Abstract Objective: To investigate whether long non-coding RNA (LncRNA) nuclear-enriched abundant transcript 1 (NEAT1) promotes hepatic fibrosis in rats with non-alcoholic fatty liver disease (NAFLD) by modulating the microRNA-150-5p (miR-150-5p)/Notch receptor 2 (NOTCH2) axis. Methods:  A NAFLD rat model was established by high-fat diet feeding, and the rats were divided into the model group, si-NC group, si-NEAT1 group, si-NEAT1+anti-NC group and si-NEAT1+anti-miR-150-5p group. Rats fed a normal diet were set as the control group, with 12 rats in each group. An automatic biochemical analyzer was used to detect the levels of high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), total cholesterol (TC), triacylglyceride (TG), aspartate aminotransferase (AST) and alanine aminotransferase (ALT). Enzyme-linked immunosorbent assay (ELISA) was adopted to measure the levels of serum interferon-γ (IFN-γ) and interleukin-4 (IL-4). Hematoxylin-eosin (HE) staining was performed to observe the morphological structure of liver tissue. Masson staining was used to observe the liver tissue fibrosis. Immunohistochemistry was used to analyze the expression of α-smooth muscle actin (α-SMA) in liver tissue. Quantitative real-time polymerase chain reaction (qRT-PCR) was applied to detect the mRNA levels of NEAT1, miR-150-5p and NOTCH2 in liver tissue, and Western blot assay was applied to determine the protein expression of NOTCH2 in liver tissue. Moreover, dual-luciferase assay was conducted to verify the targeted regulatory relationships between NEAT1 and miR-150-5p, as well as between miR-150-5p and NOTCH2. Results  Compared with the control group, the model group presented obvious structural damage of hepatic lobules, extensive hepatocellular steatosis and severe hepatic fibrosis. The levels of serum HDL-C and IFN-γ, and the level of miR-150-5p in liver tissue were lower, while the levels of LDL-C, TC, TG, AST, ALT, IL-4, the expressions of NEAT1, NOTCH2 mRNA and NOTCH2 protein, and the positive rate of α-SMA were higher (q = 36.159, 25.391, 17.323, 35.610, 39.287, 30.960, 22.244, 25.713, 27.043, 12.142, 14.532, 14.131, 29.102, P<0.05). Compared with the si-NC group, the si-NEAT1 group showed alleviated pathological damage and reduced hepatic fibrosis in liver tissue, the levels of serum HDL-C and IFN-γ, and the level of miR-150-5p in liver tissue were higher, while the levels of LDL-C, TC, TG, AST, ALT, IL-4, the expressions of NEAT1, NOTCH2 mRNA and NOTCH2 protein, and the positive rate of α-SMA were lower (q = 30.830, 18.550, 14.381, 29.938, 33.375, 24.808, 17.914, 22.907, 18.277, 10.957, 11.525, 11.747, 18.273, P<0.05). Compared with the si-NEAT1+anti-NC group, the si-NEAT1+anti-miR-150-5p group exhibited aggravated hepatic pathological damage and increased fibrosis degree, the levels of serum HDL-C and IFN-γ, and the level of miR-150-5p in liver tissue were lower, while the levels of LDL-C, TC, TG, AST, ALT, IL-4, the expressions of NOTCH2 mRNA and NOTCH2 protein, and the positive rate of α-SMA were higher (q = 24.360, 16.463, 11.439, 21.114, 27.710, 18.656, 13.950, 18.412, 15.857, 7.892, 9.875, 13.571, P<0.05), which indicated that down-regulation of miR-150-5p could reverse the ameliorative effect of the down-regulation of NEAT1 on the above indicators in NAFLD rats. Conclusion: Down-regulation of LncRNA NEAT1 can inhibit hepatic fibrosis in NAFLD rats by regulating the miR-150-5p/NOTCH2 axis.

fecal - oral, and sexual transmission, indirect transmission of pathogens through contaminated surfaces is emerging as a potential
new mode of transmission. With the continuous development of international trade, the interaction between international trade and
infectious disease outbreaks is becoming increasingly close. The cross-border transmission of epidemics through contaminated
goods has become a new challenge for port epidemic prevention and control. Understanding the survival time of pathogens on
inanimate surfaces is of great significance for port disinfection and epidemic prevention and control.Numerous studies have
described the mechanism of pathogen transmission through contaminated surfaces, and some studies have even pointed out that
some infectious disease outbreaks were caused by contaminated objects. However, there has been no systematic and in-depth
study on the survival of different pathogens on the surface of different subjects. In this paper, we systematically review studies on
the persistence of different DNA viruses such as Adenoviridae, Papillomaviridae, Herpesviridae, and Poxviridae, as well as RNA
viruses such as Reoviridae, Picornaviridae, Caliciviridae, Flaviviridae, Orthomyxoviridae, Paramyxoviridae, Filoviridae,
Coronaviridae, Retroviridae, and Hepadnaviridae on the surface of objects, and explore the possibility of cross-border
transmission through international goods, providing a reference for pandemic prevention and control at ports.

Abstract: Objective To investigate the clinical characteristics and molecular epidemiological features of Cryptococcus
isolates in HIV-positive and HIV-negative patients with cryptococcal meningitis (CM), and to provide a reference for early
recognition, diagnosis, and treatment of CM. Methods A retrospective comparative analysis was conducted on the clinical
characteristics and cryptococcal genotypes of HIV-positive and HIV-negative CM patients admitted to a tertiary hospital in
Guangzhou between March 2015 and February 2024. Antifungal susceptibility testing was performed using the ATB FUNGUS 3
system, and strain genotypes were determined by mass spectrometry and multilocus sequence typing (MLST). Results A total
of 121 patients with CM were included, comprising 102 HIV-positive patients and 19 HIV-negative patients. Compared with
the HIV-negative group, the HIV-positive group had a higher proportion of male patients (80.4% vs. 47.4%, χ2=7.680, P 均<
0.01), had a shorter interval from symptom onset to diagnosis (17 vs. 45 days, U=-3.794,P<0.01）). Laboratory findings showed
that, in the HIV-positive group, the CD4+ T-cell count, CD4+/CD8+ ratio, cerebrospinal fluid white blood cell count,
cerebrospinal fluid protein level, and cerebrospinal fluid glucose level were 12 (5, 34) cells/μL, 0.04 (0.02, 0.08), 28 (15, 61) ×
106/L, 489 (306, 833) g/L, and 2.3 (1.9, 3.1) mmol/L, respectively; all of these parameters differed significantly from those of the
HIV-negative group (U=-4.852, -5.363, -2.052, -3.552, and -3.376, respectively; all P<0.01). The 4-week and 24-week
mortality rates in the HIV-positive group were 14.7% and 28.4%, respectively, both higher than those in the HIV-negative
group (5.3% and 15.8%, respectively), though the differences were not statistically significant (both P>0.05). Molecular typing
showed that Cryptococcus neoformans VNI was the predominant molecular type in both groups (98.0% vs. 100.0%, P=0.332),
with ST5 as the predominant sequence type (81.4% vs. 94.7%). In addition, two Cryptococcus gattii (VG) isolates were
identified in the HIV-positive group, and three isolates of a novel sequence type, ST685, were detected for the first time; one
patient harboring this sequence type died at the 24-week follow-up. Conclusion Compared with HIV-negative CM patients,
HIV-positive CM patients exhibited a weaker cerebrospinal fluid inflammatory response and a relatively poorer prognosis.
Particular attention should be paid to infection with the newly identified ST685 sequence type detected in the HIV-positive
group.

