Objective  To analyze the nucleic acid screening situation of individuals at-risk for chikungunya in Hainan Province in 2025, to understand the infection patterns and screening detection efficacy among  individuals at different risk levels, and to provide a scientific basis for precise chikungunya prevention and control and optimization of nucleic acid screening strategies. Method  Conduct a special investigation on the nucleic acid screening data of at-risk populations in all medical and health institutions for Chikungunya fever prevention and control in Hainan Province in 2025. Statistical descriptive analysis was used to examine the distribution characteristics of nucleic acid screening positive rates. Chi-square tests were used to compare positive rates between groups, and a 3-day forward moving average smoothing was applied to the daily positivity rate series. Statistical analysis was performed with R software version 4.4.1, with a significance level of α= 0.05. Results  In 2025, Hainan Province conducted nucleic acid screening on 17,212 individuals at risk across 24 cases of chikungunya outbreaks (13 imported cases and 11 local cases). Among them, 38 tested positive for nucleic acid, all of whom were associated with local outbreak risks, with a positive rate of 0.24%(38/15960). Screening of 1,252 individuals associated with imported outbreaks yielded a positive rate of 0(0/1252). The infection risk varies significantly among individuals with different risk levels. The screening positive rate for locally exposed individuals during the epidemic 0.47%(19/4086) was significantly higher than that of general-risk individuals, which was 0.16% (χ2 = 10.66, P < 0.01). The positive rate among family members of cases in the local epidemic was 1.43%(3/210), significantly higher than that of other screened high-risk individuals in the community (χ2 = 6.22, P = 0.01). There were differences in detection efficiency across different screening rounds. The first round of screening reached the peak of the entire cycle (A total of 7,268 people from day 0 to day 1), with the highest initial positive rate of 0.69%(14/2037) among local Co-exposed individuals; the positive rate among local at-risk individuals significantly decreased as the screening rounds progressed, dropping to 0 from the third onward. In terms of temporal distribution, the screening scale peaked at the early stage of the epidemic (5,053 person-times) and then declined overall. Positive cases were detected from day 1 to day 7 following the identification of local cases. Conclusion  During the chikungunya outbreak, there were significant differences in the nucleic acid screening positive rates among individuals with different exposure risks and in different locations. The first round of screening was the most efficient. It is recommended that, after the discovery of an outbreak, general risk individuals and those with shared exposure be reasonably scheduled for rounds of nucleic acid testing within 4 days and 7 days, respectively, in order to achieve early detection and early control of the epidemic.

Objective: To analyze changes in influenza virus types, major influenza A subtypes, and influenza B lineages in Chinese mainland and assess structural changes in influenza activity and virus composition across periods related to the coronavirus disease 2019 (COVID-19) pandemic. Methods: Weekly laboratory influenza surveillance records for Chinese mainland from 2006 to 2025 were obtained from the World Health Organization (WHO) FluNet database. The numbers of specimens tested and influenza-positive detections were summarized, and positivity, influenza A/B types, major influenza A subtypes, and influenza B lineages were analyzed. Descriptive analyses characterized annual and seasonal patterns. Pearson chi-square tests compared virus composition across periods, with Cramer's V as the effect size. The prepandemic, pandemic, and recovery periods were defined as 2015-2019, 2020-2022, and 2023-2025, respectively. Binomial and segmented binomial logistic regression and sensitivity analyses assessed changes in influenza activity and the proportion of influenza A across these periods. The binomial distribution and the exact binomial method were used to estimate the probability of zero B/Yamagata detections and the upper limit of the B/Yamagata proportion. Results: A total of 963 weekly records were included; 8 945 580 specimens were tested, with an overall positivity of 14.31%. Annual positivity fluctuated markedly, and influenza activity was concentrated mainly at the beginning of the year. Positivity in the prepandemic, pandemic, and recovery periods was 15.80%, 7.76%, and 15.01%, respectively; the corresponding proportions of influenza A were 65.55%, 42.27%, and 84.45%. A/B type, major influenza A subtype, and influenza B lineage composition differed across the three periods (all P < 0.001). Regression analysis showed that positivity was lower during the pandemic period than during the prepandemic period (OR=0.39, 95%CI: 0.38-0.39). The comparison of positivity between the recovery and prepandemic periods differed according to the reference period used, whereas the proportion of influenza A was higher during the recovery period than during the prepandemic period (OR=1.14, 95%CI: 1.11-1.18). If the B/Yamagata proportion had remained at the prepandemic level, 64 963 detections would have been expected from 2020 to 2025, whereas none were observed. Conclusion: Influenza activity and virus composition in Chinese mainland showed marked annual and seasonal variation. Structural changes were observed across periods related to the COVID-19 pandemic: influenza activity and the proportion of influenza A decreased during 2020-2022, followed by recovery of activity and stronger influenza A predominance during 2023-2025. The persistent non-detection of B/Yamagata was statistically rare, supporting continued monitoring of influenza virus types, subtypes, and influenza B lineages.

Objective To characterize Mycoplasma pneumoniae (MP) detection, clinical manifestations, and associated factors among patients hospitalized with severe acute respiratory infection (SARI) in Yangpu District, Shanghai, and to inform MP surveillance and control. Methods Surveillance data from 2,369 SARI cases admitted to three sentinel hospitals between October 2023 and December 2025 were analyzed. Respiratory pathogens were detected using multiplex real-time PCR. Group comparisons used Pearson’s chi-square test, Fisher’s exact test, or the Cochran–Armitage trend test, with Benjamini–Hochberg adjustment. Clinical symptoms were compared between MP-only and single non-MP pathogen cases. Multivariable logistic regression was performed using complete 2024–2025 data (N=2,189), adjusting for sex, age, body temperature, antibiotic use before sampling, quarter, and year. Model diagnostics and sensitivity analyses were conducted.Results The overall MP detection rate was 10.55%, peaking at 27.59% in September 2024 and declining to 0–3.70% in 2025. MP was the sole pathogen detected in 96.40% of MP-positive cases. MP-only cases had higher proportions of vomiting, abdominal pain, and diarrhea but lower proportions of fatigue, chest tightness, and shortness of breath than single non-MP pathogen cases (all q<0.05). The odds of MP detection were higher in the 5–9-year (OR=2.91, P=0.011) and 10–17-year groups (OR=3.03, P=0.009) than in the 18–45-year group, and in patients with a body temperature ≥40 °C than in those with a temperature <39 °C (OR=1.67, P=0.041). The odds were lower in 2025 than in 2024 (OR=0.08, P<0.001).Conclusion MP detection was relatively high from the fourth quarter of 2023 through 2024 but declined markedly in 2025. Children and adolescents aged 5–17 years had higher odds of MP detection. Because clinical manifestations overlapped with those of other respiratory pathogens, pathogen testing remains necessary. Long-term, multisite surveillance is warranted to support epidemic early warning and clinical management.

Objective: To analyze the latent infection, case detection, and preventive treatment of student contacts of pulmonary tuberculosis in a high-epidemic county in Guizhou Province, as well as the influencing factors, in order to provide a scientific basis for precise tuberculosis prevention and control among students. Methods: Screening data for 84 index cases of school-associated pulmonary tuberculosis and 8,153 student contacts in the county in 2023 were collected through the Guizhou Provincial CDC Cloud Platform—School Tuberculosis Management System. Descriptive analysis was performed, and group comparisons were conducted using the chi-square test or Fisher's exact test. For latent infection, a generalized estimating equation (GEE) with school ID as the cluster variable and robust standard errors was used for multivariate analysis to control for clustering bias among students within the same school. Due to complete separation in the "primary school" group with zero detected cases, Firth-penalized likelihood logistic regression was employed for the analysis of detected cases. For preventive treatment acceptance, multivariate binary logistic regression was further conducted based on univariate analysis. Results: Among the 8,153 student contacts, the prevalence of latent tuberculosis infection was 6.61% (539/8,153), and the detection rate of active pulmonary tuberculosis was 0.69% (56/8,153). Multivariate analysis using GEE with cluster-robust standard errors revealed that general contact (OR=0.482, 95%CI: 0.336–0.692, P<0.001) and a negative microbiological test result for the index case (OR=0.476, 95%CI: 0.275–0.825, P=0.008) were independent protective factors for latent infection. Age ≥15 years (OR=1.340, 95%CI: 0.913–1.966, P=0.136) and private schools (OR=2.372, 95%CI: 0.949–5.933, P=0.065) did not reach statistical significance after cluster adjustment. Firth regression showed that private schools (OR=47.259, 95%CI: 18.265–152.707, P<0.001) and non-boarding status (OR=6.686, 95%CI: 1.422–28.960, P=0.019) were risk factors for active pulmonary tuberculosis detection, while a negative microbiological test result for the index case (OR=0.317, 95%CI: 0.114–0.718, P=0.004) was a protective factor. The absolute risk difference for case detection between private and public schools was 4.83%. Among latent infection cases, 90.91% (490/539) were eligible for preventive treatment, and the acceptance rate was 75.71% (371/490). Multivariate analysis showed that non-boarding status (OR=0.354, 95%CI: 0.205–0.613, P<0.001), a negative microbiological test result for the index case (OR=0.338, 95%CI: 0.257–0.446, P<0.001), and primary school level (OR=0.410, 95%CI: 0.186–0.903, P=0.027) were independent risk factors for refusing preventive treatment.Conclusion: Enhanced screening should focus on contacts of cases with positive pathogen indicators, with close attention to private schools (absolute risk difference: 4.83%). Health education and medication management for preventive treatment should be strengthened among key populations including non-boarding students and primary school students, in order to effectively block the transmission chain of tuberculosis in schools.

