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 gene 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.

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

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 and the serum agglutination test. 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) (χ²=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.

Objective To analyze the temporal and spatial distribution characteristics and trends of echinococcosis incidence in Xinjiang Uygur Autonomous Region (Xinjiang), this study will provide 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 was divided into multiple study phases from 2005 to 2024 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 mean incidence rates of echinococcosis varied across seasons in Xinjiang from 2005 to 2024 (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 showed an upward trend before 2017, peaked in 2017, and subsequently declined annually with localized fluctuations in Xinjiang from 2005 to 2024. Global spatial autocorrelation indicated there was a significant global spatial positive correlation in the distribution of cases, with Moran's I ranging from 0.165 to 0.227 (P<0.001). Local autocorrelation showed that the "high-high" clustering areas and hotspots were concentrated in the western counties of northern Xinjiang from 2005 to 2024. The Kriging interpolation revealed that the western counties of northern Xinjiang remained high-risk areas for echinococcosis (incidence rate>81.733/100 000), while the southern regions exhibited generally lower incidence rates. Temporal-spatial scanning identified 10 significant spatiotemporal aggregation zones, demonstrating statistical significance in both spatial and temporal clustering (P<0.05). Conclusion Echinococcosis exhibited pronounced spatiotemporal aggregation in Xinjiang, from 2005 to 2024. The incidence rate showed an initial upward trend followed by a decline. High-risk areas are concentrated in the western part of northern Xinjiang, with Bortala Mongolian Autonomous Prefecture and Yili Prefecture being key regions for prevention and control. It is suggested to strengthen surveillance in these priority areas.

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 Mg²⁺ 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.

Objective To clarify the population composition, geographical distribution, habitat types, nocturnal blood-feeding patterns and seasonal fluctuation dynamics of vector Anopheles mosquitoes in Guizhou Province in the post-malaria elimination stage, so as to provide scientific evidence for provincial malaria prevention and control as well as malaria risk assessment. Methods From 2020 to 2025, one population survey was conducted annually in July and August using mosquito light traps in at least 1/3 of all counties and districts across Guizhou Province. Meanwhile, one county or district was selected from each prefecture-level city to carry out density monitoring from June to October via double-layer human-baited tent traps. Population and density surveillance data were collected from 88 counties (districts) of the province. The χ² test was adopted for statistical analyses of indicators including population composition ratios and habitat preferences, and circular statistics were applied to estimate the peak density date of An. sinensis. Results A total of 50 644 Anopheles mosquitoes were captured at all surveillance sites across Guizhou Province from 2020 to 2025, including 48 741 An. sinensis, 5 An. anthropophagus, 88 An. minimus, and 1 810 other Anopheles species. A total of 35 087 Anopheles sinensis were collected from three habitats: outdoor areas, human dwellings, and livestock shelters. Among them, 29 117 (82.98%) were captured in livestock shelters, 4 403 (12.55%) in outdoor areas, and 1 567 (4.47%) in human dwellings. All years showed a single density peak in July except 2022, which presented bimodal peaks in June and August. Circular statistical analysis estimated that the peak date in 2022 fell on August 4th, while the peak dates of the remaining years were all distributed within July. The weighted average peak date from 2020 to 2025 was estimated as July 15. The nocturnal blood-feeding activity of Anopheles sinensis in Guizhou Province exhibited an obvious bimodal pattern, with the primary peak occurring at 19∶00-21∶00 and the secondary peak at 4∶00-5∶00. Conclusion After malaria elimination, malaria vector Anopheles mosquitoes dominated by An. sinensis are still widely distributed in Guizhou Province, with relatively high densities in some regions. Therefore, the risk of local retransmission caused by imported malaria cases will persist. To better carry out malaria prevention and control work and maintain the achievements of malaria elimination, it is necessary to continuously monitor malaria vector mosquitoes, conduct timely malaria transmission risk assessments, and implement vector control measures.

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 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.

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.