Objective   Burkholderia pseudomallei (Bp), the causative agent of melioidosis, exhibits high pathogenicity and intrinsic antibiotic resistance. However, the molecular mechanisms underlying its virulence remain incompletely understood. This study aimed to elucidate the role of caseinolytic protease P (ClpP) in the physiological functions and virulence of Bp.  Methods   This study used the auxotrophic strain (HNBP001) as the starting strain and successfully constructed the clpP gene deletion mutant (ΔclpP) and the clpP gene complementation strain (ΔclpP::clpP) using homologous recombination technology.This study systematically elucidated the multifaceted regulatory roles of the clpP gene in bacterial growth, antibiotic resistance, and virulence through growth curve analysis, antimicrobial susceptibility testing, biofilm formation assessment, and cell infection experiments.  Results   This study successfully constructed and validated the ΔclpP and ΔclpP::clpP strains. The deletion of the clpP gene led to significant alterations in key pathogenic phenotypes of the bacteria. Compared to the HNBP001, the ΔclpP mutant exhibited significantly reduced biofilm formation and motility (P<0.05).In the host cell infection experiment, the cell survival rate increased from 55% to 70%.Furthermore, the susceptibility of ΔclpP to multiple antibiotics increased significantly, with the Minimum Inhibitory Concentration (MIC) values for ceftazidime, ceftriaxone, cefotaxime, and amikacin all reduced by half compared to the baseline levels.   Conclusion   This study systematically validated the regulatory role of clpP on multiple critical phenotypes of Bp, including motility, virulence, and antibiotic resistance.The experiments demonstrated that the deletion of clpP simultaneously attenuated bacterial virulence and enhanced its sensitivity to antibiotics. This "dual-effect" characteristic highlights its unique advantage as a potential drug target.This finding suggests that clpP could serve as a crucial target for developing novel antimicrobial strategies, providing a new research direction for the prevention and control of Bp infections.

Objective: To investigate the effects of astaxanthin on liver function and liver fibrosis in hepatitis B (HBV) rats, and to analyze its relationship with the sphingosine kinase 1 (SphK1)/sphingosine 1-phosphate (S1P) pathway. Methods: HBV rat models were established and assigned into HBV group, astaxanthin-low, medium and high dose (astaxanthin-L, M, H) groups, and astaxanthin-H+SphK1 activator PMA (astaxanthin-H+PMA) groups, with 12 rats in each group. Another 12 rats were taken as the control (CK) group. The liver function indicators, fibrosis indicators, HBV markers, and inflammatory factor indicators were detected by ELISA. The pathological changes of liver tissue were detected by HE staining. Masson staining was conducted to detect liver tissue fibrosis. In addition, the protein expression was detected by Western blot. Results: The hepatocytes of rats in the CK group were arranged radially, with an orderly structure, and there was no obvious swelling or infiltration of inflammatory cells. In the HBV group, the hepatocytes of rats were randomly arranged, the nucleoli shrank, and obvious inflammatory cell infiltration and punctate necrosis occurred. Compared with the CK group, the alanine aminotransferase (ALT), aspartate aminotransferase (AST), laminin (LN), hyaluronic acid (HA), procollagen type Ⅲ (PCⅢ), hepatitis B e antigen (HBeAg), hepatitis B surface antigen (HBsAg), interleukin-2 (IL-2), interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α), Ishak scores, collagen volume fraction, and the expressions of SphK1 and S1P in the HBV group were higher (P<0.05). Compared with the HBV group, the ALT, AST, LN, HA, PCⅢ, HBeAg, HBsAg, IL-2, IFN-γ, TNF-α, Ishak scores, collagen volume fraction, and the expressions of SphK1 and S1P in the astaxanthin-L, M and H groups were lower (P<0.05). Compared with the astaxanthin-H group, the ALT, AST, LN, HA, PCⅢ, HBeAg, HBsAg, IL-2, IFN-γ, TNF-α, Ishak scores, collagen volume fraction, and the expressions of SphK1 and S1P in the astaxanthin-H+PMA group were higher (P<0.05). Conclusion: The inhibitory effect of astaxanthin on liver tissue fibrosis and its improvement on liver function in HBV rats may be achieved by inhibiting the SphK1/S1P pathway.

Abstract: Chikungunya fever is a viral infectious disease transmitted by the bite of Aedes aegypti and Ae. albopictus mos⁃
quitoes. It is widely prevalent in tropical and subtropical regions. The disease first broke out in Tanzania in 1952, and between
1952 and 2003, it was only endemc in Africa, Southeast and South Asia, however, since the outbreak of the disease in the Indi⁃
an Ocean region in 2004, the epidemic has spread to Africa, Asia, Americas, Oceania, and Europe. As of the end of 2024, indig⁃
enous epidemics of chikungunya fever had occurred in 119 countries and regions around the world, and about three-quarters of
the global population is at risk of infection by chikungunya virus (CHIKV). It has evolved into a globally rampant vector-borne
infectious disease. The continuous expansion of the distribution areas of Ae. aegypti and Ae. albopictus, along with the en⁃
hanced adability of CHIKV to these mosquito vectors, and increased transmission efficiency, are the main causes of the pan⁃
demic of the disease. Since the beginning of 2025, outbreaks of the disease have resurfaced and shown a trend of diffusion in
the Indian Ocean region, Central and South America, and Southeast Asia. Indigenous outbreak of the disease were occurred in
Guangdong Province, China, 2025. Recently, the World Health Organization has an alert, urging member countries to strength⁃
en their preparedness to deal with the epidemic. In this context, mastering the global epidemic situation of chikungunya fever,
CHIKV genotypes, and factors influencing transmission, and further strengthening surveillance of imported cases and vector Ae⁃
des mosquitoes, prevention of cross-border transmission, and local outbreaks are important tasks currently facing China. This
paper provides a review of the relevant research progress.

fever outbreak in Quanzhou City, Fujian Province, in 2025, providing empirical evidence to continuously improve the
investigation and response to locally acquired chikungunya fever outbreaks. Methods Field epidemiological investigation
methods were employed to collect case information and emergency monitoring data on mosquito vector density from a local
chikungunya fever outbreak in Fujian Province in 2025. Descriptive epidemiological methods were used to analyze the
epidemic features. WPS Office software was utilized for statistical analysis of indicators including the epidemiological
distribution of cases (time, place, and person), symptom profiles, and mosquito vector density. Results From August 24 to
September 10, 2025, a local chikungunya fever outbreak occurred in Quanzhou City, Fujian Province, with a total of 164
confirmed cases. The median interval from onset to diagnosis was 1 (1, 2) days. This outbreak exhibited a typical unimodal
distribution pattern, characterized by the coexistence of high clustering and sporadic cases. The virus demonstrated effective
transmissibility across different populations and age groups. All 164 cases were mild, with no severe or fatal cases; 23 cases
(14.02%) concurrently presented with fever, rash, and arthralgia. Following the activation of the emergency response and the
implementation of comprehensive adult mosquito control measures in risk areas, both the Breteau Index and adult mosquito
density fell below the safety threshold within 3 days, and the outbreak was effectively contained within 4 weeks.
Conclusion Chikungunya fever spreads rapidly. Establishing a more sensitive surveillance case definition is crucial for
achieving early case detection. A prompt response at the initial stage of the outbreak, together with organizing coordinated and
efficient mosquito control operations in risk areas, are core measures for successfully interrupting the transmission chain of
mosquito-borne diseases such as chikungunya fever and dengue fever.

