ObjectiveTo analyze the epidemiological and etiological characteristics of a cluster of human metapneumovirus (HMPV) infection in a kindergarten in Gongshu District of Hangzhou City in May 2024, and to provide reference for the prevention and control of similar outbreaks. MethodsAn on-site investigation was conducted using an epidemiological case investigation form. Throat swab specimens collected from cases were screened for 13 respiratory pathogens using real-time fluorescent polymerase chain reaction (PCR). For HMPV nucleic acid positive specimens, the F gene of HMPV was used as the target gene for amplification and sequencing. The sequencing results were then compared with sequences in GenBank database to determine the virus subtypes and perform phylogenetic analyses. ResultsThe outbreak occurred in a kindergarter junior class with a total of 28 preschoolers and 3 teachers and childcare workers. A total of 11 cases (10 preschoolers and 1 teacher) were identified, including 8 male cases and 3 female cases. Clinical manifestations included fever in all 11 cases (100.00%), cough in 8 cases (72.72%), catarrhal symptoms in 4 cases (36.36%), and headache in 3 cases (27.27%). All symptoms were mild, and no severe cases were observed. A total of 11 throat swab samples were collected. Real-time fluorescent PCR test results showed that 3 samples were positive for HMPV nucleic acid, 2 samples were positive for both HMPV and Streptococcus pneumoniae, and 1 sample was positive for both HMPV and rhinovirus. The sequences of the 6 HMPV nucleic acid positive specimens were amplified and analyzed using specific primers, and all were determined to be HMPV subtype A2b. The F gene fragment sequence showed the highest similarity to PV081665.1/Brazil/2024 (99.65%), and also exhibited high similarity to PP683455.1/Indonesia/2021 (99.48%), PV016275.1/Beijing/2024 (99.31%), and PV052230.1/USA/2024 (99.13%). ConclusionThis cluster of acute respiratory tract infection was caused by HMPV subtype A2b, with co-infection of rhinovirus and Streptococcus pneumoniae. The F gene fragment sequences of the HMPV in this outbreak were highly homologous to those of the A2b strains isolated from Brazil, Beijing, Indonesia, and the the United States.

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 explore the clinical characteristics of the nontuberculous Mycobacteria pulmonary dis-ease(NTMPD)patients with previous pulmonary tuberculosis(PPTB)and analyze the clinical difference from the recurrence of pulmonary tuberculosis.METHODS By means of retrospective survey,the patients who were diag-nosed with NTMPD and recurrent pulmonary tuberculosis in Wuhan Pulmonary Hospital from Mar.2021 to Oct.2023 were recruited as the research subjects,a total of 395 patients with NTMPD were enrolled in the study and were divided into the PPTB-NTMPD group with 92 cases and the NPPTB-NTMPD group with 303 cases according to the history of PPTB.The baseline data,clinical symptoms,imaging findings,underlying diseases,pulmonary diseases,and species of nontuberculous Mycobacteria(NTM)were observed and compared.Totally 92 patients with recurrent pulmonary tuberculosis were randomly screened and assigned as the recurrent pulmonary tuberculo-sis group in a 1:1 ratio by matching the PPTB-NTMPD group with the gender and age.The major clinical charac-teristics were compared between the two groups.The 92 patients with PPTB-NTMPD were divided into the 1-10 years group with 40 cases,the 10-30 years group with 37 cases,and the more than 30 years group with 15 cases according to the interval between the initial diagnosis of pulmonary tuberculosis and the diagnosis of NTMPD.The major clinical