Objective: This study aimed to identify independent risk factors associated with the occurrence of drug-resistant pulmonary tuberculosis (DR-PTB) and, based on these factors, develop and validate a nomogram risk prediction model for clinical auxiliary diagnosis, thereby providing a reference for assisting in the diagnosis of DR-PTB. Methods: Using a convenience sampling method, 329 pulmonary tuberculosis patients admitted to the Sixth People's Hospital of Zhengzhou City between June 2022 and April 2024 were selected as study subjects. Based on drug susceptibility test results, all patients were divided into a drug-resistant group (178 cases) and a non-drug-resistant group (151 cases). Multivariate logistic regression analysis was employed to identify independent risk factors, which were then used to construct a nomogram risk prediction model. The model's performance was systematically evaluated using the receiver operating characteristic (ROC) curve, calibration tests, and decision curve analysis. Results: Univariate analysis revealed statistically significant higher rates of drug resistance in patients who were male, aged ≥40 years, had a BMI ≤18.50 kg/m⊃2;, engaged in manual labor, had a history of smoking or alcohol consumption, had comorbidities such as diabetes or chronic liver disease/drug-induced liver injury, presented with cough or expectoration lasting ≥3 weeks, had fever, had pulmonary cavities, had a history of interrupted treatment, were retreatment cases, had anemia, or had hypoproteinemia. The absolute counts of total T cells and helper T cells were significantly lower in the drug-resistant group compared to the non-drug-resistant group. Multivariate logistic regression analysis identified the following as independent predictive factors for DR-PTB: engagement in manual labor (OR=13.790, 95% CI: 6.115–31.099), presence of pulmonary cavities (OR=3.397, 95% CI: 1.469–7.854), comorbidity with anemia (OR=3.423, 95% CI: 1.126–10.410), and BMI ≤18.50 kg/m⊃2;. The nomogram model established based on these factors demonstrated an area under the ROC curve (AUC) of 0.914 (95% CI: 0.879–0.948) for predictive efficacy. The Hosmer-Lemeshow goodness-of-fit test yielded c⊃2;=9.461, P=0.305 >0.05, indicating good consistency between the model's predicted probabilities and actual observations and ideal calibration. Conclusion: Manual labor, low body mass index (BMI), pulmonary cavities, and anemia are independent risk factors for the development of drug-resistant pulmonary tuberculosis. The nomogram prediction model constructed based on these factors in this study demonstrates excellent discriminatory ability and calibration, showing potential for application in clinical practice as an auxiliary diagnostic tool.

Objective To analyze the clinical characteristics of newly diagnosed tuberculosis (TB) patients in Lujiang County, Anhui Province, and explore the factors influencing their recurrence within 5 years, this study will provide a scientific basis for formulating prevention and control strategies for TB recurrence in local patients. Methods Data for this study were sourced from the "Tuberculosis Management Information System", a subsystem of "China Disease Prevention and Control Information System". Case information on newly diagnosed TB patients was extracted from 2013 to 2020, with patients who had been followed up for at least 5 years selected as the study subjects. A total of 7 923 patients were included, and basic demographic information, diagnosis and treatment information were collected, the dependent variable was "recurrence within 5 years". Descriptive statistical analysis was conducted to describe the general characteristics of the patients, and univariate analysis was used to screen potential factors influencing recurrence within 5 years. Multivariate binary logistic regression analysis was employed to identify independent factors affecting recurrence within 5 years. Results Among the 7 923 newly diagnosed TB patients, 551 (7.0%) relapsed within 5 years, with the primary initial treatment regimen being 2HRZE/4HR (5 101 cases, 64.4%) [Drug abbreviations: isoniazid (H), rifampicin (R), pyrazinamide (Z), ethambutol (E)]. Multivariate logistic regression analysis showed that, dietary habits, population classification, patient diagnosis type, initial treatment regimen, and concurrent tuberculous pleurisy were independent factors influencing recurrence within 5 years in newly diagnosed TB patients. Vegetarian diet and concurrent tuberculous pleurisy were risk factors for recurrence; student/teacher/medical staff/cadre staff, retired personnel, service personnel, extrapulmonary tuberculosis, adoption of 2HRZE/10HRE regimen, and 2HRZE/7-10HRE regimen were protective factors against recurrence (P<0.05). Conclusion Considering implementing nutritional intervention for high-risk populations, strengthening follow-up management for patients with pleurisy, and optimizing treatment options may help reduce the risk of long-term recurrence.

Abstract: Objective　To evaluate the effect of point-of-care testing (POCT) technology combined with a refined management model on improving timeliness of pathogen detection, clinical decision-making efficacy, and clinical outcomes of patients in fever clinics, provide insights for the management practices of medical institutions in China. Methods　A retrospective cohort study was conducted, including 300 patients from the fever clinic of a tertiary hospital in Beijing between October 2024 and March 2025. To control for baseline confounders, 1∶1 propensity score matching was performed on the 260 eligible patients after screening. Ultimately, 240 patients were successfully matched and divided into a POCT group (January-March 2025, n=120) and a PCR group (October-December 2024, n=120). The POCT group underwent bedside testing using the Flash 10 analyzer following standardized training, along with refined management measures; while the PCR group received conventional centralized laboratory testing. The detection efficiency [turnaround time (TAT), consultation duration], clinical decision-making efficacy (targeted treatment rate, time to medication decision-making), and clinical outcomes (7-day revisit rate, hospitalization rate) were compared between the two groups. Results　The median TAT was 0.72 (0.63, 0.75) h in the POCT group, which was significantly shorter than that in the PCR group 15.55 (10.89, 18.78) h, with a statistically significant difference (Z=-11.32, P<0.001). The median total consultation duration was 2.48 (2.25, 2.62) h in the POCT group, which was significantly shorter than that in the PCR group 17.41 (12.77, 21.78) h, with a statistically significant difference (Z=-11.89, P<0.001).  The targeted treatment rate was 90.0% (108/120) in the POCT group, which was significantly higher than that in the PCR group 65.0% (78/120) with a statistically significant difference (χ2=21.28,P<0.001); The median time to medication decision-making was 0.4 (0.3, 0.6) h in the POCT group, which was significantly shorter than that in the PCR group 2.1 (1.9, 2.4) h, with a statistically significant difference (Z=-10.75,P<0.001). The 7-day revisit rate (7.5% vs 14.2%) and hospitalization rate (1.7% vs 5.8%) were both lower in the POCT group than those in the PCR group, with statistically significant differences (χ2=4.51,P<0.05; Fisher's exact test, P<0.05). Conclusions　This integrated mode can significantly shorten pathogen detection time, improve diagnostic and therapeutic precision, and enhance patients' clinical outcomes, form a chain effect of "rapid detection-precise treatment-outcome improvement". This mode provides a practical basis for managing respiratory infectious diseases in fever clinics. Future efforts should focus on optimizing equipment throughput, strengthening community training, and enhancing emergency response capacity. 

Abstract: Objective　To explore the epidemic trends and etiological characteristics of influenza in Ningxia Hui Autonomous Region (hereinafter referred to as Ningxia) from 2015 to 2024, thereby offering a scientific foundation for formulating effective control and prevention strategies in the region. Methods　Surveillance data were obtained from China Influenza Surveillance Information System, pertaining to influenza-like illness (ILI) and pathogen detection in Ningxia from 2015 to 2024. The analysis employed descriptive epidemiological approaches, with statistical processing conducted via Excel and SPSS 26.0. Results　From 2015 to 2024, a total of 88 321 specimens from influenza-like illness cases were collected at sentinel hospitals in Ningxia, among which 9 619 tested positive for influenza, with a positive rate of 10.48%. The positive rates in different years showed statistically significant differences (χ⊃2;=1 289, P<0.001), peaking in 2019 (16.76%) and reaching the lowest in 2020 (4.16%). In terms of pathogenology, the predominant strain was H3N2 subtype of influenza A (38.70%), followed by Victoria lineage of influenza B (29.40%) and H1N1 subtype of influenza A (26.26%), with alternating variations in predominant strains across different years. Concentration analysis showed that the influenza virus transmission showed a strong seasonal pattern in Ningxia from 2015 to 2024. After analyzing with the weighted positive rate, it was discovered that the overall seasonal intensity over the decade and in more than half of the years was significantly weakened. The increase in the conclusion of "even time distribution" after correction indicated that the weighted processing helped to more objectively restore the natural time distribution pattern of the virus activity. By conducting a circular distribution of the weighted positive rate, the results suggested that the overall trend of influenza wss not significant in Ningxia. The P values of all years were greater than 0.05, indicating that there was no significant concentration pattern of influenza during the observation period, and the distribution was relatively uniform or random, with the peak of influenza virus activity concentrated from December to March of the following year. Pearson correlation analysis showed that there was no significant correlation between the concentration M value and the circular distribution γ value. (r=-0.164，P=0.65). Conclusions　From 2015 to 2024, influenza exhibited strong seasonality in Ningxia, with different viral strains circulating alternately across years, and the dominant strains were different. There were strict seasonality in 2016 and 2023, strong seasonality in 2017, 2019 and 2022, relatively strong seasonality in 2021 and 2024, some seasonality in 2015 and 2018, and uniform distribution in 2020. The peak of influenza virus activity varied slightly from year to year, but the peak of influenza virus activity was concentrated from December to March of the next year in most years. 

Abstract: Objective　To analyze the epidemiological characteristics of the first confirmed case of mpox virus Ⅰb in Changzhou City, Jiangsu Province, this study summarizes the findings and disposal process, trace the source of infection, and provide scientific evidence for prevention and control of mpox. Methods　An epidemiological investigation of mpox virus Ⅰb confirmed case was conducted in Changzhou city, Jiangsu Province on August 9, 2025. Clinical samples from the case and close contacts, such as oropharyngeal swab, rash smear and serum, and the relevant environment were taken for the detection of nucleic acid of mpox virus by real-time RT-PCR. Results　The case had a high-risk exposure history to an imported case from Tanzania in Zhejiang Province on July 28, 2025. On August 2, the case had a fever with a peak temperature of 39 ℃. On August 4, pustular rashes was developed on the genital and surrounding skin, accompanied by itching and pain. On August 9, mpox virus Ⅰb nucleic acid was detected in throat swab, rash smear and serum specimen, and the case was diagnosed with mpox. None of the 17 close contacts have developed related symptoms, and all oropharyngeal swabs and serum were tested negative. A total of 30 environmental samples were collected (17 from the hotel room, 8 from the clinic, and 5 from the car). In these samples, 13 tested positive for the mpox virus (12 from the hotel room, 1 from the car), while the other 17 tested negative. Conclusions　This was the first confirmed infection case of mpox virus Ⅰb in Jiangsu Province, which was imported from another province. The characteristics of short incubation period and heterosexual transmission suggest that it is necessary to refine the clinical management protocols and optimize the prevention and control strategies for mpox.

