The characteristics of mcr-bearing plasmids in clinical Salmonella enterica in Sichuan, China, 2014 to 2017.

Salmonella is one of the most important zoonotic pathogens and a major cause of foodborne illnesses, posing a serious global public health hazard. The emergence of plasmid-mediated mcr genes in Salmonella has greatly reduced the clinical choice of salmonellosis treatment. The aim of this study was to investigate the plasmid characteristics of mcr-positive Salmonella identified from patients in Sichuan, China during 2014 to 2017 by whole genomes sequencing. In this study, a total of 12 mcr-positive isolates (1.15%, ; mcr-1, n=10; mcr-3, n=2) were identified from 1046 Salmonella isolates using PCR. Further characterization of these isolates was performed through antimicrobial susceptibility testing, conjugation assays, whole genome sequencing, and bioinformatics analysis. The mcr-1 gene in these isolates were carried by three types of typical mcr-1-bearing plasmids widely distributed in Enterobacteriaceae (IncX4, IncI2 and IncHI2). Of note, two mcr-1-harboring IncHI2 plasmids were integrated into chromosomes by insertion sequences. Two mcr-3-bearing plasmids were IncC and IncFIB broad-host-range plasmids respectively. Genetic context analysis found that mcr-1 was mainly located in Tn6330 or truncated Tn6300, and mcr-3 shared a common genetic structure tnpA-mcr-3-dgkA-ISKpn40. Overall, we found that mcr gene in clinical Salmonella were commonly carried by broad-host plasmids and have potential to transfer into other bacteria by these plasmids. Continuous surveillance of MDR Salmonella in humans and investigation the underlying transmission mechanisms of ARGs are vital to curb the current severe AMR concern.

Ambient PM2.5 and its components associated with 10-year atherosclerotic cardiovascular disease risk in Chinese adults.

BACKGROUND: Exposure to particulate matter with aerodynamic diameters less than 2.5 µm (PM2.5) may increase the risk of 10-year atherosclerotic cardiovascular disease (ASCVD) risk. While PM2.5 is comprised of various components, the evidence on the correlation of its components with 10-year ASCVD risk and which component contributes most remains limited. METHODS: Data were derived from the baseline assessments of China Multi-Ethnic Cohort (CMEC). In total, 69,722 individuals aged 35-74 years were included into this study. The annual average concentration of PM2.5 and its components (black carbon, ammonium, nitrate, sulfate, organic matter, soil particles, and sea salt) were estimated by satellite remote sensing and chemical transport models. The ASCVD risk of individuals was calculated by the equations from the China-PAR Project (prediction for ASCVD risk in China). The relationship between single exposure to PM2.5 and its components and predicted 10-year ASCVD risk was assessed using the logistic regression model. The effect of joint exposure was estimated, and the most significant contributor was identified using the weighted quantile sum approach. RESULTS: Totally 69,722 participants were included, of which 95.8 % and 4.2 % had low and high 10-year ASCVD risk, respectively. Per standard deviation increases in the 3-year average concentration of PM2.5 mass (odds ratio [OR] 1.23, 95 % confidence interval [CI]: 1.12-1.35), black carbon (1.21, 1.11-1.33), ammonium (1.21, 1.10-1.32), nitrate (1.25, 1.14-1.38), organic matter (1.29, 1.18-1.42), sulfate (1.17, 1.07-1.28), and soil particles (1.15, 1.04-1.26) were related to high 10-year ASCVD risk. The overall effect (1.19, 1.11-1.28) of the PM2.5 components was positively associated with 10-year ASCVD risk, and organic matter had the most contribution to this relationship. Female participants were more significantly impacted by PM2.5, black carbon, ammonium, nitrate, organic matter, sulfate, and soil particles compared to others. CONCLUSION: Long-term exposure to PM2.5 mass, black carbon, ammonium, nitrate, organic matter, sulfate, and soil particles were positively associated with high 10-year ASCVD risk, while sea salt exhibited a protective effect. Moreover, the organic matter might take primary responsibility for the relationship between PM2.5 and 10-year ASCVD risk. Females were more susceptible to the adverse effect.

Association between spicy food and hypertension among Han Chinese aged 30-79 years in Sichuan Basin: a population-based cross-sectional study.

