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.

AIM To sequence the chloroplast genome of Artemisia vestita Wall.ex Bess.METHODS Based on ethnobotanical surveys,sample collection and original plant identification were carried out.The chloroplast genome was sequenced using the Illumina platform,followed by assembly and annotation.A comprehensive comparative analysis was conducted with six Artemisia species.The maximum likelihood(ML)phylogenetic tree was constructed based on the chloroplast genome sequences of A.vestita and 32 other Asteraceae species,with Leptocodon hirsutus D.Y.Hong of Campanulaceae as outgroup.RESULTS The chloroplast genome of A.vestita was 151 204 bp in length,including a small single-copy region of 18 331 bp,a large single-copy region of 82 949 bp,and inverted repeat regions of 24 962 bp,with a total GC content of 37.45％.134 genes were annotated,including 89 protein-coding genes,8 ribosomal RNA genes,and 37 transfer RNA genes.A total of 67 SSRs and 44 LSRs were detected in the chloroplast genome.Comparative analysis with closely related species of Artemisia revealed 3 highly variable genes(clpP,rpl36,ycf1)and 6 highly variable intergenic regions(trnK-UUU-matK,rps18-rpl20,rpl36-infA,rpl14-rpl16,rpl16-rpl3 and trnL-UAG-ccsA),which could serve as candidate DNA barcodes for Artemisia identification.Phylogenetic analysis showed that Artemisia formed a highly supportive monophyletic group,with A.vestita and A.gmelinii Web.ex Stechm.being closely related.CONCLUSION This study may provide fundamental data for phylogenetic analysis of Artemisia,taxonomic identification and DNA barcoding construction of Tibetan herb.

Objective:To observe the effect of sodium cantharidate assisted bevacizumab in the treatment of advanced non-small cell lung cancer patients.Methods:A total of 120 patients with advanced lung adenocarcinoma from January 2021 to December 2023 were divided into two groups,namely the experimental group and the matched group,each consisting of 60 cases.All patients were treated with AP(cisplatin+pemetrexed)chemotherapy,and bevacizumab(7.5 mg/kg)was administered intravenously at the same time as chemotherapy in the matched group,while sodium bengalate(0.5 mg)was added intravenously in the experimental group on the basis of the matched group,and the patients in the two groups were compared with each other in terms of the clinical efficacy of the two groups in a cycle of 21 d,and the changes in the indicators of the immune function,symptoms and scores of adverse reactions were also compared before and after the 4 consecutive cycles of treatment.After 4 consecutive cycles of treatment,we compared the clinical efficacy,immune function indexes,symptoms,and the incidence of adverse reactions.Results:There was no difference in disease control rate(P＞0.05),and the objective response rate was higher than the matched group(P＜0.05);Pretherapy,tthe levels of natural killer(NK)cells,CD4+/CD8+,immunoglobulin(Ig)G and IgA were different between the test and matched group(P＞0.05).Post-treatment,the levels of NK cells,CD4+/CD8+,IgG,and IgA in the experimental group increased,while there was no significant change in the matched group.The experimental group was higher than the matched group(P＜0.05);Pretherapy,there was no difference in VAS and BFI scores between the experimental group and the matched group(P＞0.05).Post-treatment,the scores of the Simplified Fatigue Scale(BFI)and Visual Analog Scale(VAS)in both groups decreased,and the experimental group was lower than the matched group(P＜0.05);The incidence of grade Ⅰ-Ⅱ gastrointestinal reactions,liver and kidney dysfunction,bone marrow suppression,leukopenia,and grade Ⅲ-Ⅳ bone marrow suppression in the experimental group were all lower than those in the matched group(P＜0.05).Conclusion:Sodium cantharidate assisted bevacizumab therapy has a significant effect on non-small cell lung cancer.Compared with single chemotherapy combined with bevacizumab,it can improve the objective remission rate of patients,improve their immune function level,alleviate pain and cancer-related fatigue symptoms,and assist in reducing the incidence of adverse reactions caused by bevacizumab and chemotherapy.

