Objective To investigate the correlation between adverse drug reactions (ADRs) of antituberculosis prophylactic therapy and microRNAs (miRNAs), and to explore the predictive value of miRNAs for ADRs by comparing miRNA expression differences before medication. Methods Newly detained female individuals in a prison in eastern China who received prophylactic therapy from October 2022 to October 2023 were enrolled as study subjects. Structured questionnaires were used to collect demographic characteristics, behavioral risk factors, and medical history. Participants were regularly followed up at baseline (baseline) and at 1, 2, and 3 months after medication to record ADRs. Control subjects without ADRs were randomly matched at a 1∶2 ratio. Serum samples were collected from both case and control groups before medication (baseline) for miRNA sequencing. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) functional enrichment analyses were performed to identify biological processes and signaling pathways associated with ADRs. Results Of the 187 eligible participants enrolled, 88 (47.06%) completed the full treatment course, with 28 cases experiencing ADRs. Thirteen participants were selected by simple random method as the case group and matched with 26 controls based on general demographic characteristics. Liver function abnormalities were the most common ADR, accounting for 84.61% (11/13). Serum samples from 36 participants (13 cases, 23 controls) were successfully analyzed. A total of 102 differentially expressed miRNAs (33 upregulated, 69 downregulated) were identified. Enriched biological processes included negative regulation of DNA-templated transcription, semaphorin-plexin signaling pathway, and mitophagy. Molecular functions enriched included sequence-specific double-stranded DNA binding, protein self-association, DNA-binding transcription factor activity, and acetylcholine receptor activity. Significant KEGG pathways included oxytocin signaling pathway, insulin secretion, cholinergic synapse, hypertrophic cardiomyopathy, and dilated cardiomyopathy pathways. Conclusions ADRs, particularly liver function abnormalities, are common in individuals receiving antituberculosis prophylactic medication. The enriched biological processes and signaling pathways provide a reference for medication evaluation and regimen adjustment.

Objective To obtain the recombinant BfrB protein of Mycobacterium tuberculosis, investigate its self-assembly and immunological properties, and provide insights for the development of novel subunit vaccines against tuberculosis. Methods The BfrB gene was cloned into the pET28a(+) vector, and the recombinant plasmid was transformed into competent E. coli cells. Following induction with IPTG at 16 ℃, the expression of recombinant BfrB protein was identified by Western blot, and the target protein was purified by affinity chromatography. The multimeric structure of BfrB was characterized by native polyacrylamide gel electrophoresis (native-PAGE) and transmission electron microscopy (TEM). Mice were immunized intramuscularly with BfrB protein three times. Serum antibody levels were detected by indirect ELISA, the proliferation of mouse splenocytes was measured by CCK-8 assay, and the levels of cytokines secreted by splenocytes were determined by double-antibody sandwich ELISA. Results The prokaryotic expression vector pET28a(+)-BfrB was successfully constructed and subsequently transformed into the expression strain for IPTG induction. Western blot analysis confirmed the successful expression of the target protein with a molecular weight of approximately 22 kDa. The purified recombinant BfrB protein was obtained by affinity chromatography with a concentration of 0.8 mg/mL. Native-PAGE demonstrated that the purified BfrB spontaneously formed high-molecular-weight multimers, and TEM observation revealed that it could self-assemble into regular nanoparticles with a diameter of 10 nm. BfrB immunization induced high levels of specific IgG in mice with a titer of 1∶51 200, and significantly promoted the proliferation of mouse splenocytes. Following stimulation with recombinant BfrB protein and Mtb protein, the levels of IFN-γ and IL-10 secreted by splenocytes were significantly elevated compared to those in the control group, and the level of IL-2 secreted by splenocytes was significantly elevated after stimulation with recombinant BfrB protein.These results indicated that BfrB protein immunization could induce a robust humoral immune response in mice, while simultaneously eliciting both Th1 and Th2 type cellular immune responses. Conclusion The recombinant BfrB protein possesses self-assembly properties and strong immunogenicity, and can induce robust humoral and cellular immune responses in mice. This study lays a theoretical foundation for the development of novel subunit vaccines against tuberculosis.

Objective To characterize epidemiological trends in incidence, mortality, and treatment of multidrug-resistant/rifampicin-resistant tuberculosis (MDR/RR-TB) in Jiangsu Province, thereby providing evidence to optimize tuberculosis prevention and control strategies. Methods Using data from the China Tuberculosis Information Management System in Jiangsu Province (2009-2023), we calculated the age-standardized incidence rate (ASIR), age-standardized mortality rate (ASMR), case-fatality rate (CFR), and treatment rate for MDR/RR-TB. Trends were described using the annual percentage change (APC) and average annual percentage change (AAPC) from Joinpoint regression models. An age-period-cohort (APC) model was applied to estimate age, period, and cohort effects on incidence and mortality. Results In 2023, ASIR (0.65 per 100 000) and ASMR (0.038 per 100 000) showed increasing trends compared with 2009, with AAPCs of 11.24% and 6.97%, respectively. Rising trends were observed in ASIR (AAPC=10.50%, P=0.002) and ASMR (AAPC=7.94%, P=0.004) among males, while CFR (AAPC=-1.82, P=0.601), female ASIR (AAPC=13.98%, P=0.056) and ASMR (AAPC=1.95%, P=0.494) showed no statistically significant trends. Southern Jiangsu exhibited a steeper increase in ASIR (AAPC=17.66%) compared to central (AAPC=4.40%) and northern Jiangsu (AAPC=14.33%). Treatment rates gradually increased to 100.00, with females (AAPC=1.90%), newly treated patients (AAPC=3.47%), and northern Jiangsu (AAPC=3.45%) showing significant increases. The APC model indicated that both incidence (net drift=8.69%) and mortality (net drift =5.64%) increased with age, peaking at 75-79 years (7.41 per 100 000) and 80-84 years (2.77 per 100 000), respectively. Period effects revealed elevated incidence (RR=1.23) and mortality (RR=1.45) risks in 2019-2023. Cohort effects demonstrated rising incidence risk with later birth cohorts, peaking in 2004 (RR=26.24), whereas mortality risk remained stable across cohorts. Conclusion Between 2009 and 2023, MDR/RR-TB incidence and mortality increased in Jiangsu Province, while the treatment rate remained high. The disease burden is particularly prominent among males, residents of Southern Jiangsu, and the elderly. Targeted interventions for these key populations are essential for future epidemic control.

