse probability of treatment weighting (IPTW) balanced confounders across groups. The primary outcome was poor prognosis (modified Rankin Scale score >2). Secondary outcomes included aneurysm rupture, ischemic stroke, compression symptoms, and VBFA-related deaths. Logistic regression estimated odds ratios (ORs) and 95% confidence intervals (CIs). Subgroup and sensitivity analyses were performed. Results: Among 1,115 patients with unruptured VBFAs, 838 (median age, 54 years; 655 men) were included. After IPTW, baseline characteristics were balanced. Median follow-up was 54 months. FD was associated with a lower risk of poor prognosis than CM (OR, 0.48 [95% CI, 0.30 to 0.77]; p=0.002), with no difference between CM and SAC. FD also reduced aneurysm rupture (OR, 0.20 [95% CI, 0.07 to 0.60]; p=0.004) and compression symptoms (OR, 0.30 [95% CI, 0.13 to 0.68]; p=0.004) versus CM. Time-to-event analyses further revealed significant differences in vertebral artery lesions and Type I–II VBFAs, whereas no significant differences were observed in basilar or vertebrobasilar junction lesions or in Type III–IV VBFAs. Conclusions Compared with CM, FD was associated with improved long-term outcomes in unruptured VBFAs, particularly in vertebral artery lesions and Type I–II VBFAs, although residual confounding cannot be excluded.

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 To gain insights into the biological characteristics of different populations of Culex tritaeniorhynchus within and around the Yellow Sea wetland from the perspective of the occurrence of resistance, we investigated the levels of resistance to deltamethrin and kdr gene mutation in the wetland and its peripheral areas. Methods Specimens were collected from Cx. tritaeniorhynchus populations at two monitoring sites in the Rare Bird National Nature Reserve and Tiaozi Ni Wetland Scenic Area, and also from two populations in Yancheng City and the Liuhe District of Nanjing, and the resistance of these mosquitoes to deltamethrin was determined using the CDC biotest bottle method. For each concentration of deltamethrin assessed, a random subset of exposed specimens was selected for amplification of the kdr gene fragment, followed by Sanger sequencing to identify and analyze resistance-associated mutations. Results The LC50 levels of deltamethrin among mosquitoes from the four populations in Luhe, Yancheng, the Rare Bird National Nature Reserve and the Tiaozi Ni Wetland Scenic Area were 2.048 5, 7.798 2, 3.473 3, and 17.695 5 mg/mL, respectively, with corresponding concentrations of deltamethrin ranging from 0.005 to 5.000,0.050 to 50.000,0.050 to 25.000 and 0.050 to 50.000 mg/mL, respectively. Furthermore, the ranges of the KT50 values were 11.76-107.43, 67.05-216.30,29.77-107.43 and 28.40-329.51 min; the 1-h knockdown rates were 34.58%-99.15%, 9.52%-43.80%, 55.09%-73.01%, and 10.09%-68.07%; and the 24-h mortality rates were 12.15%-67.52%,9.52%-79.56%,13.17%-82.21%, and 11.01%-78.99%, respectively. With respect to kdr gene mutation, we assayed a total of 63,70,59, and 57 mosquitoes for the four populations, for which we detected L1014F mutation frequencies of 14.29%, 35.00%, 20.34%, and 31.58%, respectively, with a majority of these mutations being heterozygous for resistance. In addition, five adult mosquitoes were identified has having synonymous mutations at site 1011[i. e. , AAT(asparagine)mutation to AAC(asparagine)]. Conclusions Our findings revealed the clear resistance of Cx. tritaeniorhynchus to deltamethrin in the Yancheng region of the Yellow Sea wetland, and the resistance phenotype and kdr frequency of Cx. tritaeniorhynchus in the wetland environment were comparable to those of Cx. tritaeniorhynchus in the wetland environment, thereby indicating that the resistance of different populations of Cx. tritaeniorhynchus was homogeneous under the pressure of different insecticide selection within and around the wetland. However, the underlying mechanisms need to be further studied.