Abstract: Objective　To explore the factors influencing mortality outcomes of pulmonary tuberculosis patients complicated with malignant tumors in Nanjing, and this study will provide the basis for early comorbidity screening of pulmonary tuberculosis and malignant tumors. Methods　Through the "Tuberculosis Information Management System", tuberculosis patients diagnosed and registered whose cases have been concluded were exported in Nanjing from January 2016 to December 2025. The classification and diagnosis of tuberculosis shall be defined according to the relevant standards. A case-control study was conducted, patients with tuberculosis complicated with malignant tumors were regarded as the case group, and tuberculosis patients registered and diagnosed during the same period were randomly matched with the case group at a ratio of 1:4 based on age and gender. Logistic regression model was used to analyze baseline data, clinical data and the influencing factors of tuberculosis complicated with malignant tumors, and to predict mortality outcomes for patients with tuberculosis complicated with malignant tumors. Results　A total of 340 patients were included in this study, with 270 males (79.41%) and 70 females (20.59%). Of these, 185 (54.41%) patients sought direct medical consultation, while 155 (45.59%) were referred, tracked, or from other sources. Multivariate logistic stepwise regression analysis showed that the source of patients being non-direct referral (OR=2.27, 95%CI: 1.30-3.95) was an independent risk factor for tuberculosis complicated with malignant tumors. Compared with other treatments, standard short-course regimens of 2HRZE/4HR (OR=0.36, 95%CI: 0.18-0.72) and long-term regimens (OR=0.31, 95%CI: 0.14-0.69) were protective factors for tuberculosis complicated with malignant tumors. Whereas increasing age (OR=1.08, 95%CI: 1.02-1.14) and occupation types of farmers and workers (OR=4.78, 95%CI: 1.55-14.67) increased the risk of mortality in patients with tuberculosis complicated with malignant tumors. Conclusions　Indirect referral pathways may lead to missed or delayed diagnosis of tuberculosis complicated with malignant tumors, highlighting the need to optimize referral and screening processes. Standardized regimens can significantly improve outcomes. It is urgently necessary to strengthen the prevention and control of tuberculosis among populations such as the elderly and rural workers, and to carry out early screening for co-diseases of tuberculosis and malignant tumors.

Abstract: Objective  To investigate of resistance phenotypes and mutations in kdr and ace-1 genes in wild populations of Aedes albopictus to different insecticides in Jinghong City, and we will provide theoretical support for the scientific control of dengue vectors. Methods  From July to August 2025, larvae of wild populations of Ae.albopictus were collected and raised in the urban area of Jinghong City. After determining their resistance levels to four insecticides using the adult mosquito filter paper contact tube method, the mutations of kdr and ace-1 genes were detected through direct sequencing. Results  Adult Ae.albopictus mosquitoes from Jinghong have developed varying degrees of resistance to permethrin and deltamethrin, but remained sensitive to propoxur and malathion. A total of 174 sequences were obtained at locus 1016: two alleles were identified, the wild-type GTA/V (valine) and the mutant type GGA/G (glycine), with frequencies of 74.71% and 25.29%, respectively; three genotypes were found, the wild-type homozygote V/V, the wild-type/mutant heterozygote V/G, and the mutant homozygote G/G, with frequencies of 64.37%, 20.69%, and 14.94%, respectively. A total of 174 sequences were obtained at locus 1532: two alleles were identified, the wild-type ATC/I (isoleucine) and the mutant type ACC/T (threonine), with frequencies of 99.14% and 0.86%, respectively; two genotypes were found, the wild-type homozygote I/I and the wild-type/mutant heterozygote I/T, with frequencies of 98.28% and 1.72%, respectively. A total of 174 sequences were obtained at locus 1534: 4 alleles were identified, wild-type TTC/F (phenylalanine), mutant type TGC/C (cysteine), mutant type TCC/S (serine), and mutant type TTG/L (leucine), with frequencies of 20.98%, 18.10%, 59.20%, and 1.72%, respectively; 7 genotypes were identified, wild-type homozygote F/F, wild-type/mutant heterozygote F/S, F/L, mutant heterozygote C/S, mutant homozygote L/L, C/C, and S/S, with frequencies of 5.17%, 30.46%, 1.15%, 6.32%, 1.15%, 14.94%, and 40.80%, respectively. A total of 6 types of combined mutations were identified, among which two types (V1016G + F1534S, G1016 + C1534) of combined mutations account for the higher proportion, with frequencies of 17.24% and 13.79%, respectively. There were no statistically significant differences in the mutation frequencies at loci 1016, 1532, and 1534 between resistant and sensitive phenotypes of permethrin and deltamethrin. A total of 176 sequences were obtained at locus 119, and 2 allelic genotypes were found, both encoding glycine (GGA/G and GGG/G). The wild-type GGA/G had the highest allele genotype frequency of 90.63%. The other synonymous mutation type GGG/G had a lower allele genotype frequency of 9.37%. The three genotypes: frequency of the wild-type homozygous G/G genotype was 85.80%, frequency of the synonymous mutation homozygous G/G genotype was 4.55%, and frequency of the wild/mutant heterozygous G/G (GGA/GGG) genotype was 9.65%. Conclusion  Despite the high diversity and frequency of kdr gene mutations, no significant link was established between these mutations and phenotypic resistance.. At the locus 119, only a single base mutation was observed, and it did not cause any change in the corresponding amino acid. In the context of severe pyrethroid resistance, the alternate use of carbamates or organophosphates is currently a reasonable and effective strategy choice.

Abstract：Objectives   To analyze the genetic evolution and molecular characteristics of whole genome sequences of dengue virus type 1 (DENV-1) detected in Changsha from 2019 to 2025, and this study will provide a scientific basis for dengue prevention and control. Methods   DENV-1 positive specimens from confirmed dengue cases detected were screened for whole genome sequencing in Changsha during 2019 and 2025. Based on the obtained whole genome sequences, a series of bioinformatics analyses were performed sequentially, including sequence homology alignment, calculation of nucleotide and amino acid variation rates of each gene segment, phylogenetic tree construction, and screening of amino acid mutation sites. Results   A total of 51 whole genome sequences of DENV-1 were obtained in this study, with genome coverage ranging from 98.29% to 99.20%. Phylogenetic analysis revealed that all the strains belonged to genotype I: Specifically, 45 strains in 2025 clustered together with 3 strains in 2023 belonged to the clade K.1; one strain in 2024 and another strain in 2025 belonged to the clade K.2; the strain in 2019 belonged to the clade E.1. Statistical analysis of gene variation showed that there was no significant differences in the nucleotide and amino acid variation rates among different gene segments. Amino acid mutation analysis demonstrated that all strains carried mutations V324I and V380I in the E protein, accompanied by multiple virulence-associated substitutions. Two strains harbored the N293S mutation in the NS1 protein, while the core functional domains of non-structural proteins remained highly conserved in most other strains. Conclusions   All DENV-1 strains isolated in Changsha from 2019 to 2025 belonged to Genotype I, and the evolutionary clades showed diversified characteristics. Multiple variations were observed in key amino acid sites and functional domains. These findings suggest that continuous case surveillance and early warning should be strengthened to prevent local transmission.

Abstract: Objective   To investigate the prevalence, antimicrobial resistance profiles and genetic characteristics of Staphylococcus aureus (S. aureus) in meat and seafood products in Ningbo region, this study will provide support for local food safety testing and public health protection. Methods   From April 20 to August 5, 2025, six supermarkets and eight wet markets were selected as sampling sites in the Ningbo area using convenience sampling. Meat and seafood samples were collected from sampling sites for bacterial isolation and PCR identification. The Kirby-Bauer (K-B) disk diffusion method was applied to determine the antimicrobial resistance of the isolates. Whole genome sequencing (WGS) was performed via next-generation sequencing (NGS) technology to analyze the molecular and genetic characteristics of the isolates. Results   A total of 23 strains of Staphylococcus aureus were isolated from 134 samples of fresh meat and seafood, with an overall detection rate of 17.2%. These isolates exhibited resistance to 7 antimicrobial agents, with the highest resistance rate to penicillin at 91.3% (21/23). The detection rates of multidrug-resistant (MDR) strains and methicillin-resistant Staphylococcus aureus (MRSA) were 52.2% (12/23) and 13.0% (3/23), respectively. Hemolysin genes (hlgA/B/C and hla/b) and biofilm formation genes (icaA/B/C/D) were detected in 82.6% of the isolates, while the sea enterotoxin gene was exclusively found in strains belonging to sequence type 6 (ST6). ST3055-T84 was the dominant epidemic clone (21.7%, 5/23). Phylogenetic analysis based on whole-genome sequencing single-nucleotide polymorphisms (WGS-SNP) revealed that, all five ST3055-T84 strains were derived from fresh pork (4 strains) and seafood (1 strain) obtained from different wet markets, suggesting the local epidemic of this clonal group; and the three MRSA strains were isolated from fresh pork samples collected from supermarkets and wet markets, among which 2 strains shared a closer genetic relationship. Conclusions   S. aureus strains isolated from meat and seafood exhibit multidrug resistance with a broad resistance spectrum in Ningbo, and carry multiple virulence genes. The isolation of MRSA strains in these food samples poses a potential threat to public health. Therefore, it is necessary to strengthen surveillance of S. aureus, particularly MRSA strains in this region.