Objective To preliminarily investigate the role of the Rv0305c (PPE6) protein in Mycobacterium tuberculosis (Mtb) pathogenesis through experimental and bioinformatics analyses, thereby laying the groundwork for elucidating its molecular mechanism and identifying novel targets for anti-tuberculosis drug development. Methods The Mtb Rv0305c gene knockout strain (ΔRv0305c) was constructed from the Mtb H37Rv wild-type (WT) strain using CRISPR-Cas-assisted non-homologous end joining gene editing technology. Subsequently, the complemented strain (ΔRv0305c::Rv0305c) was generated by electroporation with an Rv0305c expression vector. Bacterial morphology was examined by scanning electron microscopy. Growth curves were plotted by measuring the optical density at 600 nm (OD₆₀₀) of the bacterial cultures using an ultraviolet-visible spectrophotometer. Dynamic expression differences of inflammation-related cytokines in THP-1 macrophages infected with the three strains were analyzed by reverse transcription quantitative PCR (RT-qPCR). For bioinformatic characterization of Rv0305c, the following tools were employed: AlphaFold for protein three-dimensional (3D) structure modeling, STRING for protein-protein interaction network analysis, SignalP for signal peptide prediction, DeepTMHMM for transmembrane structure prediction, InterPro for family and domain analysis, DeepLocPro for subcellular localization prediction, and ELM for eukaryotic-like short linear motif analysis. Results The ΔRv0305c and ΔRv0305c::Rv0305c strains were successfully constructed. Deletion of the Rv0305c gene did not affect the morphology or growth rate of Mtb. However, ΔRv0305c induced significantly upregulated expression of IL-6 and TNF-α and significantly downregulated expression of IL-10 in macrophages at 2 h, 24 h, and 48 h post-infection, compared to the WT and ΔRv0305c::Rv0305c (P<0.05). Bioinformatic analyses revealed the following: Rv0305c is genomically adjacent to PPE5 and is transcribed in the same direction; the Rv0305c protein belongs to the glutamine-rich protein 2 family and contains specific domains; its predicted 3D structure model has an average confidence score of 78.19; its interaction network includes several PE/PPE family members, as well as CpnT, Rv2082, and Rv1004c; Rv0305c may contain eukaryotic-like short linear motifs; it lacks a signal peptide, is predicted to be an non-transmembrane extracellular globulin, and shows the highest probability of extracellular localization (0.721 8). Conclusion Rv0305c contributes to the suppression of the inflammatory response in Mtb-infected macrophages and is predicted to be a eukaryotic-like non-classically secreted protein.

Objective To investigate the cross-drug resistance of rifampicin (RIF) and rifabutin (RFB) in Mycobacterium tuberculosis in Yunnan province and its related mechanism, and to provide reference for RFB replacement therapy for RIF-resistant patients. Methods 348 strains of Mycobacterium tuberculosis isolated from Yunnan Province from January to December 2022 by center for Disease Control and Prevention of Yunnan Province were tested by minimum inhibitory concentration (MIC) method and the whole genome was sequenced. The cross-resistance of RIF and RFB and the mutation characteristics of resistance genes in the resistant strains were analyzed. Results Among 348 strains of Mycobacterium tuberculosis, 89 strains resistant to RIF or RFB were detected by MIC drug sensitivity test, of which 63 strains were simultaneously resistant to RIF and RFB, 25 strains were resistant to RIF but sensitive to RFB, and 1 strain was sensitive to both RIF and RFB. The cross-drug resistance rate of RIF and RFB was 70.79% (63/89). Among the 348 strains of Mycobacterium tuberculosis, 88 strains showed rpoB gene mutations by whole genome sequencing. The RIF and RFB resistance rates of the strains with mutations at positions 516, 526, and 531 of the rpoB gene were 100.00% (6/6) and 33.33% (2/6), 94.12% (16/17) and 76.47% (13/17), respectively, showing relatively high resistance rates. Conclusion In Yunnan province, approximately 30% of RIF-resistant patients can be treated with RFB. The mutations at positions 511, 516, 522, and 533 of rpoB have relatively poor correlation with RFB resistance. When these mutations are detected, RFB treatment can be selected. The mutations at positions 526 and 531 of rpoB have better correlations with RIF and RFB resistance, and when these mutations occur, the use of RIF and RFB for treatment will be affected.

Objective To systematically analyze the core influencing factors of medical consultation delay in tuberculosis (TB) patients in Huai'an City, and to construct a high-precision prediction model for medical consultation delay, thereby providing empirical evidence for optimizing TB prevention and control strategies in the agricultural areas of northern Jiangsu Province. Methods A total of 17 169 patients registered in the Tuberculosis Management Information System of Huai'an City from 2015 to 2024 were selected as the research subjects. Chi-square test and t-test were used to compare baseline and clinical characteristics. Multivariate Logistic regression analysis was applied to identify the independent influencing factors of medical consultation delay. Meanwhile, six machine learning algorithms, including Random Forest (RF), Adaptive Boosting Classification (AdaBoost), and Gradient Boosting Decision Tree (GBDT), were used to construct prediction models. Model performance was evaluated using indicators such as accuracy and Area Under the Curve (AUC). Results The medical consultation delay rate of TB patients in Huai'an City was 67.0% (11 496/17 169). Multivariate Logistic regression showed that actual medication management method (family member management vs. medical staff management: OR=1.598, 95%CI: 1.159-2.203, P=0.004), treatment mode (outpatient treatment vs. non-outpatient treatment: OR=4.129, 95%CI: 1.227-13.888, P=0.022), and symptom pattern (hemoptysis-related: OR=0.172, P<0.001; asymptomatic: OR=0.142, P<0.001) were independent influencing factors of medical consultation delay, while demographic characteristics such as age and gender had no significant effects. Among the machine learning models, GBDT model demonstrated optimal comprehensive performance, with an accuracy of 0.948 and an AUC value of 0.993, significantly outperforming other algorithms. Conclusion The actual medication management method, treatment mode, and symptom pattern are the key targets for regulating the medical consultation timing of TB patients in Huai'an City. This study successfully constructed six machine learning algorithm models, especially the prediction model based on the Gradient Boosting Decision Tree (GBDT) algorithm, which enables accurate assessment of delayed-visit risk and provides technical support for the precise prevention and control of tuberculosis in the region.