characteristics were compared among the groups.RESULTS The age was(64.21±10.71)years old in the PPTB-NTMPD group,(60.26±11.83)years old in the NPPTB-NTMPD group(t=3.020,P=0.003).The proportion of patients with body mass index less than 18.5 kg/m2 was 59.78％in the PPTB-NTMPD group,41.25％in the NPPTB-NTMPD group(x2=6.155,P=0.013);the proportion of patients with cough was 77.17％in the PPTB-NTMPD group,65.68％in the NPPTB-NTMPD group(x2=4.313,P=0.038);the inci-dence of cavitary shadow was 50.00％in the PPTB-NTMPD group,35.31％in the NPPTB-NTMPD group(x2=6.414,P=0.011);the incidence of emphysema and pulmonary bullae was 29.35％in the PPTB-NTMPD group,12.87％in the NPPTB-NTMPD group(x2=13.766,P＜0.001);the incidence of chronic obstructive pulmonary disease(COPD)was 22.83％in the PPTB-NTMPD group,14.19％in the NPPTB-NTMPD group(x2=3.875,P=0.049);the incidence of damaged lung was 9.78％in the PPTB-NTMPD group,2.97％in the NPPTB-NT-MPD group(x2=7.530,P=0.014);there were significant differences.Mycobacterium intracellulare and Myco-bacterium abscessus were the predominant species of NTM in both the PPTB-NTMPD group and the NPPTB-NT-MPD group,there was no significant difference in the distribution of NTM species between the two groups of pa-tients.The incidence of patch shadow of the PPTB-NTMPD group was lower than that of the recurrent pulmonary tuberculosis group(P＜0.05),the incidence of bronchiectatic shadow of the PPTB-NTMPD group was higher than that of the recurrent pulmonary tuberculosis group(P＜0.05).There were significant differences in the age,incidence of pleural thickening and incidence of COPD among the patients with different time intervals between ini-tial diagnosis of pulmonary tuberculosis and the diagnosis of NTMPD in the PPTB-NTMPD group(P＜0.05).CONCLUSIONS The previous pulmonary tuberculosis mainly affect the body mass index less than 18.5 kg/m2 and the post-tuberculosis pulmonary diseases such as cough,pulmonary cavity,emphysema,pulmonary bullae,COPD and damaged lung of the NTMPD patients.The NTMPD patients with previous pulmonary tuberculosis are more likely to have bronchiectasia than the patients with recurrent tuberculosis.It is necessary for the clinicians to attach great importance.

Objective:To analyze the evolution and diffusion characteristics of policies related to life expectancy(LE)and healthy life expectancy(HLE)in China from 1982 to 2024 using a biometric approach to policy analysis,revealing the patterns of policy diffusion.Methods:By retrieving databases such as PKULAW.com,We comprehensively collected 701 policy documents closely related to LE and HLE during the period(including 62 central policies and 639 local policies),the policy diffusion process was quantified in four dimensions:diffusion intensity,diffusion breadth,diffusion speed and diffusion direction by using the policy—reference network analysis method.Results:Related policy has gone through the germination period(1982-2001),the development period(2002-2010),the rapid rise period(2011-2015)and the four stages of innovation and pioneering period(2016—present).Policy diffusion is influenced by the hierarchical level of the issuing institution,policy type,and regional economic development level.Policies promulgated by central institutions exhibit stronger and broader diffusion,with guideline—type policies diffusing most widely.The diffusion rate follows a trend of"initial growth,followed by deceleration,and then a slight increase."The primary diffusion directions are vertical diffusion from central to local levels and horizontal diffusion among peers.Conclusion:The diffusion of policies related to LE and HLE is characterized by significant stages,regions and levels.

