This paper presents a systematic examination of the transition of heat stroke from a traditionally overlooked "hidden concern" to a pressing "visible threat" in the era of global warming, now characterized by broader prevalence and routine occurrence. With the rising frequency and intensity of extreme heatwaves, the population affected has extended beyond specific high‑risk occupations to the general public. Furthermore, its epidemiological profile has evolved from sporadic, isolated incidents to recurrent, widespread outbreaks, often manifesting as "cluster" events during extreme heat periods, which poses a significant challenge to public health emergency responses. Through an analysis of its worsening epidemiological trends, far‑reaching socio‑economic consequences, and the marked cost‑effectiveness of a "prevention over treatment" approach, the paper argues for elevating heat stroke prevention to a level of attention comparable to that given to tropical diseases. Drawing on the established "surveillance‑warning‑intervention‑assessment" framework used in tropical disease control, it proposes the development of an integrated prevention and response system encompassing a national monitoring and early‑warning network, tailored public education, proactive intervention for vulnerable groups, research capacity, and coordinated multi‑sector governance. Elevating heat stroke prevention and control to the height of national public health strategy, thereby promoting a shift from reactive rescue to proactive defense, and from isolated case management to systematic preparedness, is an inevitable requirement and an urgent task for mitigating the health threats of climate change, ensuring the equity of public livelihood, and achieving the goals of the "Healthy China 2030" initiative.

Exertional heat stroke (EHS) is a severe, life-threatening heat-induced illness. In-hospital management constitutes a critical component in lowering the mortality rate of EHS. In recent years, clinical practice has demonstrated that some EHS patients present with insidious onset and atypical symptoms, with rapid disease progression that predisposes them to delayed diagnosis and formidable treatment challenges, ultimately resulting in fatal outcomes in some cases of critical EHS. By analyzing EHS treatment data compiled by the Military Expert Group on Heatstroke in recent years, we have summarized ten key considerations for the in-hospital management of EHS. These include identifying atypical clinical manifestations of EHS; recognizing high-risk factors for EHS deterioration; screening for concurrent infectious diseases in EHS patients; implementing personalized fluid resuscitation during rescue efforts; preventing injury and hemorrhage during invasive procedures; determining the optimal timing for plasma exchange; adopting comprehensive cerebral protection strategies throughout the treatment course; identifying and managing myoclonus, shivering, and convulsions; and avoiding early limb rehabilitation exercises. These ten considerations serve as practical guidance for the in-hospital management of exertional heat stroke.

Objective To analyze and explore a standardized in-hospital management protocol for mass casualty heat stroke events, aiming to effectively translate the core principle of "early and rapid cooling" into coordinated mass casualty care, and to enhance the hospital's emergency response and collaborative treatment capabilities. Methods Based on the practical experience gained from managing mass heat stroke casualties at Hainan Hospital of Chinese PLA General Hospital, a standardized emergency response protocol was developed and activated. The effectiveness of this protocol was evaluated through a retrospective analysis of treatment data from a mass heat stroke event in September 2023. Results A standardized emergency response protocol was established, including establishing a three-tier organizational response and graded spatial clearance mechanism, establishing a mechanism centered on "rapid cooling" with modular deployment of active cooling devices equipped for temperature monitoring and corresponding human resources, and creating a standardized heat stroke resuscitation unit to implement an integrated early warning-triage-cooling linkage mechanism and adopt differentiated combined cooling strategies based on the severity of the patient's condition. In the mass heat stroke event in September 2023, a total of 18 cases of heat illness occurred, of which 7 cases were transported to Hainan Hospital of Chinese PLA General Hospital for emergency treatment. The 5 patients diagnosed with heat stroke were all male, aged (33.0±7.2) years, with a mean core body temperature of (40.3±0.6) °C upon admission. Following the initiation of enhanced combination cooling, the mean time from emergency department admission to the reduction of core body temperature <38.5 °C was (23.2±1.9) min, and the total time from onset to target temperature achievement was (53.0±3.7) min. The patients spent an average of (96.6±5.2) min under emergency observation before being transferred to the ICU. The mean ICU stay was (5.8±4.1) d, and the total hospital stay was (19.4±11.8) d. After a two-year follow-up, no significant residual organ dysfunction was observed in any of the patients. Conclusion The establishment and implementation of a standardized, modular, and emergency department-centered in-hospital emergency response protocol for mass heat stroke casualties can effectively enable rapid alerting, flexible resource allocation, and a front-loaded workflow of "cooling-upon-alert". This system ensures effective core temperature control within the "golden half-hour" even in mass casualty scenarios, significantly improves patient outcomes, and provides a feasible systematic solution for responding to such public health emergencies.

Objective To investigate the distribution of regulatory T cells (Tregs) in different organs/tissues and their correlation with the survival outcomes in a murine model of heat stroke, to further elucidate the potential role of Tregs in the immune dysregulation associated with heat stroke. Methods Male BALB/c mice were exposed to heat to establish a mouse model of classic heat stroke. The heat-stressed mice were allocated to the survivor group or the non-survivor group based on their survival outcomes. An additional normal control group was set up without heat exposure. At 24 h after the onset of heat stroke, core body temperature, body weight, and liver and kidney function indicators were measured in each group to assess the severity of heat stroke. Subsequently, flow cytometry was employed to quantify the proportion of Tregs among CD4⁺T cells and the number of Tregs in the peripheral blood, spleen, and liver to determine the distribution of Tregs in different organs/tissues. Additionally, the levels of Tregs in peripheral blood prior to heat exposure were determined by flow cytometry to reflect baseline levels. Results Compared with the survivor group, the non-survivor group exhibited hypothermia, more pronounced weight loss, and markedly elevated indicators of liver and kidney at 24 h post the onset of heat stroke, indicating an increased severity of heat stroke. At 24 h after heat stroke, no statistically significant differences were observed in the frequency of Tregs among CD4⁺T cells across groups in either spleen or liver. However, the numbers of Tregs in the peripheral blood, spleen, and liver of the non-survivor group were remarkably lower than those in the survivor group. Although Treg numbers of the peripheral blood, spleen, and liver in the survivor group decreased compared to controls, significant differences were only observed in the peripheral blood. Furthermore, comparable baseline levels of Tregs were observed in the peripheral blood of both survivor and non-survivor groups prior to heat exposure. Conclusion The numbers of Tregs in the peripheral blood, spleen, and liver closely correlate with heat stroke severity and prognosis; this quantitative difference is not attributable to baseline variations prior to heat exposure.