Objective To explore the relative contributions of metabolic enzyme activity and kdr mutations to pyrethroid resistance development in Aedes albopictus from Jinghong City. Methods From July to August 2025, wild populations of Aedes albopictus were collected in the urban area of Jinghong City; bioassays confirmed resistance to both permethrin and deltamethrin. The activities of three metabolic enzymes—glutathione S-transferase (GST), cytochrome P450 (CYP450), and nonspecific esterase (NSE)—were determined by ELISA, and enzyme activity ratios were calculated. Based on the CART model implemented in R language, the relative contributions of alterations in metabolic enzyme activity and kdr gene mutations to resistance development were analyzed. Results A total of 200 samples were tested. Compared with the susceptible strain, the activity ratios of GST, NSE, and CYP450 in the permethrin and deltamethrin groups ranged from 0.96 to 3.29-fold, 0.97 to 2.09-fold, and 1.05 to 2.37-fold, respectively. The median (P₂₅, P₇₅) activities of GST, NSE, and CYP450 in insecticide-exposed Ae. Albopictus were generally higher than those in the susceptible strain (Z=-5.928, -5.390, -3.930, -5.210, -5.094, -3.815, -4.920, P<0.05). The resistant/susceptible (R/S) activity ratios of GST, NSE, and CYP450 were 2.39-fold, 1.56-fold, and 1.12-fold in the permethrin treatment group, and 2.16-fold, 1.93-fold, and 2.26-fold in the deltamethrin treatment group, respectively. Enzyme activities differed significantly between the survival and death groups for both pyrethroid insecticides (Z=-4.517, -5.077, -3.075, -5.061, -2.691, -5.167, P<0.05). CART model analysis on the relative contribution of resistance mechanisms indicated that GST was the dominant variable contributing to permethrin resistance, with the contribution order of GST > CYP450 > NSE > kdr; for deltamethrin resistance, CYP450 served as the primary contributing factor, and the variable contribution order was CYP450 > GST > NSE > kdr. Conclusion Metabolic enzyme activities were generally elevated in pyrethroid-resistant populations of Aedes albopictus from Jinghong City, and three metabolic enzymes were closely associated with the resistant phenotypes to permethrin and deltamethrin. Metabolic resistance was the predominant mechanism underlying resistance to both insecticides tested, with target-site resistance playing an auxiliary role.

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 in Lujiang County, Anhui Province 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. 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 (P<0.05). Vegetarian diet and concurrent tuberculous pleurisy were risk factors for recurrence (P<0.05); 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 A vegetarian diet and concomitant tuberculous pleurisy are risk factors for relapse within five years among newly diagnosed tuberculosis patients in Lujiang County, Anhui Province. 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.

Objective To investigate the effects of meteorological factors, air pollutants, and socioeconomic factors on the incidence of pulmonary tuberculosis (PTB) in the Xizang Autonomous Region (Xizang), and to provide scientific evidence for PTB prevention and control. Methods Data on PTB cases, meteorological factors, air pollutants, and socioeconomic indicators in Xizang from 2010 to 2022 were collected. Joinpoint regression was used to describe the long-term trend of incidence, quantified by the annual percentage change (APC). Interrupted time series analysis was employed to assess the impact of non-pharmaceutical interventions (NPIs) on the trend of PTB incidence. Distributed lag nonlinear models and generalized additive models were applied to quantify the exposure-response relationships of meteorological factors and air pollutants, as well as the associations between socioeconomic factors and PTB risk, respectively. Results The average reported incidence of PTB in Xizang from 2010 to 2022 was 141.97 per 100 000 population. The incidence showed an increasing trend from 2010 to 2019 (APC=4.56%, P<0.001) followed by a decreasing trend from 2019 to 2022 (APC=-16.50%, P=0.001). The implementation of NPIs was significantly associated with the decline in incidence (regression coefficient β₃= -0.265, P<0.001). The relationship between average temperature and PTB risk was U-shaped, with a relatively high relative risk at -7.10 ℃ (RR=2.35). Relative humidity was positively associated with PTB risk. Excessive cumulative sunshine duration (RR=1.81 at 12.40 h) and elevated concentrations of SO₂ and NO₂ were also associated with increased PTB risk. A per capita GDP below approximately 30 000 yuan was associated with a higher risk of tuberculosis incidence, whereas a natural population growth rate exceeding 13‰, a population density above 324 people/km², and insufficient hospital and physician resources were each associated with an increased risk. Conclusion The incidence of PTB in Xizang is influenced by a combination of meteorological, environmental, and socioeconomic factors. Future efforts should establish a multi-level, long-term prevention and control system integrating environmental interventions and socioeconomic support.