Aim To investigate the protective effect of liraglutide(LIRA)on acetaminophen(APAP)-in-duced hepatotoxicity at the in vivo level and to reveal the underlying mechanism.Methods Forty SPF grade male C57BL/6J mice were randomly divided into the Control,LIRA(200 μg·kg-1),APAP(500 mg·kg-1),LIRA+APAP,LIRA+APAP+3-methylade-nine(3-MA,30 mg·kg-1)groups,with eight mice in each group.The mice were administered for three con-secutive days,and the materials were taken after 24 h.The general condition and body weight of mice in each group were recorded,and liver morphology was ob-served.Serum ALT and AST levels,as well as SOD ac-tivity,MDA,and GSH content in liver homogenates,were measured using biochemical assay kits.The levels of inflammatory cytokines IL-6,TNF-α,and IL-1β in serum were detected by ELISA.Liver pathological changes were assessed by HE staining,while mitochon-drial and autophagosome structures in liver tissues were observed using transmission electron microscopy.The number of PCNA-positive cells in liver tissues was e-valuated using immunohistochemical staining.The pro-tein expression levels of LC3Ⅱ,p62,Bax,Bcl-2,PC-NA,and CyclinD1 in liver tissues were determined by Western blot.Results LIRA pretreatment can im-prove the general condition of mice with acetamino-phen-induced liver injury(AILI),reduce serum ALT and AST levels,and effectively ameliorate the appear-ance and morphology of the liver as well as the patho-logical damage to liver tissue.Simultaneously,the lev-els of inflammatory cytokines IL-6,TNF-α,and IL-1βare significantly decreased;SOD activity and GSH con-tent are significantly increased,while MDA content is significantly reduced.Transmission electron microsco-py observations reveal the presence of numerous auto-phagosomes in the cytoplasm of liver tissue.Immuno-histochemical staining results indicate a significant in-crease in the number of PCNA-positive cells.Further-more,the expression of LC3Ⅱ,Bcl-2,PCNA,and Cy-clinD1 proteins in liver tissue is significantly upregulat-ed,while the expression of p62 and Bax proteins is significantly downregulated.However,after interven-tion with the autophagy inhibitor 3-MA,the aforemen-tioned protective effects of LIRA are significantly.Conclusions LIRA pretreatment can significantly im-prove liver injury in AILI mice.Its protective mecha-nism may be related to enhancing autophagy in hepato-cytes,thereby reducing oxidative stress,inflammatory response and apoptosis in liver of AILI mice.

Aim To study the therapeutic effect of al-licin on senna-induced diarrhea in mice and to explore the underlying mechanism.Methods Forty-eight C57BL/6J mice were randomly divided into six groups:control,model,loperamide positive control group(2 mg·kg-1),allicin low-dose group(6 mg·kg-1),allicin medium-dose group(12 mg·kg-1)and allicin high-dose group(18 mg·kg-1).Except for the con-trol group,the diarrhea model was induced in the other groups by intragastric administration of senna leaf ex-tract.After drug administration,several diarrhea indi-ces were measured:the rate of loose stools,diarrhea index,accumulated frequency of loose stools at differ-ent time points within 5 hours,and small intestine pro-pelling rate.Serum levels of TNF-α and IL-6 were de-tected by ELISA.Serum NO content was determined u-sing the Griess method.The activities of SOD and CAT,as well as MDA content in the ileum and colon,were measured.The pathological changes and the ex-pression of mRNA related to intestinal barrier proteins in the ileum and colon were evaluated using HE stai-ning and RT-qPCR.Results Allicin improved diar-rhea symptoms in mice induced by senna leaf.It re-duced the rate of loose stools,diarrhea index,cumula-tive number of loose stools in five hours,and the intes-tinal propulsion rate.Allicin also protected the intesti-nal mucosa,decreased serum TNF-α and IL-6 levels,and lowered MDA content in the intestines.It in-creased serum NO levels and enhanced SOD and CAT activities in the intestines.Additionally,allicin upreg-ulated the mRNA expression of AQP1,AQP4,and ZO-1 in intestinal tissues.Conclusions Allicin has a significant therapeutic effect on senna-induced diarrhea in mice.The underlying molecular mechanisms may involve anti-inflammatory and antioxidant effects,in-creased NO content,and upregulation of mRNA ex-pression of aquaporins and tight-junction proteins.