Objective To explore the role of monocyte pyroptosis in heat stroke-induced thrombosis using animal models combined with clinical samples. Methods Forty C57BL/6J mice were randomly divided into a control group, a heat stroke group, a heat stroke thrombosis group (inferior vena cava constriction + heat shock), and a sham operation group. At 3 and 72 hours post-modeling, coagulation function [four coagulation parameters, thromboelastography (TEG)], thrombus formation (ultrasound, laser speckle), and inflammatory markers [interleukin-6 (IL-6), tissue factor (TF)] were measured. Peripheral blood was collected from 32 patients with heat stroke, and changes in monocyte subsets were analyzed using flow cytometry. The pyroptotic morphology of monocytes in the thrombi was observed by transmission electron microscopy, which was combined with transcriptome sequencing, pseudotime analysis, and Western blot to explore the underlying molecular mechanism. Results The mice with heat stroke exhibited a hypercoagulable state as early as 3 hours after heat shock, peaking at 72 hours. Definite thrombus formation was observed in the inferior vena cava and intracranial venous sinuses, accompanied by massive inflammatory cell infiltration. The proportion of non-classical monocytes (CD14^-CD16⁺) significantly increased in the peripheral blood of patients with heat stroke and in mouse thrombi; these cells exhibited typical pyroptotic morphology, including membrane rupture and vesicle formation. Transcriptomic analysis revealed that the NLRP3/Caspase-1/GSDMD pyroptosis pathway was significantly activated in non-classical monocytes within the thrombus, with the upregulated expression of hypoxia-inducible factor-1α (HIF-1α) and the prothrombotic molecule thrombospondin-1 (THBS1). Pseudotime analysis suggested that classical monocytes infiltrated the thrombi and transformed into a non-classical phenotype, which highly expressed neutrophil chemokines such as CXCL2 and CXCL8. Conclusion In heat stroke, circulating monocytes transform into a non-classical phenotype (CD16⁺), which undergoes pyroptosis in the thrombotic microenvironment and may promote thrombosis formation by recruiting neutrophils.

Objective To investigate the impact of different anticoagulation initiation timings on organ function in exertional heat stroke (EHS), and to analyze the related risk factors for prognosis. Methods A retrospective case-control study was conducted, to collect clinical data of 75 EHS patients admitted to the 900th Hospital of PLA Joint Logistic Support Force between 2014 and 2024. Patients were divided into an ultra-early anticoagulation group (initiated within 6 h of EHS diagnosis) and an early anticoagulation group (initiated after 6 h of EHS diagnosis), based on the timing of initiating anticoagulation therapy after admission. Differences in D-Dimer, prothrombin time (PT), activated partial thromboplastin time (APTT), thrombin time (TT), fibrinogen (Fg), D-dimer/fibrinogen ratio (DFR), platelet count (PLT), creatinine (Cr), alanine aminotransferase (ALT), aspartate aminotransferase (AST), myoglobin (Myo), and creatine kinase (CK) were compared between the two groups of patients on the 1st, 3rd, 5th, and 7th days after admission to the EICU. Partial correlation was used to analyze the correlation between prognosis and the aforementioned indicators. Multivariate logistic regression was employed to identify independent risk factors for prognosis. Results A total of 75 patients were ultimately included, with 55 in the ultra-early group and 20 in the early group. Compared to the early group, the ultra-early group demonstrated superior improvement in coagulation function: PT, APTT, and TT showed a shortening trend (P<0.05), while Fg and PLT showed an increasing trend (P<0.05). D-Dimer levels in both groups showed an initial increase followed by a decrease, but the decline was more significant in the ultra-early group after peaking on the third day. Regarding liver function, ALT and AST improved more significantly in the ultra-early group (P<0.05). Correlation analysis revealed that, 28-day mortality was positively correlated with early (first 3 days in EICU) D-Dimer, DFR, ApacheⅡscore, incidence of DIC, Cr, ALT, AST, Myo, and PT (P<0.05), and negatively correlated with early PLT (P<0.05). Multivariate logistic regression indicated that age and ALT level on day 1 in EICU were independent risk factors for 28-day mortality. Conclusion Both ultra-early and early anticoagulation therapy can improve coagulation and liver function in EHS patients. However, the ultra-early anticoagulation strategy demonstrates superior early organ protective effects compared to early anticoagulation.