Abstract: Objective To analyze the epidemiological characteristics and influencing factors of chikungunya fever in
Nanhai District, Foshan City, Guangdong Province in 2025, and to provide a reference for the development of local prevention
and control measures. Methods A descriptive epidemiological analysis was conducted on 1 274 local cases. Chi-square
tests, non-parametric tests, correlation and regression analyses were employed to examine the temporal, spatial, and
demographic distribution, urban-rural disparities, the impact of population density, and the timeliness of case diagnosis.
Results The epidemic lasted for 130 days, peaking in July. It affected 199 villages/communities across 7 towns/subdistricts,
with Guicheng accounting for 46.94% of cases. The proportion of affected villages/communities was significantly higher in
urban communities (92.96%) than in rural communities (61.01%) (P<0.001). Local transmission predominated (94.8%), yet the
urban-rural distribution differed: locally acquired cases were more prevalent in rural communities (59.4%), whereas imported
cases were concentrated in urban communities (66.7%) (P<0.001). Case count and density both correlated strongly with
population density (r=0.860 and 0.865, P<0.05); for every 1000 people/km⊃2; increase in population density, an estimated 54
additional cases occurred. Cluster outbreaks (90 events, 73.55% of cases) were more common in urban settings (P<0.001).
Cases were predominantly male (53.38%) and aged 30–39 years. Occupationally, homemakers, the unemployed, and retirees
constituted the largest groups. The mean time from onset to diagnosis was (2.17±1.31) days. Diagnosis delay was associated with
infection acquired outside the district, diagnosis rendered outside the district, case reporting from primary-level medical
institutions, and a healthcare-seeking distance of ≥20 km (P<0.05). Mosquito vector surveillance indicated that the density of
Aedes albopictus had already exceeded the transmission risk threshold during the early phase of the epidemic (June–July).
Conclusion The epidemic showed significant spatial clustering and pronounced urban-rural disparities. High population
density was the key risk factor driving epidemic spread. Urban communities faced a higher incidence risk and a greater
frequency of cluster outbreaks, and served as the main gateway for imported cases; rural communities, by contrast, were the
principal zone of local transmission. Diagnostic delays highlighted deficiencies in cross-regional coordination, primary-care
diagnostic capacity, and healthcare accessibility. Differentiated prevention and control strategies are recommended, along with
strengthened regional joint-prevention mechanisms and primary-level capacity building.

after the emergency response to dengue and chikungunya outbreaks in Hainan Province in 2025, and to explore rational
insecticide application strategies for short-term outbreak recurrence. Methods In 2025, Aedes albopictus larvae were
collected from the core areas of dengue fever and chikungunya fever outbreaks in Haikou, Wuzhishan, and Sanya before and
after emergency response and disposal. The collected larvae were reared under laboratory conditions to the first-filial-
generation adults. Insecticide resistance was assessed by the adult contact diagnostic dose method, with adult mortality used
to evaluate resistance changes following outbreak disposal. The χ2 test was employed to analyze changes in Aedes albopictus
mortality to insecticides, and Pearson correlation analysis was performed to examine inter-insecticide associations.
Results Prior to emergency response and disposal, Aedes albopictus populations from Haikou, Sanya and Wuzhishan cities
demonstrated suspected resistance to propoxur, with97.53%、96.67% and 95.35% mortality; populations from Haikou and
Wuzhishan cities showed resistance to fenitrothion, with mortality rates of 75.32% and 70.37%; the Sanya population
demonstrated susceptibility to fenitrothion, with a mortality rate of 98.78%; populations from Haikou, Sanya and Wuzhishan
cities demonstrated resistance to deltamethrin, permethrin and lambda-cyhalothrin. Following emergency response and
disposal, resistance-associated mortality further declined across populations. In Haikou City, mortality to propoxur decreased
to 53.49%. Mortality to fenitrothion showed a significant decrease (χ2=4.32, P<0.05), while mortality to deltamethrin,
permethrin, and lambda-cyhalothrin remained unchanged. In Wuzhishan City, mortality to all tested insecticides declined, with
significant decreases observed for propoxur and permethrin (χ2=5.65 and 29.6, respectively; P<0.05 for both). In Sanya City,
the resistance level to fenitrothion increased, with mortality rising to 95.29%; mortality to propoxur, deltamethrin, permethrin,
and lambda-cyhalothrin declined, with significant decreases observed for propoxur, deltamethrin, and permethrin (χ2=7.61,
21.90, and 54.70, respectively; P<0.05 for all). A very strong positive correlation was observed between mortality to
deltamethrin and fenitrothion (r=0.789, 0.05<P=0.062<0.1). No significant correlations were observed among mortality rates to
the other insecticides (P>0.1). Conclusion Large-scale insecticide application during the epidemic response exacerbated the
resistance of Aedes albopictus to insecticides. It is recommended that, during short-term outbreak recurrence, insecticide
application be guided by resistance profiles, environmental management measures such as breeding site elimination be
strengthened, and insecticide use be strategically reduced to slow the development of resistance.

Abstract: Objective To analyze the epidemiological characteristics of confirmed chikungunya fever cases reported in
Sichuan Province in 2025, with a focus on the epidemiological features and emergency response to the first local outbreak in a
county of Sichuan Province in October, and to provide a scientific basis for the surveillance, early warning, prevention, and
control of mosquito -borne infectious diseases. Methods Case data were derived from individual records of confirmed
chikungunya fever cases with current residence in Sichuan Province reported in 2025 through the National Notifiable Infectious
Disease Reporting Information Management System, as well as epidemiological investigation reports of cases involved in the
outbreak. Data collation and statistical analysis were performed using Excel 2016 software. Results In 2025, a total of 363
confirmed cases of Chikungunya fever were reported in Sichuan Province, of which 341 (93.94%) were locally acquired. The
incidence peaked in October, accounting for 326 cases (89.81%). Nanchong City reported the highest number of cases (302,
83.20%), exhibiting a unimodal outbreak pattern. The county-level outbreak documented 282 confirmed cases, with a male-tofemale
ratio of 0.71∶1 and a mean age of 46.03±25.07years. Farmers accounted for the largest occupational group (49.65%).
Cases were mostly localized in the county's urban area, accounting for 271 cases (96.10%). The epidemic persisted for 12 days,
demonstrating a fast onset and prompt remission. The predominant clinical manifestation was arthralgia (85.11%), followed by
fever (64.54%) and rash (30.50%); only 17.02% of cases presented with the classic triad of symptoms simultaneously. Through
a four-tier coordinated command system, grid-based management, and expedited mosquito vector treatment, the Breteau Index
(BI) and net trap index (NTI) declined rapidly to safe levels, facilitating prompt containment of the epidemic. All cases received
timely treatment, with no severe cases or deaths. Conclusion The 2025 chikungunya fever epidemic in Sichuan Province
exhibited a distinct seasonal peak and pronounced local clustering. The epidemic signifies that chikungunya fever has
established local transmission in Category II inland regions of China, marked by rapid onset and unusual clinical presentations.
Enhancing early monitoring throughout the epidemic season, improving clinical recognition capacity, and establishing an
efficient emergency response system and precise mosquito vector control framework are essential for controlling mosquitoborne
infectious illnesses.

