Objective To explore the application potential of a simplified liquid chromatography-tandem mass spectrometry (LC-MS/MS) protocol for simultaneous identification and flagellar antigen typing of Escherichia coli. Methods A simplified LC-MS/MS workflow was established using a whole-genome-based proteomic bacterial identification database and a curated database containing all E. coli flagellar antigen proteins. This protocol was applied to analyze 8 reference strains and 54 clinical isolates of E. coli for concurrent identification and flagellar antigen typing. Additionally, the correlation between sample dilution factors and the number of flagellar antigen peptides detected was evaluated using three E. coli reference strains. Results The simplified LC-MS/MS protocol achieved 100% accuracy in both identification and flagellar antigen typing for all 62 E. coli strains. For the three reference strains, a strong linear relationship was observed between sample dilution factors and the number of flagellar antigen peptides detected, with correlation coefficients of 0.99, 0.97, and 0.98, respectively. Conclusion The simplified LC-MS/MS protocol enables accurate and simultaneous identification and flagellar antigen typing of E. coli. This study highlights its significant advantages in dual-analysis applications of simultaneous identification and flagellar antigen typing of E. coli, demonstrating promising prospects for microbial identification, typing, and microbiome research. LC-MS/MS is expected to become a vital tool in clinical microbiology and public health surveillance.

Objective To analyze clinical and microbiological characteristics of two cases with erysipeloid septicemia admitted to the People's Hospital of Ledong Li Autonomous County, Hainan Province, and to provide reference basis for diagnosis and treatment of erysipeloid septicemia. Methods A retrospective analysis was conducted on clinical characteristics and treatment outcomes of 2 cases with confirmed erysipeloid septicemia by blood culture who were admitted to Le Dong Li Ethnic Autonomous County People's Hospital from December 2023 to January 2024. The pathogenic bacteria were cultured using automated blood culture system and solid culture medium, and identified by MALDI-TOF MS and 16S rDNA. The sensitivity of the pathogenic bacteria to antibiotics was detected by microdilution of broth method. Results Two patients were onset in winter, both had a history of trauma infection from contact with pigs or fish. They presented with high fever, chills, chest tightness and shortness of breath. They were admitted to the hospital with the diagnosis of "fever to be investigated". Laboratory tests showed increased inflammatory indicators; Both patients' blood and wound secretions samples yielded bacterial growth. After blood culture turned positive, smear examination showed Gram-positive bacilli arranged in long filaments or short chains. After cultured on Columbia blood agar at 35 ℃ for 24 hours, the colonies were pinpoint-shaped, and became larger, round, moist, transparent, raised with neat edges and gray white after 48 hours, and a narrow hemolytic ring was formed around the colony. The gram staining of the colony smear was negative. The strains were identified as Erysipelothrix rhusiopathiae by MALDI-TOF MS and 16S rDNA sequencing; Both isolates from the patients were sensitive to penicillin, ampicillin, ceftriaxone, cefepime, meropenem, erythromycin, clarithromycin and levofloxacin. The case 1 had underlying malignant tumor, when the patient visited hospital, the infection in the surrounding tissues was already quite severe. After using amoxicillin and clavulanate potassium for 3 days, the treatment failed. The case 2 was treated with ampicillin for 8 days and achieved good results. Conclusion The clinical features of erysipeloid are atypical and easily misdiagnosed. For individuals engaged in occupations related to pigs or fish, and those who develop high fever after trauma, relevant specimens should be promptly collected for pathogen testing, and antibiotics should be used according to antimicrobial susceptibility result to improve prognosis.

Objective To explore diagnostic efficacy of individual and combined detection of mannoprotein (Mp1p), galactomannan (GM) and 1-3-β-D glucan (BDG) in the detection of patients with acquired immunodeficiency syndrome (AIDS) with Talaromycosis marneffei (TSM). Methods Peripheral blood samples were collected from 291 AIDS patients with TSM admitted to our hospital and 300 objects in health examination．The diagnostic value of Mp1p, GM and BDG individually and in combination, was analyzed for detecting TSM. The diagnostic efficacy of Mp1p, GM, BDG and their combined detection was analyzed by ROC curve. Results In individual detection, there were statistical differences in the sensitivity and specificity of Mp1p, GM and BDG (P<0.05). Compared with GM and BDG, Mp1p had the best diagnostic efficiency with the best sensitivity and specificity. In pairwise combined detection, the sensitivity of combination of Mp1p and GM was better than that of GM and BDG, and the difference was statistically significant (P<0.05); the specificity of combination of Mp1p and GM and combination of Mp1p and BDG was better than that of GM and BDG, the difference was statistically significant (P<0.05), the diagnostic efficacy of combination of Mp1p and GM was the best. The sensitivity of the tripartite combined detection was better than that of pairwise combined detection, the difference was statistically significant (P<0.05). The specificity of combination of Mp1p and GM, combination of Mp1p and BDG was better than the tripartite combined detection, the difference was statistically significant (P<0.05). There was no significant difference in the positive rate of Mp1p, GM and BDG tests in patients with CD4⁺T cell count ≤100/μL and >100/μL, respectively (P>0.05). Conclusion Mp1p, GM and BDG tests are early auxiliary diagnostic indicators of AIDS combined with TSM. The diagnostic efficiency of Mp1p was better than that of GM and BDG, and combined detection could improve the diagnostic efficiency.

