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.

Nitrofurazone(NFZ)is an antibiotic that is used as a veterinary drug in aquaculture.NFZ abuse can lead to a series of environmental and health issues,making it crucial to establish a rapid and highly sensitive method for NFZ detection.In this study,platinum nanoparticle(PtNPs)-loaded zeolitic imidazolate framework(ZIF-8)was used as a precursor,and PtNPs functionalized nitrogen doped porous carbon(NC)was obtained through pyrolysis.Pt@NC was combined with multi-walled carbon nanotubes(MWCNTs)and cast onto a glassy carbon electrode(GCE)surface to construct an electroch-emical sensor.Electrochemical tests revealed that Pt@NC/WCNT/GCE exhibited an electrochemical active area of 0.066 cm2 and a heterogeneous electron transfer rate constant(k0)of 2.03×10-3 cm/s,which were higher than other materials.Compared with the electrodes modified by other materials,the NFZ generated the highest peak current of irreversible reduction peak on the Pt@NC/WCNT/GCE electrode.In comparison with Pt@ZIF-8/WCNT/GCE,after pyrolysis and carbonization treatment,the reduction current of NFZ increased by 2.19 times,and the reduction peak potential shifted positively by 19 mV simultaneously.When compared with NC/WCNT/GCE,the PtNPs in the composite material enhanced the NFZ current by 4.25 times.Additionally,the experimental conditions for detecting NFZ using the sensor were optimized,including the carbonization temperature of Pt@ZIF-8,ratio of Pt@NC to CNT,loading amount of the modified material,and electrolyte pH.Under the optimized conditions,the sensor demonstrated a linear detection range for NFZ of 0.20-240 μmol/L,a sensitivity of 9.995 μA/((μmol/L)?cm2)and a limit of detection(LOD)of 0.06 μmol/L.The sensor exhibited excellent anti-interference capability,good reproducibility,and stability,with spiked recoveries for NFZ in water samples ranging from 94.6%to 105.6%.This study provided a novel electrochemical sensing approach for NFZ detection.

Objective To investigate the effects of extracellular vesicles(EVs)derived from synovial fluid of rheumatoid arthritis(RA)patients on angiogenesis of human umbilical vein endothelial cells,and to preliminarily explore the underlying mechanisms.Methods Synovial fluid samples of knee joint were collected from 20 patients with RA and 20 patients with osteoarthritis(OA)in this study.EVs were purified using ultracentrifugation.The morphology and size of EVs were observed by transmission electron microscopy and nanoparticle tracking analysis.CD9,CD63,cytochrome c(Cyt-c),vascular endothelial growth factor(VEGF),lysyl oxidase(LOX),matrix metalloproteinase 2(MMP2),tumour necrosis factor alpha(TNF-α),transforming growth factor beta1(TGF-β1)in EVs were detected using Western blot.Human umbilical vein endothelial cells(HUVEC)were treated with the EVs.The growth,migration and angiogenesis of HUVEC were observed by CCK8 assay,TranswellTM chamber assay,scratch test and matrigel angiogenesis assay,respectively.The effect of EVs on the PI3K/AKT pathway in HUVEC was assessed using Western blot.Results Both EVs from RA synovial fluid(RA-EVs)and OA synovial fluid(OA-EVs)were cup-shaped,mainly between 30-400 nm in diameter,expressing CD63 and CD9,but not Cyt-c.RA-EVs carried more VEGF,LOX,MMP2,TNF-α and TGF-β1 than OA-EVs.Compared with the OA-EVs intervention,RA-EVs significantly promoted the proliferation,migration,and angiogenesis of HUVECs,as well as upregulated PI3K/AKT phosphorylation.The inhibitor of PI3K suppressed angiogenesis induced by EVs.Conclusion EVs in synovial fluid of RA carried more cytokines and enzymes that related angiogenesis and inflammation.These EVs exert their pro-angiogenic effects by activating the PI3K/AKT pathway,then contributing to the pathological progression of RA.