Objective:To construct and validate a competency-oriented entrustable professional activities (EPAs)-Competency-Milestones system for assessment of laparoscopic skills (ESALS).Methods:A research team composed of 15 members from five medical institutions was established. Taking the Staged Training and Assessment of Laparoscopic Skills program as the basis for EPAs, ESALS was constructed through scoping review and behavioral event interviews and the use of the Kawakita Jiro method. ESALS was applied to evaluate 49 trainees and 10 specialists (including 5 novices and 5 experts) on basic and advanced laparoscopic skills, respectively, in comparison with two other assessment tools—the Training and Assessment of Basic Laparoscopic Techniques (TABLT) and the Global Operative Assessment of Laparoscopic Skills (GOALS). Perform Mann Whitney U test using SPSS 23.0 software, and use Spearman coefficient for correlation analysis.Results:Through reviewing 13 literuature articles and interviewing 10 senior surgeons, the ESALS was developed, comprising 10 EPAs, 8 dimensions of competency characteristics, and 5 entrustable levels. In the basic skills evaluation, the ESALS score was significantly positively correlated with the TABLT score ( R=0.881, P<0.001). In the advanced skills evaluation, significant differences were observed between the novice and expert groups in all the ESALS competency scores (all P<0.05), while no difference was detected in the GOALS score of tissue handling ability. Conclusions:ESALS provides comprehensive evaluation, detailed standards, and precise feedback, with comparable efficacy to conventional tools. Future work should focus on expanding sample size to verify its generalizability and developing an electronic tool to enhance application efficiency.

AIM To compare total sugar,total protein,total phenol,total flavonoid,calycosin-7-O-β-D-glucoside,liquiritin,lobetyolin,quercetin,isoferulic acid,hesperidin,glycyrrhizic acid contents and their antioxidant activities,hypoglycemic activities of big honey pill,small honey pill,water pill,concentrated pill,granule,mixture and decoction of Buzhong Yiqi Recipe.METHODS Anthraquinone-sulfuric acid method,Coomassie brilliant blue method,Folin-phenol colorimetry method,sodium nitrite-aluminum nitrate method and HPLC were adopted in the content determination of total sugar,total protein,total phenol,total flavonoid and seven constituents,respectively,after which the scavenging capacities,reducing powers on DPPH·free radical,ABTS+free radical,hydroxyl free radical,and inhibition capacity on α-glucosidase activity were detected.Subsequently,correlation analysis was performed.RESULTS Total sugar,total protein,total phenol and total flavonoid contents demonstrated significant differences among different dosage forms(P＜0.05,P＜0.01).Calycosin-7-O-β-D-glucoside,glycyrrhizin,codonoside and quercetin displayed the highest contents in the decoction,while those of isoferulic acid,hesperidin and glycyrrhizin were observable in the mixture.The water pill exhibited the strongest antioxidant activity,while those of the concentrated pill and mixture were weak;the big honey pill exhibited the strongest hypoglycemic activity,while that of the decoction was the weakest.Total protein,total phenol,total flavonoid and liquiritin contents displayed significant positive correlations between antioxidant activity(P＜0.05,P＜0.01),while hesperidin content displayed significant negative correlation between the latter(P＜0.05);total protein,calycosin-7-O-β-D-glucoside,codonoside and quercetin contents displayed significant negative correlations between hypoglycemic activity(P＜0.05,P＜0.01).CONCLUSION Active constituent contents and their biological activities of Buzhong Yiqi Recipe with different dosage forms exist differences,total sugar,total protein,total flavonoids,calycosin-7-O-β-D-glucoside,licorice glycoside,hesperidin,codonoside and quercetin can be taken as quality control indices for this prescription.