Objective To establish a rapid, sensitive, and efficient loop-mediated isothermal amplification (LAMP) detection method for metacercariae of Paragonimus westermani in freshwater crabs. Methods From September 2022 to December 2023, freshwater crabs were collected in Xiuning County, Anhui Province, an endemic area of Paragonimus westermani. Specific LAMP primers were designed based on the sequence of the second internal transcribed spacer (ITS2) region of P. westermani ribosomal DNA. A novel chromogenic agent, hydroxynaphthol blue (HNB), was selected and compared with the traditional calcein chromogenic agent to establish the LAMP detection method for P. westermani metacercariae. Additionally, the specificity, sensitivity, and field application effectiveness of this method were evaluated by comparing it with the PCR method. Results The established LAMP reaction system could specifically amplify the DNA of metacercariae and adult of P. westermani, with a detection limit as low as that equivalent to a single metacercariae of P. westermani. No cross-reactions were observed with Schistosoma japonicum, Toxoplasma gondii, Clonorchis sinensis, Ancylostoma duodenale and Necator americanus. The minimum nucleic acid concentration detection limit of the HNB chromogenic reagent method and its LAMP real-time turbidity meter method was 1.92×10^-13 g, while that of the calcein chromogenic reagent method, its LAMP real-time turbidity meter method and the PCR method was 1.92×10^-12 g. Among 40 field-collected freshwater crabs tested by sedimentation microscopy, LAMP (HNB, calcein), and PCR, the positive rates were 22.5% (9/40), 27.5% (11/40), and 27.5% (11/40), respectively. Among them, two microscopy-negative samples tested positive by both LAMP and PCR. There was no statistically significant difference between the results of the three methods(χ²=0.348, P>0.05). The Kappa value of the consistency test between HNB and calcein was 1. Conclusion In this study, a LAMP method for detecting P. westermani in freshwater crabs was successfully established, which provided a simple and efficient molecular detection method for the surveillance of paragonimiasis in freshwater crabs in endemic areas.

Objective To understand the current status of diagnostic delay in elderly pulmonary tuberculosis (PTB) patients in He'nan Province, analyze the factors affecting the PTB detection rates and diagnosis time, and provide a basis for formulating TB prevention and control measures. Methods Information on suspected and clinically diagnosed cases of PTB in individuals aged 65 years or older reported by non-TB prevention institutions in He'nan Province from 2016 to 2023 was collected through the "Tuberculosis Information Management System (TBIMS)", a subsystem of the "China Information System for Disease Control and Prevention". Epidemiological retrospective analysis was conducted to organize the demographic and medical data of the research subjects. The detection rate, diagnosis time, and diagnostic delay rate of PTB were calculated. Univariate log-rank test and multivariate Cox proportional hazards regression model were used to analyze the factors influencing TB diagnosis time and detection rates. Results Among the 105 458 elderly suspected and clinically diagnosed cases in He'nan Province from 2016 to 2023, 24 846 were ultimately diagnosed with PTB, with a detection rate of 23.56%. The median diagnostic time was 68 days (interquartile range: 27, 184 days), with diagnostic delay rates of 53.57% and 35.34% at 14 days and 28 days, respectively. Cox regression analysis results showed that "correction card" (HR=0.059, 95.0% CI: 0.057-0.062, M=75 d), "suspected case" (HR=0.218, 95.0% CI: 0.212-0.225, M=113 d), and "primary medical institution as the first visit unit" (HR=0.605, 95.0% CI: 0.572-0.639, M=158 d) were the top three independent influencing factors for the diagnosis time and detection rates for elderly PTB. Conclusion Diagnostic delay among elderly PTB patients in He'nan Province is serious. There is an urgent need to develop more convenient and effective diagnosis and referral processes tailored to the elderly population, improve the diagnosis capabilities of the first diagnosis institution, strengthen health education for the public, and carry out active screening relying on the basic public health services project. These measures aim to achieve early diagnosis and treatment, reduce diagnosis delay, and control the spread of TB.

Objective To evaluate the application value of targeted next-generation sequencing (tNGS) in the differential diagnosis of Mycobacterium tuberculosis complex (MTBC) and non-tuberculous mycobacteria (NTM) infections. Methods In this retrospective study, 506 samples, including sputum, bronchoalveolar lavage fluid (BALF), and other types, were collected from 466 hospitalized patients with suspected mycobacterial infection admitted to Guangzhou Chest Hospital between August 2023 and January 2025. MTBC and NTM were differentially diagnosed using tNGS and traditional diagnostic methods (smear microscopy, mycobacterial culture with species identification, and molecular biology testing), and the detection efficacy of tNGS was evaluated. Results The detection rate of MTBC using the tNGS method was 30.27%, higher than the 26.99% achieved by culture identification/molecular biology, although the difference was not statistically significant （χ²=1.28，P>0.05）. The detection rate for NTM using tNGS was 28.83%. The detection rates of MTBC through tNGS in sputum, BALF, and other samples, were 37.50%, 28.07%, and 36.84%, respectively, all consistently higher than those obtained by culture identification/molecular biology methods (35.42%, 25.13%, and 21.05%, respectively), but the differences did not achieve statistical significance （χ²=0.09，0.83，1.15，P>0.05）. Additionally, among the 148 patients identified as MTBC-positive using tNGS, 47.97% were confirmed as MTBC by mycobacterial culture identification, 4.06% as NTM, and 47.97% were culture-negative. The tNGS sequence reads in culture-positive MTBC samples were significantly higher than those in the culture-negative group（Z=-3.05，P<0.05）. The tNGS sequence reads in the smear-positive and molecular biology-positive groups were also significantly higher than those in the corresponding negative groups（Z=-4.99，-3.19，P<0.05）. Using culture identification/molecular biology as the reference standard, the sensitivity, specificity, and Kappa value of tNGS for detecting MTBC were 84.85%, 89.92%, and 0.72, respectively. Conclusion The tNGS technology demonstrates high sensitivity and rapidity for MTBC detection, providing effective support for early differential diagnosis of tuberculosis.