Objective To explore the association between the exposure to air pollutants sulfur dioxide (SO2), nitrogen dioxide (NO₂), inhalable particles (PM₁₀), fine particulate matter (PM_2.5), carbon monoxide (CO) and ozone (O₃) and the incidence risk of pulmonary tuberculosis (PTB) among students, so as to provide a scientific basis for formulating environmental risk control and prevention strategies for students' tuberculosis. Methods Basic information of student PTB patients, ambient air pollutant and meteorological data were collected from thirteen major cities in Jiangsu Province during 2014 to 2024. Generalized additive models (GAM) were employed to estimate incidence in PTB risk increase in ambient air pollutant. Sensitivity analyses included dual-pollutant models and generalized additive mixed models (GAMM) with city as a random effect to account for heterogeneity were conducted. Restricted cubic spline functions were used to fit exposure-response curves to examine the linearity of dose-response relationships. Results Between 2014 and 2024, there were 11 084 newly diagnosed PTB cases among students reported in the 13 cities in Jiangsu. Significant positive associations were observed between PTB risk and short-term NO₂ (12.41%, 95% CI: 7.07%-18.02% at lag 0 weeks), PM₁₀ (9.19%, 95% CI: 4.32%-14.29% at lag 6 weeks), PM_2.5 (9.83%, 95% CI: 5.07%-14.81% at lag 5 weeks), and O₃ (10.04%, 95% CI: 4.27%-16.12% at lag 5 weeks) per 10-μg/m³ increase. These associations remained robust in GAMM sensitivity analyses. Concentration-response curves indicated approximately linear relationships for NO₂, PM_2.5, and O₃ with the risk of PTB. There is a generally positive correlation between PM₁₀ and PTB incidence risk, but the curve remains relatively flat within the low concentration range, with more pronounced risk increases in the medium to high concentration range. Conclusions Exposure to NO₂, PM₁₀, PM_2.5, and O₃ is associated with increased PTB risk among students in Jiangsu Province. The findings highlight students as a vulnerable population to air pollution-related PTB. These findings suggest that integrating stringent air pollution control measures may contribute to PTB prevention strategies.

Objective To delineate the population structure, drug resistance profiles, and transmission dynamics of Mycobacterium tuberculosis (Mtb) in Qinghai Province, China, and to provide molecular epidemiological evidence for precision tuberculosis (TB) control strategies in this high-burden region. Methods Clinical isolates from 194 TB patients in Qinghai Province (2015-2024) were analyzed through whole-genome sequencing (WGS). Lineage classification, drug resistance-associated mutation detection, and phylogenetic reconstruction were performed using core genome single-nucleotide polymorphism (SNP) profiling to evaluate genetic clustering and recent transmission events. Results The overall drug resistance rate among Mtb strains was 30.4%, comprising mono-drug resistance (21.6%), multidrug-resistant TB (MDR-TB, 4.1%), and pre-extensively drug-resistant TB (pre-XDR-TB, 0.5%). Streptomycin resistance was most prevalent (15.5%). Phylogenetic analysis revealed Lineage 2 (L2) as the overwhelmingly dominant lineage (83.2%, 159/191), with the L2.2.1 sub-lineage being the most predominant (66.5%). WGS identified diverse mutations, including promoter region variants, nonsynonymous (amino acid-altering), and synonymous (nucleotide-changing) mutations, predominantly in rpsL, katG, and rpoB. Mutations associated with drug resistance detected by WGS showed significant correlations with clinical treatment outcomes (P<0.05). Genetic clustering analysis (using ≤12 SNPs as the genetic distance threshold) identified 16 transmission clusters comprising 57 isolates, yielding an overall clustering rate of 29.4%. Multivariate analysis demonstrated that pre-XDR-TB strains were more likely to cluster than drug-susceptible strains (P<0.05). Furthermore, isolates collected during 2015-2017 exhibited significantly higher clustering rates than those from 2023-2024. Conclusion Clonal spread of Lineage 2 dominates the Mycobacterium tuberculosis epidemiology in Qinghai Province, with high prevalence of diverse resistance mutations to first-line anti-tuberculosis drugs. Prioritizing cluster-level interventions targeting recent transmission is critical for implementing precision TB control strategies.

Objective To investigate the distribution characteristics of natural killer (NK) cells and their subsets (CD56⁺CD16⁺, CD56⁺CD16^-, CD56^-CD16⁺) in peripheral blood of patients with active pulmonary tuberculosis (ATB) and latent tuberculosis infection (LTBI), and to analyze the influence of different sputum bacterial loads on the distribution of NK cells and their subsets in peripheral blood of ATB patients. Methods Peripheral blood samples were collected from 272 subjects who met the inclusion and exclusion criteria in the Outpatient Department of Beijing Center for Disease Control and Prevention from 2024 to 2025, including 100 cases in the healthy control (HC) group, 72 cases in the ATB group, and 100 cases in the LTBI group. Percentage of total peripheral blood NK cells and proportion of each subset in total NK cells were detected by flow cytometry; in addition, 62 ATB patients positive for sputum Xpert test were divided into four groups (extremely low, low, medium, and high) according to sputum bacterial load, and differences in the distribution of NK cells and their subsets in peripheral blood were analyzed among the groups. Results The proportion of total NK cells in the LTBI group was higher than that in the HC group (q=8.498, P<0.000 1) and the ATB group (q=4.846, P=0.002 0), while there was no statistically significant difference between the HC group and the ATB group (P>0.05). Subset analysis showed that the proportion of CD56⁺CD16⁺ in the LTBI group was higher than that in the HC group (Z=5.015, P<0.000 1) and the ATB group (Z=2.402, P= 0.048 9); there was no statistically significant difference in the proportion of CD56⁺CD16^- subset between the ATB group and the LTBI group (Z=0.191 6, P>0.05), but the proportion of this subset in both groups was lower than that in the HC group (ATB vs HC: Z=4.958, P<0.000 1; LTBI vs HC: Z=5.628, P<0.000 1), with statistically significant differences. The proportion of CD56^-CD16⁺ in the LTBI group was lower than that in the HC group (Z=2.507, P=0.036 5), while there was no statistically significant difference in the proportion of this subset in the ATB group compared with the HC group and the LTBI group (P>0.05). There were no statistically significant differences in the proportions of total NK cells and their subsets in peripheral blood among ATB patients with different sputum bacterial loads (P>0.05). Conclusion The infection status of Mycobacterium tuberculosis can affect the distribution characteristics of NK cells and their subsets in peripheral blood, while there is no significant correlation between sputum bacterial load and NK cells immunophenotype in ATB patients.