Objective To investigate the epidemiological and demographic characteristics of Mycoplasma pneumoniae(Mp),respiratory syncytial virus(RSV),influenza A(FluA),and influenza B(FluB)in a single center in Minhang District,Shanghai after the end of the COVID-19 pandemic.Methods A retrospective analysis of data from Apr 2023 to Jul 2024 of patients who underwent testing for Mp,RSV,FluA,and FluB due to respiratory tract infections in Minhang Hospital,Fudan University was conducted.Differences in pathogen infections were analyzed among different seasons and age groups.Results A total of 39 103 individuals of respiratory tract infections with simultaneous testing for all four pathogens were included in this analysis,with a total detection rate of 44.7%(17 490/39 103).The detection rates were as follows:Mp 25.2%,FluA 13.1%,FluB 10.9%,and RSV 1.5%.Co-infections accounted for 5.9%,predominantly with Mp and FluA.The detection rate of Mp was>38%in children under 14 years old,gradually decreasing with age.In the child group(≤14 years),both FluA and FluB had the highest detection rates in the 7-14 years age group(FluA,16.5%;FluB,10.4%).In the adult group(≥15 years),the highest detection rates for FluA and FluB were found in the 15-24 years age group(12.5%)and the 35-44 years age group(15.9%),respectively.The detection rates of pathogens varied significantly across different months(P<0.001),with an increasing trend in the total number and overall detection rate of the four pathogens after Sept 2023(P<0.001).Conclusion After the end of the COVID-19 pandemic,there was an abnormal increase in Mp in a single center in Minhang District,Shanghai.Mp,along with influenza viruses,became the main pathogens causing respiratory tract infections.Targeted prevention and control measures based on the epidemiological characteristics of infections should be implemented to guide clinical diagnosis and treatment.

Objective To investigate the epidemiological and demographic characteristics of Mycoplasma pneumoniae(Mp),respiratory syncytial virus(RSV),influenza A(FluA),and influenza B(FluB)in a single center in Minhang District,Shanghai after the end of the COVID-19 pandemic.Methods A retrospective analysis of data from Apr 2023 to Jul 2024 of patients who underwent testing for Mp,RSV,FluA,and FluB due to respiratory tract infections in Minhang Hospital,Fudan University was conducted.Differences in pathogen infections were analyzed among different seasons and age groups.Results A total of 39 103 individuals of respiratory tract infections with simultaneous testing for all four pathogens were included in this analysis,with a total detection rate of 44.7%(17 490/39 103).The detection rates were as follows:Mp 25.2%,FluA 13.1%,FluB 10.9%,and RSV 1.5%.Co-infections accounted for 5.9%,predominantly with Mp and FluA.The detection rate of Mp was>38%in children under 14 years old,gradually decreasing with age.In the child group(≤14 years),both FluA and FluB had the highest detection rates in the 7-14 years age group(FluA,16.5%;FluB,10.4%).In the adult group(≥15 years),the highest detection rates for FluA and FluB were found in the 15-24 years age group(12.5%)and the 35-44 years age group(15.9%),respectively.The detection rates of pathogens varied significantly across different months(P<0.001),with an increasing trend in the total number and overall detection rate of the four pathogens after Sept 2023(P<0.001).Conclusion After the end of the COVID-19 pandemic,there was an abnormal increase in Mp in a single center in Minhang District,Shanghai.Mp,along with influenza viruses,became the main pathogens causing respiratory tract infections.Targeted prevention and control measures based on the epidemiological characteristics of infections should be implemented to guide clinical diagnosis and treatment.