Objective To investigate epidemiological and pathogenic characteristics of Salmonella strains responsible for foodborne outbreaks, and to provide scientific evidence for the control and prevention of food-borne diseases. Methods Surveillance data of foodborne outbreak events and 66 Salmonella isolates, were collected in Changsha from 2019 to 2023. Serotypes were determined using slide agglutination tests. Antibiotic susceptibility to 17 antibiotics was tested by the microdilution broth method. Molecular typing was performed by pulsed-field gel electrophoresis (PFGE). Multi-locus sequence typing (MLST), antibiotic resistance genes, and cgMLST were analyzed by whole genome sequencing and generating phylogenetic tree to assess genetic relatedness. Results From 2019 to 2023, a total of 13 Salmonella-related foodborne outbreaks occurred in Changsha, the majority of outbreaks occurred in catering sector (69.23%), including central kitchens, large hotels and medium and small-sized restaurants. Baked foods and egg products were the primary contaminated foods, accounting for 63.64% and 20.32% of the total outbreaks, respectively. The 66 Salmonella isolates were classified into 4 serotypes, with Salmonella enteritidis being the predominant serotype，and ST11 were the predominant sequence type. In the antibiotic susceptibility test, the strains showed high resistance rates to streptomycin (100.00%), nalidixic acid (69.70%), and ampicillin (54.55%). Using the resFinder database, 6 antibiotic resistance genes from 4 categories were predicted, and the resistance phenotypes roughly matched the identified resistance genes. PFGE typing showed 100% similarity between strains isolated from food and patients in the same outbreak. In the cgMLST phylogenetic tree, strains from the same outbreak clustered separately, indicating genomic differences and distinct origins. Conclusion Salmonella enteritidis ST11 is a common pathogen responsible for foodborne outbreaks in Changsha. These strains exhibit multi-drug resistance and carry various resistance genes. Strengthening supervision of central kitchens and the food service industry, particularly for baked goods and egg products, is essential for controlling outbreaks.

Mosquito-borne diseases, including Japanese encephalitis, malaria, dengue fever, yellow fever, and other harmful infectious diseases, are natural focal diseases transmitted by vector female mosquitoes. These diseases pose significant global public health challenges. With climate change and the rise in international exchanges, the incidence of imported mosquito-borne diseases has occasionally surged in recent years in China. Consequently, on-site rapid detection technologies are urgently needed to control the epidemic in its early stages. The emerging point-of-care testing (POCT) technology allows for the rapid acquisition of test results directly at the sampling site using portable analytical instruments and supporting reagents. This approach facilitates the rapid, efficient, and accurate diagnosis of pathogens. Currently, POCT has been applied in multiple fields such as food safety, port quarantine, and forensic inspection, and it demonstrates considerable potential for application in the early warning of mosquito-borne disease outbreaks. In recent years, research on POCT technology for mosquito-borne diseases has significantly expanded, and this paper reviews the basic concepts of POCT technology and the research progress in mosquito-borne diseases, aiming to provide a reference for peers in the field to enhance the detection of mosquito-borne diseases.

Chikungunya fever is widely distributed in tropical and subtropical regions around the world. In the past 20 years, this disease been seriously epidemic in the Indian Ocean Region, South Asia and Southeast Asia, and has led to the occurrence of imported cases at least 16 provinces (municipalities and autonomous regions) in China. Among them, indigenous epidemics have occurred in Guangdong, Zhejiang and Yunnan provinces. Chikungunya virus (CHIKV) gene sequences have been detected in serum samples from fever of unknown origin cases and mosquito specimens collected from Yunnan Province, mosquitoes from Zhejiang Province and midges from Guangxi Zhuang Autonomous Region. The epidemic strains of CHIKV in China were mainly the Indian Ocean clade lineage of the East/Central/South African genotype, and the Asian genotype. Aedes albopictus is widely distributed in central, eastern and southern China, and Ae. aegypti is distributed in some parts of Hainan, Guangdong, Yunnan and Taiwan. These two kinds of Aedes mosquitoes in China are highly susceptibility and transmissibility to CHIKV. There is still a risk of outbreak or pandemic of chikungunya fever in the distribution areas of Ae. albopictus and Ae. aegypti in China. The monitoring and management of imported cases and the control of the vector Aedes mosquitoes is an important focus of prevention and control work that needs to strengthened at present. This article reviews the epidemiology and pathogenic biology on chikungunya fever in China, which can provide a reference for the prevention and control of this disease.

