BACKGROUND AND OBJECTIVES

Municipal solid waste workers (MSWWs) are important in the city’s waste management. With these vital contributions, they face unique occupational hazards and health risks. This study aims to determine the prevalence of occupational infections, such as soil-transmitted helminth infections (STHI) and hepatitis A virus (HAV), as well as the occurrence of Helicobacter pylori infection among the MSWWs of Baguio City.

This cross-sectional analytic study collected data from volunteer MSWWs using a questionnaire to gather information on age, duration of employment, use of gloves in the workplace, and hand hygiene practices. Stool samples were obtained from participants and were analyzed for STHI using the Formalin Ether Concentration Technique (FECT). H. pylori infection was detected using the SD Bioline rapid antigen test kit on stool samples while blood samples were collected and tested for HAV antibodies using the Aria IgG/IgM rapid test kit.

Of the 44 volunteer MSWWs tested, 25 were infected with hazardous pathogens. Specifically, six workers (13.6%) were infected with STHI, four (9.1%) were infected with HAV and 15 (34.1%) were infected with H. pylori. Among those infected with STHI, Ascaris lumbricoides and Endolimax nana were the predominant species, each with a prevalence rate of 33.3%. In contrast, Blastocystis hominis and hookworm infections each had a prevalence rate of 16.7%. A significant association was found between STHI prevalence and the preference for alcohol hand rubs over hand washing, with a p-value of 0.008.

The analysis revealed a significant association between the prevalence of STHI and the preference for alcohol hand rubs over hand washing, suggesting that MSWWs may have a false sense of security regarding their hygiene practices. The findings revealed the critical importance of proper hand washing in preventing STHI. Future research should expand data collection to encompass a broader range of socio-demographic, environmental, and lifestyle factors that may influence infection rates. Additionally, including a control group of individuals not exposed to waste management could help differentiate between factors specific to waste handling and those related to other occupations. This study emphasizes the need for collaborative efforts among researchers, public health authorities, and waste management agencies to enhance the health and safety of MSWWs while addressing broader public health concerns related to waste management practices.

Objective To investigate the role and mechanism of RNA-Specific adenosine deaminase 3 (Adar3) in regulating macrophage polarization during Coxsackievirus B3(CVB3)-induced viral myocarditis (VM). Methods Bone marrow-derived macrophages (BMDM) from mice were cultured in vitro and induced into M1/M2 macrophages using interferon-gamma (IFN-γ)/lipopolysaccharide (LPS) or interleukin 4 (IL-4), respectively. The mRNA expression levels of Adar1, Adar2, and Adar3 in each group of cells were assessed by real-time quantitative PCR (qRT-PCR). Specific siRNAs targeting the Adar3 gene were designed, synthesized, and transiently transfected into M2 macrophages. The mRNA levels of M2 polarization-related marker genes-including arginase 1 (Arg1), chitinase 3-like molecule 3 (YM1/Chi3l3), and resistin-like molecule alpha (RELMα/FIZZ1)-were detected by qRT-PCR. RNA sequencing was performed to analyze the signaling pathways affected by Adar3. The expression levels of Wnt/β-catenin signaling pathway were further validated using qRT-PCR and Western blot. The adeno-associated virus overexpressing Adar3 was designed, synthesized, and injected into mice via tail vein. Three weeks later, a myocarditis mouse model was established. After an additional week, the phenotype and function of cardiac macrophages, as well as multiple indicators of VM (including echocardiography, body weight, histopathology and serology) were examined. Additionally, the protein levels of the Wnt/β-catenin signaling pathway were assessed. Results Compared to M0-type macrophages, the expression level of Adar3 was significantly increased in M2-type macrophages. After transfection of Adar3 siRNA, the mRNA levels of Arg1, YM1 and FIZZ1 in M2 macrophages were downregulated. RNA sequencing revealed 149 upregulated genes and 349 downregulated genes. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis and subsequent validation experiments indicated that Adar3 modulated the Wnt/β-catenin signaling pathway. In vivo experiments demonstrated that Adar3 overexpression alleviated the cardiac dysfunction of VM mice. The proportion of M1 macrophages in the heart decreased, while the proportion of M2 macrophages increased. At the same time, the Adar3 overexpression activated the Wnt/β-catenin signaling pathway. Conclusion Adar3 promotes macrophage polarization toward the M2 phenotype by activating the Wnt/β-catenin signaling pathway, thereby alleviating VM.
Animals ; Adenosine Deaminase/metabolism* ; Macrophages/immunology* ; Wnt Signaling Pathway/genetics* ; Myocarditis/immunology* ; Mice ; Coxsackievirus Infections/metabolism* ; Male ; Mice, Inbred BALB C ; Enterovirus B, Human/physiology* ; beta Catenin/genetics*