Objective To investigate the spatiotemporal characteristics and its associated factors of pulmonary tuberculosis (PTB) incidence in Hainan Province, providing a reference for formulating targeted control and prevention strategies. Methods Data on reported PTB cases, meteorology, socioeconomic status, and healthcare resources were integrated from all cities and counties in Hainan Province during 2014 to 2023. Five Bayesian spatiotemporal models of increasing complexity were constructed, progressively incorporating spatial and temporal effects. Bayesian inference was performed using the integrated nested Laplace approximation (INLA) algorithm, and the optimal model was selected based on the deviance information criterion (DIC), Watanabe-Akaike information criterion (WAIC), and conditional predictive ordinates (CPO) to analyze the spatiotemporal characteristics and influencing factors of PTB incidence. Results Significant spatial heterogeneity in PTB incidence risk was observed in cities and counties across Hainan Province from 2014 to 2023, with estimated relative risks (RR) ranging from 0.840 to 1.580. The overall spatial pattern showed the highest risk in the southeastern coastal region, followed by the western region, while generally lower risks in the northern and central-southern areas. From 2014 to 2023, annual relative risks fluctuated slightly around the baseline, ranging from 0.968 to 1.029. Monthly relative risks exhibited seasonal variation, with peaks in January and March (winter-spring), and lower values below the baseline in August, September (summer) and December. Spatiotemporal interaction analysis revealed an overall declining trend in PTB risk and a significant reduction in regional disparities. Analysis of influencing factors showed that, temperature (RR=0.988, 95%CI: 0.978-0.997) and atmospheric pressure (RR= 0.994, 95% CI: 0.991-0.998) were protective factors, whereas sunshine duration (RR=1.002, 95%CI: 1.001-1.003) was a risk factor. Per capita net income of rural residents was a statistically significant protective factor, although its effect size was minimal. Conclusion PTB incidence exhibits distinct spatiotemporal heterogeneity in Hainan Province, influenced by both meteorological and socioeconomic factors. It is recommended to implement differentiated control and prevention strategies, establish meteorological response mechanisms, strengthen interventions during seasonal peaks, and improve rural economic and healthcare conditions to enhance protective effects.

Objective To analyze the temporal trends in the disease burden of cysticercosis in China from 1990 to 2021 and project future trends from 2022 to 2035, thereby providing epidemiological evidence for the prevention and control of cysticercosis in China. Methods Data on age-standardized prevalence rate (ASPR) and age-standardized disability-adjusted life years rate (ASDR) of cysticercosis in China, along with their 95% uncertainty intervals (UIs), were extracted from the 2021 Global Burden of Disease (GBD) database. The Joinpoint regression model was used to analyze the changing trends of cysticercosis burden in China from 1990 to 2021. An age-period-cohort (APC) model was constructed to assess the effects of age, period, and cohort on the cysticercosis burden in China during the same period. Additionally, a Bayesian age-period-cohort (BAPC) model was applied to predict the ASPR and ASDR up to 2035. Results The ASPR of cysticercosis in China decreased by 37.16%, from 56.00 per 100 000 population (95%UI: 39.43, 75.54) in 1990 to 35.19 per 100 000 population (95%UI: 24.27, 49.00) in 2021. Similarly, the ASDR declined by 54.35%, from 18.75 per 100 000 population (95%UI: 11.94, 27.67) in 1990 to 8.56 per 100 000 population (95%UI: 4.99, 13.71) in 2021. Joinpoint analysis showed that from 1990 to 2021, both ASPR and ASDR of cysticercosis in China initially rose, then entered a period of sustained decline, followed by a recent resurgence. The APC analysis model identified age, period, and cohort were all important factors influencing the cysticercosis burden. Predictions from the BAPC model revealed an upward trend in cysticercosis burden in China from 2022 to 2035, with a more significant increase in females than in males. Conclusion While the disease burden of cysticercosis in China decreased substantially from 1990 to 2021, recent trends and future projections indicate an upward trajectory. Therefore, this highlights the need for continued and strengthened investment in medical resources to address the prevention, control, diagnosis, and treatment of cysticercosis in China.