BACKGROUND: While spicy food is believed to have cardiovascular-protective effects, its impact on hypertension remains uncertain due to conflicting findings from previous studies. This study aimed to explore the association between spicy food and hypertension in Sichuan Basin, China. METHODS: The baseline data of 43,657 residents aged 30-79 in the Sichuan Basin were analyzed including a questionnaire survey (e.g., sociodemographics, diet and lifestyle, medical history), medical examinations (e.g., height, body weight, and blood pressure), and clinical laboratory tests (e.g., blood and urine specimens). Participants were recruited by multi-stage, stratified cluster sampling in consideration of both sex ratio and age ratio between June 2018 and February 2019. Multivariable logistic regression was performed to explore the effect of spicy food on hypertension and multivariable linear regression was applied to estimate the effect of spicy food on systolic and diastolic blood pressure (SBP/DBP). RESULTS: Concerning hypertension, negative associations with spicy food consumption were observed only in females: compared to those who do not eat spicy food, the odds ratios of consuming spicy food 6-7 days/week, consuming spicy food with strong strength, and years of eating spicy food-to-age ratio were 0.886 (0.799, 0.982), 0.757 (0.587, 0.977), 0.632 (0.505, 0.792), respectively. No significant association was found in males (All P trends > 0.05). In the stratified analyses, participants in the subgroup who were 50 to 79 years old (OR, 95%CI: 0.814, 0.763, 0.869), habitually snored (OR, 95%CI: 0.899, 0.829, 0.976), had a BMI < 24 kg/m2 (OR, 95%CI: 0.886, 0.810, 0.969), had a normal waist circumference (OR, 95%CI: 0.898, 0.810, 0.997), and had no dyslipidemia (OR, 95%CI: 0.897, 0.835, 0.964) showed a significantly stronger association. For SBP, consuming spicy food had negative effects in both genders, but the effect was smaller in males compared to females: among males, the ß coefficients for consuming spicy food 1-2 days/week, weak strength, and years of eating spicy food-to-age ratio were 0.931 (-1.832, -0.030), -0.639 (-1.247, -0.032), and - 2.952 (-4.413, -1.492), respectively; among females, the ß coefficients for consuming spicy food 3-5 days/week, 6-7 days/week, weak strength, moderate strength, and years of eating spicy food-to-age ratio were - 1.251 (-2.115, -0.388), -1.215 (-1.897, -0.534), -0.788 (-1.313, -0.263), -1.807 (-2.542, -1.072), and - 5.853 (-7.195, -4.512), respectively. For DBP, only a positive association between the years of eating spicy food-to-age ratio and DBP was found in males with ß coefficient (95%CI ) of 1.300 (0.338, 2.263); Little association was found in females (all P trends > 0.05), except for a decrease of 0.591 mmHg ( 95%CI: -1.078, -0.105) in DBP among participants who consumed spicy food 1-2 days/week, compared to those who did not consume spicy food. CONCLUSION: Spicy food may lower SBP and has an antihypertensive effect, particularly beneficial for women and individuals with fewer risk factors in the Sichuan Basin. Spicy food consumption may decrease DBP in women but increase it in men. Further multicenter prospective cohort studies are needed to confirm these findings.

Two a posteriori dietary patterns are associated with risks of hyperuricemia among adults in less-developed multiethnic regions in Southwest China.

Diet is a major determinant of hyperuricemia, but little is known about the association between diet and hyperuricemia in less-developed multiethnic regions (LMERs). This study aimed to assess the hypothesis that dietary patterns (including 2 a-priori and 3 a posteriori dietary patterns) are associated with hyperuricemia in LMERs. The 2 a priori dietary patterns were the Dietary Approaches to Stop Hypertension (DASH) and the alternative Mediterranean diet (aMED). We derived 3 a posteriori dietary patterns from the China Multi-Ethnic Cohort study by principal component factor analysis. Given that those dietary patterns had high geographic discrimination, they were named the Sichuan Basin (economically developed industrial region), Yunnan-Guizhou Plateau (agricultural region), and Qinghai-Tibet Plateau (nomadic region) dietary pattern, respectively. We performed a logistic regression model to investigate associations between dietary patterns and hyperuricemia with potential confounders identified by the directed acyclic graph. Comparing the highest with the lowest quintiles, the Sichuan Basin dietary pattern was positively associated with hyperuricemia, whereas the Yunnan-Guizhou Plateau dietary pattern showed an inverse association with hyperuricemia. For the Qinghai-Tibet Plateau dietary pattern, DASH, or aMED, our results did not show a significant association with hyperuricemia. The results were robust among stratified analyses and different analyzing strategies. The dietary patterns showed great diversity in Southwest China. Both DASH and aMED, which many studies widely recommended, did not show expected beneficial effects on preventing hyperuricemia. However, the plant-based protein dietary pattern in the Yunnan-Guizhou Plateau showed a beneficial association with the risk of hyperuricemia, which can provide insights for dietary guidance in LMERs.