Objective To observe the value of 68Ga-DOTA-NOC PET/CT for diagnosing pheochromocytoma(PCC)and paraganglioma(PGL).Methods Thirty-eight patients with suspected or confirmed PCC/PGL who underwent 68 Ga-DOTA-NOC PET/CT were retrospectively enrolled,among them 20 cases underwent 131I-metaiodobenzylguanidine(MIBG)SPECT/CT during the same period.The value of 68Ga-DOTA-NOC PET/CT for diagnosing PCC/PGL at individual and lesion levels were analyzed and compared to the results of 131I-MIBG SPECT/CT.Results Among 38 cases,there were 20 cases of PCC,14 cases of PGL,1 case of adrenocortical carcinoma and 3 cases of benign adrenal hyperplasia.The sensitivity,specificity,positive predictive value,negative predictive value and accuracy of 68 Ga-DOTA-NOC PET/CT for diagnosing PCC/PGL in all 38 cases was 87.88％(29/33),60.00％(3/5),93.55％(29/31),42.86％(3/7)and 84.21％(32/38),respectively.Totally 188 lesions were detected in 34 cases,with detection rate of 89.95％(188/209).For 20 patients who underwent both 2 kinds examinations,the detection rate of bone,lymph node,liver,lung metastases and the overall lesions of 68Ga-DOTA-NOC PET/CT were all higher than those of 131I-MIBG SPECT/CT(all P＜0.05).No significant difference of diagnostic accuracy of PCC/PGL was found between 68 Ga-DOTA-NOC PET/CT and 131I-MIBG SPECT/CT(P＞0.05).Conclusion The value of 68 Ga-DOTA-NOC PET/CT for diagnosing PCC/PGL was comparable to that of 131I-MIBG SPECT/CT,but the former showed higher detection rate of metastases,hence being helpful to staging and risk stratification of PCC/PGL.

OBJECTIVE To explore the clinical characteristics of the nontuberculous Mycobacteria pulmonary dis-ease(NTMPD)patients with previous pulmonary tuberculosis(PPTB)and analyze the clinical difference from the recurrence of pulmonary tuberculosis.METHODS By means of retrospective survey,the patients who were diag-nosed with NTMPD and recurrent pulmonary tuberculosis in Wuhan Pulmonary Hospital from Mar.2021 to Oct.2023 were recruited as the research subjects,a total of 395 patients with NTMPD were enrolled in the study and were divided into the PPTB-NTMPD group with 92 cases and the NPPTB-NTMPD group with 303 cases according to the history of PPTB.The baseline data,clinical symptoms,imaging findings,underlying diseases,pulmonary diseases,and species of nontuberculous Mycobacteria(NTM)were observed and compared.Totally 92 patients with recurrent pulmonary tuberculosis were randomly screened and assigned as the recurrent pulmonary tuberculo-sis group in a 1:1 ratio by matching the PPTB-NTMPD group with the gender and age.The major clinical charac-teristics were compared between the two groups.The 92 patients with PPTB-NTMPD were divided into the 1-10 years group with 40 cases,the 10-30 years group with 37 cases,and the more than 30 years group with 15 cases according to the interval between the initial diagnosis of pulmonary tuberculosis and the diagnosis of NTMPD.The major clinical characteristics were compared among the groups.RESULTS The age was(64.21±10.71)years old in the PPTB-NTMPD group,(60.26±11.83)years old in the NPPTB-NTMPD group(t=3.020,P=0.003).The proportion of patients with body mass index less than 18.5 kg/m2 was 59.78％in the PPTB-NTMPD group,41.25％in the NPPTB-NTMPD group(x2=6.155,P=0.013);the proportion of patients with cough was 77.17％in the PPTB-NTMPD group,65.68％in the NPPTB-NTMPD group(x2=4.313,P=0.038);the inci-dence of cavitary shadow was 50.00％in the PPTB-NTMPD group,35.31％in the NPPTB-NTMPD group(x2=6.414,P=0.011);the incidence of emphysema and pulmonary bullae was 29.35％in the PPTB-NTMPD group,12.87％in the NPPTB-NTMPD group(x2=13.766,P＜0.001);the incidence of chronic obstructive pulmonary disease(COPD)was 22.83％in the PPTB-NTMPD group,14.19％in the NPPTB-NTMPD group(x2=3.875,P=0.049);the incidence of damaged lung was 9.78％in the PPTB-NTMPD group,2.97％in the NPPTB-NT-MPD group(x2=7.530,P=0.014);there were significant differences.Mycobacterium intracellulare and Myco-bacterium abscessus