Objective To develop an early diagnostic prediction model for rhinovirus (RV) pneumonia in children using routine hematological indicators and basic clinical characteristics based on machine learning algorithms, and to evaluate its predictive performance, aiming to improve early identification and diagnostic accuracy of this disease. Methods A total of 4 444 hospitalized children from the pediatric departments of six tertiary Class A hospitals in Gansu Province between October 2023 and September 2024 were enrolled as study subjects. Among them, 869 children diagnosed with pneumonia and RV-positive were assigned to the case group, while 3 575 children diagnosed with pneumonia but RV-negative served as the control group. A total of 27 feature variables were collected for each participant, including 23 routine hematological parameters and 4 clinical characteristics (age, gender, address, and admission time). Subsequent statistical analyses were performed to compare these variables. Nine machine learning algorithms were employed to construct early diagnostic prediction models using R Studio 4.2.1 software, including random forest, decision tree, naive Bayes, logistic regression, XGBoost, support vector machine (SVM), neural network (NN), k-nearest neighbors (KNN), and extremely randomized trees (ERT). Internal cross-validation was applied to evaluate model performance. Results Among the nine machine learning models compared, the random forest model demonstrated the best predictive performance, with an AUC of 0.636 (95%CI: 0.594-0.678). The PR-AUC, accuracy, specificity, sensitivity, precision, and F1 score were 0.317 (95%CI: 0.265-0.377), 0.745 (95%CI: 0.719-0.770), 0.854 (95%CI: 0.832-0.876), 0.295 (95%CI: 0.234-0.361), 0.330 (95%CI: 0.263-0.396), and 0.311 (95%CI: 0.251-0.369), respectively. The SHAP plot revealed that the top 5 most important feature variables were hematocrit (HCT), lymphocyte percentage (LYMPH%), white blood cell count (WBC), eosinophil absolute count (EO), and monocyte percentage (MONO%). Conclusion The early diagnostic prediction model constructed using machine learning approaches based on hematological data and basic clinical information may assist medical staff in making rapid clinical decisions during the diagnosis and treatment of rhinovirus pneumonia in children, and provide data support for disease prevention and control.

Objective To analyze the distribution characteristics and pathogen carrying situation of free-living tick in Hefei, this study will provide a reference basis for the prevention and control tick-borne diseases. Methods In May and October 2025, free-living ticks were collected from 90 sampling sites using the flagging method in Hefei, and morphological identification was performed using a stereomicroscope. The carriage of Bandavirus dabieense (DBV), Coxiella burnetii (Cb), Borrelia burgdorferi (Bb), Spotted fever group Rickettsia (SFGR), Anaplasma phagocytophilum (APH), and Ehrlichia chaffeensis(E. chaffeensis) were detected in ticks using real-time fluorescent PCR technology. Descriptive statistics such as tick density, composition ratio, and positive tube rate were used, and statistical analysis was performed using t- tests, rank-sum tests, tests and others. Results A total of 3 885 ticks were captured, of which 3 673 were Haemaphysalis longicornis, accounted for 94.54%. The total tick density in May [2.389/(flag·100 m)] was lower than that in October [3.269/(flag·100 m)], and the difference was statistically significant (H=3.043, P=0.002). The tick density in different regions from high to low were as follows: Changfeng [3.257/(flag·100 m)], Feidong[2.972/(flag·100 m)], Lujiang [2.963/(flag·100 m)], Chaohu [2.797/(flag·100 m)], Main Urban Area [2.400/(flag·100 m)], Feixi [2.368/(flag·100 m)], with no statistically significant difference (H=6.265, P=0.281). Tick density in different habitat from high to low were as follows: scenic areas [3.136/(flag·100 m)], forest land [2.869/ (flag·100 m)], farmlands [2.713/(flag·100 m)], barren grassland [2.512/(flag·100 m)], and the difference was statistically significant (H=5.575, P=0.134). Etiological detection of 572 ticks showed that, the positive tube rate for SFGR, DBV, Cb and Bb were 47.03%(269/572), 0.70%(4/572), 0.52%(3/572) and 0.35%(2/572), respectively. APH and E.chaffeensis were not detected. Conclusion H. longicornis is the dominant tick species in Hefei, with higher densities in scenic areas and forest land, carrying pathogens such as DBV, Cb, and Bb, among which SFGR has a high carrying rate. It is recommended to strengthen comprehensive prevention and control measures such as environmental sanitation, vector control, and public health education in areas with high tick density, and to enhance prevention and control of tick-borne diseases.