Objective To investigate the protective effect of butyrate preconditioning on intestinal barrier injury in mice with classical heat stroke (CHS) and explore its potential mechanism based on transcriptomic analysis, so as to provide a theoretical basis for the development of butyrate as a therapeutic agent for heat stroke. Methods C57BL/6J mice were randomly divided into four groups: control group (CON), butyrate group (NaB), heat stroke group (HS), and butyrate preconditioning+heat stroke group (NaB+HS), with 10 mice in each group. Mice in the NaB and NaB+HS groups were intragastrically administered sodium butyrate solution for 2 consecutive weeks. A mouse model of CHS was then established, and core body temperature and survival rate were monitored. Intestinal pathological damage and intestinal permeability were detected by HE staining and FITC-dextran assay, respectively. The expression of tight junction proteins (ZO-1, Occludin) was measured by Western blot, and the intestinal ultrastructure was observed by transmission electron microscopy (TEM). Ileal tissues were collected for transcriptome sequencing; KEGG and GO enrichment analyses were performed on differentially expressed genes (DEGs), with results verified by quantitative reverse transcription polymerase chain reaction. Results Butyrate preconditioning significantly slowed the elevation of core body temperature (P<0.001) and improved the survival rate of HS mice (from 60% to 100%, P=0.029). It also alleviated intestinal mucosal structural damage induced by heat exposure and reduced intestinal permeability. Western blot and TEM results showed that butyrate preconditioning significantly upregulated the expression of ZO-1 and Occludin proteins in the intestines of HS mice (P<0.05) and maintained the integrity of tight junction structures. Transcriptomic analysis identified 2 136 DEGs between the HS and CON groups, primarily enriched in immune and inflammation-related pathways. In contrast, 1 429 DEGs were found between the NaB+HS and HS groups, primarily involved in metabolic regulation and vascular permeability regulation. qRT-PCR verification confirmed that the expression changes of key DEGs, including Gpr182, Lyve1, and Ddit4, were consistent with the sequencing results. Conclusion Butyrate preconditioning protects intestinal barrier function in mice with CHS by alleviating intestinal injury and enhancing the expression of tight junction proteins. Its mechanism may be associated with the regulation of immune-metabolic balance, providing a theoretical basis for developing butyrate as a preventive intervention strategy for CHS.

Objective To construct a heat tolerance assessment scale and systematically evaluate its reliability, validity, and quantifying heat tolerance levels and application value in screening for individuals at high risk of heat stroke. Methods Based on a systematic review of theories related to heat stress physiology and heat acclimatization, and with reference to diagnosis and prevention guidelines of exertional heat illness in the US and UK militaries, a preliminary framework of a heat tolerance assessment scale was formed through literature analysis, item pool construction, and expert consultation. The scale consists of four dimensions (subjective perception, physiological sensation, physical performance, and training load) comprising a total of 13 items. The Delphi expert consultation method was used to conduct multiple rounds of evaluation and revision of the scale items to ensure their scientific validity and practical feasibility. Subsequently, a questionnaire survey was conducted among 383 military personnel from Fujian and Hebei provinces. Exploratory factor analysis (EFA) and confirmatory factor analysis (CFA) were performed to examine the structural validity of the scale, and internal consistency indices were used to assess its reliability. A further questionnaire survey was carried out among 1 451 military personnel from Hainan, Jiangxi, and Xinjiang. Receiver operating characteristic (ROC) curve analysis was applied to evaluate the predictive validity of the scale. Results The Kaiser-Meyer-Olkin value of the questionnaire was 0.792, and the Bartlett test χ²=2 892.832 (P<0.001), EFA extracted 4 common factors with a cumulative variance contribution rate of 76.9%. The fit indices of CFA were good (χ²/df=1.836, RMSEA=0.047, CFI=0.983, TLI=0.977). The overall Cronbach's α was 0.832, and the α value for each dimension was >0.75. ROC analysis demonstrated that the scale exhibited a good discriminative validity for identifying the risk of exertional heat stroke (AUC=0.789) and an excellent predictive validity for determining superior heat tolerance levels (AUC = 0.894), with corresponding cutoff values of 38.5 and 45.5, respectively. Conclusion The heat tolerance assessment scale has good reliability and validity, and can be used as an effective tool for assessing the heat tolerance levels of military personnel and screening for individuals at high risk of exertional heat stroke.

Objective To evaluate clinical value of heat stroke severity score system (HSSS) in assessment of disease severity and predicting prognosis in patients with heat stroke (HS), to provide a basis for accurate use of HSSS. Methods A retrospective analysis was conducted on the clinical data of 143 patients with heat stroke admitted to the 908th Hospital and the 900th Hospital of the Joint Logistics Support Force of Chinese People's Liberation Army from March 2022 to October 2024. Patients were stratified into mild group (n=110), moderate group (n=23), and severe group (n=10), based on their HSSS scores. Demographic characteristics, acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ), sequential organ failure assessment (SOFA), and Glasgow coma scale (GCS) scores, as well as clinical indicators including blood routine, coagulation function, liver and kidney function, lactate levels, and mortality rates, were compared among groups. Receiver operating characteristic (ROC) curve analysis was performed to evaluate the predictive efficacy of HSSS for mortality， multivariate Cox regression model was employed to calculate hazard ratios (HR). Results The mortality rate in the severe group was 90.0%, significantly higher than in the mild and moderate groups (both 0, P<0.001). Organ injury patterns analysis revealed that three-system involvement was the most common (40 cases, 28.0%), followed by four-system involvement (36 cases, 25.2%). As HSSS scores increased, APACHE Ⅱ scores, SOFA scores, and lactate levels increased, while GCS scores decreased; the severity of coagulation disorders and liver/kidney injury also worsened (P<0.05). Correlation analysis showed a strong positive correlation between HSSS and SOFA scores (r=0.942, P<0.001), as well as APACHE Ⅱ scores (r=0.816, P<0.001). ROC analysis demonstrated that the AUC of HSSS for predicting death was 0.998(95%CI:0.992-1.000), which was higher than that of APACHE Ⅱ score (AUC=0.948, Z=2.384, P=0.017), and comparable to that of SOFA score (AUC=0.985, P=0.093). At an optimal cutoff of 20 scores, sensitivity was 100.0% and specificity was 93.4%. Survival analysis indicated that patients with HSSS >20 had a significantly increased mortality risk (HR=2.06, 95%CI:1.11-3.84, P=0.02), and significantly lower cumulative survival compared to those with scores ≤20 (Log-rank P<0.001). Conclusion The HSSS can accurately assess disease severity and reliably predict adverse outcomes in patients with heat stroke.