Objective To characterize the seroprevalence of IgM antibodies against eight common respiratory pathogens and delineate their epidemiological features among patients with respiratory tract infections (RTIs) at a tertiary care hospital in Taiyuan. Methods A single-center, retrospective observational study was conducted. We included 26,148 first-time test records from outpatients, emergency cases, and inpatients with RTIs between January 2015 and February 2025. Serum samples were analyzed for IgM antibodies against adenovirus, Chlamydia pneumoniae, influenza A and B viruses, Legionella pneumophila, Mycoplasma pneumoniae, parainfluenza virus, and respiratory syncytial virus. Differences across year, month, age and sex were compared. Multivariable binary logistic regression models were fitted with overall positivity and pathogen-specific IgM positivity as dependent variables.The ordered yearly trend of overall positivity was assessed using the Cochran-Armitage trend test. Joint effects of sex and age group were evaluated using logistic regression models with interaction terms. Monthly patterns were visualized by heatmaps and interpreted as seasonal distributions, and correlations were assessed by Spearman rank analysis.Results The overall IgM seropositivity rate for the eight pathogens was 49.11% (12,840/26,148), consisting of 20.89% (5,462/26,148) single infections and 28.22% (7,378/26,148) co-infections. The specific positivity rates were ranked as follows: influenza B virus (34.33%), influenza A virus (29.16%), Mycoplasma pneumoniae (12.04%), Legionella pneumophila (5.99%), Chlamydia pneumoniae (2.38%), parainfluenza virus (1.56%), respiratory syncytial virus (0.85%), and adenovirus (0.69%). Significant differences were observed across years ( χ2 for trend=2,965.807, P<0.001), with the positivity rate peaking in 2024 and being lowest in 2016. Positivity rates for individual pathogens also differed significantly across years (P<0.05). Differences between months were statistically significant (Wald χ2=326.829, P<0.001), with November showing the highest positivity rate (58.95%) and May the lowest (40.37%).The overall positivity rate differed significantly across age groups (Wald χ2=1,078.489, P<0.001), being lowest in infants (<1 year, 8.19%) and highest in children aged 6-14 years (68.74%), followed by those aged 15-24 years (63.97%) and then children aged 3-5 years (60.79%). Females had a higher overall positivity rate than males (51.87% vs. 46.64%, Wald χ2=50.894, P<0.001).Conclusion Patients with respiratory infections in the Taiyuan region exhibited a generally high overall positivity rate for IgM antibodies, with a notable proportion of compound-positive cases. The predominant pathogens were influenza B and A viruses.Significant differences were observed in terms of year, month, age, and gender, indicating the need to implement preventive measures targeting high-risk periods for infection (November–December) and key populations.The findings are more suitable for sero-epidemiological surveillance , and caution is needed when extrapolating to all respiratory tract infection patients.

Abstract: Objective     To analyze the epidemiological changes and clinical characteristics of pertussis infection in children in this region and provide the up-to-date basis for its diagnosis, prevention and control. Methods     A retrospective analysis was conducted on pertussis nucleic acid test data and clinical information of patients under 18 years of age who visited the Children's Hospital of Hebei Province due to suspected pertussis infection from August 2022 to December 2025. Epidemiological and clinical characteristics, such as the population and seasonal distribution of pertussis, were statistically analyzed. Results     The detection rate of pertussis varied across different years. The detection rate of pertussis was the highest in 2022 (37.98%, 602/1 585) and the lowest in 2025 (3.77%, 84/2 228), and the difference was statistically significant (χ²=815.82, P<0.001). The detection rate of pertussis in summer 2022 was higher than that in autumn and winter. The detection rate of pertussis was the highest in winter and the lowest in spring in 2023. Subsequently, the peak detection rates of pertussis in 2024 and 2025 were both concentrated in spring and summer. The differences in detection rates among the various seasons in 2022, 2023, and 2024 were statistically significant (χ²=22.23, χ²=53.31, χ²=852.33,  all P<0.001). In 2022, the pertussis detection rate was the highest in the group under 6 months. In 2024 and 2025, the detection rate of pertussis was the highest in the group aged 7 to < 18 years old, followed by the group under 6 months old. The differences in detection rates among age groups in 2022, 2024, and 2025 were all statistically significant (χ²=27.22, χ²=538.74, χ²=44.16, all P<0.001). The detection rate of pertussis in hospitalized children (14.01%, 751/5359) was lower than that in outpatients (34.04%, 5 237/15 386), and the difference was statistically significant (χ²=776.12, P<0.001). Among the outpatients with pertussis, the group aged 7 to < 18 years old had the largest number, while among the inpatients with pertussis, the group under 6 months old had the largest number. The most common clinical symptoms among hospitalized children with pertussis were paroxysmal cough (64.58%, 485 cases), facial flushing after coughing (68.97%, 518 cases), and nocturnal cough (49.67%, 373 cases). Among the 404 hospitalized children, 263 cases were simultaneously detected with other respiratory pathogens, and the co-infection rate was 65.10%. The pathogen with the highest co-infection detection rate was rhinovirus (33.17%, 134/404).  Except for post-tussive vomiting, there were no statistically significant differences between the simple pertussis group and the co-infection group in terms of paroxysmal coughing, nocturnal coughing, inspiratory whoop at the end of coughing, wheezing, facial flushing after coughing, fever, and the duration of coughing (all P>0.05). Among all the children with pertussis, 260 children had no history of pertussis vaccination (4.34%), and 5 728 children had a history of immunization (95.66%). Conclusions   The epidemiological characteristics of pertussis are continuously evolving under the influence of multiple factors. In 2025, the detection rate decreased significantly, and the proportion of older children increased. Medical institutions should continuously enhance their surveillance and diagnostic capabilities.

Abstract: Objective  To investigate the distribution characteristics and potential clinical value of QFT-Plus antigen-stimulated CD8⁺ T lymphocyte responses in different states of tuberculosis, this study will provide a new auxiliary basis for early differentiation, disease severity assessment, and immune monitoring of tuberculosis. Methods  The clinical data of 3 624 patients after QFT-Plus test who were admitted to the Third People's Hospital of Shenzhen in 2023 were retrospectively analyzed, including 925 patients diagnosed with active tuberculosis (ATB) through culture, 1 425 latent tuberculosis infections (LTBI), and 1 274 healthy controls. The differences of TB1-Nil, TB2-Nil, and TB2-TB1 were compared among the three groups. Patients diagnosed with ATB were reclassified into low, moderate, and high bacterial load groups based on their sputum smear results. The positive rate of QFT-Plus was compared across these different bacterial load groups. Additionally, differences in the distribution of TB1-Nil, TB2-Nil, TB2-TB1, as well as CD4+ and CD8+ T lymphocytes were analyzed among the groups, and their correlations were examined. Results  QFT-Plus TB2TB1 was higher in the ATB patients [0.260(0, 1.000)IU/mL] than that of the LTBI group [0.190 (-0.030, 0.670) IU/mL] and control group [0 (-0.010, 0.010) IU/mL], with statistically significant differences（Z=4.903，P=0.027）. The positive rate of QFT-Plus in the high bacterial load group (94.850%) was higher than that in the low bacterial load group (81.490%) and the medium bacterial load group (82.610%), with a statistically significant difference (χ⊃2;=14.713, P<0.001). There was a statistically significant difference in TB2-TB1 among different bacterial load groups (H=12.685, P=0.002). TB2-TB1 was positively correlated with the bacterial load of Mycobacterium tuberculosis (MTB) (r=0.117, P<0.001), and the proportion of CD8+T lymphocytes among total lymphocytes (r=0.088, P=0.029). Conclusion  QFT-Plus specific CD8⁺T lymphocyte response（TB2-TB1） not only can distinguish ATB and LTBI, but also provides certain clinical reference value for monitoring the progress of tuberculosis and indicating the severity of the disease.

Abstract: Objective  Using quantitative nucleic acid testing for Mycobacterium tuberculosis as an effective tool for screening tuberculosis infection, this study examines factors affecting the positive detection rate in suspected TB patients, and evaluates inadequacies in detection efficiency and patient numbers for pulmonary tuberculosis(PTB) and extra-pulmonary tuberculosis (EPTB). It also assesses the impact of these factors on developing in vitro diagnostic (IVD) products. Methods  The detection outcomes of the tuberculosis molecular diagnosis initiative in "Quantitative Nucleic Acid Detection for Mycobacterium tuberculosis (TB-PCR)" were systematically analyzed at a hospital from January 2018 to October 2022. The study compared variations in the positive detection rates influenced by four factors: sample type (respiratory vs. non-respiratory sources), test frequency (once, twice, and three times or more), statistical method (initial laboratory tests vs. all laboratory tests), and severity of illness (outpatients, general inpatients, and intensive care unit patients). Additionally, the study examined the differences in distribution of copy numbers and its associated trends of target gene IS6110. Results  A total of 8 441 patients with respiratory tract origins and 2 211 patients with non-respiratory tract origins were included in the study. Among outpatients, general inpatients, and intensive care unit patients, the positive detection rate for samples originating from the respiratory tract was significantly higher than that from non-respiratory tract origins（χ⊃2;=166.175，P<0.001；χ⊃2;=52.026，P<0.001；χ⊃2;=82.421，P<0.001）. The number of patients with testing frequencies of 1, 2, and 3 or more were 6 563, 1 401, and 477, respectively. The positive detection rates of the three "test frequency types" demonstrated statistical significance exclusively among outpatients and general inpatients（χ⊃2;=48.897，P<0.001；χ⊃2;=23.935，P<0.001）. However, no statistical significance was observed among intensive care unit patients (χ⊃2;=5.384，P=0.068). A total of 8 441 patients were included in the analysis of initial laboratory test and all laboratory test. Among the three types of patients, there was no statistically significant difference in positive detection rates between the two statistical methods （χ⊃2;=2.054，P=0.152；χ⊃2;=2.086，P=0.149；χ⊃2;=2.668，P=0.102）；Among the samples from respiratory tract and non-respiratory tract, there were 3 752 cases and 1 393 cases, 3 242 cases and 464 cases, 1 447 cases and 35 cases among the three types of patients, respectively. Under fixed conditions (test frequency or statistical method), the positive detection rates were statistically significant of the three types of patients (χ⊃2;=307.882，P<0.001；χ⊃2;=28.679，P<0.001；χ⊃2;=26.574，P<0.001；χ⊃2;=325.894，P<0.001；χ⊃2;=37.571，P<0.001；χ⊃2;=103.216，P<0.001）; The copy number of IS6110 in the patients exhibited a statistically significant upward trend correlating with the deterioration of their condition (J-T value=6.795, P<0.001). Conclusion  The positive detection rate can be influenced by three factors: sample type, testing frequency, and disease severity; The detection rate and sensitivity of EPTB are significantly lower than those of PTB, there is an urgent need for the development of more advanced testing methods. Outpatients and general inpatients should undergo testing a minimum of two to three times, however, there is an observed issue of inadequate testing among patients. Increasing the testing frequency is crucial for individuals suspected of tuberculosis; The influence of sample type and illness severity on the positive detection rate holds substantial significance for guiding the research and development of IVD products.

