Objective:To establish a digital reverse transcription polymerase chain reaction（dRT-PCR）detection method for hepatitis E virus（HEV）using nanoplates，and to provide technical reference for HEV monitoring in shellfish by combining virus enrichment pretreatment methods.Methods:The annealing temperature，primer and probe concentrations of HEV dRT-PCR were optimized，and the specificity of the method was evaluated；the sensitivity of this method for detecting HEV in water samples and oyster extracts was compared. The inhibition rate and recovery rate of HEV detection in artificially contaminated oyster samples were calculated，commercially available oyster samples were tested，and compare them with real-time fluorescence quantitative RT-PCR（qRT-PCR）method.Results:The optimized annealing temperature for HEV dRT-PCR was determined to be 60 ℃，and the final concentrations of primers and probes were 0.4 μmol/L，0.4 μmol/L，and 0.2 μmol/L，respectively，indicating good specificity. The sensitivity of both methods for detecting HEV RNA in water samples was higher than that in oyster extracts. The recovery rates of HEV in oyster specimens contaminated with HEV fecal suspension by dRT-PCR and qRT-PCR were 18.76% and 18.36%，respectively，with no statistically significant difference（ P>0.05）；the inhibition rates were 17.26% and 9.58%，respectively，with statistically significant differences（ P<0.05）；55 commercially available oyster samples were tested，and both methods detected HEV RNA positivity in the same sample. Conclusion:The dRT-PCR method established in this study，combined with “proteinase K digestion，PEG/NaCl precipitation，and chloroform/n-butanol extraction” pretreatment，has a good recovery effect on HEV in shellfish food containing a large amount of PCR inhibitors，and can achieve absolute quantification. It has certain application value in monitoring and risk assessment of HEV in shellfish food.

Human immunodeficiency virus（HIV）entry inhibitors are important in preventing HIV infection by specifically targeting viral envelope proteins（gp120/gp41）or host receptors（CD4/CCR5/CXCR4）to inhibit HIV entry into host cells. This article systematically reviews the research progress of HIV entry inhibitors in recent years，and some of the novel inhibitors such as Fostemsavir，Ibalizumab，and bifunctional fusion inhibitors（2P23-iMab）have shown better effects in inhibiting multidrug-resistant strains. Although great progress has been made in the research and development of HIV entry inhibitors，there are still a series of challenges including drug resistance，drug limitation，target toxicity，which limit their widespread clinical application. Future research directions should focus on multi-target combination strategies，broad-spectrum neutralizing antibody development，and long-term design. In summary，the analyses on the research progress，challenges and future development directions of HIV entry inhibitors will provide a reference for the further research and development and optimization of HIV entry inhibitors，and then promote the development of AIDS prevention and control.

Latent human immunodeficiency virus（HIV）infection remains the principal barrier to functional cure. Proviruses integrate into multiple immune cell types and persist long term，creating a heterogeneous reservoir. Accurate identification of latent infection and assessment of proviral intactness and function are prerequisites for progress toward cure. Here we compare key reservoir assays and their trade-offs，including HIV DNA quantification by quantitative PCR（qPCR）and droplet digital PCR（ddPCR），detection of replication-competent virus by the quantitative viral outgrowth assay（QVOA），and inducible transcription/protein readouts such as Tat/Rev induced limiting dilution assay（TILDA）and viral protein spot assay（VIP-SPOT）. We also outline applications of spatial transcriptomics and in situ hybridization（RNAscope/DNAscope）for anatomic localization and functional analysis. We also summarize advances in experimental models，spanning in-vitro systems（primary T-cell and myeloid latency models）and in-vivo platforms（humanized mice and rhesus macaque SHIV models）used for mechanism studies and intervention testing. Looking ahead，coordinated use of these orthogonal tools can improve estimates of reservoir size，inducibility，and tissue distribution，and provide a practical platform for translational studies toward functional cure.

Exosome play a crucial role in human immunodeficiency virus（HIV）infection. They share high similarities with HIV virions in physicochemical properties and secretion mechanisms：both the formation and secretion of exosome and the budding of HIV depend on the endosomal sorting complex required for transport（ESCRT）complex and VPS4 protein，with their membrane structures being phospholipid bilayers and comparable ranges of diameter and density. These similarities pose challenges to related research. Exosome participate in HIV infection through multiple mechanisms，including promoting HIV entry，inhibiting HIV replication，activating HIV latent reservoirs，and engaging in host antiviral immune responses. In terms of clinical applications，the potential of exosome as biomarkers or vaccine carriers has attracted attention，showing translational prospects. In summary，exosome influence the progression of HIV infection through complex mechanisms，and their application prospects in diagnostic markers，vaccine development，and therapeutic targets merit in-depth exploration.