Objective:To investigate the host and vector composition of pestis and the epidemic situation of pestis among animals in the border port areas of Longchuan County, Yunnan Province.Methods:In September 2022, a survey was conducted on rodents and their surface parasitic fleas in the border port areas of Longchuan County, according to three habitat types: residential areas, agricultural areas, and forest-shrubbery areas. Samples of murine animals organs and their surface parasitic fleas were collected for isolation and identification of Yersinia pestis. Blood samples of plague indicating animals, such as murine animals, dogs and cats were collected, and serum plague F1 antibody was detected by indirect hemagglutination assay. Meanwhile, a retrospective investigation was conducted on the occurrence of self dead rats, sick rats, and suspected cases in the local area from 2018 to 2022 through interviews with farmers. Results:A total of 168 murine animals belonging to 3 orders, 5 families, 10 genera, and 11 species were captured in three habitats in Longchuan County. Among them, the capture rate in residential areas was 5.00% (30/600), with Rattus tanezumi and Suncus murinus as dominant species, with a composition ratio of 50.00% (15/30). The capture rate in agricultural areas was 9.67% (122/1 262), with Rattus tanezumi and Suncus murinus as dominant species, with a composition ratio of 50.82% (62/122) and 44.26% (54/122), respectively. The capture rate in the forest-shrubbery areas was 6.25% (16/256), with Hylomys suillus and Eothenomys eleusis as dominant species, with a composition ratio of 37.50% (6/16) and 31.25% (5/16), respectively. Among the captured murine animals, 20 individuals carried 52 parasitic fleas, belonging to 2 species of 2 genera and 2 families. The total flea infection rate was 11.90% (20/168), and the total flea index was 0.31 (52/168). The dominant specie was Xenopsylla cheopis (90.38%, 47/52). The flea infection rate in residential areas was 33.33% (10/30), and the flea index was 1.23 (37/30). The flea infection rate in agricultural areas was 7.38% (9/122), and the flea index was 0.11 (14/122). The flea infection rate in the forest-shrubbery areas was 6.25% (1/16), and the flea index was 0.06 (1/16). The samples of murine animals and their parasitic fleas obtained were isolated and cultured by Yersinia pestis, and the results were negative. A total of 144 serum samples from murine animals, dogs and cats were separated, and no F1 antibody against pestis was detected. According to interviews and investigations, no abnormal situations such as a large number of self dead rats, sick rats, and suspected cases were found from 2018 to 2022. Conclusions:No plague epidemic has been found in the border port areas of Longchuan County recently. The main host of plague, Rattus tanezumi, and the main vector, Xenopsylla cheopis, remain the dominant species in this area.

The blood products industry,both domestically and internationally,exhibits distinct features in product research,development,and technological innovation.International companies possess extensive expertise in developing immunoglobulins,coagulation factors,and recombinant plasma protein products,demonstrating continuous advancements-particularly in specific immunoglobulin development,long-acting formulation optimization,and manufacturing process improvements.In recent years,Chinese enterprises have also achieved notable progress in related fields,especially in immunoglobulin process refinement and the development of novel recombinant coagulation factor products.However,there remains significant scope for improvement in areas such as the application of recombinant protein technologies,efficient utilization of plasma resources,and the adoption of advanced manufacturing techniques.Additional challenges include the accumulation of patented technologies,the supply of critical raw materials,and access to comprehensive epidemiological data.Driven by ongoing advances in gene recombination technologies,innovations in drug delivery systems,digital transformation,and the rise of personalized medicine,the blood products industry is poised for broader development prospects.To foster sustained and stable domestic industry growth and enhance global competitiveness,Chinese blood product enterprises should intensify their technological accumulation,upgrade manufacturing processes,and optimize plasma resource utilization.