Objective  To isolate viruses from human H9N2 infection cases and concurrent environmental positive samples in Shiyan City in 2024, conduct whole-genome sequencing and analysis, evaluate the cross-species transmission risk of the virus, and provide evidence for local prevention and control.Methods  Whole-genome next-generation sequencing was carried out on one human-derived strain and four environmental strains. Phylogenetic analysis, homology analysis, key amino acid mutation analysis of the HA/NA genes, as well as temporal divergence and population dynamics analysis of the HA gene, were conducted. Results  All five strains belonged to the H9N2 Beijing/1/94 lineage. The nucleotide and amino acid homologies of the HA gene were 98.25%-99.87% and 99.03%-100%, respectively; for the NA gene of the three strains, the nucleotide and amino acid homologies were 99.57%-99.93% and 99.57%-100%, respectively. Analysis showed that the time to the most recent common ancestor (TMRCA) of the HA gene of 201 H9N2 strains circulating in China from 2020 to 2025 was 1982.74, with an evolutionary rate of 4.11×10⁻⊃3; substitutions/site/year (95% confidence interval: 3.695×10⁻⊃3;–4.537×10⁻⊃3;); the divergence time of the HA gene of the current strains was 2021.61. Population dynamics analysis revealed a stable population size from 1982 to 2015, followed by a gradual increase after 2015, an explosive growth in 2022, a peak in 2023, and a rapid decline thereafter. The HA cleavage site motif was PSRSSR↓GLF in all strains (low-pathogenicity feature). Mutations in the receptor-binding domain included S158N/D, S/F/A160N, H183N, E190V, Q226L, and Q227M. Compared with classic reference strains, the 105NGT and 218NRT sites were deleted, while a new 313NCS site was added, resulting in a total of seven conserved potential N-glycosylation sites. Three strains showed a deletion of TEI at positions 63–65 in the NA protein stalk, and mutations K368N and D369S in the erythrocyte-binding site; no mutations were found at neuraminidase inhibitor (oseltamivir and zanamivir) resistance sites.Conclusion This human infection originated from exposure in live poultry markets. The circulating strains have acquired key mutations that enhance human-type receptor binding ability, showing significant genetic evolution. It is recommended to strengthen routine surveillance and disinfection of live poultry markets, continuously conduct whole-genome monitoring of human and environmental strains, closely track mutations and drug resistance dynamics, and provide timely early warnings.

Objective To analyze the molecular epidemiological characteristics of 46 Listeria monocytogenes strains isolated from clinical cases, associated family members, and food sources in Hainan Province during 2020–2025; to clarify their antimicrobial resistance, genomic features, and potential traceability relationships; and to provide data support for surveillance and source tracing of listeriosis in Hainan Province. Methods A total of 46 L. monocytogenes isolates collected from 2020 to 2025 were included, comprising 13 isolates from clinical patients, 4 isolates from patients’ family members, and 29 foodborne isolates. Whole-genome sequencing was performed for all isolates, and their epidemiological characteristics were systematically analyzed. Bioinformatic methods were used to predict serogroups and sequence types, screen virulence and antimicrobial resistance genes, and conduct single-nucleotide polymorphism analysis, so as to reveal the population structure and molecular epidemiological characteristics of the isolates. Results From 2020 to 2025, a total of 13 listeriosis cases were reported in Hainan Province, with a hospitalization rate of 100% and a case-fatality rate of 53.85%. Patients aged over 60 years accounted for 38.46% of the cases. The main clinical manifestation was bacteremia/septicemia (61.54%), followed by intrauterine infection (23.08%). Fever was the most common symptom (92.31%), and blood and cerebrospinal fluid were the main clinical specimens. Among all food-borne isolates, poultry meat was the most common source, followed by edible fungi and aquatic products. Among the 46 isolates, one foodborne isolate showed dual resistance to tetracycline and erythromycin, one clinical isolate showed intermediate susceptibility to ciprofloxacin, and the remaining isolates were susceptible to all tested antibiotics. Whole-genome analysis of the 46 isolates showed that 15 isolates belonged to lineage I, among which clinical isolates accounted for 53.33%, while 30 isolates belonged to lineage II, among which food-borne isolates accounted for 70.00%. A total of 19 sequence types were identified, including one novel sequence type assigned to clonal complex 7, and these isolates were classified into 15 clonal complexes. Serogroup IIa was predominant, accounting for 54.35% of the isolates. All isolates carried LIPI-1; 6 isolates carried LIPI-3, 6 carried LIPI-4, 27 carried SSI-1, and 5 carried SSI-2. Two ST87 isolates carried the antimicrobial resistance genes msrD, mefA, and tetM. ST8 and ST87 were the predominant sequence types, both of which were detected in food-borne and clinical isolates. Isolates from Haikou City covered most of the identified sequence types. SNP analysis showed no SNP differences among four isolates from maternal-neonatal vertically transmitted infection cases. Only 3 SNPs differed between one ST8 clinical isolate and one ST8 foodborne isolate, and only 1 SNP differed between one ST87 clinical isolate and one ST87 foodborne isolate from the same year, indicating high genetic relatedness; however, direct epidemiological contact evidence was lacking in all instances. Conclusion Although the number of reported listeriosis cases in Hainan Province was limited, the case-fatality rate was high, and the proportion of patients aged over 60 years was notable. Poultry meat was the food category with the highest contamination rate. ST8 and ST87 were the predominant sequence types in food-borne and clinical isolates, respectively. It is suggested that relevant regulatory authorities should strengthen the whole chain monitoring of high-contamination foods such as poultry meat, and health authorities should strengthen the traceability of ST8, ST87 and other key types. This study elucidates the epidemiological and genomic characteristics of L. monocytogenes in Hainan Province, provides baseline data for establishing and improving surveillance systems for food contamination and listeriosis, and offers important reference value for the precise prevention and control of listeriosis.

almonella can be classified into two types: typhoidal Salmonella and non-typhoidal Salmonella. The former primarily causes invasive infections, whereas the latter often leads to self-limited diarrhea, with a small proportion of patients developing disseminated extraintestinal infections. Invasive non-typhoidal Salmonella infection typically presents as a nonspecific febrile illness rather than gastrointestinal symptoms, is associated with higher morbidity and mortality compared to noninvasive infections, and is difficult to differentiate from other causes of febrile illnesses. Infection caused by invasive non-typhoidal Salmonella is emerging as a new global health burden. This article analyzes the disease characteristics, clinical manifestations, diagnosis and treatment course, and laboratory findings of two cases of multisystem infection caused by non-typhoidal Salmonella, aiming to provide therapeutic experience for the clinical management of such infections. Epidemiological data, clinical features, diagnostic methods, and therapeutic regimens of the two cases were collected and analyzed with reference to the literature. The data indicated that neither patient had a clear history of unclean food intake. Despite different modes of disease onset, both patients were ultimately diagnosed with disseminated Salmonella infection based on clinical manifestations and ancillary examinations. During treatment, effective therapeutic regimens were selected according to the extent of multisystem involvement and patients'drug tolerance, and the patients achieved favorable prognoses. Salmonella antimicrobial resistance is climbing year by year, posing challenges to clinical practice. Early diagnosis and treatment are crucial determinants of disease outcomes.

Objective  To analyze the temporal and spatial distribution characteristics and trends of echinococcosis incidence in Xinjiang, providing reference for its prevention and control. Methods  Based on the reported incidence of echinococcosis in counties, cities (prefectures), and districts of Xinjiang from 2005 to 2024, the time trends were analyzed using the Joinpoint regression model. The period from 2005 to 2024 was divided into multiple study phases using annual percentage change (APC) and turning points. Spatial autocorrelation analysis was employed to explore the spatial clustering of echinococcosis outbreaks in Xinjiang. Kriging interpolation technology was utilized to predict spatiotemporal distribution hotspots. Spatiotemporal scanning analysis was conducted using SaTScan software, and the results were visualized with ArcGIS 10.8 software. Results  The average incidence rates of echinococcosis in Xinjiang from 2005 to 2024 varied across seasons (F=5.853, P=0.001),with winter and spring being the main seasons for incidence. The Joinpoint regression model analysis revealed that the reported incidence rates of echinococcosis in Xinjiang from 2005 to 2024 showed an initial upward trend followed by a decline. Global spatial autocorrelation indicated significant global positive spatial correlation in case distribution, with Moran's I ranging from 0.165 to 0.227 (all P<0.001). Local autocorrelation showed that the "high-high" clustering areas and hotspots from 2005 to 2024 were concentrated in the western counties of northern Xinjiang. The Kriging interpolation revealed that the western counties of northern Xinjiang remain high-risk areas for echinococcosis (incidence rate>81.733/100 000), while the southern regions exhibit generally lower incidence rates. Temporal-spatial scanning identified 10 significant spatiotemporal aggregation zones, demonstrating statistical significance (allP<0.05) in both spatial and temporal dimensions. Conclusion:  From 2005 to 2024, echinococcosis in Xinjiang exhibited pronounced spatiotemporal aggregation. The incidence rate showed an upward trend before 2017, peaked in 2017, and subsequently declined annually with localized fluctuations. High-risk areas are concentrated in the western part of northern Xinjiang, with Bortala Prefecture and Yili Prefecture being key regions for prevention and control. Strengthened surveillance in these priority areas is recommended.

Abstract: Objective    This study aims to investigate the clinical and immunological characteristics affecting lung injury in patients with previously untreated pulmonary tuberculosis (TB) and to identify independent indicators related to lung injury. Methods    Patients with previously untreated pulmonary TB were divided into mild and severe lung injury groups according to structural destructive changes on chest imaging. Clinical data, laboratory examinations, and flow cytometry results were collected. Chi-square test and Mann‑Whitney test were used for univariate intergroup comparisons. Multivariate Logistic regression analysis was further performed to adjust for confounding factors and screen independent correlated factors for lung injury severity. Results    Univariate analysis showed that the proportion of TB patients complicated with other pulmonary diseases, diabetes mellitus, and positive sputum bacteriology was significantly higher in the severe lung injury group than in the mild group. The levels of peripheral blood C-reactive protein (CRP), platelet (PLT), and absolute monocyte count (MONO#) were significantly elevated; lymphocyte percentage (LYM%) was decreased; the proportion of granzyme B⁺ NK cells showed a significant difference compared with the mild group. Multivariate logistic regression analysis demonstrated that being complicated with other pulmonary diseases, diabetes mellitus, positive bacteriology, and PLT count were independent correlated factors for lung injury severity (P<0.05). After adjusting for the above confounding factors, the correlations of CRP, LYM%, MONO#, and granzyme B⁺ NK cell percentage with lung injury severity were no longer statistically significant (P>0.05). Conclusion    The lung injury severity in patients with previously untreated pulmonary TB is closely associated with being complicated by other pulmonary diseases, having diabetes mellitus, exhibiting positive sputum bacteriology, and PLT levels; these metrics can serve as independent indicators for evaluating lung injury severity. The correlations of CRP, LYM%, MONO#, and granzyme B⁺ NK cells may be affected by confounding factors, thus lacking independent evaluation value.