BACKGROUND: Japan lacks comprehensive reports on the nationwide voluntary vaccine coverage. The effectiveness of public subsidies in promoting vaccination has not been fully investigated. Therefore, we aimed to estimate the nationwide coverage of voluntary vaccines, compare it with that of national immunization program (NIP)-included vaccines, and investigate the effectiveness of public subsidies. METHODS: We obtained nationwide monthly vaccine market data for rotavirus, Haemophilus influenzae type b (Hib), diphtheria, tetanus toxoid, acellular pertussis, inactivated poliovirus (DTaP-IPV), and mumps vaccines; estimated recipient numbers; and calculated coverage as the proportion of children from October 2011 to March 2022. Regarding the NIP-included vaccine, we compared vaccine coverage calculated from nationwide annual market data with that estimated by World Health Organization (WHO)/United Nations Children's Fund (UNICEF), using Bland-Altman analysis. RESULTS: The estimates of Hib and DTaP-IPV vaccine coverage derived from market data were slightly higher than the WHO/UNICEF estimates, with mean differences of 0.05 (95% CI: 0.02-0.07) for Hib and 0.03 (95% CI: 0.01-0.05) for DTaP-IPV. The coverage of the rotavirus vaccine gradually increased long before the implementation of national subsidies, reaching 0.9 in 2020. Hib vaccine coverage had already achieved 1.0 by January 2012. The coverage of the DTaP-IPV vaccine was approximately 0.6-0.8 in 2013, reaching 1.0 in 2014. The coverage of mumps vaccine increased gradually from 2011 to 2021. CONCLUSIONS Despite the possibility of overestimation, our estimates may serve as a valuable surrogate for actual vaccine coverage in Japan. An increasing trend in rotavirus and mumps vaccine coverage was observed when these vaccines were categorized as voluntary. Although vaccination policies differ from country to country, it would be beneficial to share findings on the impact of subsidies in Japan with other countries.
Japan ; Humans ; Vaccination Coverage/statistics & numerical data* ; Infant ; Immunization Programs/statistics & numerical data* ; Rotavirus Vaccines ; Haemophilus Vaccines ; Child, Preschool ; Poliovirus Vaccine, Inactivated ; Vaccines

OBJECTIVES: To observe the therapeutic effect of spermine in neonatal mouse models of severe hand, foot and mouth disease (HFMD) caused by enterovirus 71 (EV71) infection and explore its therapeutic mechanism in light of regulation of macrophage GBP5/NLRP3 inflammasome pathway. METHODS: Neonatal BALB/c mice (3-5 days old) were divided into control group, EV71 infection group and Spermine treatment group. The mice in the latter two groups received an intraperitoneal injection of 50 μL EV71 suspension (1×10⁶ TCID50 of EV71), followed 3 days later by intraperitoneal injection of 50 μL PBS or 100 μmol/L spermine. GBP5, NLRP3, CXCL10, and TNFSF10 expressions in heart, liver, lung and kidney tissues of the mice were detected using Western blotting and qPCR, and tissue pathologies and macrophage infiltration were assessed with HE staining and immunohistochemistry. In cultured THP-1 and RAW264.7 cells, the effects of EV71 infection, GBP5 siRNA transfection and treatment with spermine or eflornithine on GBP5, NLRP3, CXCL10, and TNFSF10 mRNA expressions were investigated using qPCR. RESULTS: In the neonatal mice, EV71 infection resulted in multiple organ damage, macrophage infiltration and activation of the GBP5/NLRP3 pathway, and spermine treatment significantly improved tissue injuries, reduced macrophage infiltration, and down-regulated the expressions of GBP5, NLRP3 and the inflammatory factors in the infected mice. In THP-1 and RAW264.7 cells, EV71 infection caused significant upregulation of GBP5, NLRP3, CXCL10, and TNFSF10 expressions, which were obviously lowered by spermine treatment. In THP-1 cells, treatment with eflornithine significantly suppressed the reduction of GBP5, NLRP3, CXCL10, and TNFSF10 expressions induced by GBP5 siRNA transfection. CONCLUSIONS Spermine suppressed EV71 infection-induced inflammatory responses by inhibiting GBP5-mediated NLRP3 inflammasome activation, suggesting a new strategy for treatment of severe HFMD.
Animals ; NLR Family, Pyrin Domain-Containing 3 Protein ; Mice ; Macrophages/metabolism* ; Enterovirus A, Human ; Mice, Inbred BALB C ; Inflammasomes/metabolism* ; Spermine/therapeutic use* ; Animals, Newborn ; Humans ; Enterovirus Infections ; Hand, Foot and Mouth Disease/drug therapy* ; RAW 264.7 Cells ; Chemokine CXCL10/metabolism*

