Exposure of Danio rerio to environmental sulfamethoxazole may contribute to neurobehavioral abnormalities via gut microbiome disturbance.

The neurotoxic effects and mechanisms of low-dose and long-term sulfamethoxazole (SMZ) exposure remain unknown. This study exposed zebrafish to environmental SMZ concentrations and observed behavioral outcomes. SMZ exposure increased hyperactivity and altered the transcript levels of 17 genes associated with neurological function. It impaired intestinal function by reducing the number of intestinal goblet cells and lipid content. Metabolomic results indicated that the contents of several lipids and amino acids in the gut were altered, which might affect the expression levels of neurological function-related genes. Metagenomic results demonstrated that SMZ exposure substantially altered the composition of the gut microbiome. Zebrafish receiving a transplanted fecal microbiome from the SMZ group were also found to exhibit abnormal behavior, suggesting that the gut microbiome is an important target for SMZ exposure-induced neurobehavioral abnormalities. Multi-omics correlation analysis revealed that gut micrometabolic function was related to differential gut metabolite levels, which may affect neurological function through the gut-brain-axis. Reduced abundance of Lefsonia and Microbacterium was strongly correlated with intestinal metabolic function and may be the key bacterial genera in neurobehavioral changes. This study confirms for the first time that SMZ-induced neurotoxicity in zebrafish is closely mediated by alterations in the gut microbiome.

Influence of Online Merging Offline Method on University Students' Active Learning Through Learning Satisfaction.

Students' active learning behavior determines learning performance. In post-COVID-19 period, Online Merging Offline (OMO) method become a common way of university students' learning. However, at present, there are few studies in active learning behavior in the OMO mode. Combined with learning satisfaction and Technology Acceptance Model (TAM), this paper proposes an Online Active Learning (OAL) Model to predict the influencing factors of college students' active learning behavior and then analyzes the differences between OMO model and pure online model by multi-group analysis (MGA) based on the model. The designed questionnaire was distributed, and a total of 498 valid questionnaires were collected. Using SmartPLS to analyze partial least squares structural equation modeling (PLS-SEM) and MGA, it is found that: (1) there are differences in the influencing factors of active learning between OMO and pure online model; the moderating effect of learning complaint in OMO mode is not established, and social isolation and age does not affect active learning in OMO mode; (2) learning quality, perceived ease of use, expectation, perceived usefulness, and social isolation indirectly affect active learning through learning satisfaction in both OMO model and pure online model; (3) learning satisfaction is an important mediating variable affecting active learning; and (4) learning complaints will negatively regulate the relationship between learning satisfaction and active learning only in pure online model. According to these findings, the paper provides theoretical and practical implementation suggestions implications for OMO teaching and OAL to ensure the expected learning outcome.

How to Implement Game-Based Learning in a Smart Classroom? A Model Based on a Systematic Literature Review and Delphi Method.

Game-based learning (GBL) can allow learners to acquire and construct knowledge in a fun and focused learning atmosphere. A systematic literature review of 42 papers from 2010 to 2020 in this study showed that the current difficulties in implementing GBL in classrooms could be classified into the following categories: infrastructure, resources, theoretical guidance, teacher's capabilities and acceptance of GBL. In order to solve the above problems, the study constructs a technology enhanced GBL model, from the four parts of learning objective, learning process, learning evaluation, and smart classroom. In addition, this study adopted the Delphi method, inviting a total of 29 scholars, experts, teachers and school managers to explore how to implement GBL in smart classrooms. Finally, the technology enhanced GBL model was validated and the utilization approaches were provided at the conclusion part.

Neonatal hand, foot, and mouth disease due to coxsackievirus A6 in Shanghai.