Objective To assess the disease and economic burden of dengue fever patients in Hainan Province in 2019, and to compare the economic costs per disability-adjusted life year (DALY) across provinces in China, thereby providing a scientific basis for quantifying the disease burden of dengue fever and formulating region-specific strategies for disease prevention, control, and resource allocation. Methods A retrospective analysis was conducted using clinical data from 330 confirmed dengue fever cases at Hainan General Hospital in 2019 to estimate the disease and economic burden for all 368 reported cases in the province. Furthermore, a systematic literature search was performed to compile data on the economic burden of dengue fever from other provinces in China, calculate the economic cost per DALY for each province, and compare the differences across regions. Results Among the 330 dengue fever patients, the average DALY burden was 0.03 person-years. The M (P₂₅, P₇₅) of the total economic burden was 1 822.11 (162.61, 2 601.74) yuan, with the M (P₂₅, P₇₅) of the direct economic burden at 1 513.03 (162.61, 2 144.84) yuan and the M (P₂₅, P₇₅) of the indirect economic burden at 219.28 (0, 438.56) yuan. The total DALY burden of dengue cases in Hainan Province in 2019 was 11.33 person-years. The estimated total economic burden was 670 536.48 yuan (95%UI: 575 537.30-751 654.70), comprising direct economic burden of 556 795.04 yuan (95%UI: 488 810.70-626 089.40) and indirect economic burden of 80 695.04 yuan (95%UI: 53 797.92-107 592.20). The estimated economic cost per DALY for dengue cases in the entire province was 59 182.39 yuan. Among provinces studied for dengue fever economic burden in China (Guangdong, Zhejiang, Guangxi, Yunnan, and Chongqing), Guangdong Province exhibited the highest economic cost per DALY and per capita economic burden. Conclusion The economic burden of dengue patients in Hainan Province is substantial. Measures such as expanding health insurance coverage and improving cross-regional insurance settlement are recommended to alleviate the direct economic burden on patients. Given the significant variations in economic burden across provinces in China, dengue prevention and compensation policies should be tailored to regional characteristics to optimize resource allocation.

Objective To investigate the species composition of Culicoides and the prevalence of bluetongue virus (BTV), epizootic hemorrhagic disease virus (EHDV), Tibet orbivirus (TIBOV), and Palyam serogroup virus (PALV) in Jiangcheng County, Yunnan Province, in autumn. Methods Culicoides were collected from cattle sheds in Jiangcheng between September and October 2022. UV light traps were set inside cattle sheds overnight, and the captured insects were taken back to the laboratory in a low-temperature environment. Species were identified under a stereomicroscope based on wing spot morphology. Representative Culicoides were submitted for DNA extraction, and their mitochondrial cytochrome C oxidase subunit I (COI) gene was sequenced and aligned using NCBI BLAST. A phylogenetic tree was constructed to analyze the genetic evolution relationship; qRT-PCR was performed to detect nucleic acids of BTV, EHDV, PALV, and TIBOV in the samples. Results Species such as Culicoides tainanus, Culicoides sumatrae, Culicoides spiculae, Culicoides clavipalpis, Culicoides palpifer, and Culicoides parahumeralis were identified. Their COI gene sequences shared over 98% similarity with reference sequences of corresponding species, and they clustered together in the phylogenetic tree, indicating close genetic relationships. Three Culicoides exhibited morphological characteristics similar to those of Culicoides palpifer and Culicoides parahumeralis; however, their COI gene sequences showed the highest similarity (approximately 82.99%) to C. kingi and C. parahumeralis, respectively. On the phylogenetic tree, these three specimens formed branches distinct from both Culicoides palpifer and Culicoides parahumeralis, but all belong to the subgenus Trithecoides evolutionary cluster. Among 81 pools of Culicoides, one pool was positive for TIBOV, and one was positive for PALV. Conclusion Culicoides tainanus, Culicoides sumatrae, Culicoides spiculae, Culicoides clavipalpis, Culicoides palpifer, and Culicoides parahumeralis are distributed in Jiangcheng County during autumn. Unidentified or poorly understood species exist within the Trithecoides subgenus population. Additionally, Culicoides in Jiangcheng County carry TIBOV and PALV, providing baseline data on the species composition and virus carriage of Culicoides in this area.

Objective To initially understand the infection situation of Bartonella in rodents in Yuxi area, Yunnan Province, and to provide a scientific basis for the comprehensive control and prevention of Bartonella in Yuxi area. Methods In 2024, samples were collected in 8 counties (cities and districts) of Yuxi. Fluorescence PCR was used to determine the negative and positive of the samples. Samples with Ct values less than 30 were selected and amplified rpoB gene fragments by PCR. The amplified rpoB gene fragment sequence was determined by Sanger sequencing, the phylogenetic was constructed using the neighbor-joining method, and phylogenetic analysis was performed. Results A total of 215 organ tissue samples from 215 rodents were collected, from 5 species of rodents and belonging to 2 genera, and Rattus norvegicus were the dominant species (74.42%, 160/215). The overall positive rate of Bartonella in these animal tissue samples was 30.09% (65/215); Among them, the positive rate of Mus musculus was 30.00% (3/10), that of Rattus flavipectus samples was 26.19% (11/42), and that of Rattus norvegicus was 31.25% (50/160). There was no statistically significant difference in the positive rates of Mus musculus, Rattus flavipectus, and Rattus norvegicus (χ²=0.404，P>0.05). Bartonella was detected positive in all 8 counties (cities and districts), and the difference in the positive rates in different regions were statistically significant (χ²=23.088，P< 0.01). The positive rate in Huaning was the highest, 60.00% (18/30), and that in Chengjiang was the lowest, 13.33% (6/45). The rpoB gene was amplified and determined on 19 positive samples with Ct values less than 30. After sequence alignment analysis, a total of 4 species of baltonella were detected, including 10 strains of Bartonella tribocorum, 2 strains of Bartonella elizabathae, 1 strain of Bartonella phoceensis and 6 strains of unidentified Bartonella. Conclusions There were at least 4 species of Bartonella in rodents in Yuxi, and B. tribocorum was the main prevalent species.