The bioactivity-guided isolation of potentially active natural products has been widely utilized in pharmaceutical discovery. In this study, by screening fungal extracts against coxsackievirus B3 (CVB3), three new aspochalasins, templichalasins A‒C (1‒3), along with six known aspochalasins (4‒9) were isolated from an active extract derived from the endophytic fungus Aspergillus templicola LHWf045. Compound 1 features a unique 5/6/5/7/5 pentacyclic ring system, while compounds 2 and 3 possess unusual 5/6/6/7 tetracyclic skeletons. Their structures were characterized through extensive spectroscopic analyses, electronic circular dichroism (ECD) calculations, and single-crystal X-ray diffraction analysis. Additionally, we demonstrated that compound 4 can be readily converted into compounds 1‒3 under mild acidic conditions and proposed a plausible mechanism for this conversion. Bioactivity evaluation of compounds 1‒9 against CVB3 revealed the inhibitory effects of all compounds against the virus. Notably, compound 9 exhibited superior antiviral activity, surpassing the commercial drug ribavirin in selectivity index (SI) value.
Antiviral Agents/isolation & purification* ; Aspergillus/chemistry* ; Molecular Structure ; Enterovirus B, Human/drug effects* ; Endophytes/chemistry* ; Cytochalasins/isolation & purification* ; Drug Discovery ; Humans

Senecavirus A (SVA) is a major viral pathogen causing disease in pigs, and effective monitoring of SVA infection is critical for disease control. In this study, we aimed to develop a reliable ELISA method for rapidly detecting neutralizing antibodies against SVA. We used HEK293F cells to express an SVA-specific porcine Fab antibody and verified the biological activity of the Fab antibody by indirect ELISA, immunofluorescence assay, virus neutralization test, and Western blotting. The Fab antibody was biotinylated and used as a competitive antibody to establish a competitive ELISA (C-ELISA) for detecting neutralizing antibodies against SVA. We then evaluated the C-ELISA in terms of sensitivity, specificity, repeatability, and result agreement rate with the VNT. The results showed that we successfully prepared an SVA-specific porcine Fab antibody, which showed high affinity for SVA. We named this antibody 1M33Fab and designated it as Bio-1M33Fab after biotin labeling. The assay conditions were optimized as follows: the coating concentration of SVA particles being 1 μg/mL, the working concentration of Bio-1M33Fab being 0.5 μg/mL, the optimal serum dilution of 1:10, and the optimal dilution of enzyme-labeled avidin being 1:30 000. At a percent inhibition (PI) of 47%, the assay demonstrated the highest sensitivity (96.88%) and specificity (100%), with no cross-reactivity observed with the positive sera of major porcine viral diseases. The intra-assay coefficient of variation ranged from 1.12% to 7.34%, while the inter-assay coefficient of variation ranged from 1.10% to 8.97%, indicating good repeatability. In the detection of 224 clinical pig serum samples, C-ELISA and VNT showed a result agreement rate of 93.75%. In conclusion, we successfully develop a C-ELISA method for detecting neutralizing antibodies against SVA by using a porcine-derived Fab antibody, which lays a foundation for the development of detection kits.
Animals ; Swine ; Antibodies, Neutralizing/immunology* ; Enzyme-Linked Immunosorbent Assay/methods* ; Immunoglobulin Fab Fragments/immunology* ; Antibodies, Viral/immunology* ; Picornaviridae/immunology* ; Humans ; HEK293 Cells ; Swine Diseases/diagnosis* ; Picornaviridae Infections/diagnosis*