Objective To investigate cadmium levels in major dietary foods and assess the risk of dietary cadmium exposure among residents in Hainan Province, thereby providing a theoretical basis for regional food safety decision-making. Methods Cadmium concentrations detected in dietary samples collected from 20 cities and counties across Hainan Province from 2014 to 2025 were statistically analyzed. A point estimate method combined with Monte Carlo simulation was applied to evaluate residents' dietary cadmium exposure levels and their distributions. The margins of safety (MOS) approach and the health risk assessment model recommended by the United States Environmental Protection Agency (USEPA) were employed to assess potential health risks. Results Among 1 726 samples covering 14 categories of major dietary foods, the overall detection rate of cadmium was 76.1% (1 313/1 726), with detected levels ranging from not detected (ND) to 12.400 mg/kg. The exceedance rate was 7.4% (108/1 469), with edible fungi recording the highest exceedance rate at 25.0% (10/40). The mean cadmium content across all food samples was 0.500 mg/kg. Relatively higher mean cadmium levels were found in tea, crabs and shellfish, laver, crustaceans, edible fungi, and poultry and livestock offal, at 1.568, 1.153, 1.027, 0.832, 0.725, and 0.104 mg/kg, respectively. Sensitivity analysis indicated that the sensitivity coefficient of cadmium content in rice was 7.47×10^-1, followed by a sensitivity of 2.35×10^-1 for the intake of rice. The estimated average monthly cadmium exposure for residents in Hainan was 23.090 μg/(kg·BW), accounting for 92.4% of the provisional tolerable monthly intake (PTMI). The mean MOS value was 1.08. The main dietary sources of cadmium were rice (57.73%), aquatic foods (23.05%), and vegetables (10.12%), together accounting for 90.90% of total cadmium exposure. The estimated individual average carcinogenic risk and the individual carcinogenic risk at high exposure level (P₉₅) were 6.14×10^-5/year and 2.00×10^-4/year, respectively. Conclusion Cadmium content in some foods in Hainan Province exceeds the permissible limit. The overall risk of dietary cadmium exposure among residents is relatively low; however, the high-exposure population may face certain health risks from dietary cadmium exposure. Monitoring of cadmium content in major foods such as rice, aquatic products, and vegetables should be strengthened.

Objective To investigate characteristics of whole-genome and mitochondrial genome and phylogenetic relationships of Biomphalaria straminea (B. straminea) in Shenzhen, provides a scientific basis for the prevention and control of Schistosomiasis 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 enriched in neuroplasticity and development-related pathways, including the glutamatergic synapse, thyroid hormone signaling, retrograde endocannabinoid signaling, and Wnt 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.

Objective This study aims to investigate the clinical and immunological characteristics affecting lung injury in patients with newly diagnosed pulmonary tuberculosis, to identify independent indicators associated with lung injury, and to provide a basis for early risk stratification and prognostic assessment of tuberculosis-related lung injury. Methods Patients with previously untreated pulmonary TB at Beijing Chest Hospital Capital Medical University, from 2023 to 2024, 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.

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, analyze the epidemiological features of NoV genotypes and the potential recombinant-associated transmission risks, and further investigate the structural and functional alterations of the gut microbiota in NoV-infected children, as well as the potential associations between different genotype infections and gut microbiota changes. Results Dual-gene genotyping indicated that the samples contained two major genogroups, GⅠ and GⅡ, among which GⅠ.7[P7] （12, 40.0%） and GⅡ.4[P16] （8, 26.7%） were the predominant infection types. Furthermore, the various genotypic strains exhibited close genetic relatedness to isolates from the United States, Shanghai (China), Hong Kong (China), and Guangdong (China). 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 acids （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.