Objective To investigate the antibiotic resistance spectrum and the distribution patterns of resistance genes of Vibrio cholerae in Shaanxi Province from 2021 to 2024, providing a scientific basis for clinical treatment and public health prevention and control. Methods A total of 149 Vibrio cholerae strains reported on the National Pathogenic Bacteria Identification Network from 2021 to 2024 were collected. Antimicrobial susceptibility testing was conducted using the microbroth dilution method (MIC). Genomic DNA was extracted from the strains using a bacterial genomic DNA extraction kit, and whole-genome sequencing was performed on the Illumina MiniSeq platform. Resistance genes were predicted and queried using the AMRFinderPlus 3.11.26 program integrated in Seqsphere 10.0.5 software. Resistance rates to various antimicrobial agents were calculated, and multidrug resistance (MDR) profiles were analyzed. A co-occurrence matrix of drug resistance genes was generated, and network analysis was conducted using Gephi 0.8.2. Results The highest resistance rate was observed for ampicillin (49.66%), followed by nalidixic acid (48.32%) and azithromycin (16.11%). All isolates were sensitive to tigecycline. The multi-drug resistance rate was 41.61% (62/149), with turtle samples exhibiting the highest rate at 76.74% (33/43). Among the turtle isolates, 27.91% showed resistance to ≥6 antimicrobial classes and 48.84% to 3-5 classes, significantly higher than other sample types. Whole genome analysis predicted quinolone resistance genes in 60 strains (40.27%), followed by β-lactamase genes in 25 strains (16.78%). Discrepancies existed between genotypic and phenotypic resistance profiles. Co-occurrence network analysis revealed that floR and aph(3'')-Ib genes exhibited the strongest co-occurrence relationship (weight=12), forming the core hub of the network. The gyrA_S83I mutation, conferring quinolone resistance, showed cross-co-occurrence with multiple β-lactamase genes. Conclusion Vibrio cholerae in the environment of Shaanxi Province exhibits high antimicrobial resistance, and the turtle farming system is the key to the spread of drug resistance. Chloramphenicol and aminoglycoside resistance genes may undergo synergistic transmission, and plasmid-mediated horizontal transfer of resistance genes exists. Establishing a surveillance network for antibiotic use in aquaculture and strictly controlling non-therapeutic antibiotic applications in aquaculture are imperative.

Objective To analyze the characteristics of β-lactam (ceftriaxone, imipenem) resistance genes in Nocardia farcinica using whole-genome sequencing (WGS) technology, investigate the underlying resistance mechanisms, and provide a molecular basis for precise clinical medication in N. farcinica infections. Methods Nocardia strains were from clinical samples at The Second Hospital of Hebei Medical University, Hebei Chest Hospital, and Beijing Chest Hospital between October 2018 and December 2024. Strains were identified to the species level as N. farcinica using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and 16S rRNA gene sequencing. Antimicrobial susceptibility testing (AST) for ceftriaxone and imipenem was performed via the microbroth dilution method, and strains were divided into resistant and susceptible groups. Twenty resistant strains underwent WGS; genome assembly was conducted using SMRT Link v5.0.1 software, and genomic characteristics were analyzed with tools including GeneMarkS, Repeat Masker, and tRNAscan-SE. Resistance genes were identified by aligning with resistance databases (ARDB, CARD), and their accuracy was verified by PCR. Results A total of 105 Nocardia strains were isolated and identified, among which 58 (55.2%) were N. farcinica, mainly isolated from lower respiratory tract specimens (82.8%, 48/58). AST results showed that 65.5% (38/58) of N. farcinica strains were resistant to ceftriaxone, and 41.4% (24/58) were resistant to imipenem. Eventually, 20 strains dually resistant to both antibiotics were selected for sequencing. WGS results revealed an average N50 value of 125.6 kb, a mean sequencing depth of 50.2×, an average of 6 320 predicted coding genes per strain, a mean repetitive sequence content of 3.2%, an average number of non-coding RNAs (tRNA, rRNA, sRNA) of 128, and an average of 18 genomic islands per strain. Resistance gene analysis indicated that all 20 resistant strains carried the bla_FAR-1 gene (encoding class A β-lactamase), and 18 strains harbored the RbpA/MtrA/vanR-O gene cluster. PCR validation showed 100.00% consistency (20/20) between the bla_FAR-1 gene and ceftriaxone resistance phenotype, and 90.00% consistency (18/20) between the RbpA/MtrA/vanR-O gene cluster and imipenem resistance phenotype. Conclusion N. farcinica exhibits high resistance rates to ceftriaxone and imipenem. The bla_FAR-1 gene is the key determinant of ceftriaxone resistance, while the RbpA/MtrA/vanR-O gene cluster may be involved in imipenem resistance mechanisms. WGS can effectively characterize resistance genes in N. farcinica, providing molecular evidence for the precise clinical selection of β-lactam antibiotics.

Objective To preliminarily evaluate the immunoprotective efficacy and potential as a vaccine candidate of the 41.5 kDa recombinant cysteine protease of Clonorchis sinensis (rCsCP). Methods The signal peptide-truncated 41.5 kDa CsCP protein was expressed in E. coli BL21. Purified protein combined with Freund's adjuvant was used to immunize BALB/c mice, with PBS and PBS+Freund's adjuvant groups serving as controls. Immunization was administered every 2 weeks for a total of 3 times. Serum samples were collected before each immunization and 2 weeks after the last immunization. Specific IgG and its subclasses (IgG1/IgG2a) antibody titers were measured by enzyme-linked immunosorbent assay (ELISA). Two weeks after the last immunization, mice were infected with C. sinensis metacercariae. Liver tissues were taken at 0, 2, and 4 weeks post-infection for hematoxylin-eosin (HE) staining, Masson's trichrome staining, and immunohistochemistry. Splenocytes were isolated to measure the levels of interferon-γ (IFN-γ), interleukin-2 (IL-2), IL-4, and IL-10 in culture supernatants. Results After rCsCP immunization, specific IgG, IgG1, and IgG2a antibody titers continuously increased, with IgG antibody titer reaching 1:25 600 two weeks after the last immunization, predominantly IgG1. Compared with PBS and PBS+adjuvant groups, the rCsCP immunization group showed significantly reduced inflammatory cell infiltration and peribiliary collagen deposition in liver tissue (P<0.005). Immunohistochemistry indicated decreased expression of α-smooth muscle actin (α-SMA) and type I collagen (Collagen-I) in liver tissue of the rCsCP immunization group (P<0.005). Cytokine detection revealed that two weeks after the last immunization (i.e., 0 weeks post-infection), the secretion levels of IFN-γ, IL-2, and IL-10 in the rCsCP immunization group were significantly higher than those in the PBS and PBS+adjuvant groups (P < 0.05), but decreased rapidly after infection. However, IL-4 levels increased continuously from the 2nd week post-infection, rising to more than 20 times that of the PBS and PBS + adjuvant groups by the 4th week (P<0.005), suggesting that rCsCP activated a mixed Th1/Th2 response dominated by Th2. Conclusion Immunization with 41.5 kDa rCsCP can significantly reduce liver inflammation and fibrosis injury caused by C. sinensis infection. Its protective mechanism may be related to the induction of high-titer specific antibodies and a Th2-dominant mixed Th1/Th2 immune response.