Abstract： Objective　To deeply analyze the epidemiological characteristics and trends of hepatitis E among the elderly population in eastern China, providing scientific data support for developing regional hepatitis E prevention and control strategies. Methods　Incidence data on hepatitis E among individuals aged 60 years and above in eastern China from 2008 to 2022 were retrieved from China Disease Control and Prevention Information System. Descriptive methods were used to analyze the epidemiological distribution of the disease. Joinpoint regression analysis was performed to examine the temporal trends in reported incidence and age-standardized incidence rates. An age-period-cohort model was conducted to assess the independent effects of age, period, and birth cohort. Results　The average annual age-standardized reported incidence rate (ASRIR) of hepatitis E among the elderly population was 5.42/100 000 in eastern China, which was higher in males (7.81/    100 000) than in females (3.17/100 000). From 2008 to 2022, the ASRIR of hepatitis E in the elderly showed a downward trend in eastern China, with an average annual percentage change (AAPC) of -3.33% (-5.22%~1.41%, P=0.003), and -4.19%         (-6.01%~2.33%, P<0.001) for males and -1.50% (-4.56%~1.67%, P=0.351) for females, respectively. The age-period-cohort analysis showed that the risk of hepatitis E decreased with age in the elderly in eastern China, with RR declining from 1.28 (95%CI: 1.17-1.41) in the 60-<65 years old group to 0.45 (95%CI: 0.38-0.53) in the ≥85 years old group. The period effect showed incidence of hepatitis E in the elderly showed a continuous downward trend from 2008 to 2022, with RR decreasing from 1.06 (95%CI: 0.99-1.13) in 2008-2012 to 0.70 (95%CI: 0.65-0.75) in 2018-2022. The cohort effect demonstrated the later the birth cohort, the lower the risk, with the higest RR in the 1923-1927 birth cohort (RR=2.88, 95%CI: 2.01-4.13) and the lowest in the 1953-1957 cohort (RR=0.63, 95%CI: 0.55-0.72). Conclusion　From 2008 to 2022, the ASRIR of hepatitis E among the elderly population showed a declining trend in eastern China. It is necessary to strengthen the monitoring of the changing trend of hepatitis E in the elderly male, and take targeted prevention and control measures to actively respond to the epidemic of hepatitis E in the elderly.

Abstract： Objective　To analyze the investigation and disposal process of the first local chikungunya fever(CHIKF) case in Shanghai, and to provide reference for the diagnosis, treatment and prevention of CHIKF. Methods　Epidemiological investigation was adopted for the case survey. Viral nucleic acid was detected by multiplex real-time fluorescent quantitative PCR. Risk areas of the epidemic foci were demarcated, and Aedes density monitoring was conducted using Breteau index method, double net trap method and BG-Trap method. Results　The case presented symptoms such as fever, arthralgia, conjunctival hyperemia and rash; blood routine showed decreased white blood cell count and increased C-reactive protein. Laboratory test indicated that the Chikungunya virus nucleic acid was positive (Ct value: 21.92), and gene sequencing results showed that the virus strain was consistent with that of the imported case in the same community, both belonging to the East/Central/South African genotype. From October 10th, 2025 to the end of the epidemic, the mosquito vector density reached the safe level [Breteau index<5, tent trapping index<0.9 mosquitoes/(tent⋅hour)] in both the core area and the warning area of the epidemic site. The epidemic was finally ended on October 31st. Conclusion　The case is the first local CHIKF case in Shanghai. Early detection and isolation of the case, efficient implementation of mosquito vector monitoring and disinfection, and multi-department joint prevention and control are the keys to effectively curbing the local epidemic of CHIKF. 

Abstract: Objective　To verify the diagnostic efficacy of Mycobacterium tuberculosis nucleic acid colloidal gold technology (PCR colloidal gold assay) for detecting Mycobacterium tuberculosis in sputum samples of clinical pulmonary tuberculosis patients, to evaluate the diagnostic value of PCR colloidal gold assay for tuberculosis. Methods　Sputum samples from 239 active pulmonary tuberculosis patients registered in four designated tuberculosis medical institutions were collected in Nantong City between July 1, 2024, and May 31, 2025. The same sputum samples were tested for Mycobacterium tuberculosis using sputum smear microscopy, GeneXpert, and PCR colloidal gold assay. The differences in positive rates were compared among the three methods for different types of patients, the diagnostic efficacy of PCR colloidal gold assay was evaluated, with GeneXpert results and clinical comprehensive diagnosis as reference standards. Results　Among the 239 pulmonary tuberculosis patients, 111 cases (46.44%) were diagnosed with pulmonary tuberculosis (pathogen-positive), and 128 cases (53.56%) were clinically diagnosed (pathogen-negative). The positive rate was 30.96% (74/239) of sputum smear microscopy, was 43.93% (105/239) of GeneXpert method, and was 51.05% (122/239) of PCR colloidal gold assay. There was a statistically significant difference in positive detection rates of Mycobacterium tuberculosis among at least two methods (P<0.001). Further analysis revealed that in the overall patients, positive detection rates for Mycobacterium tuberculosis by GeneXpert method and PCR colloidal gold assay were higher than that by sputum smear microscopy, with statistically significant differences, but no statistically significant difference was observed between the GeneXpert method and PCR colloidal gold assay. Using GeneXpert detection as the standard, the sensitivity of the PCR colloidal gold assay and smear microscopy were 96.19%(95%CI: 90.53%-98.95%) and 66.67%(95%CI:56.80%-75.57%), respectively; the specificity were 84.33%(95%CI:77.05%-90.03%) and 97.01%(95%CI:92.53%-99.18%), respectively; the positive predictive values were 82.79%(95%CI:75.07%-88.93%) and 94.59%(95%CI:86.76%-98.51%), respectively; and the negative predictive values were 96.58%(95%CI:91.56%-99.06%) and 78.79%(95%CI:71.89%-84.70%), respectively. The overall consistency between PCR colloidal gold assay and GeneXpert was good (Kappa=0.791), significantly higher than that of smear microscopy (Kappa=0.658). With clinical diagnosis results as the reference standard, the sensitivity of the PCR colloidal gold asasay was 95.50%(95%CI：89.97%-98.54%), the specificity was 87.50%(95%CI：80.50%-92.73%), the concordance was 91.21%(95%CI：86.77%-94.61%), and the Kappa value was 0.825(95%CI：0.758-0.892). Conclusion　The PCR colloidal gold assay showed high consistency with clinical diagnosis, and equivalent comparable diagnostic efficacy to GeneXpert, which can improve the positive detection rate of sputum samples from suspected pulmonary tuberculosis patients. The method offers convenience, visual readout, low cost, simple in steps, and have low requirements for laboratory conditions, making it suitable for widespread application in primary medical institutions as an early clinical pulmonary tuberculosis diagnostic method.