Abstract: Objective   To investigate characteristics of whole-genome and mitochondrial genome and phylogenetic relationships of Biomphalaria straminea in Shenzhen, this study provides a scientific basis for the prevention and control of S. mansoni. Methods   Thirty B. straminea snails were collected from agricultural areas in Longgang and Pingshan districts, Shenzhen, from August to December, 2023. Whole-genome resequencing and mitochondrial genome assembly were performed to detect single nucleotide polymorphisms (SNPs), insertions and deletions (Indels), and structural variations (SVs), followed by gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) enrichment analyses. The 16S rDNA sequence was amplified, and phylogenetic trees were constructed. Results   Multiple types of genomic variations, including SNPs and Indels, were identified, along with significant chromosomal translocations. GO and KEGG enrichment analysis revealed that these variations were predominantly enriched in cellular localization, substance transport functions, and pathways such as glutamatergic synapse, cell cycle, and Hippo/Notch signaling. The mitochondrial genome exhibited a circular structure, with a codon usage bias toward TTA. The ND4, ND2, and ND6 genes were under positive selection, whereas ND1, cytochrome b (CYTB), cytochrome oxidase subunit genes COX1 and COX3 were under purifying selection. Overall nucleotide diversity was low, with the highest levels observed in the COX1 and ND2 genes. The 16S rDNA sequences showed high homology, and the strains clustered with Brazilian strains in phylogenetic analysis, confirming their South American origin. Conclusion   There is a close phylogenetic relationship between B. straminea from Shenzhen, China and B. straminea Brazil. Genome-wide associated variants were predominantly significantly enriched in functions related to cellular localization and substance transport, as well as in neuroplasticity and developmental pathways such as the glutamatergic synapse and Hippo/Notch signaling pathways. The mitochondrial genes ND4, ND2, and ND6 displayed strong positive selection signals (Ka/Ks > 1), suggesting adaptive adjustments in oxidative phosphorylation-related metabolism.

Abstract:  Objective    To understand the phylogenetic characteristics of Norovirus (NoV)and the gut microbiota features in pediatric patients in Anhui Province from 2023 to 2024,so as to provide a theoretical basis for exploring microbiota targeted intervention strategies for NoV prevention and control. Methods    Samples were collected from diarrhea surveillance sentinel hospitals during 2023–2024. NoV nucleic acid was extracted and subjected to whole-genome sequencing; 30 positive samples with a Ct value <30 were selected as the NoV infection group (NoV group), and 15 fecal samples from healthy children were included as the control group (HC group). Structural and functional alterations in the gut microbiota of the NoV group were characterized and compared with those of the HC group using 16S rRNA amplicon sequencing. Results    Dual-gene genotyping indicated that the samples contained two major genogroups, GI and GII, among which GI.7[P7] (40.0%) and GII.4[P16] (26.7%) were the predominant infection types. Furthermore, the various genotypic strains exhibited close genetic relatedness to isolates from the United States, Shanghai, Hong Kong, and Guangdong. Gut microbiota diversity analysis showed that α-diversity significantly decreased and β-diversity significantly deviated from the healthy state in the NoV group, reflecting reduced intra-community diversity and altered inter-group community structure, respectively. Among the differential genera, only the genus Dorea was significantly enriched in the NoV group, while various potential short-chain fatty acid (SCFA)-producing bacteria, such as Faecalibacterium, Veillonella, and Anaerostipes, showed a significant reduction. Functional prediction analysis indicated that in the NoV group, the relative abundances of functional genes associated with substance transmembrane transport, DNA repair, carbohydrate metabolism, glycolysis, and antioxidant stress response were significantly upregulated, while those related to iron acquisition were significantly downregulated. In the correlation analysis between genotypes and microbiota differences, the microbiota structures of different genotype groups were significantly separated, with obvious differences between GI.7[P7] and GII-related genotypes, and the differential microbiota were still dominated by Dorea and various potential SCFA-producing bacteria. Conclusion    Among the 30 children with NoV infection, GI.7[P7] and GII.4[P16] were predominant. The local circulating strains were closely related to those reported both domestically and internationally, suggesting that they are more likely represent the sustained transmission of existing epidemic lineages in this region or transmission following importation. Compared with healthy children, NoV infection is closely associated with gut microbiota dysbiosis, manifested as decreased diversity and evenness, a reduction in potential SCFA-producing bacteria, and shifts in certain metabolic pathways. These findings further suggest that different NoV genotypes may differentially influence the patterns of microbiota alterations, providing a basis for understanding NoV-associated intestinal microecological disturbances and their potential intervention targets.

Abstract: Objective    To establish a single-tube duplex recombinase-aided isothermal nucleic acid amplification (RAA) assay for the simultaneous detection of Giardia lamblia and Cryptosporidium. We also evaluated its analytical sensitivity and specificity to support rapid field detection of these two pathogens in drinking water. Methods    Conserved regions of the Giardia lamblia rRNA tandem repeat unit gene and the Cryptosporidium cowp gene were selected as target sequences. Specific primers and fluorescent probes were designed, and a dual-channel single-tube assay was developed. Competitive performance was assessed using mixtures of the two target plasmids at different concentration combinations. Specificity was evaluated with genomic DNA from Clonorchis sinensis, Angiostrongylus cantonensis, Spirometra mansoni, Toxoplasma gondii, Necator americanus, Echinococcus granulosus, and Echinococcus multilocularis. Sensitivity was assessed using serially diluted recombinant plasmids, genomic DNA from the two target parasites, and DNA extracted from varying numbers of cysts/oocysts. Results    The optimal primer/probe sets were GIA-F3/R1/P2 for Giardia lamblia and CRY-F2/R1/P1 for Cryptosporidium. The optimal primer/probe volumes were 0.95 μL (10 mmol/L) each for the forward and reverse primers and 0.55 μL (10 mmol/L) for the probe of Giardia lamblia. For Cryptosporidium, the optimal volumes were 1.55 μL (10 mmol/L) each for the forward and reverse primers and 0.55 μL (10 mmol/L) for the probe. The optimal Mg⊃2;⁺ input was 7.5 μL magnesium acetate (280 mmol/L). The optimal reaction condition was 39 ℃ for 20 min. Under these conditions, both targets were amplified simultaneously in a single tube, while the negative control showed no amplification. In the competition assay, a high-concentration target (200 copies/μL) did not inhibit amplification of the other target at a low concentration (20 copies/μL). Specificity testing showed positive signals only for the target pathogens. All non-target parasitic DNA samples were negative. The limit of detection (LOD) was 1 copy/μL for both targets when recombinant plasmids were used as templates. When genomic DNA was used, the LOD was 0.001 ng/μL. When DNA extracted from different numbers of cysts/oocysts was used, the LODs were 5 cysts/100 μL for Giardia lamblia and 5 oocysts/100 μL for Cryptosporidium. Conclusion    A single-tube duplex fluorescent RAA assay for the simultaneous detection of Giardia lamblia and Cryptosporidium was successfully established. The assay demonstrated rapid reaction kinetics, high sensitivity, strong specificity, and stable dual-target amplification, providing a methodological basis for the development of a rapid nucleic acid detection platform for these two pathogens in drinking water. 

Abstract: Objective　An outbreak of cutaneous anthrax occurred in Yuanmou County in 2024. This study aimed to systematically analyze its aetiology, transmission chain and risk factors to provide a scientific basis for formulating precise and efficient prevention and control strategies. Methods　The study was carried out by the combination of field epidemiological investigation and laboratory detection. Case interviews were conducted to trace onset time, clinical symptom progression, exposure history and close contact information; All close contacts and co-exposed individuals were comprehensively traced to assess risk of infection; The source investigation of the dead animals was carried out to clarify its origin, breeding, onset and disposal; Environmental exposure assessment was carried out to identify high-risk exposure areas. Blood from the case and co-butchers, lesion exudate of the case, dead cow samples (beef, bovine blood water, etc.), and environmental soil samples were collected for combined detection of Bacillus anthracis nucleic acid, antigen, antibody and isolation culture. Comprehensive interventions such as case isolation, contact management, key area disinfection, were implemented, according to Technical Specifications for Anthrax Prevention and Control.Results　The infection source of this cutaneous anthrax outbreak was a dead diseased cow, with 1 confirmed case was reported. A total of 25 specimens were collected, of which 9 samples were tested for Bacillus anthracis nucleic acid, antigen and antibody, and 2 samples were detected for positive, and 1 strain of Bacillus anthracis was isolated from the 25 samples. Epidemic-related bovine products were harmlessly disposed by incineration and deep burial; key areas such as butchering site, cattle pen, abandoned water cellar, patient's residence, were fully disinfected. No new cases were found during 14-day health monitoring of co-exposed individuals. Conclusion　This was a sporadic cutaneous anthrax case caused by unprotected butchering and consumption of an anthrax-dead cattle. Standardized comprehensive prevention and control measures (source control, case treatment, contact management, environmental disinfection) effectively blocked epidemic transmission, confirming their pertinence and effectiveness. In the future, animal quarantine and health education should be strengthened to eliminate the risk of similar exposure.

Abstract： Objective　This study aims to evaluate the consistency of AIMTB, a novel IGRA based on fluorescence immunochromatography, with the currently widely recognized method QuantiFERON-TB Gold plus (QFT-plus) in diabetic patients, and to provide an efficient and suitable solution for active tuberculosis screening in this population. Methods　A cross-sectional study designed was conducted, patients with diabetes through the National Basic Public Health Services platform were selected as the research subjects. General information, diabetes duration, fasting blood glucose level and other data were collected. Venous blood samples were collected simultaneously, and AIMTB and QFT-plus were used to detect tuberculosis infection respectively. Furthermore, the consistency was observed after stratification by diabetes duration and fasting blood glucose levels. Results　A total of 191 diabetic patients were enrolled, after excluding 3 patients with inactive pulmonary tuberculosis, 188 patients were finally included, among whom 100 were males and 88 were females. Patients aged over 60 years accounted for 92.55% (174/188). After excluding 2 cases with indeterminate results, the consistency analysis showed that the overall agreement rate was 95.16% (Kappa=0.84, 95%CI: 0.73-0.94) between AIMTB and QFT-plus. Stratified by diabetes duration <5 years and ≥5 years, the consistency rates were 95.83% (Kappa=0.84, 95%CI: 0.70-0.97) and 94.44%(Kappa=0.83, 95%CI: 0.67-0.99), respectively. After stratification by fasting blood glucose level ≤7.0 mmol/L and >7.0 mmol/L, the agreement rates were 93.44% (Kappa=0.78, 95%CI: 0.57-0.98) and 96.00% (Kappa=0.87, 95%CI: 0.75-0.98), respectively. There was a statistically significant difference in γ-interferon levels among the groups (P<0.05). Conclusion　AIMTB demonstrates good agreement with QFT-plus for detecting TB infection in diabetic patients, and this consistency is not affected by diabetes duration or fasting blood glucose levels, showing superior stability. Considering the advantages of AIMTB such as low cost and simple operation, it can be considered as one of the preferred methods for screening tuberculosis infection among diabetic patients in primary medical institutions.