The orthobunyavirus is a major group of mosquito-borne viruses，mainly prevalent in the Americas，Europe，and Africa. It can cause severe human diseases（e.g.，encephalitis，hemorrhagic fever）and livestock abortions. Reverse genetics，as a powerful tool for viral genome research，has been widely applied to study viral gene functions，develop vaccines，and screen antiviral drugs. This review systematically presents the evolutionary history and applications of the reverse genetics system for orthobunyaviruses，providing critical insights into studies on the infection and transmission mechanisms of these viruses，as well as the development of novel vaccines and therapeutic agents.

Reduning is a Chinese patent medicine made from three traditional Chinese herbs： Artemisia caruifolia Buch.-Ham. ex Roxb.， Lonicera japonica Thunb. and Gardenia jasminoides J. Ellis. It has the effect of clearing heat and detoxifying and is widely used in the treatment of respiratory diseases. This article collates the main components of 70 kinds of heat-toxin，including iridoids，lignans，etc. By reviewing the domestic and foreign literature related to influenza and heat-toxin in recent years，this article reviews chemical composition，pharmacological effects，clinical applications and current predicaments of Reduning，and further discusses the differences between the "multi-component-multi-pathway" of heat-toxin and the single-target mechanism of the drug.

Objective To evaluate the viral composition in respiratory samples from hospitalized children with acute respiratory infections in Yancheng，Jiangsu province，between January 2024 and June 2025 using metagenomic next-generation sequencing（mNGS）.Methods:A total of 247 respiratory specimens，including throat swabs，sputum，and bronchoalveolar lavage fluid，were collected from 247 hospitalized children in Yancheng，Jiangsu province，during the study period. The viral composition in these samples was detected and analyzed using mNGS technology.Results:Among the 247 cases，there were 136 cases of pneumonia（including 4 cases of severe pneumonia），45 cases of bronchopneumonia，27 cases of upper respiratory tract infection，and 39 cases of bronchitis. The ages of the patients ranged from 41 days to 14 years，with a median age of 3 years. High-throughput sequencing results showed that the positive detection rate for viruses in the respiratory samples was 89.88%（222/247），The detection rate of viruses causing respiratory tract infections was 69.23%（171/247），With RNA viruses detected in 71.65%（177/247）of the cases and DNA viruses in 43.32%（107/247）. The top three viruses detected were rhinovirus（24.29%，60/247），human herpesvirus 7（HHV-7）（21.86%，54/247），and influenza A H1N1（20.24%，50/247）. The common detected viruses that cause respiratory infections are rhinovirus（24.29%），influenza A virus H1N1（20.24%），RSV（17.81%），metapneumovirus（10.53%），enterovirus（6.88%），parainfluenza virus（6.07%），bocavirus（6.07%），coronavirus（2.43%），and adenovirus（2.43%）. In children with pneumonia，the top two respiratory viruses detected were rhinovirus（19.85%，27/136）and influenza A H1N1（16.91%，23/136）. In children with bronchopneumonia，the main viruses detected were rhinovirus（31.11%，14/45）and respiratory syncytial virus（24.44%，11/45）. In children with upper respiratory tract infections，the main viruses detected were influenza A H1N1（33.33%，9/27）and rhinovirus（25.93%，7/27）. In children with bronchitis，the main viruses detected were rhinovirus（30.77%，12/39）and respiratory syncytial virus（25.64%，10/39）. Among the 4 cases of severe pneumonia，the viruses detected were influenza A H1N1（2/4），respiratory syncytial virus（1/4），and bocavirus（1/4）.Conclusion:mNGS analysis revealed a high positive detection rate of respiratory viruses（89.88%）in hospitalized children with acute respiratory infections in Yancheng，Jiangsu，from January 2024 to June 2025，with influenza A H1N1 and rhinovirus being the most predominant.