BACKGROUND: Evidence of hand, foot, and mouth disease (HFMD) in neonates is limited. The aim of this study was to evaluate the clinical symptoms, pathogens, possible transmission routes, and prognosis of neonatal HFMD in Shanghai. METHODS: This was a case-control study based on the HFMD registry surveillance system. All neonates and infected family members were enrolled between 2016 and 2017 in Shanghai. Neonates with HFMD were followed for at least half a year. Detailed questionnaires, medical history, and physical examination were recorded. Routine blood examination, liver and renal function, immunophenotypes of peripheral blood lymphocytes (CD3, CD4, and CD8 T-cells; NK cells), immunoglobulin (Ig) M, IgG, and IgA, and cytokine interleukin (IL-1ß, IL-2R, IL-6, IL-8, IL-10, and TNF-α) levels were measured. All rectal swab specimens were collected and genotyped for enterovirus, and phylogenetic analysis based on the VP1 sequences of coxsackievirus A6 (CV-A6) was performed to investigate molecular and evolutionary characteristics. T-test or nonparametric test was used to evaluate the differences. Logistic analysis was applied to calculate the risk of clinical manifestations in the group of HFMD neonates and their paired siblings. RESULTS: There were 16 neonates among the 12,608 diagnosed patients with HFMD, accounting for 0.13%. All neonatal infections were transmitted by other members of the family, mainly the elder siblings, and were caused by CV-A6. CV-A6 was the emerging and predominant causative agent of HFMD in Shanghai. None of the neonates with HFMD experienced fever, onychomadesis, or severe complications. However, two elder sibling patients showed lethargy, and one developed hypoperfusion. In the elder siblings with HFMD, the proportion of white blood cells was generally higher than in neonates with HFMD. The immunologic function of the neonates with HFMD was basically normal. The levels of inflammatory markers were higher in both neonates and elder siblings with HFMD compared to age-matched controls. The clinical symptoms receded about 1 week after onset. None of the neonates had sequelae. CONCLUSIONS: In our study, CV-A6 infection in neonates was benign, but had the character of family clustering. Due to the two-child policy in China, elder siblings may be the main route of HFMD transmission.

Clinical characteristics and managements of severe hand, foot and mouth disease caused by enterovirus A71 and coxsackievirus A16 in Shanghai, China.

BACKGROUND: Hand, foot and mouth disease (HFMD) is a transmissible infectious disease caused by human enteroviruses (EV). Here, we described features of children with severe HFMD caused by EV-A71 or coxsackievirus A16 (CV-A16) in Shanghai, China. METHODS: Severe EV-A71 or CV-A16 caused HFMD children admitted to the Xinhua Hospital from January 2014 and December 2016, were recruited retrospectively to the study. Symptoms and findings at the time of hospitalization, laboratory tests, treatments, length of stay and residual findings at discharge were systematically recorded and analyzed. RESULTS: Of 19,995 children visited clinic service with probable HFMD, 574 children (2.87%) were admitted, 234 children (40.76%) were confirmed with EV-A71 (90/574) or CV-A16 (144/574) disease. Most (91.02%) of the patients were under 5 years. Initial clinical symptoms of EV-A71 and CV-A16 cases were: fever > 39 °C in 81 (90%) and 119 (82.63%), vomiting in 31 (34.44%) and 28 (19.44%), myoclonic twitching in 19 (21.11%) and 11(7.64%), startle in 21 (23.33%) and 20 (13.69%), respectively. Serum levels of interleukin-1ß (IL-1ß), IL-2, IL-6, IL-8, interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α) were significantly upregulated in severe HFMD subjects. Forty-seven children (20.08%) treated with intravenous gamma globulin (IVIG) showed decreased duration of illness episodes. All children were discharged without complications. CONCLUSIONS: EV-A71 and CV-A16 accounted 40.76% of admitted HFMD during 2014 to 2016 in Xinhua Hospital. IVIG appeared to be beneficial in shortening the duration of illness episodes of severe HFMD.

Virome profiling of rodents in Xinjiang Uygur Autonomous Region, China: Isolation and characterization of a new strain of Wenzhou virus.

Rodents, as the most diverse and widest distributed mammals, are a natural reservoir of many zoonotic viruses. However, little is known about the viral diversity harbored by rodents in China. Here we performed viral metagenomic analyses of 314 wild rodents covering 7 species, sampled in North-western China. We also conducted a systematic virological characterization of a new Wenzhou virus (WENV) isolate, QARn1, from a brown rat (Rattus norvegicus). Full genomic and phylogenetic analyses showed that QARn1 is a previously unidentified strain of Wenzhou mammarenavirus and forms a new branch within the Asian clade. Experimental infection of Sprague-Dawley rats with QARn1 did not present overt pathology, but specific humoral immune responses developed and mild hemorrhage and immunocyte infiltration of the lungs and thymus were observed. These observations have expanded the geographic distribution of WENV to Central Asia, and further confirm that brown rats are natural hosts of Wenzhou virus.