Heat stroke (HS) is a life-threatening condition triggered by exposure to extreme heat or strenuous exercise, characterized by significantly elevated core temperature, central nervous system dysfunction, and multiple organ failure. Accumulating evidence underscores that mitochondrial injury plays a pivotal role in the pathogenesis of HS. Sustained hyperthermia and oxidative stress lead to mitochondrial membrane depolarization, impaired adenosine triphosphate (ATP) synthesis, and excessive reactive oxygen species (ROS) accumulation, ultimately activating apoptosis and inflammatory cascades, resulting in organ damage. Mitochondrial autophagy (mitophagy), a selective mitochondrial quality control mechanism, exhibits a typical “double-edged sword” effect in HS: moderate activation facilitates the elimination of damaged mitochondria, alleviates oxidative stress, and preserves cellular homeostasis, whereas excessive mitophagy results in energy depletion and cell death. Current research indicates that the PINK1/Parkin pathway, BNIP3/NIX receptor-mediated signaling pathway, and AMPK/mTOR axis are key regulators of mitophagy under heat stress conditions, and the activity balance among these pathways determines whether mitophagy exerts a protective or detrimental role. Overall, mitophagy in heat stroke-associated multiorgan dysfunction carries both defensive significance and potential pathogenic implications. In-depth exploration of its molecular regulation and its roles in heat stroke-induced multiorgan dysfunction holds promise for unveiling novel therapeutic insights and guiding the development of targeted interventions for heat stroke.

Heat stroke (HS), as the terminal stage of heat-related diseases, is characterized by an uncontrolled cascade of inflammation, oxidative stress, and programmed cell death that underlies its high mortality rate. Recent studies have revealed that ferroptosis, pyroptosis, and autophagy dysregulation constitute the key interrelated pathological mechanisms. Specifically, ferroptosis is characterized by abnormal accumulation of ferrous ions (Fe²⁺), which directly leads to cell damage mainly by catalyzing lipid peroxidation; pyroptosis is driven by heat stress-activated NOD-like receptor thermal protein domain associated protein 3 (NLRP3) inflammasome and cysteinyl aspartate specific proteinase-1 (caspase-1), mediating cell lysis and release of large quantities of pro-inflammatory factors through gasdermin D (GSDMD) pore formation, thereby amplifying systemic inflammation; autophagy plays a dual role, exerting a protective function in the early stage but capable of being cleaved into pro-apoptotic fragments under sustained stress. These processes do not exist in isolation but are intertwined and synergistically amplified, collectively laying the molecular foundation for multiple organ failure. Based on the understanding of this interaction network and the treatment experience from other organ injury models, future intervention strategies may focus on stage-specific, precise targeting: inducing protection by targeting ferroptosis and autophagy in the early stage, and strongly inhibiting inflammasome activation in the late stage. This article reviews the complex interaction network formed by "ferroptosis-inflammation-autophagy dysregulation".

In recent years, the increasing frequency of extreme weather events worldwide has driven a sustained rise in the incidence of heat stroke. As a fatal emergency that can lead to multiple organ failure, heat stroke involves highly complex pathological mechanisms. The gut microbiota, serving as the largest microecosystem in the human body, has been found to play a key role in the occurrence and development of heat stroke. This article systematically reviews the bidirectional interaction between heat stroke and the gut microbiota. On the one hand, the sudden elevation of core body temperature, intestinal ischemia/reperfusion injury, and systemic inflammatory response triggered by heat stroke can disrupt the integrity of the intestinal barrier, leading to microbial ecological dysbiosis. On the other hand, the dysregulated gut microbiota further amplifies heat stroke-induced organ damage through several mechanisms: exacerbating intestinal barrier damage, generating harmful metabolites such as trimethylamine N-oxide (TMAO) and hydrogen sulfide, and perturbing the central nervous system through the "gut-brain axis", thereby establishing a vicious cycle. Based on this, this paper further explores potential therapeutic strategies targeting the intestinal microecology, including the preconditioning protective effects of heat acclimatization, microencapsulation technology, nanotechnology, mucosal adhesion technology, and synbiotic preparations, aiming to provide a new theoretical basis and direction for the prevention and treatment of heat stroke.

This report describes a case of exertional heat stroke (EHS) with epilepsy as the primary presenting symptom, to provide insights for the recognition and management of this atypical presentation EHS. A 21-year-old male patient developed sudden impaired consciousness and recurrent generalized tonic-clonic seizures during a 3-kilometer run under conditions of high temperature and humidity, accompanied by grade 0 limb muscle strength and multiple organ dysfunction. Despite the administration of rapid cooling and conventional anti-epileptic treatment, the seizure episodes persisted. On day 23 of hospitalization, a regimen combining sodium valproate and gabapentin was initiated, which resulted in the complete cessation of seizures within four days. The patient subsequently regained limb muscle strength, resumed normal ambulation, and was discharged following a full recovery. The presentation of EHS with epilepsy as main manifestation is exceptionally rare in clinical practice. Seizures can represent the primary or even sole central nervous system manifestation, which frequently leads to misdiagnosis as primary epilepsy or other neurological disorders, and delays critical interventions such as cooling. This analysis in combination with this case, explores the pathological mechanisms, therapeutic challenges, and clinical management strategies for seizure control in EHS.