Objective To investigate the effect of immune response induced by mosquito saliva protein (MSP) in Aedes albopictus on dengue virus 2(DENV-2) infection in type I interferon receptor-deficient (IfnaR^-/- ) mice. Methods Female mosquitoes of Aedes albopictus (Guangzhou strain) were used to bite to immunize IfnaR^-/- mice once a week for four consecutive weeks, and observe their blood saturation, and immune sera were obtained three weeks after the last bite. And anti-MSP antibodies in mouse serum were detected by Western blot. The titer and subclasses of anti-MSP IgG were measured using an indirect ELISA. 10⁷ TCID₅₀/mL of DENV2 was microinjected into the mosquito thorax using a Nanoject Ⅱ automated nanoliter injector (Drummond). Real-time quantitative reverse transcription PCR (qRT-PCR) was used to detect DENV-E gene levels in mosquito salivary glands at different time points. IfnaR^-/- mice with/without pre-immunized were infected by DENV-2-infected female mosquitoes biting. Viral residues in mosquitoes were detected by qRT-PCR from immunized/unimmunized groups. Five days after infection by mosquito bite, DENV-E gene in different tissues of immunized/non-immunized mice were evaluated through qRT-PCR; Live viruses in mouse brain were detected by the virus plaque forming assay. Results Western blot revealed that mouse serum contained specific IgG antibodies of a variety of MSPs. Indirect ELISA assays showed anti-MSP IgG potency was up to 1∶800, predominantly IgG1 and maintained for 21 days. DENV-E gene levels in mosquito salivary glands reached a replication plateau 10 days after thoracic injection. The qRT-PCR results after the completion of the bite showed the amount of residual virus in the mosquito salivary glands, and it was hypothesized that the initial infections were similar in the two groups of mice. DENV-E gene in the spleen, liver and brain, as well as live virus in brain of the immunized mice were significantly lower than those of the non-immunized group (P<0.000 1, P<0.001 3). Conclusions The adaptive immunity induced by Aedes albopictus mosquito saliva might have the potential to inhibit DENV2 virus transmitted by mosquitoes in mice, providing a new pathway to explore the immunogen from mosquito saliva against arbovirus infection.

Objective To investigate the prevalence of Leptospira carriage in rodents in three Cities/Counties of Yunnan, and to detect the seroprevalence of Leptospira antibodies in dog serum. Methods From July to August 2019, natural villages were selected for field rodent trapping in Lianghe County, Mangshi City, and Mile City in Yunnan Province. Following morphological identification of the murine species, kidney tissue was collected for DNA extraction. Conventional PCR was employed to amplify the target gene fragment of the SecY gene of Leptospira. Sequencing was conducted on the positive samples, and the obtained sequences were subjected to homology alignment and phylogenetic tree construction. Femoral arterial blood was collected from dogs in the survey area, and serum samples were collected. Indirect enzyme-linked immunosorbent assay (ELISA) was used to detect IgG antibodies against Leptospira in dog serum. The chi-square (χ²) test was applied to analyze the serum Leptospira antibody status of dogs in relation to various influencing factors. Results A total of 490 rodents were captured. Leptospira carriage was detected through PCR in Rattus tanezumi and Rattus sladeni in Lianghe County, with a carrier rate of 0.41%. No case was detected in Mangshi City and Mile City. Homologous evolution analysis showed that both Leptospira strains were pathogenic Leptospira borgpetersenii. In addition, 305 dog serum samples were collected, among which 16 tested positive for Leptospira IgG antibodies, resulting in a positive rate of 5.25%. Conclusion Both rodents and dog serum in the three Cities/Counties of Yunnan Province were found to be infected with Leptospira, posing a potential risk of disease to humans. It is essential to enhance surveillance and implement preventive and control measures.

Objective In order to discover the characteristics of HCV spontaneous clearance in co-infected individuals with human immunodeficiency virus (HIV) and hepatitis C virus (HCV), identify the influencing factors of HCV spontaneous clearance, and compare the differences between HCV spontaneous clearance individuals and HCV-RNA positive patients, it is of clinical significance for guiding the screening and treatment of HCV-RNA in patients co-infected with HIV and HCV. Methods From January 2022 to May 2024, HIV/HCV co-infected patients who met the inclusion criteria in Yunnan Provincial Infectious Disease Hospital were selected as the research subjects. Untreated HCV-RNA negative patients were selected as the experimental group (clearance group), and untreated HCV-RNA positive patients were selected as the control group (positive group). The demographic, clinical characteristics, treatment, and laboratory testing indicators of patients in two groups were collected to calculate ability of HCV spontaneous clearance. Logistic regression analysis was used to explore influencing factors of HCV spontaneous clearance after HIV and HCV co-infection, and the laboratory indicators and liver function damage of clearance group and positive group were compared. Results A total of 773 HIV/HCV co-infected individuals were collected, including 70 cases in the clearance group and 78 cases in the positive group (excluding 3 cases with incomplete data). The spontaneous clearance rate of HCV was 9.1%(70/773). Univariate analysis showed statistical significance (P<0.05) for age, antiretroviral treatment regimen, and duration of antiviral treatment ≥ 16 years. Multivariate regression analysis showed that use of protease inhibitor regimens (OR=0.228, 95%CI: 0.064-0.810) was a factor that hindered the spontaneous clearance of HCV, while antiviral treatment duration ≥16 years (OR=8.587, 95%CI: 1.854-39.775) became a factor that promoted spontaneous clearance of HCV. Comparing laboratory indicators between positive group and clearance group, it was found that positive group had higher levels of ALT, AST, APRI score, FIB-4 index mean, and abnormality rate than the HCV spontaneous clearance group (P<0.001). Conclusions The spontaneous clearance rate of HCV in HCV co-infected HIV patients is reduced, and the use of ART regimen containing protease inhibitors and antiviral treatment for ≥ 16 years are independent influencing factors for HCV spontaneous clearance. HCV spontaneous clearance patients have a milder degree of liver function damage than HCV positive patients.