Genome-wide linear B-cell epitopes of enterovirus 71 in a hand, foot and mouth disease (HFMD) population.

BACKGROUND: Enteroviruses cause hand, foot and mouth disease (HFMD). The host B-cells recognize the viral proteins and provoke humoral responses. Deciphering the B-cell responses to the viral epitopes helps diagnosis and vaccine development. OBJECTIVES: The objective of the present study was to investigate for the first time the landscape of genome-wide linear B-cell epitopes of enterovirus 71 in HFMD population. STUDY DESIGN: The peptides encompassing the entire coding region of EV71 were chemically synthesized and displayed on a microarray. The peptide microarray was used to screen serum samples from an HFMD population, including EV71-, CAV10-, CAV16- and CAV6-infected patients. We identified the dominant epitope-containing-peptides (DECPs) that react with the sera of more than 20% of the HFMD population and the common DECPs that cross-react with the sera from other enteroviruses-infected population. RESULTS: Ten DECPs reacting with IgM and 9 DECPs reacting with IgG antibodies were identified, of which, 6 IgM and 5 IgG common DECPs cross-reacted with the sera from other enteroviruses. Some DECPs preferentially reacted with IgG or IgM antibodies and some epitope-antibody interactions correlated with the severity of HFMD. CONCLUSIONS: We uncovered the DECPs and the common DECPs among a group of enteroviruses in HFMD population and found that some epitope-antibody reactions were associated with the outcome of HFMD. These data may guide developing vaccines against the enteroviruses and help the diagnosis and prognosis of HFMD.

A novel recombinant lineage's contribution to the outbreak of coxsackievirus A6-associated hand, foot and mouth disease in Shanghai, China, 2012-2013.

Since late 2012, coxsackievirus A6 (CVA6) has gradually become the predominant pathogen responsible for hand-foot-mouth disease (HFMD) in several provinces of China. A total of 626 patients diagnosed with HFMD in Shanghai, China from January 2012 to September 2013 were enrolled in this study. Of these, 292 CVA6 infected cases were subjected to clinical analyses. Whole-genome sequencing, recombination and phylogenetic analyses were also performed. A recombinant CVA6 monophyletic lineage was found during an outbreak of CVA6-associated HFMDs in Shanghai, China in November 2012, and accounted for 21.9% (64/292) of the CVA6 strains during the study period. Recombination analyses showed that the 2C gene of the novel CVA6 virus was probably derived from a coxsackievirus A4 (CVA4) strain circulating in the population. Clinical observation showed that this recombinant CVA6 virus led to a more generalized rash than did the non-recombinant CVA6 virus. This newly emerged CVA6 lineage was associated with a considerable proportion of HFMD cases from 2012 to 2013 in Shanghai, and poses a potential threat to public health.

Genome sequence of a novel recombinant coxsackievirus a6 strain from shanghai, china, 2013.

A novel recombinant coxsackievirus A6 (CVA6) strain was isolated during a coxsackievirus A6 outbreak in Shanghai, China, in 2013. Genomic sequence and similarity plot analysis showed that the novel CVA6 strain shared higher similarity with a recent CVA4 strain rather than the recent CVA6 strain in the 2C and 3' untranslated regions (UTRs).

Severe enterovirus type 71 nervous system infections in children in the Shanghai region of China: clinical manifestations and implications for prevention and care.