Objective To elucidate the prevalence and genotypic distribution of spotted fever group rickettsiae (SFGR) in ticks from the Daxing'anling forest region of Hulunbuir City, and to provide an evidence base for control and prevention of tick-borne spotted fever. Methods From April to September 2025, host-ttached ticks were collected by manual removal and questing ticks by flag-dragging in the Oroqen Autonomous Banner, Genhe City, and Yakeshi City of Hulunbuir. Tick species were identified based on morphological characteristics, and SFGR infection was detected using real-time fluorescence quantitative PCR. Positive samples were sequenced and analyzed, and molecular phylogenetic trees were constructed. Results A total of 716 ticks were collected, including 469 Dermacentor silvarum and 247 Ixodes persulcatus. The overall positive rate of SFGR in ticks was 75.18%, with positive rates of 86.05% and 58.18% in D. silvarum and I. persulcatus, respectively, and the difference was statistically significant between tick species (χ²=13.958, P<0.05). The positive rates of SFGR in Oroqen, Genhe and Yakeshi were 78.26%, 68.57%, and 77.11%, respectively, with no statistically significant difference among different regions (χ²=1.102, P=0.577). Two SFGR species were detected: Rickettsia heilongjiangensis and Rickettsia raoultii. Phylogenetic analysis based on the ompA gene fragment showed that the detected R. heilongjiangensis sequences were closely related to R. heilongjiangensis (AF179362.2) and strain HL-93 (AF179364.1), while the R. raoultii sequences were closely related to the Russian strain Rickettsia sp. DnS14 (AH009130.2) and the Jilin strain Rickettsia sp. JL-02 (AY093696.1), with sequence identities ranging from 98.46% to 100.00%. Conclusion Dermacentor silvarum are the predominant tick species in the Daxing'anling forest region of Hulunbuir City, and Ixodes persulcatus is distributed in Yakeshi and Genhe. The tick-borne SFGR carrying rate is relatively high, and presence of pathogenic R. heilongjiangensis and R. raoultii in local tick populations suggests that surveillance of tick-borne spotted fever should be strengthened in this region, clinicians' capacity for recognition and diagnosis of relevant clinical manifestations should be enhanced, and conduct scientific protection education and publicity for the public.

Objective To investigate the distribution of Bacillus anthracis bacteriophages in the soil of the anthrax endemic focus in Yuanmou County, Yunnan Province, and to explore their potential role and application prospects in anthrax epidemic prevention and control. Methods Based on previously identified epidemic locations, five representative epidemic sites were selected for soil sample collection and testing. Transport blanks and non-epidemic site controls were established concurrently. Soil samples were collected using stratified random sampling. With extraction blanks included as controls, anthrax phage isolation, purification, and general characterization were performed through double-layer plate assays, electron microscopy, and lysis spectrum analysis. Results A total of 88 soil samples were collected from 5 epidemic sites, yielding 10 isolated anthrax phages with a positivity rate of 11.36%. The positivity rates in Yuanma Town and Jiangyi Township were 50.00% and 20.00%, respectively. The differences in phage positivity rates among sampling sites were statistically significant, with Yuanma Town's rate (50.00%) significantly higher than other townships (Fisher's exact test, P<0.05). Spearman's rank correlation analysis revealed a statistically significant association between epidemic site positivity rates and cumulative case numbers in previous years (2007-2013) (r=1.000, P<0.001). The phages isolated in this study exhibited strong lytic activity against both the anthrax vaccine strain A16R and clinical strains. Some phages also demonstrated lytic activity against Bacillus cereus. Conclusion Bacillus anthracis phages are prevalent in the soil of the anthrax epidemic focus in Yuanmou County, with positivity rates showing significant township-level variation. These can serve as a supplementary indicator of the risk of the epidemic focus independent of case number. It is recommended that townships with a phage-positive rate >20.00 % be classified as "green surveillance areas" for routine monitoring. Historical epidemic sites with a 0% positive rate should be designated as high-risk areas, with a focus on inspecting abandoned carcasses of dead animals. The LP202 strain isolated in this county completely lysed 8 clinical strains from Yunnan Province at 37 ℃, demonstrating potential for both identification and ecological load reduction. However, temperature-dependent cross-reactions need to be corrected before their use for preliminary screening of suspicious colonies at the grass-roots level. This finding provides a valuable reference for exploring environmental anthrax monitoring in natural endemic foci.

Objective To analyze the serotypes, antimicrobial resistance profiles, virulence gene carriage, molecular typing, and other characteristics of Vibrio cholerae isolates from case surveillance and food surveillance samples in Suzhou City from 2016 to 2024, providing a basis for the assessment of the pathogenic risk of Vibrio cholerae and clinical medication guidance. Methods Serotypes 29 Vibrio cholerae strains were identified by serum agglutination and validated by fluorescent PCR. Antimicrobial susceptibility testing was performed using the broth microdilution method. Based on whole-genome sequencing results, genomic characteristics including virulence genes and molecular types were analyzed. Results Among the 29 strains of Vibrio cholerae, the resistance rates to polymyxin E, ampicillin, and ertapenem were the highest, at 82.76%, 72.41%, and 62.07%, respectively. The multi-drug resistance rate reached 79.31% (23/29); however, all strains showed high sensitivity to tetracycline-class antibiotics, with 100.00% sensitivity to tigecycline and a resistance rate of only 3.45% to tetracycline. The results showed that 29 strains of Vibrio cholerae were predominantly non-O1/non-O139, accounting for 86.21% (25/29), while 3 strains of O139 and 1 strain of O1 were detected. The ctx gene encoding cholera toxin was not detected in any of the strains, but the tcp and zot genes were detected in one strain, and the hlyA gene was detected in all strains. Type Ⅲ secretion system (T3SS)-related virulence genes were detected in 48.27% of the strains. All the strains carried at least 7 genes related to type Ⅵ secretion systems (T6SS); among them, the carrying rates of vgrG-2 and vgrG-3 were 89.66% and 10.34%, respectively. Both multilocus sequence typing (MLST) results and core genome multilocus sequence typing (cgMLST) results exhibited high polymorphism, with no obvious dominant type. Conclusion From 2016 to 2024, the serotypes of Vibrio cholerae in Suzhou City were predominantly non-O1/non-O139 types, none of which carried the cholera toxin gene. The molecular types were diverse, and they showed high sensitivity to tetracycline antibiotics, but the phenomenon of multidrug resistance was prevalent.