Isoflavones which mainly distributed in leguminous plants have plenty of health benefits.Isoflavone synthase(IFS)is a membrane-associated cytochrome P450 enzyme(CYP450)which carries out the unique aryl-ring migration and hydroxylation.So far,few crystal structures of plant P450s have been obtained.We determined the crystal structure of IFS from Medicago truncatula at 1.9 ? by MAD method using a selenomethionine substituted crystal and conducted molecular docking and mutagenesis study.The structure of IFS complexed with imidazole exhibits the helix Ⅰa-loop-helix Ⅰβ motif which cor-responds to helix Ⅰ of other P450s.Compared with structures of common P450s,IFS/imidazole structure contains an extra domain,i.e.,the γ-domain.The structure reveals a homodimer in which the γ-domain of one molecule interacts with the β-domain of another.The plane of heme group makes an angle of ap-proximately 40° with the helix Ⅰa-loop-helix Ⅰβ motif.Molecular docking combined with mutagenesis study suggested that Trp-128 and Asp-300 might play important roles in substrate binding and recogni-tion.Phe-301,Ser-303 and Gly-305 from the helix Ⅰa-loop-helix Ⅰβ motif may play important roles in the aryl-ring migration.These novel structural features reveal insights into the unique reaction mechanism of IFS and provide a basis for engineering IFS in leguminous crops for health purpose.

Isoflavones which mainly distributed in leguminous plants have plenty of health benefits.Isoflavone synthase(IFS)is a membrane-associated cytochrome P450 enzyme(CYP450)which carries out the unique aryl-ring migration and hydroxylation.So far,few crystal structures of plant P450s have been obtained.We determined the crystal structure of IFS from Medicago truncatula at 1.9 ? by MAD method using a selenomethionine substituted crystal and conducted molecular docking and mutagenesis study.The structure of IFS complexed with imidazole exhibits the helix Ⅰa-loop-helix Ⅰβ motif which cor-responds to helix Ⅰ of other P450s.Compared with structures of common P450s,IFS/imidazole structure contains an extra domain,i.e.,the γ-domain.The structure reveals a homodimer in which the γ-domain of one molecule interacts with the β-domain of another.The plane of heme group makes an angle of ap-proximately 40° with the helix Ⅰa-loop-helix Ⅰβ motif.Molecular docking combined with mutagenesis study suggested that Trp-128 and Asp-300 might play important roles in substrate binding and recogni-tion.Phe-301,Ser-303 and Gly-305 from the helix Ⅰa-loop-helix Ⅰβ motif may play important roles in the aryl-ring migration.These novel structural features reveal insights into the unique reaction mechanism of IFS and provide a basis for engineering IFS in leguminous crops for health purpose.

Objective:To construct and validate a competency-oriented entrustable professional activities (EPAs)-Competency-Milestones system for assessment of laparoscopic skills (ESALS).Methods:A research team composed of 15 members from five medical institutions was established. Taking the Staged Training and Assessment of Laparoscopic Skills program as the basis for EPAs, ESALS was constructed through scoping review and behavioral event interviews and the use of the Kawakita Jiro method. ESALS was applied to evaluate 49 trainees and 10 specialists (including 5 novices and 5 experts) on basic and advanced laparoscopic skills, respectively, in comparison with two other assessment tools—the Training and Assessment of Basic Laparoscopic Techniques (TABLT) and the Global Operative Assessment of Laparoscopic Skills (GOALS). Perform Mann Whitney U test using SPSS 23.0 software, and use Spearman coefficient for correlation analysis.Results:Through reviewing 13 literuature articles and interviewing 10 senior surgeons, the ESALS was developed, comprising 10 EPAs, 8 dimensions of competency characteristics, and 5 entrustable levels. In the basic skills evaluation, the ESALS score was significantly positively correlated with the TABLT score ( R=0.881, P<0.001). In the advanced skills evaluation, significant differences were observed between the novice and expert groups in all the ESALS competency scores (all P<0.05), while no difference was detected in the GOALS score of tissue handling ability. Conclusions:ESALS provides comprehensive evaluation, detailed standards, and precise feedback, with comparable efficacy to conventional tools. Future work should focus on expanding sample size to verify its generalizability and developing an electronic tool to enhance application efficiency.