Objective To analyze the epidemiological characteristics of pulmonary tuberculosis in Wuhan from 2014 to 2023, and to provide a scientific basis for formulating and improving tuberculosis prevention and control planning strategies. Methods Descriptive statistical analysis was carried out on the information data of pulmonary tuberculosis (TB) cases registered and managed in Wuhan from 2014 to 2023 in China Disease Control and Prevention Information System, and the spatial, temporal, and demographic distribution characteristics of pulmonary tuberculosis patients were analyzed. Results From 2014 to 2023, a total of 58 131 active pulmonary tuberculosis patients were registered in Wuhan, with an annual registered incidence rate of 49.06/100 000 and an annual decline rate of only 4.70%. Meanwhile, the proportion of bacterial-positive pulmonary tuberculosis has been increasing year by year, with an annual growth rate of 4.66%. The male-to-female ratio for registered cases was 2.12∶1, with significantly higher incidence rates among males than females. The highest proportion of cases occurred in the 50 to <70 years (36.85%) and 20 to <40 years (31.75%) age groups, while the registered incidence rate had two peaks in the age groups of 17-29 years and 60-85 years. Annual peaks in TB registration occurred between April and June, while troughs were observed from December to February of the following year, showing a certain seasonality The patient's occupations mainly included domestic service, household chores, unemployed individuals, farmers, retirees, commercial service personnel, and students, accounting for 35.56%, 15.10%, 14.83%, 6.07% and 5.85% of the total number of patients, respectively. The registered incidence rate in rural districts was higher than that in urban districts, with the top five districts being Caidian District (71.52/100 000), Huangpi District (59.43/100 000), Xinzhou District (58.63/100 000), Qiaokou District (55.56/100 000), and Jiangxia District (55.23/100 000), recording a total of 15 318 cases, accounting for 26.35% of the city's cases. Local spatial autocorrelation analysis showed that the high-risk hotspots for tuberculosis in Wuhan were mainly distributed in Baibuting Community, Erqi Community, Yijiadun Community, Liujiaoting Community, Heping Street Community, Changqian Street Community, Xingou Town, Yuxian Town, Laoshan Town, Zhuzi Town, Jinkou Town, Wulijie Town, Wuhu Street, Panlong Economic Development Zone, as well as Fenghuang Town, Daoguanhe Street, and other places. Conclusion The decline rate of the tuberculosis epidemic in Wuhan over the past decade has not reached expectations, with relatively high incidence rates observed in economically backward regions and vulnerable groups with low living incomes. The health administrative departments should adopt localized prevention strategies and innovative technologies to strengthen active screening and latent infection interventions targeting high-risk areas and populations.

Objective To investigate effects of azithromycin on the pupation period of Ae. aegypti and Ae. albopictus, analyze changes in related metabolic pathways and differential expression of key genes, and preliminarily explore regulatory mechanism of azithromycin on the pupation cycle of these two mosquito species. Methods Multiple concentration gradients of azithromycin solution were used to treat both mosquito species. Pupation time, pupation rate and other growth indicators were regularly observed and recorded. Statistical methods including survival analysis and chi-square test were employed to analyze pupation differences. Transcriptome sequencing was performed on differentially treated and control samples, pathways and key genes associated with the pupation cycle of both mosquito species were validated by combine with RT-qPCR. Results Exposure to 19.600 μg/L azithromycin significantly shortened the pupation time of both mosquito species (P<0.01), while 0.196 μg/L and 1.960 μg/L azithromycin showed no significant effects (P>0.05). After treatment with 19.600 μg/L azithromycin, 695 differentially expressed genes (DEGs) were identified in Aedes aegypti; GO enrichment analysis revealed 32 functional categories, and KEGG enrichment showed alterations in pathways including protein processing in endoplasmic reticulum. In Ae. albopictus, 984 DEGs were identified, with GO enrichment showing 35 categories and KEGG enrichment indicating changes in oxidative phosphorylation and other pathways. RT-qPCR validation demonstrated that in Ae. aegypti, genes AAEL013350, AAEL020291, AAEL023117 and AAEL027089 in the endoplasmic reticulum protein processing pathway were upregulated in pupae, while AAEL023595 was downregulated. In Ae. albopictus,gene-LOC109404801, gene-LOC109428450 and gene-LOC115257348 in the oxidative phosphorylation pathway were upregulated in pupae. Conclusion This study confirms that 19.600 μg/L azithromycin can significantly shorten the pupation period of Ae. aegypti and Ae. albopictus, potentially through promoting metabolic pathways such as endoplasmic reticulum protein processing and oxidative phosphorylation, along with regulating the expression of related genes.

Objective To investigate density, distribution, population composition, and seasonal fluctuations of mosquito in different habitats in Nanchang city of Jiangxi province, and to provide a scientific basis for effective control and prevention of mosquitoes and mosquito-borne infectious diseases. Methods Mosquito-borne disease vector monitoring was carried out in habitats such as residential areas, parks, hospitals, farmers' households and livestock sheds in Nanchang city, 2018-2023 by using mosquito traps. Monitoring was conducted twice a month from March to November, once in the first ten days and once in the last ten days of each month. The monitoring results were then summarized and analyzed. The rank sum test for completely randomized design (Krusal-Wallis H) was adopted to compare the differences in mosquito density among different years, different months and different habitats. The chi-square test was used to compare the differences in the composition ratio of mosquitoes among different years. Results A total of 269 587 adult mosquitoes were captured, and the overall average density was 46.34 mosquitoes per lamp per night. The fluctuation of mosquito density showed a unimodal trend as a whole, with the peak in June. The average mosquito density showed a downward trend among different years, but the difference was not statistically significant (H=1.890, df=5, P=0.864). There were statistically significant differences in mosquito density among different months (H=47.448, df=8, P<0.001), and there were also statistically significant differences in mosquito density among different habitats (H=87.876, df=4, P<0.001). Culex pipiens quinquefasciatus was the dominant mosquito species in residential areas, parks and hospitals, while Culex tritaeniorhynchus was the dominant mosquito species in farmers' households and livestock sheds. The period from June to September each year was the peak period for adult mosquito density, and there was a significant decline in October. The mosquito density in livestock sheds 470.15 mosquitoes per lamp per night was significantly higher than that in other habitats. Over the past six years, the proportions of Culex pipiens quinquefasciatus and Aedes albopictus increased year by year, the proportion of Anopheles sinensis first increased and then decreased (with the highest proportion in 2021), the proportion of Culex tritaeniorhynchus decreased year by year, and the proportion of Armigeres subalbatus fluctuated at a low level. Conclusions The overall density of mosquito vector in Nanchang City shows a decreasing trend year by year, with Cx. pipiens quinquefasciatus and Cx. tritaeniorhynchus are the dominant mosquito species in the local area. The peak period of mosquito density is from June to September, and livestock sheds and farmers are high-risk areas for mosquito infestation. In order to prevent outbreak or spread of mosquito-borne infectious diseases, it is necessary to continue to strengthen ecological monitoring of key mosquito vector areas and take effective control measures in time based on monitoring results.