Objective To analyze the clinical characteristics and prognosis of Pseudomonas aeruginosa peritoneal dialysis-associated peritonitis(PaeP).Methods Peritoneal dialysis-associated peritonitis(PDAP)patients who were followed up in the nephrology outpatient department of a hospital from January 2019 to December 2020 were analyzed retrospectively.According to bacterial culture results,patients were divided into the PaeP group and non-PaeP group.Clinical characteristics of PaeP patients and antimicrobial susceptibility testing results of Pseudomonas aeruginosa were analyzed,clinical manifestations,laboratory test results,and prognosis of two groups of patients were compared.Results A total of 124 peritoneal dialysis patients were included in analysis,164 cases of peritoni-tis occurred,16 cases were in the PaeP group and 148 in the non-PaeP group.11 patients developed 16 episodes of Pseudomonas aeruginosa infection,accounting for 8.9％of PDAP patients.Among them,4 patients had peritoneal dialysis catheter exit-site infection,with 5 recurrence cases,1 case cured,1 case died,and 9 cases were extubated.Among the extubated patients,1 withdrew dialysis,3 were recovered to peritoneal dialysis after hemodialysis,5 changed to permanently hemodialysis,with a technical failure rate of 54.5％.Compared with the non-PaeP group,patients in the PaeP group had a shorter dialysis time(13.83±4.92 vs 38.53±35.77 months).During the infection period,C-reactive protein levels were higher(96.61±6.17 vs 45.87±44.65 mg/L),while albumin levels were lower(25.62±4.42 vs 29.46±8.25 g/L).At the onset of infection,the proportion of polymorphonuclear cells in perito-neal dialysis fluid was relatively higher.On the 5th day of treatment,the negative conversion rate of white blood cell count in peritoneal dialysis fluid was relatively low.Differences were all statistically significant(all P＜0.05).The cure rate of patients in the PaeP group was lower than that in the non-PaeP group,the technical failure rate was higher than that in the non-PaeP group,both with statistically significant differences(both P＜0.05).There was no statistically significant difference in the mortality between two groups of patients(P＞0.05).Conclusion PaeP patients have severe clinical manifestations,poor clinical treatment prognosis,high recurrence and extubation rates.For patients with repeated episodes,resetting and replacing the tunnel after extubation is an effective means to re-duce technical failures.

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 explore the risk factors of suicidal ideation in bereavement callers of Beijing psy-chological support hotline.Methods:Bereavement callers from 2022-2023 hotline callers who were assessed for grief reactions were included in this study.Callers were assessed for current suicidal ideation by asking them if they had suicidal ideation within two weeks.The general demographic data of bereavement callers were collected,and the distress,hope and suicide risk factors of bereavement callers were assessed by hotline interview.The depression de-gree of bereavement callers was assessed by diagnostic screening scale for depression.Logistic regression analysis was used to explore the risk factors of suicidal ideation.Results:Seventy-seven point two percent(1 028/1 332)of callers reported suicidal ideation.Those who were over 25 years old(OR=0.50),with 10-16 years of education(OR=0.59),with over 16 years of education(OR=0.28),and had a high sense of hope(OR=0.41)were less likely to have suicidal ideation.Those with previous suicide history(OR=2.70),high levels of distress(OR=1.72),and high levels of depression(OR=2.86)were more likely to have suicidal ideation.Conclusion:A history of previous suicide,high levels of distress,and high levels of depression among bereavement callers were risk fac-tors for suicidal ideation.

AIM To sequence the chloroplast genome of Artemisia vestita Wall.ex Bess.METHODS Based on ethnobotanical surveys,sample collection and original plant identification were carried out.The chloroplast genome was sequenced using the Illumina platform,followed by assembly and annotation.A comprehensive comparative analysis was conducted with six Artemisia species.The maximum likelihood(ML)phylogenetic tree was constructed based on the chloroplast genome sequences of A.vestita and 32 other Asteraceae species,with Leptocodon hirsutus D.Y.Hong of Campanulaceae as outgroup.RESULTS The chloroplast genome of A.vestita was 151 204 bp in length,including a small single-copy region of 18 331 bp,a large single-copy region of 82 949 bp,and inverted repeat regions of 24 962 bp,with a total GC content of 37.45％.134 genes were annotated,including 89 protein-coding genes,8 ribosomal RNA genes,and 37 transfer RNA genes.A total of 67 SSRs and 44 LSRs were detected in the chloroplast genome.Comparative analysis with closely related species of Artemisia revealed 3 highly variable genes(clpP,rpl36,ycf1)and 6 highly variable intergenic regions(trnK-UUU-matK,rps18-rpl20,rpl36-infA,rpl14-rpl16,rpl16-rpl3 and trnL-UAG-ccsA),which could serve as candidate DNA barcodes for Artemisia identification.Phylogenetic analysis showed that Artemisia formed a highly supportive monophyletic group,with A.vestita and A.gmelinii Web.ex Stechm.being closely related.CONCLUSION This study may provide fundamental data for phylogenetic analysis of Artemisia,taxonomic identification and DNA barcoding construction of Tibetan herb.