Objective To investigate the long-term seasonal variation of Anopheles dirus density in Wuzhishan City, Hainan Province, and to analyze its association with environmental factors, including meteorological factors, normalized difference vegetation index (NDVI), and land use patterns, providing evidence for malaria re-transmission risk prevention and control. Methods Surveillance data on An. dirus density was systematically collected from the monitoring site in Maoyang Town, Wuzhishan City, Hainan Province, during 2001-2024, along with meteorological, NDVI, and land use data. Descriptive analysis was first conducted to characterize the seasonal patterns of An. dirus density, followed by Spearman correlation to assess the influence of environmental factors on mosquito density. The distributed lag non-linear model (DLNM) was then applied to explore the non-linear lagged effects of key environmental variables. Results During 2001-2024, the average An. dirus density was 0.66 mosquitoes/person-night, with a seasonal peak in March-June and an exceptional high value of 3.83 mosquitoes/person-night in May 2020. Spearman correlation showed significant positive associations of mosquito density with monthly mean minimum temperature (r_s=0.40, P<0.001), maximum temperature (r_s=0.31, P<0.001), and mean temperature (r_s=0.28, P<0.001), a negative association with water body proportion (r_s=-0.39, P<0.001), a weak positive association with forest proportion (r_s=0.28, P<0.001), and no significant association with NDVI (r_s=-0.09, P>0.05). DLNM results indicated that mosquito density risk was highest at minimum temperatures of 18-22 °C, with effects concentrated within 0-1.5 months; precipitation of 0-100 mm increased risk (maximum RR=1.46) with stronger effects at 1-3 months lag; and NDVI<0.76 was associated with increased risk (RR=1.10-1.40). Conclusions Environmental factors are closely related to An. dirus density in Wuzhishan City. Temperature, rainfall, NDVI, and water body proportion were the main influencing factors on An. dirus density and should be incorporated into mosquito surveillance and risk assessment systems to provide a scientific basis for malaria re-transmission risk monitoring in the region.

Objective To investigate the structural and compositional disparities in gut microbiota between children with idiopathic short stature (ISS) and those of normal stature, to explore the potential role of gut microbiota in the pathogenesis and furnish a theoretical foundation for further uncovering the underlying mechanisms of ISS and for developing targeted interventions of ISS. Methods Twenty-two children aged 4.0-7.5 years and diagnosed with ISS who visited the Department of Pediatrics, Haikou Affiliated Hospital of Central South University Xiangya School of Medicine, between January 2023 and July 2024, were enrolled as the ISS group, while fifteen age-matched healthy children with normal height were recruited as the control group. Clinical data and fecal samples were collected from all participants, and the composition of their gut microbiota was analyzed using 16S rDNA sequencing. Results No significant difference in alpha diversity was observed between the two groups (P>0.05), whereas beta diversity showed a statistically significant difference (R=0.109, P<0.05). At the phylum level, the dominant phyla in both groups were Firmicutes, Bacteroidota, and Proteobacteria. At the genus level, the predominant genera in the ISS group were Bacteroides (29.72%), Faecalibacterium (10.89%), Escherichia-Shigella (5.92%), Phascolarctobacterium (3.09%), and Alistipes (2.49%). In contrast, the predominant genera in the control group were Bacteroides (26.61%), Faecalibacterium (9.04%), Escherichia-Shigella (4.41%), Bifidobacterium (2.93%), and Megamonas (2.92%). LEfSe analysis identified 7 dominant taxa in the ISS group and 6 dominant taxa in the control group at the family and genus levels. At the family level, the dominant taxon in the ISS Group was Veillonellaceae, while those in the control group were Selenomonadaceae, Proteaceae, and Morganellaceae. At the genus level, the dominant taxa in the ISS Group were Veillonella, Hydrogenoanaerobacterium, Hungatella, Erysipelatoclostridium, Oscillibacter, and Dialister, whereas those in the control group were Megamonas, Subdoligranulum, and Morganella. Genus-level co-occurrence network analysis indicated that the ISS group exhibited a more simplified network structure compared to the control group. Conclusion The dominant gut microbiota taxa in ISS children differ significantly from those in healthy controls, characterized by dysbiosis and increased abundance of taxa associated with inflammation, suggesting that gut microbiota may play a crucial role in the onset and progression of ISS.

Objectives To present the epidemiological characteristics, management process, and transmission chain of the first confirmed case of mpox virus subclade Ⅰb in Chongqing, and to provide reference for the control and prevention of mpox outbreaks nationwide, particularly for the early detection, investigation, and management of female cases. Methods A field epidemiological investigation was conducted for the mpox case confirmed in Chongqing on 6th August, 2025. Clinical and environmental samples were collected from the case and contacts. Real-time fluorescent PCR was used for nucleic acid detection of mpox virus. Results The patient was a 25-year-old female international student who presented with fever and sore throat, followed consecutively by vesiculopustular eruptions on the hands, face, and external genitalia. Throat swab and vesicular fluid samples from the case tested positive for mpox virus nucleic acid (Ct value 20.0), and genetic sequencing confirmed subclade Ⅰb. Epidemiological investigation revealed that the case had a travel history in G Province from 15th to 27th July 2025, and had multiple close contact (kissing and hugging) with a foreign national who had experienced a rash. After returning to Chongqing, she developed fever and sore throat on 1st-3rd August; self-medication brought no relief on 2nd August. On 4th August a vesiculopustular rash appeared and she attended the gynaecology department of a local hospital. On 6th August, she was seen in the dermatology department of the same hospital, where vesicular fluid and a throat swab were collected for testing. Four environmental samples from the case's residence were positive (Ct values 28.95-33.90). Seven general contacts were identified, none of whom developed secondary infections. The case presented with mild symptoms and has recovered following isolation and treatment. Conclusion This study reports the first confirmed case of mpox virus subclade Ⅰb in Chongqing, transmitted through non-sexual close contact. It highlights the need to strengthen health monitoring of international arrivals and close contacts, enhance healthcare staff's ability to recognize mpox, and maintain vigilance against atypical transmission routes.