Objective To investigate the virulence and antimicrobial resistance characteristics of non-O1/non-O139 Vibrio cholerae in sewage environments of Xuhui District, Shanghai, and to provide scientific basis for cholera prevention and control in this area. Methods Non-O1/non-O139 Vibrio cholerae isolates were isolated from sewage samples collected at vegetable markets in Xuhui District, Shanghai, between April 2024 and September 2025. O1 and O139 serotypes were identified using the serum agglutination method. Biochemical identification was performed using the VITEK 2 Compact automated biochemistry analyzer and Gram-negative bacteria identification cards. Detection of ctxA/B virulence genes was performed using quantitative real-time polymerase chain reaction. Antimicrobial susceptibility testing was conducted via broth microdilution method. Whole genome sequencing and bioinformatics analysis was uesed to predict antimicrobial resistance and virulence genes in the isolates. Results A total of 345 sewage samples were collected, with 57 (16.52%) positive for non-O1/non-O139 Vibrio cholerae. The detection rate of non-O1/non-O139 Vibrio cholerae showed statistically significant temporal variation (χ²=48.598，P<0.001), with the highest rate (70.00%, 7/10) observed in July 2025. Antimicrobial susceptibility testing revealed that the isolates showed the highest resistance rate to cefazolin (70.18%, 40/57) among the 57 isolates, while all isolates were susceptible to azithromycin (100.00%, 57/57). The 57 isolates harbored 19 resistance genes across 7 classes (β-lactams, sulfonamides, tetracyclines, aminoglycosides, quinolones, aminopenicillanides, and disinfectants/preservatives), with β-lactam resistance genes exhibiting the highest carriage rate (54.39%, 31/57). Twenty-nine virulence genes across four categories (hemolysin, RTX toxin, type Ⅱ secretion system, and type Ⅵ secretion system) were detected in the 57 isolates. All strains carried the epsH and epsK virulence genes. Conclusion This study isolated 57 non-O1/non-O139 Vibrio cholerae strains exhibiting significant temporal variation. These strains demonstrated high sensitivity to azithromycin, harbored a diverse array of resistance genes, and possessed virulence genes covering toxin production, transport, release, and injection processes, warranting close attention.

Healthy carriage of Neisseria meningitidis (Nm) is a prerequisite for the development of invasive meningococcal disease (IMD), but the relationship between carriage and disease onset does not constitute a simple direct correlation. This review systematically summarizes the epidemiological characteristics of global Nm healthy carriage, highlighting significant spatiotemporal heterogeneity and complex serogroup distribution patterns. By comparing the effects of different vaccine types (polysaccharide, conjugate, and protein-based vaccines) on both carriage and disease, this review reveals the limitations of polysaccharide vaccines in reducing carriage despite their ability to lower disease incidence, the advantage of conjugate vaccines in achieving herd immunity by reducing carriage, and the constraints of serogroup B vaccines in this regard. Furthermore, this review discusses the multidimensional considerations for integrating complex influencing factors into immunization strategy development, particularly the challenge of cost-effectiveness analysis in the context of low disease incidence. Future prevention and control strategies should shift from a sole focus on disease control toward precision interventions based on dynamic surveillance and transmission dynamics modeling.

Objective To determine the source of infection, transmission pathway, and etiological characteristics of the first reported infant case of Brucella canis infection in Zhejiang Province, and to provide a scientific basis for precise prevention and control of pet-associated brucellosis in non-pastoral and urbanized regions. Methods A detailed source-tracing investigation was conducted for a confirmed infant case in Jiaxing using field epidemiological and environmental health methods. Laboratory testing employed both smooth and rough Brucella antigens for serological screening and confirmation. The isolate obtained from the child's blood was subjected to Gram staining, biochemical identification, species-specific PCR, and whole-genome sequencing (WGS). Molecular source tracing was performed through multilocus sequence typing (MLST) and phylogenetic tree analysis. Results The patient, an 11-month-old infant, initially presented with high fever accompanied by respiratory tract infection and convulsions. Serological screening revealed a weakly positive antibody response to the smooth antigen test, while antibodies against the specific rough antigen were positive with a titer of 1∶200. Whole-genome sequencing of the isolated strain JX-01 demonstrated high homology with the reference Brucella canis strain ZJ-1, confirming its identification as Brucella canis. Epidemiological investigation revealed that a free-roaming pet dog in the household had a history of abnormal abortion and tested strongly positive for Brucella antibodies. Comprehensive assessment indicates that this outbreak was highly likely caused by the child's close contact with an infected dog and its potentially contaminated environment. Conclusion Brucella canis infection is highly prone to being missed in routine clinical serological screening. Subclinically infected companion animals and contaminated environments pose potential exposure risks to non-occupational populations. Clinical practice should improve the differential diagnosis of atypical cases. Relevant sectors should implement the "One Health" concept, deepen cross-sector collaboration and pathogen molecular tracing, and strengthen public health education on responsible pet ownership and disease prevention.