Abstract: Objective　To establish a single-tube PCR assay based on multicolor melting curve analysis (MeltPlus MTBDR) for the simultaneous detection of the Mycobacterium tuberculosis complex (MTBC) and resistance to rifampicin (RIF) and isoniazid (INH), and this study aims to evaluate its analytical performance and clinical applicability. Methods　A one-tube MeltPlus MTBDR assay was developed using multicolor melting curve technology. The assay combines multiplex asymmetric PCR with melting temperature (Tm) discrimination across different fluorescence channels to enable simultaneous detection of MTBC and RIF- and INH-associated resistance genes. The upper limit of detection and specificity were evaluated using reference strains of Mycobacterium tuberculosis. Respiratory specimens were collected from patients with suspected pulmonary tuberculosis and tested using acid-fast bacilli smear microscopy, liquid culture, GeneXpert MTB/RIF, and MeltPlus MTBDR. Liquid culture and phenotypic drug susceptibility testing were used as reference standards, the sensitivity, specificity, and consistency of each detection method were analyzed. Results　The detection limits of MeltPlus MTBDR assay for MTBC, RIF, and INH targets were approximately 65 CFU/mL, 684 CFU/mL, and 783 CFU/mL, respectively. No cross-reactivity was observed with non-tuberculous mycobacteria or other common respiratory pathogens, indicating high analytical specificity. Using liquid culture as the reference standard, the sensitivity and specificity of MeltPlus MTBDR for MTBC detection were 97.06% and 79.21%, respectively, which were highly consistent with the culture results (Kappa=0.763). Compared with phenotypic drug susceptibility testing, the sensitivities of RIF and INH resistance detection were 96.15% and 93.75%, respectively, and the specificities were 99.39% and 98.11%, respectively. Conclusions　The one-tube PCR-based multicolor melting curve MeltPlus MTBDR assay enables simultaneous detection of MTBC and RIF- and INH-associated resistance genes in a single reaction. The assay demonstrates high sensitivity, good specificity, and high consistency with reference methods. As it can be performed on conventional real-time PCR platforms, this assay represents a useful supplementary tool for the rapid molecular diagnosis of tuberculosis and drug-resistant tuberculosis 

Abstract: Objective    This study aimed to analyze the infection status and temporal trends of brucellosis in livestock and high-risk populations in the Altun Mountains of the Tibetan Plateau from 2020 to 2025, and to explore the dynamic relationship between human and animal infection risks in the context of agricultural intensification. Methods    High-risk populations participating in brucellosis surveillance from 2020 to 2025 were included in this study. Serum antibodies were detected using the Rose Bengal Plate Test (RBPT) and the Serum Agglutination Test (SAT). Temporal trend analysis, comparisons before and after agricultural intensification, and occupational stratification analyses were conducted to evaluate changes in infection risk. Concurrently, local animal brucellosis surveillance data were collected, and Spearman correlation analysis was used to assess the associations between infection levels in livestock and seropositivity rates across different human population groups. Results    A total of 1 194 serum samples were collected from high-risk populations. The overall seropositivity rate for brucellosis during the study period was 4.44% (53/1,194). From 2020 to 2025, the seropositivity rate showed a significant increasing trend (Cochran–Armitage trend test: Z = 4.828, P < 0.001), reaching a peak in 2022 at 8.17% and subsequently fluctuating within the range of 3% and 6%. Nationally, the number of reported brucellosis cases during the same period also showed an initial increase followed by a decrease. Logistic regression analysis with interaction terms showed that the farming model had a moderating effect on the relationship between sheep infection rates and human infection risk (OR=0.78, 95%CI: 0.62–0.98). Following the commencement of large-scale livestock enterprise operations in 2023, no statistically significant change was observed in the overall seropositivity rate. However, occupational stratification analysis revealed that the seropositivity rate among traditional livestock farmers (7.58%, 26/343) was significantly higher than that among employees of intensive livestock enterprises (2.71%, 6/221) (χ⊃2; = 5.070, P = 0.024). Conclusion    The risk of Brucella infection in the Altun Mountains region fluctuates in a pattern consistent with the national epidemic rhythm, and livestock infection levels are more closely associated with the infection risk in populations engaged in traditional farming practices. Differences in livestock production patterns may influence exposure structures. It is recommended that, alongside the promotion of livestock industry transformation, continuous surveillance and targeted interventions for high-risk populations in traditional farming be strengthened; however, the long-term effects of such measures require further validation through extended follow-up studies.

Abstract   Objective    To establish a multiplex PCR system targeting species-specific genes of six clinically prevalent pathogenic Mycobacterium species, and to verify its amplification specificity and effectiveness in reference strains and clinical isolates, thereby laying an experimental foundation for the development of molecular diagnostic products for Mycobacterium species identification. Methods    Genomic DNA was conventionally extracted from six clinically common Mycobacterium reference strains (Mycobacterium tuberculosis, Mycobacterium avium, Mycobacterium intracellulare, Mycobacterium abscessus, Mycobacterium kansasii, and Mycobacterium fortuitum) and two other common pathogenic bacterial standard strains (Staphylococcus aureus and Escherichia coli). Four genes, including 16S rRNA, hsp65, rpoB, and 16S-23S internal transcribed spacer (ITS), were selected as the target genes. Species-specific primers were designed using Primer Premier 5.0 software. A multiplex PCR assay incorporating all four target genes was optimized and established using the reference strains, and subsequently validated across the six target strains and two common pathogenic bacteria (S. aureus and E. coli) to assess specificity. The system was then applied to 48 clinical isolates obtained from Beijing Chest Hospital. All amplified products were sequenced for confirmation, and the successful amplification rates were calculated to evaluate the performance of this panel. Results    A quadruplex PCR system targeting the 16S rRNA, hsp65, rpoB, and 16S–23S ITS genes was successfully established for the six most clinically common Mycobacterium reference strains, achieving 100% amplification efficiency. No amplification was detected in the two non-target control strains or the negative control, and subsequent sequencing enabled unambiguous differentiation of all six Mycobacterium species. Furthermore, among the 48 DNA templates extracted from clinical Mycobacterium isolates, all four expected bands were successfully amplified in all samples, with an amplification success rate of 100% (48/48). Sequencing of the amplification products further confirmed accurate discrimination of the six Mycobacterium species. Conclusion    A multiplex PCR system targeting species identification genes of Mycobacterium was established in this study, enabling simultaneous and specific amplification of four target genes used for species identification. The system demonstrated reliable amplification performance for both reference strains and clinical isolates of six clinically common pathogenic Mycobacterium species, providing important technical support for rapid Mycobacterium species identification and the development of related molecular diagnostic technologies.

Vaccination is one of the public health interventions currently used to reduce the disease burden associated with respiratory viral infections. Using influenza viruses and SARS-CoV-2 as representative examples, this article reviews research advances in the epidemiological characteristics, disease burden, lineage replacement, and adaptive evolution of respiratory viruses in the context of vaccination. It further discusses the implications of vaccination strategies for mitigating viral transmission. Influenza and COVID-19 vaccines can significantly reduce the risks of infection, severe illness, hospitalization, and death. However, their effectiveness in preventing infection and inducing durable immune protection is influenced by viral antigenic drift, immune escape, prior infection history, vaccine-strain matching, age, and other factors. Vaccination not only directly reduces the associated disease burden but may also shape viral evolutionary trajectories by altering the population immunity landscape arising from both vaccination and natural infection. Further efforts to optimize immunization programs, advance the development of broadly protective and mucosal vaccines, and improve equitable global access to vaccines will help inform long-term strategies for the prevention and control of respiratory viral infections.