were the predominant species of NTM in both the PPTB-NTMPD group and the NPPTB-NT-MPD group,there was no significant difference in the distribution of NTM species between the two groups of pa-tients.The incidence of patch shadow of the PPTB-NTMPD group was lower than that of the recurrent pulmonary tuberculosis group(P＜0.05),the incidence of bronchiectatic shadow of the PPTB-NTMPD group was higher than that of the recurrent pulmonary tuberculosis group(P＜0.05).There were significant differences in the age,incidence of pleural thickening and incidence of COPD among the patients with different time intervals between ini-tial diagnosis of pulmonary tuberculosis and the diagnosis of NTMPD in the PPTB-NTMPD group(P＜0.05).CONCLUSIONS The previous pulmonary tuberculosis mainly affect the body mass index less than 18.5 kg/m2 and the post-tuberculosis pulmonary diseases such as cough,pulmonary cavity,emphysema,pulmonary bullae,COPD and damaged lung of the NTMPD patients.The NTMPD patients with previous pulmonary tuberculosis are more likely to have bronchiectasia than the patients with recurrent tuberculosis.It is necessary for the clinicians to attach great importance.

Sepsis is a life-threatening organ dysfunction disease caused by a dysregulated host response to infection.It has com-plex pathophysiological changes,and in severe cases,it can rap-idly develop into septic shock and multiple organ dysfunction or multiple organ failure.At present,the pathological mechanism of sepsis and its induced organ dysfunction is complex and the in-fluencing factors are numerous.So far,there is still a lack of specific and effective treatment strategies.RNA modify-N6-methyladenosine(m6 A)is one of the most common post-tran-scriptional modifications on eukaryotic RNAs.It is involved in the regulation of the occurrence and development of a variety of inflammatory diseases,including sepsis,and even multiple organ dysfunction induced by sepsis by affecting the metabolism of RNAs.It includes cardiac dysfunction,acute lung injury(ALI)and acute kidney injury(AKI).Therefore,this article will dis-cuss the effect of m6A modification on the function of immune cells,and its important role in sepsis and its induced multiple or-gan dysfunction diseases by regulating inflammatory signals,py-roptosis,mitochondrial damage and ferroptosis.This will provide new therapeutic targets and strategies for the clinical prevention and treatment of sepsis and its induced multiple organ dysfunc-tion diseases.

Bovine rotavirus(BRV)is an important pathogen causing diarrhea in calves.To understand the genomic charac-teristics and genetic variations in bovine rotavirus,and to further enrich data on the biological characteristics of rotavirus,we aimed to amplify 11 gene segments of the isolated and cultured G6P[1]bovine rotavirus BLL strain,perform whole genome se-quencing,and analyze the molecular characteristics.MEGA7.0 and DNAMAN software were used for homology and typing a-nalysis,and the whole genome phylogenetic tree was constructed to analyze genetic evolution relationships.The complete geno-type of the BLL strain was G6-P[1]-I2-R2-C2-M2-A3-N2-T6-E2-H3.Phylogenetic analysis of the VP7 and VP4 genes of the BLL strain showed that the VP7 gene had the highest homology with RVA/Cow-wt/HB01/China/2021,and the VP4 gene of the BLL strain was in the same branch as RVA/Human-tc/ISR/Ro8059/1995.From the sequence alignment of VP8*amino acids,the sialic acid domain of the BLL strain was found to be similar to that in other P[1]strains,but different from those in other types of strains,except for residue 189,which was the same as that in Ro8059 but different from that in other strains.The results suggested that the BLL strain might potentially infect humans.Therefore,continued monitoring and study of the biological characteristics of this strain are necessary to provide more information and evidence supporting further research on the cross-species transmission of group A rotavirus in China.