Objective To investigate the clinical features and risk factors for tuberculosis associated immune reconstitution inflammatory syndrome (TB-IRIS) in AIDS patients, in order to improve the diagnosis and treatment level of this disease. Methods This retrospective study included AIDS patients with tuberculosis who were admitted to a designated AIDS hospital in Guangzhou between June 2020 and February 2024. Patients were divided into TB-IRIS and Non-IRIS groups based on whether they developed IRIS after initiating antiretroviral therapy (ART). Baseline clinical data at admission, IRIS onset data, and ART regimens were collected to analyze the clinical features and baseline risk factors of TB-IRIS. Results A total of 222 AIDS patients with tuberculosis were included, with 58(26.1%) developing TB-IRIS. The median time of IRIS onset was 17(11, 29) days. Key symptoms included recurrent fever (89.7%), worsening respiratory symptoms (43.1%), and superficial lymphadenopathy (19.0%). Compared to the Non-IRIS group, the TB-IRIS group had higher baseline levels of tumor necrosis factor-α and interferon-γ (1.4 pg/mL vs. 0.8 pg/mL, P=0.024; 8.1 pg/mL vs. 2.9 pg/mL, P=0.008). The ART regimens showed no significant differences between the groups (P>0.05); however, the interval between anti-tuberculosis therapy (ATT) initiation and ART initiation was shorter in the TB-IRIS group than that in the Non-IRIS group (10 vs. 14 days, P=0.014), with 60.4% developing IRIS within 2 to 3 weeks of ART initiation. At IRIS onset, the TB-IRIS group showed a significantly higher CD4⁺T lymphocyte count than the Non-IRIS group (143 cells/μL vs. 94 cells/μL, P=0.021), as well as a more significant increase in both CD4⁺T cell count and CD4⁺/CD8⁺ ratio from baseline compared to the Non-IRIS group (91 cells/μL vs. 37 cells/μL, P<0.001; 0.14 vs. 0.07, P=0.006). Multivariate analysis indicated that baseline CD4⁺T lymphocyte count below 50 cells/μL and an ATT-to-ART interval of less than 10 days were associated with a higher likelihood of developing IRIS (OR=4.02, P=0.029; OR=2.06, P=0.044). Conclusion AIDS patients with tuberculosis who experience recurrent tuberculosis symptoms or rapid immune reconstitution within 2 to 3 weeks of ART initiation should be monitored for possible IRIS. For HIV-positive patients should avoid progressing to advanced stage before ART is initiated, and patients with combined TB should be treated with anti-tuberculosis therapy for 10 days before ART is initiated, and these measures can help to reduce the incidence of IRIS.

Objective This study aims to investigate whether 1,25（OH）₂D₃ reduces inflammatory factor IFN-α and IFN-β secretion in peripheral blood mononuclear cells (PBMC) of mice induced by dsDNA by inhibiting cGAS/STING/IFN-I signaling, and further explore the possibility of 1,25（OH）₂D₃ as new target for treatment of systemic lupus erythematosus (SLE). Methods A total of ten 8-week-old female C57BL/6 mice were anesthetized with 10% chloral hydrate, and blood was procured from the abdominal aorta to isolate peripheral blood mononuclear cells (PBMCs). These cells were subsequently divided into two groups. Group 1, designated as the dsDNA induction group, was subjected to different concentrations of dsDNA (0, 20, 40, and 80 μg/L), with the 0 μg/L dsDNA serving as the control group. This step was conducted to screen for the optimal dosage of dsDNA action. Group 2, PBMCs were initially treated with the inducible dsDNA at a concentration of 40 μg/L, followed by treatment with 11,25（OH）₂D₃ at concentrations of 10^-8, 10^-7, and 10^-6 μmol/L. The blank group consisted of PBMCs from mice treated with 0 μg/L of dsDNA without 1,25（OH）₂D₃ treatment, while the control group comprised PBMCs from mice treated with 40 μg/L of dsDNA without 1,25（OH）₂D₃ treatment. After 48 hours, the cell viability of PBMCs in each group was assessed using CCK8 assay. The secretion levels of IFN-α and IFN-β in the PBMCs of each group were determined by enzyme-linked immunosorbent assay (ELISA). The expression levels of pTBK1, pSTING, and pIRF3 in PBMCs of mice in each group were detected by Western blot. The study aimed to observe the impact of 1,25（OH）₂D₃ on cGAS/STING/IFN-Ⅰ signaling and the expression and secretion of IFN-α and IFN-β. Results dsDNA dose-dependently induced a significant reduction in cell viability of PBMCs (P<0.01), with a more SIGNIFICANT decrease in cell viability observed at a concentration of 40 μg/L. dsDNA dose-dependently enhanced the secretion of the inflammatory factors IFN-α and IFN-β (P<0.01). Compared to control group, the expression levels of cGAS, pTBK1, pSTING, and pIRF3 were up-regulated in PBMCs of mice(P<0.05). In the group of PBMCs treated with 1,25（OH）₂D₃ and dsDNA, 1,25（OH）₂D₃ dose-dependently led to a significant increase in the cell viability of dsDNA-treated PBMCs (P<0.01), and a decrease in the secretion of the inflammatory factors IFN-α and IFN-β (P<0.01). Compared to the control group, the expression levels of cGAS, pTBK1, pSTING, pIRF3 and IFN-Ⅰ were down-regulated in dsDNA-treated PBMCs upon addition of 1,25（OH）₂D₃ (P<0.05). Conclusion 1,25（OH）₂D₃ negatively regulates dsDNA activation of the cGAS/STING/IFN-Ⅰ signaling pathway and decreases the secretion of inflammatory factors IFN-α and IFN-β in PBMCs.