Background and Objectives: Municipal solid waste workers (MSWWs) are important in the city’s waste management. With these vital contributions, they face unique occupational hazards and health risks. This study aims to determine the prevalence of occupational infections, such as soil-transmitted helminth infections (STHI) and hepatitis A virus (HAV), as well as the occurrence of Helicobacter pylori infection among the MSWWs of Baguio City. Methods: This cross-sectional analytic study collected data from volunteer MSWWs using a questionnaire to gather information on age, duration of employment, use of gloves in the workplace, and hand hygiene practices. Stool samples were obtained from participants and were analyzed for STHI using the Formalin Ether Concentration Technique (FECT). H. pylori infection was detected using the SD Bioline rapid antigen test kit on stool samples while blood samples were collected and tested for HAV antibodies using the Aria IgG/IgM rapid test kit. Results: Of the 44 volunteer MSWWs tested, 25 were infected with hazardous pathogens. Specifically, six workers (13.6%) were infected with STHI, four (9.1%) were infected with HAV and 15 (34.1%) were infected with H. pylori. Among those infected with STHI, Ascaris lumbricoides and Endolimax nana were the predominant species, each with a prevalence rate of 33.3%. In contrast, Blastocystis hominis and hookworm infections each had a prevalence rate of 16.7%. A significant association was found between STHI prevalence and the preference for alcohol hand rubs over hand washing, with a p-value of 0.008. Conclusion The analysis revealed a significant associat ion between the prevalence of STHI and the preference for alcohol hand rubs over hand washing, suggesting that MSWWs may have a false sense of security regarding their hygiene practices. The findings revealed the critical importance of proper hand washing in preventing STHI. Future research should expand data collection to encompass a broader range of socio-demographic, environmental, and lifestyle factors that may influence infection rates. Additionally, including a control group of individuals not exposed to waste management could help differentiate between factors specific to waste handling and those related to other occupations. This study emphasizes the need for collaborative efforts among researchers, public health authorities, and waste management agencies to enhance the health and safety of MSWWs while addressing broader public health concerns related to waste management practices.
Human ; Hepatitis A virus (HAV) ; Helicobacter pylori

OBJECTIVE: To comprehensively examine the molecular epidemiological characteristics of human rhinovirus (HRV) in Beijing. METHODS: A total of 7,151 children and adults with acute respiratory tract infections (ARTIs) were recruited from 35 sentinel hospitals in Beijing between 2018 and 2022. Their respiratory samples were obtained, and epidemiological and clinical data were collected. Nucleic acid testing for 11 respiratory pathogens, including HRV, was performed on the specimens. We sequenced VP4/VP2 or 5'UTR of HRV isolates to identify their genotypes using phylogenetic analyses. RESULTS: HRV was detected in 462 (6.5%) cases. A total of 105 HRV genotypes were successfully identified in 359 (77.7%) specimens, comprising 247 (68.8%) with HRV-A, 42 (11.7%) with HRV-B, and 70 (19.5%) with HRV-C. No predominant genotype was observed. HRV was prevalent year-round with two weak peaks in spring and autumn. HRV detection declined gradually between 2018 and 2022, with seven genotypes disappearing and five genotypes emerging. HRV detection rate decreased by age without resurge among old people. HRV-C was more common among children aged less than 5 years with severe community-acquired pneumonia compared to HRV-A and HRV-B. Adults infected with HRV-B had higher rates of hospitalization, intensive care unit admission, and complications than those infected with HRV-A and HRV-C. CONCLUSION HRV epidemics in Beijing were highly dispersed in genotypes, which probably resulted in a high prevalence of HRV infection, attenuated its seasonality, and made it more difficult to establish effective population immunity.
Humans ; Rhinovirus/classification* ; Beijing/epidemiology* ; Child ; Child, Preschool ; Adult ; Genotype ; Male ; Adolescent ; Picornaviridae Infections/virology* ; Female ; Middle Aged ; Infant ; Retrospective Studies ; Young Adult ; Prevalence ; Aged ; Respiratory Tract Infections/epidemiology* ; Phylogeny ; Genetic Variation