T-Shaped Aptamer-Based LSPR Biosensor Using Ω-Shaped Fiber Optic for Rapid Detection of SARS-CoV-2.

SARS-CoV-2, especially the variant strains, is rapidly spreading around the world. Rapid detection methods for the virus are crucial for controlling the COVID-19 epidemic. Herein, a localized surface plasmonic resonance (LSPR) biosensor based on Ω-shaped fiber optic (Ω-FO) was developed for dual assays of SARS-CoV-2 monitoring. Due to its strong ability to control the orientation and density, a new T-shaped aptamer exhibits enhanced binding affinity toward N proteins. After being combined on the fiber optic surface, the T-shaped aptamer sensitively captured N proteins of SARS-CoV-2 for a direct assay. Further, core-shell structured gold/silver nanoparticles functionalized with a T-shaped aptamer (apt-Ag@AuNPs) can amplify the signal of N protein detection for a sandwich assay. The real-time analytical feature of the dual assays endows time-dependent sensitivity enhancement behavior, which provides a guideline to save analytical time. With those characteristics, the LSPR biosensor has been successfully used to rapidly identify 39 healthy volunteers and 39 COVID-19 patients infected with the ancestral or variant SARS-CoV-2. With the help of simple pretreatment, we obtain a true negative rate of 100% and a true positive rate of 92.3% with a short analysis time of 45 min using the direct assay. Further, the LSPR biosensor could also broaden the detection application range to the surface of cold-chain foods using a sandwich assay. Thus, the LSPR biosensor based on Ω-FO was demonstrated to have broad application potential to detect SARS-CoV-2 rapidly.

[Epidemiological Characteristics of Hand, Foot, and Mouth Disease and the Effect of EV71 Vaccination in Chengdu from 2012 to 2020].

Objective: To analyze the epidemiological characteristics of hand, foot, and mouth disease (HFMD) in Chengdu from 2012 to 2020, to make comparison in order to examine the changes in incidence before and after vaccination was introduced, and to provide basis for the prevention of HFMD in the future. Methods: Descriptive epidemiological methods were adopted to analyze the incidence, mortality and rate of severe cases of HFMD cases reported in Chengdu from 2012 to 2020. Results: From 2012 to 2020, the cumulative cases of HFMD reported in Chengdu were as many as 279216, of which, there were 2201 severe cases and 16 deaths. The incidence increased every other year, reaching 326.43 per 100000 person-years, the highest ever, in 2018. The rate of severe cases and mortality had shown a decreasing trend since 2016. A total of 11892 cases of EV71, CoxA16 and other enteroviruses were detected in the laboratory, accounting for 14.8%, 18.8% and 66.3%, respectively. Since 2016, HFMD cases caused by EV71 virus infection had shown an overall decreasing trend, cases caused by Cox A16 virus infection had increased every other year, and cases caused by other enteroviral infections had shown an overall increasing trend. The reported cases were mainly concentrated in children aged 0-5 years (92.1%), with those in the age group of 1-2 years reporting the highest number of cases. For children of different ages, male patients always outnumber female patients. The geographic distribution showed that the areas with high HFMD incidence were always located in the central part of Chengdu City, and the three districts with the highest incidence growth rate were Qingbaijiang District, Shuangliu District, and Longquanyi District. Temporal distribution of HFMD cases showed an obvious bimodal distribution, with most of the cases concentrated in May through August and October through December of each year. The number of new cases reached the highest (12309 cases) in July 2018. Conclusion: While continuing to promote EV71 vaccination in the future, Chengdu also needs to pay more attention to viral infection serotypes other than EV71 and Cox A16 and conduct research on multivalent vaccines against a variety of enteroviruses. The focus of prevention and control can be placed on areas with high population density, large floating populations, large numbers of agriculture-related communities, and insufficient individual awareness of hygiene. For the second circle of Chengdu city, the disinfection of agriculture-related communities should be strengthened, and information sessions or other health education activities could be organized for individuals and daycare facilities with low awareness of the importance of hygiene. In addition, more attention should be given to the prevention and control of HFMD in the high incidence seasons.

[Investigation of Virulence Variation of Enterovirus 71 Strains during Thermal Adaptive Evolution].