Objective To analyze the overall diagnostic delays and influencing factors among elderly pulmonary tuberculosis patients aged ≥65 years in Guizhou Province from 2017 to 2024, providing a reference basis for formulating prevention and control measures for this population. Methods Demographic and treatment-related information of elderly pulmonary tuberculosis patients aged ≥65 years in Guizhou Province was collected and screened from the Tuberculosis Management Information System subsystem, a subsystem of the Chinese Disease Prevention and Control Information System. Joinpoint regression models and binary logistic regression models were employed to analyze the variation trends and influencing factors of overall diagnostic delays in this population. Results From 2017 to 2024, the overall diagnostic delay for elderly pulmonary tuberculosis patients in Guizhou Province was 23 (9, 59) days, with a patient delay rate of 62.9%, showing an overall downward trend (AAPC=-3.8, P<0.05, 95% CI: -4.7 to -2.9). The health system delay time was 0 (0，6 ) days, with a health system delay rate of 12.3%, showing an overall upward trend (AAPC=16.7, P<0.05, 95% CI=12.2 to 22.8). Among the nine cities and prefectures under jurisdiction, Tongren City had the highest patient delay rate at 70.4% (5 531/7 858), while Guiyang City had the highest health system delay rate at 26.7% (1 808/6 765). Multivariate analysis indicated that age ≥80 years and non-agricultural occupations may be protective factors against patient delay. Being an ethnic minority, having non-local household registration, having other comorbidities, having combined extrapulmonary tuberculosis, and being passively detected may be risk factors for patient delay. Being male, having a non-agricultural occupation, having non-local household registration, having positive microbiological results, being a retreatment case, having other comorbidities, and having combined extrapulmonary tuberculosis may be risk factors for health system delay. The presence of patient delay may be a protective factor against health system delay. Conclusion Among elderly pulmonary tuberculosis patients aged ≥65 years in Guizhou Province, overall diagnostic delay remains prevalent, though showing a gradual decline over time. Health system delay remains at a relatively low level compared to other western regions, yet it demonstrates a rising trend over time. Priority attention should be directed toward cities and prefectures with high overall diagnostic delays, with optimization of tuberculosis referral mechanisms and medical resource allocation. Targeted interventions should be implemented for elderly patients who are farmers, have non-local household registrations, are male, or have comorbidities. Innovative health education approaches should be adopted to enhance health literacy and proactive healthcare-seeking awareness among these high-risk groups, thereby further reducing overall diagnostic delay in Guizhou Province.

Objective To investigate the expression changes and functional roles of S100 calcium-binding protein A4 (S100A4) in liver fibrosis induced by Echinococcus multilocularis (Em) infection, providing experimental evidence for elucidating the molecular mechanisms of Em-induced liver fibrosis and theoretical insights for potential anti-fibrotic therapeutic targets. Methods A mouse model of Em infection was established, and liver tissues were analyzed at 1, 3, and 6 months post-infection. Histopathological injury and dynamic changes in collagen deposition were evaluated using hematoxylin and eosin (H&E) and Sirius Red staining, respectively. The expression levels of S100A4, alpha-smooth muscle actin (α-SMA), and collagen type I alpha 1 chain (COL1A1) were detected by Western blot, immunohistochemistry (IHC), and quantitative real-time PCR (qPCR). Immunofluorescence staining was performed to identify the cellular source of S100A4. Spearman correlation analysis was used to assess the relationship between S100A4 expression and the severity of liver fibrosis, as well as markers of hepatic stellate cell (HSC) activation. Results H&E staining revealed progressively aggravated liver tissue damage with the advancement of infection in Em-infected mice. Sirius Red staining results showed that the collagen deposition area gradually increased at 1, 3, and 6 months after infection [positive area (μm²): 2 058.00±220.30, 3 685.00±617.20, 5 465.00±552.60 at 1, 3, and 6 months post-infection, respectively; all P<0.05]. IHC, Western blot, and qPCR analyses consistently demonstrated that the expression of HSC activation-related proteins and genes, specifically α-SMA and COL1A1, was significantly upregulated at all time points post-infection and increased progressively with the duration of infection (IHC showed α-SMA positive area (μm²): 3 938.00±481.90, 5 661.00±339.30, 7 554.00±655.60; and COL1A1 positive area (μm²): 1 833.00±167.90, 4 397.00±528.50, 7 457.00±171.50 at 1, 3, and 6 months, respectively; all P<0.05). Western blot showed α-SMA expression levels of 0.71±0.08, 0.73±0.04, and 1.32±0.06, and COL1A1 expression levels of 0.59±0.08, 0.71±0.09, and 1.10±0.09 at 1, 3, and 6 months post-infection, respectively (all P<0.05). qPCR showed Acta2 expression levels of 1.77±0.19, 3.29±0.58, and 5.43±0.91, and Col1a1 expression levels of 1.71±0.18, 3.27±0.68, and 4.84±0.76 at 1, 3, and 6 months post-infection, respectively (all P<0.05). Furthermore, compared with the sham-operated group, both protein and mRNA levels of S100A4 in the perilesional liver tissue were significantly upregulated at all time points in the infected group (IHC showed S100A4 positive area (μm²): 549.80±56.95, 1 347.00±158.60, and 2 227.00±54.90 at 1, 3, and 6 months, respectively; Western blot showed S100A4 expression levels of 0.61±0.10, 0.77±0.13, and 1.09±0.10; and qPCR showed S100A4 expression levels of 3.50±0.76, 7.61±1.48, and 16.82±2.80, respectively; all P<0.05). Immunofluorescence analysis confirmed the co-localization of S100A4 with the macrophage marker F4/80 in Em-infected liver tissues. Spearman correlation analysis revealed significant positive correlations between S100A4 expression levels and the expression of α-SMA, COL1A1, and the collagen deposition area (r=0.89, 0.88, and 0.91, respectively, all P<0.05). Conclusion S100A4 may play a critical role in the initiation and progression of Em-induced liver fibrosis, highlighting its promise as a novel therapeutic target.