Objective To analyze the epidemiological characteristics, spatiotemporal distribution patterns, and risk factors for local transmission of chikungunya cases in Shenzhen City in 2025, providing scientific evidence for precise prevention and control of mosquito-borne diseases in megacities. Methods Analyzed the sociodemographic and spatiotemporal distribution of cases; used Spearman's cross-correlation and negative binomial regression to examine the lagged association between incidence and population inflow.Global and local spatial autocorrelation analyses together with hotspot shift analysis were conducted to explore street-level spatial clustering and evolutionary characteristics. Logistic regression models were used to identify risk factors for local transmission of imported cases. Results In 2025, a total of 818 chikungunya fever cases were reported in Shenzhen, comprising 313 imported cases and 505 locally acquired cases.Imported cases were mainly from Foshan and Guangzhou in Guangdong Province (125 cases, accounting for 39.94%), and Guangxi Zhuang Autonomous Region was the predominant source outside Guangdong Province (59 cases, 18.85%).The male-to-female ratio was 1:0.6, with cases predominantly occurring among working-age adults aged 20-49 years (65.6%). By occupation, workers accounted for 28.1% of all cases.The cases occurred from July to December, with an epidemic curve showing a unimodal distribution. The peak of onset was observed on October 11 (54 cases, accounting for 6.6% of the total).The incidence of chikungunya fever in Shenzhen showed a 6-day lagged association with population inflow (Spearman r=0.164, P=0.0387). For every 10,000 increase in population inflow, the daily number of cases after 6 days increased by 4.2% (IRR=1.042, 95%CI: 1.026-1.060).Cases exhibited significant spatial clustering. The global Moran's I value was 0.3329 (Z = 4.94, P < 0.001) for imported cases and 0.2534 (Z = 3.59, P < 0.001) for locally acquired cases, respectively.Over time, the epidemic hotspots in the city shifted from the peripheral areas toward the central urban area. Residence in urban villages was an independent risk factor for local transmission (OR=4.472, 95%CI: 1.928–11.739), while arthralgia symptoms (OR=0.459, 95%CI: 0.224–0.940)and self-medication (OR = 0.26, 95% CI: 0.04-0.93, likelihood ratio test P=0.037) were protective factors. Conclusion The chikungunya fever epidemic in Shenzhen was driven by population inflow from the Pearl River Delta region, forming high-risk clusters in central urban areas. Residence in urban villages significantly increases the risk of local transmission and should be prioritized for prevention and control. Arthralgia and self-medication reduce transmission, but mild cases pose a higher occult risk, necessitating enhanced health education and surveillance. It is recommended to establish cross-regional collaborative surveillance, focus on vector control in urban villages, and strictly implement mosquito bite prevention and isolation measures for mild cases.

Objective  To analyze the changing trend and spatio-temporal distribution characteristics of the reported incidence rate of pulmonary tuberculosis complicated with diabetes in Jiangsu Province, and to guide the formulation of scientific prevention and control strategies and measures. Methods  Medical record data of pulmonary tuberculosis complicated with diabetes cases in Jiangsu Province from 2018 to 2023 were collected from the "Tuberculosis Management Information System" in the "China Information System for Disease Control and Prevention" to describe the characteristics of temporal, population and spatial distribution. Joinpoint 4.9.1.0 software was used to analyze the changing trend of the reported incidence rate, Geoda 1.22.0 software was applied to conduct spatial autocorrelation analysis at the county level, and SaTScan 10.3.3 software was used for spatio-temporal scanning analysis. Results  A total of 6781 PTB-DM cases were reported in Jiangsu Province from 2018 to 2023, with an average annual reported incidence rate of 1.34 per 100,000 population. The reported incidence rate showed an upward trend (AAPC=12.66%, P<0.05). The regions with the highest annual reported incidence rates were Gaochun District of Nanjing (6.79 per 100,000), Hanjiang District of Yangzhou (5.28 per 100,000), and Guangling District (4.84 per 100,000). Among PTB-DM patients, males accounted for 81.39% (5,519/6,781); the highest proportion was in the ≥70-year-old age group at 28.05% (1,902/6,781); and farmers constituted the largest occupational group at 55.42% (3,758/6,781). The results of global spatial autocorrelation analysis showed that except for 2022, the reported incidence rates of PTB-DM in other years showed significant positive spatial correlation (Moran's I: 0.066-0.287, P<0.05). The results of local autocorrelation analysis showed that there were high-high clustering areas in the reported incidence rate of PTB-DM from 2018 to 2023, mainly distributed in southern and central Jiangsu. The results of spatiotemporal scanning analysis showed that there were 13 clustering areas of two types from 2018 to 2023. Among them, one type of clustering area covered 28 counties and districts, distributed in Nanjing City, Wuxi City, Zhenjiang City, Changzhou City, Taizhou City and Yangzhou City, with the clustering time being 2021-2023. Conclusion  From 2018 to 2023, the reported incidence rate of PTB-DM in Jiangsu Province showed an upward trend, and men, the elderly and farmers are the key populations for prevention and control. The reported incidence of PTB-DM has spatial aggregation, and the hot spots are mainly distributed in southern and central Jiangsu.

Abstract: Objective　To establish a rapid and efficient liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for screening and quantitative analysis of 12 target compounds in biological samples from patients suspected of anticoagulant rodenticide poisoning. Methods    A total of 100 μL serum or urine samples were directly extracted using a mixture of methanol and acetonitrile. The separation was performed on an Atlantis T3 column (150 × 2.1 mm, 3 μm) with a gradient elution using ammonium acetate solution (5 mM) and acetonitrile. Mass spectrometry analysis was conducted in the negative electrospray ionization mode with multiple reaction monitoring (MRM). The method was comprehensively validated for linearity, sensitivity, accuracy, and precision, and it was applied to the clinical detection of a critically ill patient in late pregnancy. Results    Method validation demonstrated good linearity (r≥0.997) for all 12 anticoagulant rodenticides within a range of 0-400 μg/L. The limits of detection (LOD) and quantification (LOQ) were 0.1-0.5 μg/L and 0.3-1.7 μg/L, respectively. Spiked recoveries for blood and urine samples were 79%-129% and 67%-118%, respectively, with relative standard deviations (RSDs) below 11%, indicating satisfactory accuracy and precision. When applied to clinical cases, the method rapidly detected a concentration of warfarin as high as 1 664 μg/L in the mother's serum and 360 μg/L in the newborn's serum. The maternal-neonatal serum concentration ratio and their respective blood-urine distribution ratios were consistent with the pharmacokinetic characteristics of warfarin. Conclusion    The developed LC-MS/MS method features minimal sample volume, simple pretreatment, fast analysis, high sensitivity, and good accuracy, making it suitable for emergency toxicological screening. The detection of high warfarin concentrations in both maternal and neonatal serum, alongside clinical symptoms and coagulation tests, confirmed warfarin poisoning. This method proved effective for rapid diagnosis and therapeutic monitoring, providing reliable technical support for clinical management and emergency responses to anticoagulant rodenticide poisoning.