Objective To investigate the mortality and risk factors of HIV-infected patients complicated with progressive multifocal leukoencephalopathy(PML)to inform the outcomes of these patients.Methods The clinical data of people living with HIV related PML who were treated at Chongqing Public Health Medical Treatment Center from January 1,2019 to December 31,2023 were retrospectively analyzed.Kaplan-Meier method was used for survival analysis.Log-rank test was performed.Cox regression model was used for multivariate analysis.Results A total of 42 people living with HIV related PML were enrolled,including 34 males(81.0％).The median age was 44.5(36.3,51.0)years.PML was confirmed in 76.2％(32/42)of patients,and possible PML was diagnosed in 23.8％(10/42)of patients.The median time to diagnosis was 32.5(14.8,58.3)days.The median CD4+T cell count was 69.0(39.8,112.0)cells/μL.The median HIV viral load was 5.05(2.62,5.77)log10 copies/mL.The overall mortality rate was 54.8％(23/42)for patients with HIV related PML.CD4+T cell count ≤50 cells/μL and initial modified Rankin scale(mRS)score ≥4 points were independent risk factors for the overall mortality of people living with HIV related PML(P＜0.05).Conclusions People living with HIV related PML have a high mortality rate.Low CD4+T cell count and high initial mRS score were independent risk factors for the mortality of patients with HIV related PML.Currently,no definitive and effective drug treatment is available for PML.Early detection,diagnosis and initiation of antiretroviral therapy may improve the outcomes of patients.

Purpose To evaluate the diagnostic value of 18F-fluorodeoxyglucose(18F-FDG)PET/CT in differentiating primary lung cancer(PLC)from metastatic pulmonary breast cancer in breast cancer patients with solitary pulmonary lesion(SPL).Materials and Methods This retrospective analysis included 58 breast cancer patients with SPL in the Cancer Hospital of the Chinese Academy of Medical Sciences from January 2015 to August 2024.All patients had pathologically confirmed breast cancer with SPL and underwent PET/CT scans,including 36 cases of PLC and 22 cases of metastatic pulmonary breast cancer.Differences in clinical pathological features,high resolution CT morphological,and PET metabolic characteristics were analyzed.Multivariate Logistic regression was utilized to investigate independent predictive factors for SPL diagnosis.Results Statistically significant differences were observed between lesion density,nodule-lung interface,bubble sign,vascular convergence sign and pleural indentation sign(χ2=9.420,7.000,8.487,all P<0.05).PET/CT metabolic analysis revealed a significant difference in maximum standardized uptake value(SUVmax)between the two groups(Z=4.613,P<0.001).Multivariate analysis showed that bubble sign,pleural indentation sign and lower SUVmax were independent predictors for PLC.With an SUVmax threshold of 2.5,the area under the receiver operating curve(AUC)was 0.862,with a sensitivity of 90.9%,specificity of 77.8%and an accuracy of 82.8%.Combined analysis of high resolution CT and SUVmax demonstrated the highest diagnostic efficacy,with an area under the curve of 0.937,sensitivity of 90.9%,specificity of 94.4%and accuracy of 93.1%.Conclusion The presence of bubble sign,pleural indentation and SUVmax<2.5 suggests a high likelihood of PLC.Combined high resolution CT imaging features and PET metabolic characteristics significantly improve diagnostic accuracy for SPLs in breast cancer patients.

Objective:Analysis of the composition of pathogen spectrum and prevalence characteristics in throat swabs of patients with acute respiratory infections (ARI) in Yucheng city, Henan province, from March to June 2023.Methods:After 1 153 throat swabs were collected from ARI patients in Yucheng, 18 respiratory pathogens were tested using a real-time fluorescence quantitative polymerase chain reaction (qPCR) method. The characterization of pathogens spectrum was analyzed.Results:A total of 1 153 throat swabs from ARI patients were collected from March to June 2023 in Yucheng, including 171 outpatients and 982 hospitalized patients. A total of 244 positive samples for common respiratory pathogens were detected (at least one pathogen per sample was detected). The total detection rate of respiratory pathogens was 21.16%, and the top three detection rates were, in descending order, human bocavirus (HBoV), enterovirus (EV), and human parainfluenza virus (HPIV). The main detection month for pathogens was May, with a detection rate of 42.3% (60/142). The main respiratory pathogens detected are HBoV, EV, and HPIV. The detection rate of the age group under 1 year old was the highest, at 25.1% (49/195), mainly consisting of HBoV, respiratory syncytial virus (RSV), and HPIV. The main clinical manifestations of respiratory pathogen-positive patients were fever and cough, and the clinical diagnosis was mainly lower respiratory tract infection, all of which were hospitalized patients.Conclusions:The respiratory pathogens in ARI patients were mainly HBoV, EV, and HPIV from March to June, 2023 in Yucheng. The peak of the epidemic was in May, mainly infecting children under 5 years of age.