Objective This study aimed to characterize the genetic variation characteristics and evolutionary features of Human parainfluenza virus type 2 (HPIV-2) in Yunnan, whole-genome sequencing and molecular characterization analysis were conducted with an HPIV-2 strain (PIV2/Kunming/0621/2024) isolated in Yunnan. Methods One HPIV-2-positive sample was identified through influenza-like illness (ILI) surveillance in Yunnan in 2024. LLC-MK2 cells were inoculated to isolate the virus and observe the cell lesions. Viral nucleic acid detection was carried out by fluorescence quantitative PCR. Viral RNA was extracted using magnetic bead-based methods, reverse-transcribed to cDNA, and subjected to library preparation. Whole-genome sequencing was performed using the Illumina MiSeq platform. Reads were assembled using CLC Genomics Workbench 23.0.1, aligned to the reference strain HPIV2/JAP/Toshiba/1990, and analyzed for sequence homology and key mutations. Phylogenetic relationships were inferred using MEGA 7.0, and potential N-linked glycosylation sites were predicted using NetNGlyc 1.0. Results The complete genome of PIV2/Kunming/0621/2024 (GenBank accession no. PQ621733) was obtained, with 15 563 bp in length, and encoding six structural proteins (N, P, M, F, HN, and L), consistent with typical HPIV-2 genome. Phylogenetic analysis clustered the strain within clade C, closely related to a 2017 Beijing isolate. Compared with the prototype strain, the HN protein contained 31 amino acid substitutions and an additional putative N-linked glycosylation site at position 316. Multiple substitutions and glycosylation alterations were also identified in the F, NP, P, M, and L structural proteins, including novel glycosylation motifs at position 99 (NISA) in F, 485 (NPSV) in NP, and 146 (NMSQ) in P. Conclusion This study provides the first complete genome sequence of an HPIV-2 isolate from Yunnan, revealing novel mutations and glycosylation site variations in structural proteins. Phylogenetic analysis indicates that this strain belongs to clade C that has been prevalent in recent years, the findings enhance understanding of HPIV-2 molecular evolution and may inform future surveillance, vaccine development, and functional studies.

Chikungunya virus (CHIKV), an arbovirus transmitted by Aedes mosquitoes, is rapidly spreading across Asia, Africa, and South America, triggering multiple large-scale outbreaks and posing a serious threat to global public health security. Since the clinical manifestations of CHIKV infection are similar to those of dengue fever, Zika virus disease, and other illnesses, accurate and rapid diagnostic techniques are of great significance for disease differentiation and epidemic prevention and control. This review outlines recent progress in CHIKV detection, including real-time quantitative PCR, isothermal amplification, CRISPR-assisted detection, and rapid diagnostic tests, with emphasis on their advantages and limitations in terms of sensitivity, specificity, and field application feasibility. Furthermore, research findings on CHIKV whole-genome sequencing and genotyping are summarized, elaborating the epidemiological characteristics of different genotypes and their role in viral tracing, mutation monitoring, and vaccine development.

Globally, human diseases caused by non-tuberculous mycobacteria (NTM) are increasingly prevalent, with clinical manifestations ranging from skin lesions to systemic diseases, and the prognosis of patients is relatively poor. These bacteria are widely distributed in natural environments, leading to extremely common exposure of humans to them. In view of the above characteristics, accurate identification and characterization of the NTM, as well as the formulation of appropriate subsequent treatment regimens, remain major challenges at present. The treatment of NTM diseases is complex and requires the long-term combined use of multiple drugs, which not only results in high treatment costs but also often causes adverse reactions in patients. Traditional Chinese Medicine (TCM), as traditional Chinese medicine, has a long history and rich experience in the treatment of infectious diseases. In recent years, growing attention has been paid to TCM, and research data have shown that TCM possesses certain potential in the treatment of NTM diseases. It can improve clinical efficacy, alleviate patients' symptoms, reduce adverse reactions of Western medicines, and reverse NTM drug resistance, etc. This article explores the epidemiological characteristics, current status of drug resistance, antibiotic treatment, and application of TCM in the treatment of NTM diseases, aiming to provide more insights and approaches for clinical treatment, improving the treatment level of this disease, and benefiting more patients.

Objective To prevent the occurrence and cross-border spread of major epidemics such as plague in border areas both domestically and internationally. This study reviews the discovery of natural plague foci along the China-Kazakhstan border historically and the recent event of the first isolation of plague bacteria in the Altai Mountain area of the China-Mongolia border, providing a reference for future monitoring. Methods From the perspective of the history of science and technology, historical documents were reviewed and analyzed, combined with the latest survey data to examine the discovery, transmission risks, and early warning mechanisms of plague in the border areas of China-Kazakhstan and China-Mongolia ports and their surrounding regions. Results In the surrounding areas of the four ports along the China-Kazakhstan and China-Mongolia borders, 9 plague-positive sites were discovered successively for the first time,2009-2018. A total of 74 batches of 27 types of disease vectors, including rats, fleas, ticks, cockroaches, bats, and wild animals, were intercepted in the port clearance areas. The risk of cross-border transmission of pathogens carried by disease vectors in the border areas and the epidemic dynamics of major infectious diseases such as plague both domestically and internationally warrant alert. Conclusion To address the threat of cross-border transmission of major infectious diseases such as plague originating from natural epidemic foci in the the port clearance zones and the border line areas, advanced monitoring and early warning identification technologies for blocking cross-border epidemic transmission should be further developed. Moreover, the application and refinement of the 'five-level monitoring barrier' theory and practice for major infectious diseases like plague are crucial to eliminate outbreaks in their nascent stages, both abroad and at the border.