Abstract: Objective    Arylalkylamine N-acetyltransferase (aaNAT) plays a critical role in regulating various physiological processes in insects, including cuticle sclerotization, limb pigmentation, and oviposition. In Aedes aegypti, the aaNAT gene family has expanded into 13 members, categorized into three subfamilies: canonical insect-type, polyamine NAT-like, and mosquito-specific. Among these, the functional roles of the "polyamine NAT-like" subfamily remain insufficiently understood. Given that polyamines are essential molecules involved in core life processes such as insect development and reproduction, this study focuses on the uncharacterized member aaNAT3 within this subfamily, aiming to elucidate its biochemical properties and spatiotemporal expression patterns, thereby providing insights into the functional roles of this subfamily. Methods    The soluble recombinant aaNAT3 protein was obtained through prokaryotic expression, and an in vitro enzymatic activity assay system was established to screen for substrates. Molecular docking of aaNAT3 with active substrates was performed using AutoDock Vina 1.5.7 software. Quantitative real-time PCR (qPCR) was employed to analyze the expression profile of aaNAT3 across different developmental stages and in various tissues of male and female adult mosquitoes. Results    In vitro substrate screening demonstrated that aaNAT3 could not catalyze traditional arylalkylamines such as dopamine, 5-hydroxytryptamine, or L-DOPA. However, it exhibited catalytic activity toward cadaverine, spermine, spermidine, and C₉–C₁₂ long-chain alkyldiamines, including 1,9-diaminononane, 1,10-diaminodecane, 1,11-diaminoundecane, and 1,12-diaminododecane. Molecular docking results indicated that aaNAT3 exhibits high binding affinity for these active substrates, with binding energies ranging from -4.7 to -5.0 kcal/mol, consistent with the in vitro enzymatic activity data. qPCR analysis revealed that aaNAT3 expression was highest overall during the pupal stage and exhibited sex-specific expression patterns in adults: primarily expressed in the cuticle of males and predominantly enriched in the hind legs and wings of females. Conclusion    aaNAT3 is a polyamine-preferring acetyltransferase in Aedes aegypti with a unique substrate spectrum. Its high expression during the pupal stage and sex-specific expression pattern in adults suggest that this enzyme may play a key regulatory role in mosquito pupal development and sex-specific physiological functions of adults, providing new molecular targets and theoretical basis for novel mosquito behavioral intervention strategies based on developmental stages and sex differences.

Abstract: Objective    To investigate the impact of gene silencing of Peptidoglycan recognition proteins (PGRP) in Aedes aegypti larvae on their survival rate following infection with the entomopathogenic fungus Beauveria bassiana, using RNA interference (RNAi) technology. Additionally, the role of PGRP in the innate immunity of Aedes aegypti was explored. Methods    Real-time quantitative PCR (RT-qPCR) was employed to analyze the expression levels of PGRP-LE and PGRP-S1 genes in Aedes aegypti larvae after fungal infection. The relative expression levels of these genes at different developmental stages were also measured. Double-stranded RNA (dsRNA) was used to silence PGRP-LE and PGRP-S1 in the larvae, after which the larvae were challenged with Beauveria bassiana to evaluate the effects of PGRP-LE and PGRP-S1 silencing on mosquito survival. Results    After RNAi-mediated silencing of PGRP-LE and PGRP-S1 in Aedes aegypti, a significant enhancement in the pathogenicity of Beauveria bassiana was observed. The survival rates of the gene-silenced groups dropped sharply to 2.4% and 0%, respectively, compared with 16.67% in the dsGUS control group, with a significant difference between the groups (P<0.05). Conclusion    This study successfully silenced the expression of PGRP-LE and PGRP-S1 genes in Aedes aegypti through RNAi, effectively impairing the mosquito's immune defense. As a result, the biocontrol efficacy of Beauveria bassiana was significantly increased, achieving a markedly lethal effect on Aedes aegypti larvae.

Abstract: Objective   To analyze the characteristics of viral load variations and their influencing factors among patients during the chikungunya fever outbreak in Guangdong Province in 2025, providing a scientific basis for precise assessment and the formulation of intervention and management measures. Methods  Data of Chikungunya fever cases in Guangdong Province from July 18 to November 5, 2025, were extracted from the China Disease Control and Prevention Information System and the Guangdong  Provincial Field Epidemiological Investigation System. The cycle threshold (Ct value) obtained from real-time fluorescence quantitative RT-PCR testing of chikungunya virus nucleic acid in case samples was analyzed to examine its variation characteristics over the course of the disease and the influencing factors. Results   A total of 4 409 patients were included in the analysis, with an average age of 39.41±19.98 years and an average Ct value of 25.06±5.52. The distribution of Ct values showed an inverted U-shaped pattern across different age groups. In the multivariate analysis, elderly individuals (≥65 years) had lower Ct values than those aged 18–64 years, and the difference was statistically significant (P<0.001). Children (<6 years) also had lower Ct values compared with the 18–64 age group, but the difference was not statistically significant (P=0.093). Ct values gradually increased over time after disease onset, with a more gradual increasing trend in elderly and pediatric populations. Multivariate regression analysis indicated that elderly patients (≥65 years) had lower Ct values (β = -3.21, 95% CI: -3.68 to -2.73), while those from the eastern Guangdong (β = 0.82, 95% CI: 0.37 to 1.28) and those with a longer interval from disease onset to detection (β = 1.14, 95% CI: 1.05 to 1.24) had higher Ct values. After adjusting for basic characteristics, fever (β = -3.06, 95% CI: -4.09 to -2.04) and arthralgia (β = -2.64, 95% CI: -3.60 to -1.69) were associated with lower Ct values, while rash (β = 1.60, 95% CI: 0.78 to 2.41) was associated with higher Ct values. Conclusion   The viral load of Chikungunya patients in Guangdong Province in 2025 varied across different disease courses, age groups, and symptomatic populations, which can be integrated into infection risk assessments. Special attention should be given to elderly and pediatric patients, and timely intervention should be provided for patients presenting with fever and arthralgia.

Abstract: Objective    To evaluate the diagnostic value of nanopore targeted next-generation sequencing (tNGS) of tissue specimens combined with pathological testing for paucibacillary tuberculosis. Methods    A total of 81 patients with suspected tuberculosis were enrolled. Using the final definitive diagnosis as the standard, the positive rates of tNGS, pathology, GeneXpert, TB-DNA, Mycobacterium tuberculosis culture, and acid-fast staining were compared; the diagnostic efficacies of tNGS, histopathology, and the combined detection were evaluated. Results    Among the 81 cases, 67 were ultimately confirmed as active tuberculosis. Based on definitive diagnosis and regarding two categories of tuberculosis pathological diagnosis as positive indicators, the positive rates of tNGS, pathology, GeneXpert, TB-DNA, Mycobacterium tuberculosis culture, and acid-fast staining were 85.07%, 70.15%, 36.67%, 21.88%, 8.89%, and 7.14%, respectively. The differences between tNGS and other methods were statistically significant (P<0.05). The combination of tNGS and pathology showed a higher diagnostic value compared to individual methods, with an area under the curve of 0.899, sensitivity of 94.03%, specificity of 85.71%, and Kappa value of 0.755. An increase in tNGS read count was significantly associated with a higher proportion of tuberculosis diagnoses, and the final diagnostic composition varied significantly across different read count ranges (χ⊃2;=22.352, P=0.0004). As the number of detected Mycobacterium tuberculosis sequences increased in the pathological tissues, the proportions of necrotic lesions and hyperplasia gradually rose; necrotic changes were most prominent in the group with sequence reads ≥10 000 reads, showing statistically significant differences (χ⊃2;=15.338, P=0.009 0). Conclusion    When detecting Mycobacterium tuberculosis in tissue lesions, tNGS demonstrates higher sensitivity than traditional methods. The combination of tNGS and pathology exhibits superior diagnostic efficacy. The read count of tNGS serves as a promising trend indicator for supporting tuberculosis diagnosis.