Objective Dengue fever patients are frequently complicated with myocardial injury, while the clinical characteristics and prognostic impacts of this complication remain unclear. This study aimed to investigate the clinical characteristics and risk factors of dengue-associated myocardial injury during the epidemic peak of dengue fever in Foshan area. Methods A retrospective cohort study was conducted, enrolling patients diagnosed with dengue-associated myocardial injury at The First People's Hospital of Foshan. Myocardial injury was defined as cardiac troponin I or T (cTnI/cTnT) levels exceeding the 99th percentile upper reference limit, accompanied by abnormal findings on electrocardiogram, echocardiography, or elevated B-type natriuretic peptide (BNP). Systemic parameters and clinical outcomes were compared between patients with and without myocardial injury. Multivariate logistic regression analysis was performed to identify independent risk factors. Results A total of 55 patients with dengue fever were enrolled in this study, including 35 cases in the myocardial injury group and 20 cases in the non-myocardial injury group. Procalcitonin levels were significantly elevated in the myocardial injury group (Z=-2.297, P<0.05), whereas no statistically significant differences were observed in interleukin-6 (IL-6, Z=-1.370, P>0.05), high-sensitivity C-reactive protein (hs-CRP, Z=-0.860, P>0.05), and ferritin (Z=1.585, P>0.05) levels. Patients with myocardial injury exhibited a markedly higher median cardiac troponin level (565.0 vs. 9.5 ng/L, P<0.01), and a decreased left ventricular ejection fraction (LVEF: 54.0% vs. 66.0%, P<0.01), as well as significantly higher rates of ICU admission (83% vs. 15%, P<0.001) and mortality (17.1% vs. 0, P<0.01). Advanced age (OR=1.13, 95% CI: 1.03-1.17) and male gender (OR=3.15, 95% CI: 1.73-13.54) were independent risk factors for the development of dengue-associated myocardial injury. Multivariate logistic regression further demonstrated that the risk of ICU admission in patients with myocardial injury was 8.92-fold higher than that in patients without myocardial injury (OR=8.92, 95% CI: 1.65-48.33, P<0.05). Conclusion Dengue-associated myocardial injury is significantly associated with adverse outcomes, with elderly and male patients being particularly vulnerable. These findings highlight the need for enhanced early monitoring and intervention in high-risk populations.

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.

Invasive non-typhoidal Salmonella infection usually presents as a non-specific febrile illness without typical gastrointestinal symptoms. It is associated with higher morbidity and mortality than non-invasive infection, and is difficult to differentiate from other febrile diseases. This study retrospectively analyzed the clinical data of 2 cases of multi-system infection caused by non-typhoidal Salmonella. Combined with literature review, we summarized the disease characteristics, clinical manifestations, diagnosis and treatment courses, and laboratory findings.Neither patient had a definite history of unclean dietary exposure, and their onset patterns differed. One patient presented with fever, dyspnea and arthralgia, and was admitted to Cangzhou Central Hospital on November 15, 2025. The condition progressed rapidly with multi-system dysfunction, and the patient was subsequently transferred to the Department of Infectious Diseases from the Intensive Care Unit. The other patient sought medical care at several hospitals in Cangzhou for fever and gastrointestinal symptoms, and was finally admitted to Cangzhou Central Hospital on December 1, 2025. Elevated C-reactive protein and procalcitonin were detected after admission, and pulmonary inflammatory changes were observed on imaging. Disseminated Salmonella infection involving the digestive, respiratory, skeletal and other systems was ultimately confirmed based on clinical manifestations and auxiliary examinations including mNGS. Anti-infective regimens were selected according to multi-system involvement of non-typhoidal Salmonella and patient tolerability during treatment, and both patients achieved favourable outcomes. The rising antimicrobial resistance of Salmonella poses growing clinical challenges. Early diagnosis and timely treatment are critical to improving patient prognosis.