Objective To systematically describe the epidemiological trend and spatiotemporal distribution characteristics of bacteriologically positive pulmonary tuberculosis (PTB) in Qinghai Province, providing a reference for TB prevention and control. Methods Reported incidence data of bacteriologically positive PTB cases in Qinghai Province from 2015 to 2024 were extracted from the infectious disease surveillance module of the China Information System for Disease Control and Prevention. The Joinpoint regression model was used to analyze the temporal trend of the reported incidence rate. Spatial autocorrelation analyses along from 2020 to 2024. Second-level clusters (2 clusters) covered 10 counties/districts. One secondary cluster encompassed the entire Huangnan Prefecture, as well as Xunhua, Hualong, Minhe, and Ping'an District in Haidong City, and Guinan and Guide Counties in Hainan Prefecture, spanning from 2020 to 2024. The other secondary cluster was located in Maduo County of Golog Prefecture, spanning from 2015 to 2018. Third-level clusters (5 clusters) were distributed across parts of Golog, Huangnan, Hainan, Yushu, Haibei, and Haixi Prefectures, covering a total of 16 counties/districts, with cluster times spanning various years. Conclusions The reported incidence rate of bacteriologically positive PTB in Qinghai Province showed a significant increasing trend, with obvious spatial clustering and regional disparities. Clusters were mainly distributed in Yushu and Golog Prefectures in southern Qinghai, as well as parts of Haixi, Hainan, and Huangnan Prefectures. It is necessary to strengthen active screening among key populations in southern Qinghai, ensure comprehensive case management, continue to improve bacteriological detection capacity, vigorously conduct TB health education, and gradually reduce TB incidence. with spatiotemporal scan analysis were employed to explore spatial clustering. Results From 2015 to 2024, a total of 20 198 bacteriologically positive PTB cases were reported in Qinghai Province, with an average annual reported incidence rate of 33.95 per 100 000 The reported incidence rate showed a significant upward trend (AAPC=9.99%, 95%CI: 5.02%~15.20%, P=0.001). Global spatial autocorrelation results indicated that Moran's I values for each year ranged from 0.062 to 0.584. With the exception of 2015, the values from 2016 to 2024 were statistically significant (P<0.05), demonstrating a spatially clustered distribution. Local spatial autocorrelation results revealed an uneven distribution of the reported incidence rate across Qinghai Province from 2015 to 2024. High-high clusters were identified in each year, primarily concentrated in related counties/cities of Yushu Prefecture and Golog Prefecture in southern Qinghai. Spatiotemporal scan analysis detected a total of 8 statistically significant spatiotemporal clusters (all P<0.05), categorized into three classes. The first-level cluster (1 cluster) covered 15 counties/cities, including the entire areas of Yushu and Golog Prefectures, Xinghai and Tongde Counties in Hainan Prefecture, and Dulan County in Haixi Prefecture, spanning from 2020 to 2024. Second-level clusters (2 clusters) covered 10 counties/districts. One secondary cluster encompassed the entire Huangnan Prefecture, as well as Xunhua, Hualong, Minhe, and Ping'an District in Haidong City, and Guinan and Guide Counties in Hainan Prefecture, spanning from 2020 to 2024. The other secondary cluster was located in Maduo County of Golog Prefecture, spanning from 2015 to 2018. Third-level clusters (5 clusters) were distributed across parts of Golog, Huangnan, Hainan, Yushu, Haibei, and Haixi Prefectures, covering a total of 16 counties/districts, with cluster times spanning various years. Conclusions The reported incidence rate of bacteriologically positive PTB in Qinghai Province showed a significant increasing trend, with obvious spatial clustering and regional disparities. Clusters were mainly distributed in Yushu and Golog Prefectures in southern Qinghai, as well as parts of Haixi, Hainan, and Huangnan Prefectures. It is necessary to strengthen active screening among key populations in southern Qinghai, ensure comprehensive case management, continue to improve bacteriological detection capacity, vigorously conduct TB health education, and gradually reduce TB incidence.

The role of antiviral therapy in influenza now extends beyond shortening symptom duration to the early identification of high-risk patients, reduction of the risk of disease progression, antiviral resistance surveillance, and mitigation of transmission risk. Recent guidelines, randomized controlled trials, systematic reviews, and antiviral susceptibility surveillance indicate that neuraminidase inhibitors remain the mainstay of treatment. Cap-dependent endonuclease inhibitors targeting the polymerase acidic protein can be used for the early treatment of non-severe influenza and for post-exposure prophylaxis; however, the overall benefit of their routine combination with neuraminidase inhibitors in hospitalized patients with severe influenza has not been established. The detection of resistance-associated substitutions alone should not be regarded as evidence of treatment failure or as an indication to switch therapy. Such findings should be interpreted together with viral load, duration of viral shedding, immune status, clinical progression, and transmission risk. Influenza treatment should move beyond comparisons between drug classes toward dynamic management guided by virological feedback. Rapid testing and risk stratification should inform treatment initiation, whereas virological reassessment and resistance monitoring should guide treatment duration and regimen adjustment, with public health measures implemented when warranted.

Objective    To investigate the prevalence, antimicrobial resistance, and genomic characteristics of Campylobacter isolates from human and poultry in Hongkou District, Shanghai. Methods    Fecal samples from diarrheal patients in sentinel hospitals and retail poultry meat samples in Hongkou District were collected for Campylobacter detection from 2023 to 2024. The broth microdilution method was used to determine the antimicrobial resistance to 11 antimicrobial compounds. Whole genome sequencing (WGS) was performed to analyze the carriage of antimicrobial resistance and virulence genes, and conduct multilocus sequence typing (MLST). The baseline characteristics and antimicrobial resistance phenotypes between the sequenced and unsequenced isolates were compared. Results    A total of 376 diarrheal samples were collected, from which 40 Campylobacter strains were detected, yielding a detection rate of 10.64%. These included 32 Campylobacter jejuni (C. jejuni) and 8 Campylobacter coli (C. coli). From 240 poultry meat samples, 67 Campylobacter strains were detected, yielding a detection rate of 27.92%, which comprised 31 C. jejuni, 35 C. coli, and 1 Campylobacter lari strain. Among the 107 Campylobacter isolates, the resistance rates were highest for nalidixic acid (97.20%), ciprofloxacin (97.20%), and tetracycline (89.72%), with a multidrug resistance rate of 57.01%. The multidrug resistance rate was significantly higher in C. coli than in C. jejuni, and higher in poultry-derived isolates than in human-derived isolates (both P<0.05). No statistically significant differences were observed in species composition or major antimicrobial resistance phenotypes between the sequenced (n=77) and unsequenced (n=30) isolates (P>0.05), suggesting that sample loss did not introduce systematic bias. The resistance genes conferring resistance to quinolones resistance gene gyrA T86I mutation (100.00%, 77/77), tetracycline resistance gene tet(O) (74.02%, 57/77), β-lactams resistance gene bla_OXA-193 (50.65%, 39/77), and the antibiotic efflux pump regulator gene cmeR (62.34%, 48/77) were the most frequently detected among 77 Campylobacter isolates. A total of 124 virulence genes were detected in the 77 isolates; C. jejuni (80-109 types) carried a greater variety and number of virulence genes than C. coli (50-71 types) (P<0.001). MLST analysis of 47 C. jejuni isolates identified 39 sequence types (STs), with clonal complex CC-21 being relatively common and the overall ST distribution being highly diverse. Among 30 C. coli isolates, 15 STs were identified, all belonging to the CC-828 clonal complex, and 6 novel STs were discovered. Conclusion    This study provides the first comprehensive insight into the highly diverse ST profiles, the concerning prevalence of antimicrobial resistance, the complex composition of resistance and virulence genes, and the potential risk of cross-species transmission of Campylobacter strains from both human and poultry sources in Hongkou District, thereby offering key data for the precise detection and control of foodborne Campylobacter in this region.

 Objective   To explore the immunoefficacy of measles, mumps and rubella (MMR) combined attenuated live vaccine among recruits in a theater command and analyze the influence of demographic characteristics on immune response, so as to provide evidence for optimizing the prevention and control strategy of respiratory infectious diseases among recruits. Methods   A total of 210 male recruits aged 18–25 years from four stations (Beijing, Shijiazhuang, Qinhuangdao, and Luoyang) were enrolled. All subjects received a subcutaneous injection of 0.5 mL MMR vaccine. Venous serum samples were collected before vaccination (Day 0) and 28 days after vaccination. Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of measles, mumps and rubella IgG antibodies. The antibody positive rate, seroconversion rate, and geometric mean titer (GMT) were calculated, and the effects of different stations, household registration types and educational backgrounds on immune efficacy were analyzed. Results   Before vaccination, the positive rates of measles, mumps and rubella IgG antibodies were 95.24%, 68.57% and 70.95%, with GMT of 728.99, 214.95 and 21.60 IU/mL, respectively. There were no statistically significant differences in baseline antibody levels among recruits from different stations or with different household registration types and educational backgrounds (P>0.05). After vaccination, the GMT of the three antibodies increased to 2 707.74, 1 192.92 and 80.41 IU/mL, the positive rates reached 100.00%, 98.57% and 89.05%, and the seroconversion rates were 1.43%, 57.14% and 35.24%, respectively, which were significantly higher than those before vaccination (P<0.001). After vaccination, there were no statistically significant differences in antibody GMT, positive rate, or seroconversion rate among recruits from different stations or with different household registration types and educational backgrounds (P>0.05). Conclusion   MMR vaccine demonstrates good immunogenicity in the recruit population and can effectively elevate the levels of measles, mumps and rubella antibodies, thereby filling the immune gap. No significant differences in immune response are observed among recruits from different stations or with different household registration types and educational backgrounds, indicating stable and consistent vaccine immunoefficacy. These findings can provide a reliable reference for the military to uniformly formulate recruit immunization programs.

Abstract: Objective    To express and identify the Epstein-Barr virus (EBV) fusion protein EBNA1-LMP1 using the vaccinia virus /T7 system, providing an experimental basis for the development of novel EBV vaccines. Methods    Primers were designed based on the DNA sequence information of EBV nuclear antigen 1 (EBNA1) and latent membrane protein 1 (LMP1). The EBNA1 and LMP1 single genes were obtained by PCR amplification using EBNA1-pCXWB and LMP1-pMV261 plasmids as templates, respectively. The fusion gene EBNA1-LMP1 was constructed by overlap extension PCR through linker sequences. The resulting EBNA1-LMP1 fusion gene was inserted into the pBluescript-II-sk(+) plasmid to construct the recombinant plasmid EBNA1-LMP1-pBlue, which was identified by restriction enzyme digestion, PCR, and sequencing. BHK-21 cells were infected with recombinant vaccinia virus VTF7-3 and subsequently transfected with the recombinant plasmid EBNA1-LMP1-pBlue using Lipofectamine 2000. After an obvious cytopathic effect was observed, the total proteins were extracted from the collected BHK-21 cells. The molecular mass of the expressed protein was initially evaluated by SDS-PAGE, and the antigen specificity of the expressed protein was identified by Western blot analysis using specific antibodies for EBNA1 and LMP1. Results    The EBNA1-LMP1 fusion gene was successfully obtained through overlap extension PCR, with a size of approximately 2 000 bp, which was consistent with the expected design. The recombinant plasmid EBNA1-LMP1-pBlue was successfully constructed and confirmed by double restriction enzyme digestion, PCR, and sequencing. SDS-PAGE revealed that the molecular mass of the expressed protein from the EBNA1-LMP1-pBlue recombinant plasmid was approximately 73 kDa, matching the theoretical size of the EBNA1-LMP1 fusion protein. Western blot demonstrated that the protein was specifically recognized by both anti-EBNA1 and anti-LMP1 antibodies, respectively, indicating correct expression of the EBNA1-LMP1 fusion protein. Conclusion    The EBNA1-LMP1 fusion protein was successfully expressed using the recombinant vaccinia virus system. The protein retains critical antigenic epitopes of both EBNA1 and LMP1, providing important candidate antigens and a preliminary experimental foundation for the subsequent development of novel bivalent EBV vaccines.