Objective:To analyze the epidemiological characteristics of rabies in China during 2024 to understand the patterns of disease transmission and provide a scientific basis for optimizing prevention strategies and advancing the rabies elimination process.Methods:Data on the national rabies epidemic in 2024 and case-specific information from national surveillance sites were collected from China's National Notifiable Disease Reporting System（NNDRS）. Descriptive epidemiological methods were used to analyze the disease trends and epidemic characteristics.Results:In 2024，China reported 167 human rabies cases across 20 provincial-level administrative divisions（PLADs），resulting in 148 fatalities. This represents a 39.17% increase from 2023（120 cases）and the highest annual incidence recorded since 2019. The epidemic exhibited a distinct geographic clustering in central and southern China，with four provinces（Henan，Guangxi，Hunan，and Hubei）accounting for 62.87%（105/167）of nationally reported cases. October（20 cases）and November（20 cases）recorded the highest monthly incidence，while February had the lowest（2 cases）. Farmers constituted the primary affected demographic（80.84%），with a marked male predominance（male-to-female ratio：2.15∶1）-higher than the 2023 ratio（1.86∶1）. Cases spanned all age groups，predominantly affecting middle-aged and older adults. Analysis of 54 cases revealed dogs as the primary exposure source（93.88%），with domestic dogs constituting 75.00% of these.The laboratory diagnosis rate of rabies cases in 2024 was 43.71%（73/167）.Conclusion:After 16 consecutive years of decline，China's rabies epidemic rebounded in 2024，primarily driven by deficiencies in domestic dog management and weaknesses in post-exposure prophylaxis. Key recommendations include enhancing canine vaccination coverage in high-risk areas，implementing targeted health education for farmers and middle-aged and older adults，improving standardized post-exposure treatment protocols，and optimizing integrated surveillance-alert-response systems，ultimately advancing the achievement of rabies elimination goals.

Objective:To prepare a monoclonal strain of mumps virus of genotype F，which is intended to be applied as a national standard strain，and to provide corresponding virus strain resources for the surveillance，prevention and control of mumps.Methods:The plaque purification technique was used to pick monoclonal strains on Vero/hSLAM cells. The biological characteristics and stability of the monoclonal virus strains were evaluated by observing the cytopathic effect of the monoclonal virus strains，morphological observation of virological characteristics，determination of virus titer and whole-genome sequencing，and mycoplasma detection.Results:After infection of Vero/hSLAM cells，the monoclonal strain of mumps virus constructed after plaque purification could cause cell fusion. Under the negative staining electron microscope，the virus particles were spherical，with a diameter between 100 and 200 nm，and had an obvious envelope. The virus titer of the 2 nd to 6 th generations was between 10 6 and 10 7 TCID 50/ml. Through whole-genome sequencing and phylogenetic analysis，the full length of the virus genome sequence was obtained as 15 384 bp，and it was confirmed that the monoclonal virus strain was of genotype F. The nucleotide difference rate between the 2 nd and 6 th generations was 0.013%. Conclusion:The monoclonal strain of mumps virus constructed in this experiment has typical cytopathic effects and typical morphological structures. The virus has good activity and stable genetic characteristics，and can be used as a candidate strain for application as a national standard strain. It can be used for reagent research and development，evaluation of immunization effects，and other subsequent work.

Objective:To characterize the etiological and genetic features of pediatric influenza outbreaks in Changzhou between 2021 and 2024，with the goal of informing evidence-based prevention strategies and guiding effective management of influenza outbreaks in school settings.Methods:During the period of 2021 to 2024，throat swabs of influenza-like cases from school outbreaks in Changzhou were collected. These samples underwent real-time reverse transcription-polymerase chain reaction（RT-PCR）testing and virus isolation. Epidemiological data were integrated to conduct pathogenetic analysis. The HA genes of isolated strains were amplified and sequenced to perform genetic characterization.Results:Between 2021 and 2024，a total of 256 influenza outbreaks were reported in schools in Changzhou. A total of 3 201 specimens were collected，of which 2 245 were tested positive for influenza viruses，resulting in a positivity rate of 70.13%. The outbreak season was primarily concentrated from December to February each year，with settings predominantly distributed in primary schools（accounting for 73.83%）. The predominant epidemic strains were influenza A viruses，including 118 outbreaks caused by H1N1 and 104 by H3N2. A total of 74 influenza virus strains were successfully isolated from positive specimens，and sequencing of the hemagglutinin（HA）gene was completed. Phylogenetic analysis revealed that certain B/Victoria lineage strains（e.g.，B/Changzhou/01/2021）clustered closely with the vaccine strain B/Austria/3594/17（bootstrap support：99%）. Among influenza H1N1 strains，multiple isolates from 2023—2024 clustered within the same major branch as A/Victoria/4897/2022（bootstrap support：100%）. In contrast，the H3N2 strains exhibited a complex evolutionary pattern，showing variable genetic distances to vaccine strains from different years（e.g.，A/Massachusetts/18/2022，A/Darwin/6/2021）；some isolates were closely related to vaccine strains，while others were more distantly related and scattered across the phylogenetic tree.Conclusions:The influenza outbreak situation in schools was severe and has significant public health implications. Continuous surveillance is essential，and preventive strategies should be promptly adjusted based on the epidemiological and genetic characteristics of circulating strains.