Objective:To analyze the association of peripheral blood systemic immune-inflammation index(SII),neutrophil-to-lymphocyte ratio(NLR),platelet-to-lymphocyte ratio(PLR)with disease severity and progno-sis in elderly patients with chronic pulmonary heart disease(CPHD).Methods:A total of 180 elderly CPHD pa-tients admitted in Xuancheng Central Hospital between September 2021 and January 2023 were enrolled as case group.Healthy volunteers who simultaneously underwent physical examinations in our hospital were selected as con-trol group(n=50).According to the 28d prognosis,the case group was divided into death group(n=45)and sur-vival group(n=135).Levels of peripheral blood SII,NLR and PLR were compared among above-mentioned groups;Spearman correlation analysis was used to analyze the association of above indexes with cardiac function class and prognosis in these patients.Multivariate Logistic regression analysis was used to analyze risk factors for death in these patients.The predictive value of SII,NLR,and PLR for death in elderly CPHD patients was ana-lyzed using receiver operating characteristic(ROC)curve.Results:Compared with those in control group,those in the case group had significant higher levels of peripheral blood SII,NLR and PLR(P＜0.001 all).Compared with NYHA class Ⅱ group and class Ⅲ group,those in class Ⅳ group had significant higher levels of peripheral blood SII[(1759.87±179.43)vs.(1148.33±121.57)vs.(1392.44±146.36)],NLR[(8.65±0.89)vs.(7.14±0.75)vs.(7.76±0.81)],PLR[(152.45±16.79)vs.(125.29±13.46)vs.(138.77±13.58)];and levels of peripheral blood SII,NLR,PLR in class Ⅲ group were significantly higher than those of class Ⅱ group(P＜0.001 all).Com-pared with patients in survival group,those in death group had significant higher levels of peripheral blood SII[(1723.86±189.65)vs.(1296.81±142.33)],NLR[(8.24±0.89)vs.(7.63±0.78)],PLR[(148.75±15.26)vs.(134.41±14.58)](P＜0.001 all).Spearman correlation analysis indicated that the levels of peripheral blood SII,NLR and PLR were significant positively correlated with the severity and poor prognosis(r=0.336～0.432,P＜0.05 or＜0.01;r=0.319～0.504,P＜0.05 or＜0.01)in elderly CPHD patients.Multivariate Logistic regression analy-sis indicated that peripheral blood SII,NLR,PLR and smoking were independent risk factors for death(OR=1.024～9.514,P＜0.05 or＜0.01)in elderly CPHD patients.ROC curve indicated that area under curve(AUC)of combination of SII,NLR and PLR predicting death in elderly CPHD patients was 0.979(95％CI 0.946～0.995),significantly higher than those of each single detection[SII:0.847(95％CI 0.786～0.896),NLR:0.832(95％CI 0.769～0.883),PLR:0.881(95％CI 0.825～0.925),Z=3.988,4.386,4.217,P＜0.01 all].The nomogram calibration curve and decision curve showed good consistency and net benefit of the model.Conclusion:Peripheral blood SII,NLR and PLR are associat-ed with the severity and prognosis of elderly CPHD patients,and have certain predictive value for patient's prognosis.