Objective To explore the influencing factors of liver injury in patients with newly diagnosed pulmonary tuberculosis combined with type 2 diabetes mellitus (PTB-T2DM) during anti-tuberculosis treatment, and we investigated the influencing factors of adverse prognosis in patients with liver injury, in order to provide reference for reducing the incidence of liver injury and improving prognosis. Methods A retrospective cohort study was conducted to continuously enroll 170 inpatients with PTB-T2DM who developed liver injury during anti-tuberculosis treatment and met the enrollment criteria in Guangyuan Mental Health Center from November 2018 to November 2023 recorded as the liver injury group, and 340 PTB-T2DM patients who completed anti tuberculosis treatment during the same period without liver injury were matched 1∶2 according to gender and age as the control group. Medical date and related biochemical indicators of all the enrolled patients were retrospectively collected, and the influencing factors of liver injury during anti-tuberculosis treatment of patients with PTB-T2DM were analyzed by Cox proportional hazards regression model. After 8 months of follow-up, the liver injury group was divided into two subgroups based on prognosis, and the influencing factors of adverse prognosis with liver injury were analyzed. Results Compared with the control group, the fasting blood glucose level in the liver injury group was significantly increased, while the albumin to globulin ratio and hemoglobin levels were significantly decreased (P<0.05). After adjusting for the confounding factors, we found education, history of drinking, albumin globulin ratio, diabetes course and anti tuberculosis drugs were independent influencing factors for the risk of liver injury during anti tuberculosis treatment of newly diagnosed PTB-T2DM patients. The incidence of adverse prognosis was 14.12%, and multivariate Cox proportional hazards regression analysis showed that poor glycemic control, delayed diagnosis, delayed presentation, irregular medication, identification of liver injury ≥8 weeks, and a long interval between the onset of gastrointestinal symptoms and/or renal dysfunction and discontinuation of tuberculosis drugs were independent risk factors for adverse prognosis after liver injury in patients with newly diagnosed PTB-T2DM (P<0.05). Conclusion There are relatively high incidence of adverse prognosis in patients with newly diagnosed PTB-T2DM during anti tuberculosis treatment, which is influenced by multiple factors, especially poor glycemic control, delayed diagnosis, identification of liver injury ≥8 weeks, and a longer intervals from the onset of gastrointestinal symptoms to discontinuation of tuberculosis medication are associated with a higher risk of adverse prognosis after liver injury.

Objective To analyze the registration of pulmonary tuberculosis (PTB) patients aged 65 and above (elderly patients) in Guizhou from 2013 to 2022, and to provide scientific basis for the formulation of PTB prevention and control strategies in the elderly population. Methods The case information of elderly PTB patients aged ≥65 year-old in Guizhou Province who were registered from 2013 to 2022 in the sub-system "Tuberculosis Management Information System" of "China Disease Prevention and Control Information System" was collected including gender, age, nationality, occupation classification, detection method, etiological diagnosis, etc. The data of detection, registration and treatment of elderly patients with PTB were analyzed, and compared to other age groups. χ² test was used for the comparison between two groups, and χ² trend test was used for the time change trends of the registration rate. Results A total of 73 462 elderly patients with TB were registered in Guizhou from 2013 to 2022, accounting for 19.26% of the total registered cases for the whole age groups in the same period. The average annual registration rate of elderly patients was significantly higher than that of the non-elderly patients (χ²= 34 874.00, P<0.001). The PTB registration rate of elderly patients decreased at an average annual rate of 1.5%, which was lower than that of non-elderly patients by 4.7% (χ²_trend=2 409.80, P<0.001). The male to female ratio of elderly patients was about 1.50∶1, and that of non-elderly patients was about 1.98∶1. The average annual registration rate of elderly male patients was significantly higher than that of elderly females (χ²=4 716.80, P<0.001), non-elderly males (χ²=16 470.00, P<0.001) and female patients (χ²=11 277.00, P<0.001). The proportion of elderly Han patients increased at an average annual rate of 6.2% (χ²_trend=1 168.45, P<0.001), compared to the 4.5% in the ethnic minorities (χ²_trend=293.55, P<0.001). The elderly patients were primarily in stable occupation or retired, peasants, and mainly concentrated in Zunyi, Bijie and Tongren. The proportion of retreatment in the elderly patients was significantly higher than that in the non-elderly patients. The etiological positive ratio in elderly patients was 47.18%, which was significantly higher than that in non-elderly patients. The main detection method in elderly patients was referral (42.34%), and the lowest detection method was active screening (0.10%). The delay rate for diagnosis in elderly patients was significantly higher than that in non-elderly patients. The delay rate of elderly patients was significantly higher than that of non-elderly patients. The success rate of treatment was 84.88% in elderly patients, which was significantly lower than 92.24% in the non-elderly patients. Conclusion The epidemic situation of PTB in the elderly population in Guizhou is serious. It is suggested to improve the active screening rate of PTB in the elderly and the treatment success rate of confirmed patients, so as to effectively control the epidemic situation of TB in the elderly.