Abstract: Objective    To establish a universal real-time reverse transcription polymerase chain reaction (RT-PCR) method for rapid and specific detection of all lineages of the Chikungunya virus (CHIKV). Methods    Specific primers and  TaqMan probes were designed and synthesized targeting the major CHIKV lineages, including the Asian genotype, the East-Central-South African (ECSA) genotype—Central African clade, the ECSA genotype—Indian Ocean lineage, and the West African genotype. A real-time RT-PCR detection system was subsequently established. The specificity, repeatability, reproducibility, and limit of detection (LOD) of the method were systematically evaluated and validated. Finally, the developed assay was applied to clinical samples for validation. Results    A pan-lineage real-time RT-PCR assay for CHIKV detection was successfully developed. The sensitivity of the assay reached 200 copies/mL for all three major CHIKV lineages. The coefficients of variation (CVs) for repeatability were 1.51%, 0.95%, 0.97%, and 0.69%, and those for reproducibility were 0.76%, 1.57%, 1.07%, and 1.08%, demonstrating excellent repeatability and reproducibility. Specificity testing showed no cross-reactivity with dengue virus (serotypes 1–4), Zika virus, or yellow fever virus genes, confirming good specificity. In a prospective clinical evaluation of 1 740 samples, the assay correctly identified all 24 CHIKV-positive samples, with a detection rate of 100% (24/24) and Ct values ranging from 18 to 35. Notably, no false positives were detected in the remaining 1 716 CHIKV-negative samples, which comprised a challenging panel of 64 dengue virus-positive cases, 76 malaria-positive cases, and 1 576 other negative controls. This demonstrates the method's excellent clinical sensitivity and specificity. Conclusion   The real-time RT-PCR method established in this study enables rapid, sensitive, and specific detection of CHIKV, and is suitable for early clinical diagnosis of CHIKV infection.

Abstract: Objective    To investigate the diversity of species, distribution characteristics, and genetic evolutionary relationships of mosquitoes in Guizhou Province, a mosquito population survey and phylogenetic analysis were conducted across multiple regions, aiming to provide a scientific basis for the prevention and control of mosquito-borne infectious diseases in the area. Methods    Female mosquitoes were collected using methods such as ultraviolet light traps, BG-Trap CO₂ mosquito traps, and the human landing catch method. Specimens were morphologically identified, and representative samples, as well as individuals difficult to identify, were selected for tissue DNA extraction. A fragment of the mitochondrial cytochrome c oxidase subunit I (COI) gene was amplified by PCR and sequenced. Multiple sequence alignment was performed using BioEdit 7.0 software, and a phylogenetic tree was constructed using MEGA 12.0 software to analyze the genetic relationships and geographical origins of different mosquito species. Results    A total of 20 739 female mosquitoes were collected, primarily from cattle sheds (12 980) and pigsties (6 785). Combined morphological and molecular identification revealed 10 species belonging to 4 genera. Armigeres subalbatus (43.7%), Anopheles sinensis (29.4%), and Culex tritaeniorhynchus (21.7%) were the dominant species across the province. Geographically, Anopheles sinensis, Culex quinquefasciatus, and Armigeres subalbatus were widespread throughout the province, while Culex tritaeniorhynchus was distributed in all regions except Liupanshui. The remaining species (For example, Lutzia vorax and Culex bhutanensis were collected in Anshun City, while Anopheles minimus and Culex murrelli were collected in Qiannan Prefecture. Aedes vexans npponii was recorded in both Anshun City and Qiannan Prefecture.) were only collected in specific areas. Phylogenetic analysis indicated that the COI sequences of Anopheles sinensis, Armigeres subalbatus, Aedes albopictus, Culex quinquefasciatus, and Culex murrelli clustered with reference strains from China, suggesting that these species have formed stable populations in Guizhou Province and other domestic regions. Lutzia vorax and Anopheles minimus not only clustered with domestic strains but also fell within the same clade as foreign strains. In contrast, Aedes vexans npponii and Culex bhutanensis clustered with geographical strains from countries such as Japan, South Korea, Vietnam, and Bhutan, respectively, indicating their presence and formation of small local populations, which may suggest a risk of introduction from abroad. Conclusion    Mosquito species in Guizhou Province are relatively diverse and widely distributed, with clearly defined dominant species. Lutzia vorax is a newly recorded species in Guizhou Province, and Culex bhutanensis is a newly recorded species in China. The potential introduction of certain species from abroad may increase the transmission risk of mosquito-borne diseases.

Abstract: The Expert Consensus on the Construction Standards of Heat Stroke Medical Rescue Systems (2025) represents the first guiding document in China to systematically construct a full-process rescue system for heat stroke. In the context of increasing risks of heat stroke under global warming, this consensus aims to “reduce incidence rate, improve treatment success rates, and enhance long-term prognosis” by proposing an integrated prevention and control strategy covering “prevention-pre-hospital emergency care-in-hospital treatment-rehabilitation-chronic disease management.” Its core components focus on four key dimensions: promoting the shift of the prevention threshold forward to "heat not causing illness"; strengthening pre-hospital emergency care with "golden half-hour" rapid cooling; establishing a multidisciplinary in-hospital treatment system; and incorporating rehabilitation and chronic disease management into the standardized pathway for the first time. The consensus further clarifies standardized construction requirements, including a four-tier rescue network structure, the setup of dedicated rescue units, and the establishment of heat tolerance testing laboratories, with the goal of achieving systematic, standardized, and homogeneous heat stroke prevention and treatment. The implementation of this consensus provides critical guidance for addressing health threats posed by extreme climate events and enhancing national public health emergency response capabilities.

Abstract: Objective    To obtain near-full-length HIV sequences using near-full-length gene amplification combined with next-generation sequencing, and to identify their recombination forms. Methods    From two newly reported HIV-1 infected individuals in Liaoning Province in 2023 harboring potential URFs, near-full-length viral genomes were amplified from target samples using semi-genomic amplification technology. Near-full-length HIV genome sequences were acquired by next-generation sequencing (NGS), which underwent recombination analysis using SimPlot and BootScan. Recombination fragments were validated through segmented phylogenetic trees. Results    Two full-length sequences were ultimately obtained with lengths of 9 261 bp and 9 073 bp, covering all HIV structural genes. Phylogenetic and recombination analyses revealed that both sequences (LNJZ230022 and LNDL230042) possessed an HIV-1 subtype B backbone. LNJZ230022 sequence incorporated CRF07_BC and CRF01_AE gene fragments, with recombination occurring in gag and env regions and breakpoints at positions 803, 1 147, 2 120, 6 323, 7 758, and 9 274 (HXB2). LNDL230042 was inserted by CRF07_BC gene fragments, with recombination regions in vpr and env and breakpoints at 779, 5 608, 5 870, 6 745, 7 541, and 9073 (HXB2). Sample LNDL230042 exhibited an N88T mutation at position 88 in the PR region, classified as moderate resistance, and an F53L mutation at position 53, indicating low-level resistance. Sample LNJZ230022 exhibited no resistance mutations. Conclusion    The two novel HIV-1 URFs identified in newly infected individuals in Liaoning Province reflect the complex co-circulation of subtype B, CRF07_BC, and CRF01_AE strains. By employing NGS to assemble full-length HIV genomes, we successfully identified two URFs from HIV-infected individuals in Liaoning and conducted detailed characterization of their recombination patterns. This approach captured large-scale recombination events in the env gp120 region, providing a critical molecular foundation for understanding recombination and immune escape in locally circulating strains. This study not only expands our understanding of HIV-1 genetic diversity and recombination trends in Liaoning Province but also demonstrates the feasibility and importance of using NGS to obtain near-full-length sequences for precise monitoring of novel circulating recombinant HIV-1 strains.