Objective To investigate the infection status and genotypes of Wolbachia in Aedes albopictus in Shanghai. Methods From July to September 2024, adult and larval Aedes albopictus were collected from 16 districts of Shanghai, covering greenbelts (parks, etc.), construction sites, residential areas, schools, and hospitals. The Wolbachia surface protein (wsp) was amplified by PCR to determine the Wolbachia infection status. Data were statistically analyzed using SPSS 22.0 software. Furthermore, group A and group B specific primers were used for amplification, and MEGA 10.0 software was employed for the phylogenetic analysis of Wolbachia infection types. Results The average infection rate of Wolbachia in Aedes albopictus in Shanghai was 78.79%. The positive rate in urban areas (85.90%) was higher than that in suburban areas (73.26%), and the difference was statistically significant (χ²=56.519, P<0.001). The positive rate in female Aedes albopictus (91.08%) was higher than that in males (83.12%), the positive rate in adult mosquitoes (87.25%) was higher than that in larvae (61.88%), and both differences were statistically significant (χ²=22.794 and 205.506, respectively; P<0.001). Among the five habitat types, the positivity rate of Wolbachia in Aedes albopictus ranged from 75.11% to 82.53%, with no statistically significant difference (χ²=8.606, P=0.072). Among the infected specimens, 2.96% were infected with wAlbA alone, 36.01% with wAlbB alone, and 61.03% were co-infected with both wAlbA and wAlbB. Genotyping and phylogenetic analysis revealed that the Wolbachia types infecting Aedes albopictus included the wAlbA subgroup of group A, as well as the wAlbB and wPip subgroups of group B. Conclusion Aedes albopictus in Shanghai commonly carries Wolbachia, predominantly infected with groups A and B. Further exploration is warranted on how to implement Wolbachia-based biological control.

Objective To understand the epidemiological pattern of human adenovirus in influenza-like cases in Guizhou Province in 2024, and further analyze the hexon/penton base/fiber genes characteristics of human adenovirus. Method A total of 10 823 influenza-like illness cases were collected in Guizhou Province in 2024. The tests for 11 common pathogens, including human adenovirus (HAdV), rhinovirus (HRV), severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), influenza virus (IFV), respiratory syncytial virus (RSV), parainfluenza virus (HPIV), common coronavirus (HCoV), metapneumovirus (HMPV), enterovirus (EV), bocavirus (HBoV), and Mycoplasma pneumoniae (MP) were conducted by multiplex fluorescence quantitative PCR. The test data were analyzed by χ² test or Fisher test using SPSS 22. HAdV positive samples with Ct values ≤30 were screened for Hexon/Penton base/Fiber genes sequencing and evolutionary analysis. Result Among the 10 823 cases in Guizhou Province, the total detection rate of HAdV was 4.56% (493/10 823), ranking the fourth among pathogens. There were statistically significant differences in the detection rates among different genders, age groups and seasons (P≤0.05). The detection rate was higher in males (4.99%), and the infection rate was the highest in school-aged children (5-<15 years old) (8.79%). Summer was the high-incidence season. The mixed infection rate was 1.64%, with HAdV and HRV mixed infection being the main type (0.28%). Genotyping showed that among the 33 HAdV strains, there were 20 HAdV-B strains (16 strains of type B3 and 4 strains of type B21), 9 HAdV-C strains (2 strains of type C1, 3 strains of type C2 and 4 strains of type C5), 3 potential recombinant viruses composed of HAdV-C1 and C2, and 1 potential recombinant virus composed of HAdV-C2 and C6. Conclusion HAdV is one of the main pathogens of influenza-like cases in Guizhou Province in 2024. The prevalence of HAdV is mainly type B3 in Guizhou Province, and there may be a phenomenon of genetic recombination. Attention should be paid to control and prevention of school-age children and mixed infections.