Objective To explore the predictive efficacy of the carbon dioxide trap lamp method in monitoring the control level of mosquito density in urban residential environments. Methods Data on Aedes albopictus density were collected in residential areas of Dongcheng District, Beijing, using the human landing catch method and the carbon dioxide mosquito trap lamp method from July to September in 2017-2019. The receiver operating characteristic (ROC) curve was used to investigate the predictive efficacy of the carbon dioxide trap lamp method under varying mosquito density control levels and to determine the cut-off values corresponding to these levels. Results Among the 26 monitoring events conducted using the human landing catch method, the mosquito density failed to meet the required standard in 16 events. The highest absolute mosquito density recorded using the carbon dioxide mosquito trap lamp method reached 30.00 mosquitoes·(lamp·h)^-1. In the residential areas of Dongcheng District, the peak activity of Aedes albopictus mainly appeared in August. Due to abnormal climate changes, the mosquito peak in 2017 was delayed to mid-September. The areas under the ROC curve (AUC) for the carbon dioxide trap lamp method at control levels of A (landing index≤0.5), B (landing index≤1.0), and C (landing index≤1.5) were 0.919 (sensitivity 76.2%, specificity 100.0%, cut-off value 2.50 mosquitoes·(lamp·h)^-1，P<0.01), 0.865 (sensitivity 72.2%, specificity 87.5%, cut-off value 3.00 mosquitoes·(lamp·h)^-1，P<0.01), and 0.859 (sensitivity 75.0%, specificity 90.0%, cut-off value 3.13 mosquitoes·(lamp·h)^-1，P<0.01), respectively, with predictive performance at the A level being better than at the B and C levels. Conclusion The carbon dioxide trap lamp method demonstrated good predictive ability for different control levels of mosquitoes in urban residential areas, and its quantitative evaluation criteria can provide technical guidance and decision-making support for practical mosquito prevention and control work as well as the monitoring and early warning of mosquito-borne infectious diseases.

Objective To analyze the whole genome sequence of a GⅠ type norovirus in Qinghai region, understand its genetic structure and evolutionary characteristics, and we accumulated molecular epidemiological basic data on GⅠ type norovirus infection in our province. Methods Using Qinghai Women and Children's Hospital as a monitoring SITE, we conducted virus diarrhea monitoring for hospitalized children under 5 years old with diarrhea according to the requirements of the "National Viral Diarrhea Monitoring Plan" (2021 edition) in 2023. We used fluorescence quantitative PCR to detect case samples and screened a GⅠ type Norovirus (QH2023/23-24) with Ct value ≤ 30 for whole genome sequencing. The sequencing results were analyzed using phylogenetic tree analysis, homology analysis, and amino acid variation site analysis. Results The positivity rate of GⅠ type norovirus in 2023 was 3.33% (6/180), and phylogenetic analysis showed that QH2023/23-24 strains of GⅠ type norovirus were GⅠ.3[P13] type. The VP1 and RdRp regions of this virus strain are closely related to the PP584624.1 isolated in Thailand in 2023. Compared with the PP584624.1 isolated in Thailand in 2023, the amino acid sequence of the capsid VP1 region has one site change, M2V; and compared with the PQ632217.1 isolated in Zhejiang China in 2019, it has three site changes, M2V, T27A, and I237L. Conclusion For the first time, whole genome sequencing has been used to identify the genotype of GⅠ type Norovirus in our province. Sequence analysis shows that the virus strain (QH2023/23-24) is GⅠ.3[P13], which is the dominant strain prevalent in China. This study provides a sequence reference for study of the molecular evolution characteristics of norovirus in China. In the future, it is necessary to strengthen the monitoring of norovirus in the local area and provide theoretical basis for epidemic control and prevention.

Objective An investigation and analysis were conducted on a dengue fever outbreak that occurred at a construction site in Bao'an District, Shenzhen City in September 2024, to explore the application effectiveness of novel vector surveillance and insecticide resistance detection. Methods Dengue case data were gathered through epidemiological investigations, while mosquito density was monitored using the novel MS-300 vector surveillance technology and compared with the conventional breteau index (BI) method. The resistance of Aedes albopictus to pyrethroid insecticides was assessed by identifying mutations at position 1534 of the voltage-gated sodium channel (VGSC) gene. Results The epidemic persisted for 40 days, with 126 cases reported and an incidence rate of 6.94%. Dengue virus type Ⅱ (DENV-2) was identified as the main pathogen. Mosquito vector monitoring in the core area of the construction site revealed a declining trend in the number of mosquitoes captured daily by BI (from 18.00) and MS-300 (3 mosquitoes per day) starting from September 8. The 24-hour dynamic monitoring data of MS-300 demonstrated a double-peak pattern of mosquito activity at the epidemic site, with peak periods concentrated in the periods of 7:30-8:00 and 16:00-19:00. Detection of the VGSC gene at position 1534 locus in Aedes albopictus larvae collected from the epidemic site indicated two mutations, TGC (F1534C) and TCC (F1534S), suggesting resistance of the mosquito population to pyrethroid insecticides. Conclusion The preliminary application of MS-300 technology at this construction site enables online real-time, dynamic monitoring of mosquito vector activity. Combined with the detection of mosquito resistance, it provides a scientific basis for optimizing the prevention and control strategies for dengue fever at the grassroots level, constructing a more effective dengue fever prevention and control mechanism.