The aim of this study was to compare the immune responses of C57BL/6 mice immunized with two pathogens, foot-and-mouth disease virus (FMDV) and Senecavirus A (SVA), and to provide clues for revealing the regulatory mechanisms of acquired immunity. Inactivated and purified FMDV and SVA antigens were used to immunize C57BL/6 mice respectively, and the mice immunized with PBS were taken as the control. The percentages of Th1 and Th2 cells in the spleen lymphocytes of mice in each group were analyzed by flow cytometry at 14 and 28 days after immunization. RNA-Seq was performed for the spleen. Mouse macrophages were stimulated with the antigens in vitro to examine the expression of the differentially expressed genes (DEGs) screened out. The results showed that 14 days after immunization, there was no significant difference in the magnitude of the Th1/Th2 immune response elicited by the FMDV and SVA antigens. After 28 days, the magnitudes of the Th1 and Th2 immune responses elicited by the SVA antigen were higher than those elicited by the FMDV antigen. RNA-Seq revealed two common DEGs, Rsad2 and Tspan8, between the two immunization groups, which indicated that the two genes may be involved in the activation of the Th1/Th2 immune responses by FMDV and SVA antigens. FMDV and SVA antigens stimulated macrophages to secrete interleukin (IL)-12 and IL-33 in vitro, and the expression of Tspan8 and Rsad2 was consistent with the RNA-Seq results. The expression of Rsad2 was regulated by type I interferons (IFNα, IFNβ). In this study, we obtained the DEGs involved in the immune responses to the two antigens in mouse spleen, which provides a molecular basis for investigating the immune response mechanisms induced by FMDV and SVA.
Animals ; Foot-and-Mouth Disease Virus/genetics* ; Mice ; Spleen/cytology* ; Mice, Inbred C57BL ; Antigens, Viral/genetics* ; Transcriptome ; Th1 Cells/immunology* ; Immunization ; Viral Vaccines/immunology* ; Th2 Cells/immunology* ; Foot-and-Mouth Disease/immunology* ; Interleukin-33/genetics* ; Female ; Macrophages/immunology* ; Picornaviridae

Objective:b> To understand the infection status of Enterovirus (EV) in cases of acute respiratory infections (ARIs) in Luohe City, Henan Province from 2017 to 2021, and analyze the prevalence and type composition of EV in ARIs. Methods:b> From October 2017 to May 2021, pharyngeal swab samples were collected from 1 828 patients with ARIs in Luohe Central Hospital and the clinical epidemiological data of these cases were also collected. EV-positive samples were identified by Quantitative Real-time Polymerase Chain Reaction (qPCR). The 5'-untranslated region (5'UTR) was amplified by Reverse Transcription-Polymerase Chain Reaction (RT-PCR). The results of 5'UTR region were initially typed by Enterovirus Genotyping Tool Version 1.0. Based on the typing results, the full-length of VP1 region was amplified by RT-PCR. The EV typing was identified again by VP1 region. Results:b> Among 1 828 cases of ARIs, 56.7% (1 036) were males. The median (Q₁, Q₃) age was about 3 (1, 5) years. Patients under 5 years old accounted for 71.6% (1 309 cases). Among all cases, a total of 71 EV-positive samples were identified by qPCR, with a detection rate of 3.88% (71/1 828). The EV detection rates for men and women were 3.28% (34/1 036) and 4.67% (37/792), without statistically significant differences (χ²=2.32, P=0.14). The EV detection rates for 2 to <6 years, 6 months to <2 years, 6 to <10 years, and <6 months were 6.29% (48/763), 3.00% (18/600), 2.52% (4/159), and 1.67% (1/60) (χ²=27.91, P<0.001). The EV detection rate was 0.92% (3/326) in autumn and winter of 2017. The EV detection rates were 1.18% (6/508), 2.47% (12/485) and 8.31% (34/409) in each year from 2018 to 2020, with an increasing trend year by year(χ²_trend=29.76, P<0.001). The main prevalent seasons were summer and autumn. The detection rate in spring of 2021 was 4.00% (4/100). A total of 12 types were identified and classified as CVA2, CVA4, CVA5, CVA6, CVA10, CVB3, CVB5, E5, E11, E30, PV-1, and EV-D68. The types of CVA2, CVA10, CVA6, and CVB3 were the dominant phenotypes. In 59 sample of EV typing, the main clinical manifestation was upper respiratory tract infection (36/59, 61.01%). The dominant types detected in upper respiratory tract infections were CVA10 (10/36, 27.78%), CVA6 (9/36, 25.00%) and CVB3 (8/36, 22.22%). The dominant type detected in lower respiratory tract infections was CVA2 (7/19, 36.84%). Conclusion:b> In Luohe City, Henan Province from 2017 to 2021, EV infection in ARIs cases has clear seasonal and age-specific patterns, and the dominant types of upper and lower respiratory tract infections are different.
Male ; Female ; Humans ; Enterovirus/genetics* ; 5' Untranslated Regions ; Enterovirus Infections/epidemiology* ; Phenotype ; Antigens, Viral/genetics* ; Respiratory Tract Infections/epidemiology* ; Phylogeny