OBJECTIVE: To evaluate the anti-hepatocellular carcinoma (HCC) activity of total alkaloids from Gelsemium elegans Benth. (TAG) in vivo and in vitro and to elucidate their potential mechanisms of action through transcriptomic analysis. METHODS: TAG extraction was conducted, and the primary components were quantified using high-performance liquid chromatography (HPLC). The effects of TAG (100, 150, and 200 µg/mL) on various tumor cells, including SMMC-7721, HepG2, H22, CAL27, MCF7, HT29, and HCT116, were assessed. Effects of TAG on HCC proliferation and apoptosis were detected by colony formation assays and cell stainings. Caspase-3, Bcl-2, and Bax protein levels were detected by Western blotting. In vivo, a tumor xenograft model was developed using H22 cells. Totally 40 Kunming mice were randomly assigned to model, cyclophosphamide (20 mg/kg), TAG low-dose (TAG-L, 0.5 mg/kg), and TAG high-dose (TAG-H, 1 mg/kg) groups, with 10 mice in each group. Tumor volume, body weight, and tumor weight were recorded and compared during 14-day treatment. Immune organ index were calculated. Tissue changes were oberseved by hematoxylin and eosin staining and immunohistochemistry. Additionally, transcriptomic and metabolomic analyses, as well as quatitative real-time polymerase chain reaction (RT-qPCR), were performed to detect mRNA and metabolite expressions. RESULTS: HPLC successfully identified the components of TAG extraction. Live cell imaging and analysis, along with cell viability assays, demonstrated that TAG inhibited the proliferation of SMMC-7721, HepG2, H22, CAL27, MCF7, HT29, and HCT116 cells. Colony formation assays, Hoechst 33258 staining, Rhodamine 123 staining, and Western blotting revealed that TAG not only inhibited HCC proliferation but also promoted apoptosis (P<0.05). In vivo experiments showed that TAG inhibited the growth of solid tumors in HCC in mice (P<0.05). Transcriptomic analysis and RT-qPCR indicated that the inhibition of HCC by TAG was associated with the regulation of the key gene CXCL13. CONCLUSION TAG inhibits HCC both in vivo and in vitro, with its inhibitory effect linked to the regulation of the key gene CXCL13.
Animals ; Apoptosis/drug effects* ; Liver Neoplasms/genetics* ; Carcinoma, Hepatocellular/genetics* ; Humans ; Alkaloids/therapeutic use* ; Gelsemium/chemistry* ; Cell Line, Tumor ; Cell Proliferation/drug effects* ; Mice ; Xenograft Model Antitumor Assays

Hearing loss is the most prevalent disabling disease. Cochlear implantation（CI） serves as the primary intervention for severe to profound hearing loss. This consensus systematically explores the value of genetic diagnosis in the pre-operative assessment and efficacy prognosis for CI. Drawing upon domestic and international research and clinical experience, it proposes an evidence-based medicine three-tiered prognostic classification system（Favorable, Marginal, Poor）. The consensus focuses on common hereditary non-syndromic hearing loss（such as that caused by mutations in genes like GJB2, SLC26A4, OTOF, LOXHD1） and syndromic hereditary hearing loss（such as Jervell & Lange-Nielsen syndrome and Waardenburg syndrome）, which are closely associated with congenital hearing loss, analyzing the impact of their pathological mechanisms on CI outcomes. The consensus provides recommendations based on multiple round of expert discussion and voting. It emphasizes that genetic diagnosis can optimize patient selection, predict prognosis, guide post-operative rehabilitation, offer stratified management strategies for patients with different genotypes, and advance the application of precision medicine in the field of CI.
Humans ; Cochlear Implantation ; Prognosis ; Hearing Loss/surgery* ; Consensus ; Connexin 26 ; Mutation ; Sulfate Transporters ; Connexins/genetics*

Radiation-induced thrombocytopenia (RIT) faces a perplexing challenge in the clinical treatment of cancer patients, and current therapeutic approaches are inadequate in the clinical settings. In this research, oxymatrine, a new molecule capable of healing RIT was screened out, and the underlying regulatory mechanism associated with magakaryocyte (MK) differentiation and thrombopoiesis was demonstrated. The capacity of oxymatrine to induce MK differentiation was verified in K-562 and Meg-01 cells in vitro. The ability to induce thrombopoiesis was subsequently demonstrated in Tg (cd41:enhanced green fluorescent protein (eGFP)) zebrafish and RIT model mice. In addition, we carried out network pharmacological prediction, drug affinity responsive target stability assay (DARTS) and cellular thermal shift assay (CETSA) analyses to explore the potential targets of oxymatrine. Moreover, the pathway underlying the effects of oxymatrine was determined by Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses, Western blot (WB), and immunofluorescence. Oxymatrine markedly promoted MK differentiation and maturation in vitro. Moreover, oxymatrine induced thrombopoiesis in Tg (cd41:eGFP) zebrafish and accelerated thrombopoiesis and platelet function recovery in RIT model mice. Mechanistically, oxymatrine directly binds to toll-like receptor 2 (TLR2) and further regulates the downstream pathway stimulator of interferon genes (STING)/nuclear factor-kappaB (NF-κB), which can be blocked by C29 and C-176, which are specific inhibitors of TLR2 and STING, respectively. Taken together, we demonstrated that oxymatrine, a novel TLR2 agonist, plays a critical role in accelerating MK differentiation and thrombopoiesis via the STING/NF-κB axis, suggesting that oxymatrine is a promising candidate for RIT therapy.