BACKGROUND: Severe outbreaks of enterovirus 71 (EV71) have been on the rise in the Western Pacific region, including in China since 2007. We describe features of pediatric patients admitted with severe disease EV71 to a tertiary care hospital during the 2010 ongoing outbreak in the Shanghai region. METHODS: The Shanghai EV71 outbreak was studied prospectively from January 2010 to December 2012. To be eligible, children had to be 1 month to <14 years of age, admitted with confirmed EV71 infection to Xinhua Hospital and have severe illness needing Pediatric Infectious Diseases Service care with symptoms/signs of high prolonged fever and/or very unwell general appearance and/or neurological findings but no underlying medical condition. Clinical, laboratory, treatment discharge and follow-up data were recorded and analyzed. RESULTS: Of 26, 829 children presenting with possible EV71 disease, 2.9% (767) were admitted, more from less developed areas than from Shanghai proper. Of these, 29.2% (224) had severe enough EV71 for study enrolment, 4 excluded for underlying disease. Of 220 enrolled, 40.5% (89) had neurological involvement, 10.5% (23) needed pediatric intensive care unit care and 3% (7) died. Factors associated with severe disease were young age, male gender, high white count, prolonged high fever, high glucose, high C-reactive protein and neurological findings. The majority with neurological involvement and all who died were from the countryside. Early intravenous gamma globulin intervention had better survival. No survivor had persistent sequelae 2 months after discharge. CONCLUSIONS: This study confirms the morbidity for EV71 disease in China, especially for patients from the countryside versus Shanghai proper. The potential impact of public education, an EV71 vaccine, and the potential benefits versus harms of IVIG treatment are discussed.

[Efficacy and safety of ribavirin aerosol in children with hand-foot-mouth disease].

OBJECTIVE: To evaluate the efficacy and safety of ribavirin aerosol in children with hand-foot-mouth disease (HFMD). METHODS: A randomized, double-blind, placebo-controlled trial was performed. A total of 119 children with mild HFMD were randomly divided into an observed group (n=59) and a control group (n=60). In the observed group, ribavirin aerosol was given four times within the first hour, followed by once every other hour for the remaining time of the day and day 2; from days 3 to 7, it was given 4 times per day, with 2-3 sprays every time, for 7 days. In the control group, placebo was given in the same way as in the observed group. Additionally, both groups used oral antiviral liquid. The scores of clinical symptoms including oral ulcer, skin rash, nasal congestion, runny nose, sneezing, cough, and fever before and after treatment were recorded to evaluate treatment outcomes. Throat swabs were taken before treatment and 5-7 days after treatment to measure viral load by RT-PCR and to compare the negative conversion rate between the two groups. RESULTS: Fifty-seven patients in the observed group and 56 patients in the control group were tested according to the original research design. After 5-7 days of treatment, the observed group had a significantly higher overall negative conversion rate of enterovirus than the control group (P<0.01). The overall marked response rate and overall response rate of the observed group were 89% and 89%, respectively, significantly higher than those of the control group (29% and 43%). During treatment, there were no adverse reactions such as dizziness, vomiting, and notable decreases in hemoglobin, white blood cells, and platelets in the two groups. CONCLUSIONS: Ribavirin aerosol can be effectively and safely used for treating mild HFMD. With low dosage and few adverse reactions, it holds promise for clinical application.

Quality of death certificates in Quemoy, Taiwan.

BACKGROUND AND PURPOSE: The age-adjusted mortality rate on the island of Quemoy was the lowest of any county in Taiwan in the years 2000-2001. However, the island's rate of mortality due to cardiovascular diseases fluctuates widely. This fluctuation suggests that Quemoy may be issuing inaccurate death certificates. METHODS: To assess the quality of death certificates from Quemoy, 123 (15.3%) medical records for 800 deaths between 1994 and 1998 were reviewed by 3 medical specialists. The underlying cause of death from the original death certificate was compared to the underlying cause of death determined by 3 reviewers based on review of all available medical records. RESULTS: The agreement index for all causes of death was 72.4%. Neoplasms had the highest sensitivity and positive predictive value for correct determination of underlying cause of death. Cardiovascular diseases had higher sensitivity, but lower positive predictive value than respiratory diseases. Neoplasms were under-reported by 25.5% but cardiovascular diseases were over-reported by 34.3%. CONCLUSIONS: Therefore, mortality statistics in Quemoy, which are based on death certificate data, may underestimate the frequency of neoplasms and overestimate cardiovascular diseases as underlying causes of death. Our findings also suggest that researchers should exercise considerable caution when using death certificate data to determine cause of death in etiologic studies, especially in neoplasms and cardiovascular diseases.