Chikungunya fever, as an important arthropod-borne infectious disease, is intrinsically linked to cross-border transmission and population mobility. This article systematically reviews research progress in China on the association between overseas labor export and the risk of chikungunya virus importation. It is found that China's overseas labor export shows a significant regional concentration feature, with Asia (especially Southeast Asia) and Africa being the main destinations, jointly accounting for approximately 80% of the total. Concurrently, Guangdong Province, as the main entry point for returning laborers, shows high concentration of laborers returning from Asia and Africa. Based on the global chikungunya fever epidemic data from 2020 to 2024 and traceability results from a 2025 outbreak in Guangdong Province, it is suggested that the return of overseas laborers is a key route for virus importation. Due to high exposure risk, delayed detection during entry quarantine, and delayed medical treatment upon returning home, laborers from chikungunya fever-endemic areas such as Southeast Asia and Africa have become a critical vulnerable link in the importation and transmission chain of arthropod-borne diseases in China, especially in Guangdong Province. In the future, it is necessary to strengthen multi-departmental collaboration and multi-source data integration, enhance the quarantine and epidemiological research on overseas laborers at ports of entry, and establish an arthropod-borne infectious disease importation risk assessment system based on the domestic and overseas mobility patterns, thereby providing scientific support for the effective prevention and control of local outbreaks of arthropod-borne infectious diseases.

In recent years, the increasing frequency of extreme weather events worldwide has driven a sustained rise in the incidence of heatstroke. As a fatal emergency that can lead to multiple organ failure, heatstroke 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 heatstroke. This article systematically reviews the bidirectional interaction between heatstroke 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 heatstroke can disrupt the integrity of the intestinal barrier, leading to microbial ecological dysbiosis. On the other hand, the dysregulated gut microbiota further amplifies heatstroke-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 heatstroke.

Abstract：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 and preliminarily validate the interaction between the regulatory receptor IR7a, which is involved in the blood-feeding behavior of Aedes aegypti, and two co-receptors, IR25a and IR76b, within the Ionotropic Receptors (IRs) family. Furthermore, this study aims to preliminarily purify the IR7a co-receptor complex protein for further structural characterization. Methods  Spatiotemporal gene expression profiles of AaegIR7a, AaegIR25a, and AaegIR76b were investigated using real-time quantitative PCR (qPCR). Recombinant plasmids carrying different tags were constructed for IR7a and the two co-receptors, IR25a and IR76b. The IR7a-IR25a and IR7a-IR76b complexes were co-expressed using a mammalian expression system. The target proteins were purified via affinity chromatography and gel filtration chromatography, and their interactions were detected by Western Blot (WB). Finally, negative-stain electron microscopy was used to preliminarily characterize the structures of the two complexes. Results  AaegIR7a, AaegIR25a, and AaegIR76b showed high expression levels in male Aedes aegypti, and their expression decreased in female mosquitoes after blood feeding. All three genes were highly expressed in key sensory tissues, including the proboscis, antennae, and appendages, with similar expression patterns observed for the co-receptor genes AaegIR25a and AaegIR76b. Size-exclusion chromatography demonstrated protein-protein interactions between in vitro co-expressed IR7a and IR25a, as well as between IR7a and IR76b. Negative-stain electron microscopy revealed that both the IR7a-IR25a and IR7a-IR76b complexes possess structural features characteristic of ionotropic receptors. Conclusion  The specific expression of AaegIR7a, AaegIR25a, and AaegIR76b in key developmental stages and tissues associated with blood-feeding behavior suggests that IR7a may function by forming an ionotropic receptor complex with the IR25a and IR76b co-receptors to regulate the preference between nectar and blood feeding. Their high expression in sensory organs, such as the proboscis and antennae, indicates a potential for synergistic action. In vitro protein interaction studies confirmed specific interactions between IR7a and IR25a/IR76b, with preliminary observations indicating structural similarities to ionotropic glutamate receptors. This work establishes a methodological foundation for further structural elucidation of the complex and provides initial clues for deciphering the molecular mechanisms underlying host and blood recognition in Ae. aegypti.

 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 protective effect of butyrate preconditioning on intestinal barrier injury in mice with classical heatstroke (CHS) and explore its potential mechanism based on transcriptomic analysis. Methods    C57BL/6J mice were randomly divided into four groups: control group (CON), butyrate group (NaB), heatstroke group (HS), and butyrate preconditioning + heatstroke 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 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 analyze the molecular genetic characteristics of Japanese Encephalitis Virus (JEV) isolated from Anopheles sinensis in Xingren City, Guizhou Province, in 2023. 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 RTPCR and sequencing, followed by genetic evolution, amino acid variation, and protein structure analyses using MEGA, DNAStar, and SWISSMODEL. 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 SA14142, 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 virulencerelated 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 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°C, 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 evaluate the clinical value of the Heatstroke Severity Score System (HSSS) in assessing disease severity and predicting prognosis in patients with heatstroke (HS). Methods: A retrospective analysis was conducted on the clinical data of 143 patients with heatstroke. Patients were stratified into mild (score 1-10, n=110), moderate (score 11-20, n=23), and severe (score ≥21, n=10) groups 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 counts, coagulation function, liver and kidney function, lactate levels, and mortality rates, were compared among groups. Spearman’s rank correlation test was used to analyze the relationship between HSSS and SOFA/APACHE II scores. Receiver Operating Characteristic (ROC) curve analysis was performed to evaluate the predictive efficacy of HSSS for mortality, calculating the Area Under the Curve (AUC) and optimal cutoff value. Survival curves were generated using the Kaplan-Meier method, and a Cox proportional hazards model was employed to identify independent risk factors for mortality in heatstroke patients. Age, SOFA score, APACHE II score, and HSSS score were incorporated into the multivariate Cox regression model to calculate hazard ratios (HRs) and their 95% confidence intervals (CIs). Additionally, the distribution patterns of combined organ system injuries were characterized. Results: The mortality rate in the severe group was 90.0%, significantly higher than in the mild and moderate groups (both 0.0%, P < 0.001). Analysis of organ injury patterns revealed that three-system involvement was the most common (40 cases, 28.0%), with the combination of thermoregulatory dysfunction (hyperthermia), central nervous system (CNS) dysfunction, and renal impairment being the most frequent. This was followed by four-system involvement (36 cases, 25.2%), predominantly involving hyperthermia, CNS dysfunction, renal impairment, and hepatic injury. As HSSS scores increased, APACHE II scores, SOFA scores, and lactate levels significantly increased, while GCS scores significantly decreased; the severity of coagulation disorders and liver/kidney injury also markedly worsened (P < 0.05). Correlation analysis showed a strong positive correlation between HSSS and SOFA (r = 0.942, P < 0.001) and APACHE II scores (r = 0.816, P < 0.001). ROC analysis demonstrated that the AUC of HSSS... was 0.986... The AUC of HSSS was significantly higher than that of APACHE II (Z=2.384, P=0.017) and comparable to SOFA (P=0.093). at an optimal cutoff of 20 points, sensitivity was 100% 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 < 0.0001). Conclusion: The HSSS can accurately assess disease severity and reliably predict adverse outcomes in patients with heatstroke.