Objective To analyze the molecular genetic characteristics of Japanese encephalitis virus (JEV) isolated from Anopheles sinensis in Xingren City, Guizhou Province, in 2023, supplement the JEV surveillance data for Guizhou Province, and evaluate the possible effects of viral genetic variation on its virulence and vaccine protection efficacy. Methods Mosquito specimens were collected from different habitats in six cities/prefectures of Guizhou Province from July to September 2023. Virus isolation was performed using C6/36 and BHK-21 cells. The complete genome was obtained by RT-PCR and sequencing, followed by genetic evolution, amino acid variation, and protein structure analyses using MEGA, DNAStar, and SWISS-MODEL. Results One JEV strain (GZ23M09) was isolated from a pool of Anopheles sinensis collected in Xingren City, Qianxinan Prefecture. Phylogenetic analysis indicated that the strain belongs to the GIb genotype and is closely related to recent epidemic strains from Guizhou, Ningxia, Sichuan, Guangxi, and other regions. Compared with the vaccine strain SA14-14-2, multiple amino acid variations were identified in the E protein, including seven key sites associated with neurovirulence, such as E138 and E176. Additionally, a V206A substitution at site E206 was observed, which was a unique variation not detected in other Guizhou or domestic epidemic strains included in the comparison. Structural prediction revealed that three important antigenic epitopes in domain Ⅲ(E337-E345, E377-E382, and E397-E403) maintained conformational consistency with the vaccine strain. Conclusion This study reports the first isolation of a GIb genotype JEV from Anopheles sinensis in Xingren City, Qianxinan Prefecture, Guizhou Province. Analysis of the isolate GZ23M09 reveals its dual molecular characteristics: the presence of virulence-related mutation risks alongside highly conserved core neutralizing epitopes. These findings provide important evidence for assessing local viral evolution, transmission risks, and vaccine protection efficacy.

Objective To investigate the effect of N⁶-methyladenosine (m⁶A) demethylase ALKBH5 on dengue virus (DENV) replication to provide a theoretical basis for elucidating the epigenetic transcriptional regulatory mechanisms between DENV and the host. Methods Western blot analysis was performed to examine changes in ALKBH5 protein levels in Huh-7 cells following DENV infection. Small interfering RNA (siRNA) and a lentivirus-mediated overexpression system were then employed to construct ALKBH5-knockdown and overexpressing cell models, respectively. Western blot, RT-qPCR, and plaque assays were subsequently performed to measure DENV NS3 protein expression levels, extracellular DENV RNA levels, and the titers of released infectious viral particles in these cell models after DENV infection. Transcriptome sequencing was performed to screen differentially expressed genes and downstream signaling pathways regulated in ALKBH5-overexpressing cells infected with DENV. Results DENV infection significantly reduced ALKBH5 protein expression in Huh-7 cells (P<0.05). Knockdown of ALKBH5 resulted in decreased intracellular DENV NS3 protein expression (P<0.05), reduced extracellular DENV RNA levels, and decreased titers of infectious viral particles (P<0.01). In ALKBH5-overexpressing cells, DENV NS3 protein expression increased (P<0.05), and extracellular DENV RNA levels, along with released infectious viral particles, increased (P<0.05). Transcriptome sequencing identified 606 differentially expressed genes between the ALKBH5 overexpression group (OE-ALKBH5) and the control group (OE-Control). Compared with the OE-Control group, the OE-ALKBH5 group had 376 upregulated genes and 230 downregulated genes, which were primarily enriched in PI3K/Akt, NF-kappa B, and extracellular matrix-receptor interaction pathways. Conclusion ALKBH5 positively regulates DENV replication in Huh-7 cells; overexpression of ALKBH5 promotes DENV replication, whereas knockdown of ALKBH5 suppresses DENV proliferation. This study reveals a regulatory role for ALKBH5 in DENV infection, offering a novel direction for further elucidating m⁶A modification-mediated host antiviral mechanisms.

Objective To investigate the diversity and composition of viruses carried by Culex tritaeniorhynchus, the dominant mosquito species in certain areas of southeastern Yunnan Province, and to provide a scientific basis for the surveillance, prevention, and control of local arboviral diseases. Methods From July to August 2023, mosquitoes were collected from diverse habitats, including farmhouses, pig pens, and cattle sheds in Kaiyuan City and Huaning County. Following morphological identification, four sample pools of female Culex tritaeniorhynchus were constructed (50 mosquitoes per pool, with two biological replicates per region). Total RNA was extracted from the samples and subjected to metavirome sequencing using the Illumina NovaSeq 6000 platform. Bioinformatics methods were employed for data quality control, sequence alignment, viral annotation, relative abundance analysis, and phylogenetic studies. Results A total of 5 640 adult mosquitoes were collected, with Culex tritaeniorhynchus being the dominant species (71.28%). A total of 41 400 325 viral reads were obtained, from which 2 067 viral contigs were assembled. In total, 104 viruses belonging to 28 viral families were identified. The viral composition was dominated by insect-specific viruses (61.60%), followed by vertebrate-related viruses (26.90%). Both mosquito sample pools from the two regions contained viral families such as Coronaviridae, Flaviviridae, Herpesviridae, Solinviviridae, Mimiviridae, and unclassified viruses. Japanese encephalitis virus (JEV) was detected in all four sample pools, and phylogenetic analysis indicated that it belonged to genotype I. Compared with the vaccine strain SA14-14-2, 103 amino acid differences were observed; however, the key antigenic regions of the E protein remained completely identical. Additionally, other mosquito-associated viruses such as Quang Binh virus, Hubei mosquito virus 2, Wuhan mosquito orthophasmavirus 2, and Menghai rhabdovirus were detected. Conclusion This study preliminarily reveals the diversity of viruses carried by Culex tritaeniorhynchus in Southeastern Yunnan, confirms the presence of important arboviruses such as JEV, identifies a variety of insect-specific viruses and unclassified viruses. The results highlight the need for continuous monitoring of viral evolutionary dynamics and further research to clarify viral transmission mechanisms and public health risks.