Objective: To study the virulence variation of enterovirus 71 (EV71) during thermal adaptive evolution, providing references for the prevention and control of the EV71-related hand, foot and mouth disease. Methods: Parental strains and thermal-adapted strains originating from EV71 sibling strains (lineage #100 and #101) were used for plaque assay validation, CCK-8 cytotoxicity experiment, and host proteomics studies after Vero cell infection. Plaque morphology and cell inhibition rate of the viral strains were obtained. Mass spectrometry was used to examine and analyze the functions of proteins that were differential expressed in the host cells. Results: Plaque morphology variation was found only in the heat-adapted strain of lineage #101. Increase in cell inhibition rate was observed in all the thermal-adapted strains, but the amount of increase varied in different strains. According to the results of clustering analysis and principal component analysis, after infection of Vero cells, the host cell protein profile of the heat-adapted strains was similar to that of the parental strains and the host cell protein profile of cold-adapted strains was similar to that of cell-adapted strains. It showed that 500 kinds of proteins presented inter-group difference in their expression, with 239 kinds being up-regulated proteins and 261 being down-regulated. The function of the up-regulated proteins were related to post-translational protein modification, while the functions of the down-regulated proteins were related to SRP-dependent cotranslational protein translocation/targeting to membrane and retrograde protein transport. Conclusion: Virulence variations of enterovirus 71 may accompany thermal adaptive evolution, but its mechanism of action still awaits further investigation.

Association of Dietary Approaches to Stop Hypertension diet and Mediterranean diet with blood pressure in less-developed ethnic minority regions.

OBJECTIVE: We aimed to investigate the association of Dietary Approaches to Stop Hypertension (DASH)-style diet and Mediterranean-style diet with blood pressure in less-developed ethnic minority regions (LMERs). DESIGN: Cross-sectional study. SETTING: Dietary intakes were assessed by a validated food frequency questionnaire. Dietary quality was assessed by the DASH-style diet score and the alternative Mediterranean-style diet (aMED) score. The association between dietary quality and blood pressure was evaluated using multivariate linear regression model. We further examined those associations in subgroups of blood pressure level. PARTICIPANTS: A total of 81433 adults from the China Multi-Ethnic Cohort (CMEC) study were included in this study. RESULTS: In the overall population, compared with the lowest quintile, the highest quintile of DASH-style diet score was negatively associated with systolic BP (coefficient: -2.78, 95% CI: -3.15 to -2.41; P-trend<0.001), while the highest quintile of aMED score had a weaker negative association with systolic BP (coefficient: -1.43, 95% CI: -1.81 to -1.05; P-trend<0.001). Both dietary indices also showed a weaker effect on diastolic BP (coefficient for DASH-style diet: -1.06, 95% CI: -1.30 to -0.82; coefficient for aMED: -0.43, 95% CI: -0.68 to -0.19). In the subgroup analysis, both dietary indices showed a stronger beneficial effect on systolic BP in the hypertension group than in either of the other subgroups. CONCLUSION: Our results indicated that the healthy diet originating from Western developed countries can also have beneficial effects on blood pressure in LEMRs. DASH-style diet may be a more appropriate recommendation than aMED as part of a dietary strategy to control blood pressure, especially in hypertensive patients.

Incidence, Etiology, and Environmental Risk Factors of Community-Acquired Pneumonia Requiring Hospitalization in China: A 3-Year, Prospective, Age-Stratified, Multicenter Case-Control Study.

Background: Community-acquired pneumonia (CAP) is a leading infectious cause of hospitalization and death worldwide. Knowledge about the incidence and etiology of CAP in China is fragmented. Methods: A multicenter study performed at 4 hospitals in 4 regions in China and clinical samples from CAP patients were collected and used for pathogen identification from July 2016 to June 2019. Results: A total of 1674 patients were enrolled and the average annual incidence of hospitalized CAP was 18.7 (95% confidence interval, 18.5-19.0) cases per 10000 people. The most common viral and bacterial agents found in patients were respiratory syncytial virus (19.2%) and Streptococcus pneumoniae (9.3%). The coinfections percentage was 13.8%. Pathogen distribution displayed variations within age groups as well as seasonal and regional differences. The severe acute respiratory syndrome coronavirus 2 was not detected. Respiratory virus detection was significantly positively correlated with air pollutants (including particulate matter ≤2.5 µm, particulate matter ≤10 µm, nitrogen dioxide, and sulfur dioxide) and significantly negatively correlated with ambient temperature and ozone content; bacteria detection was opposite. Conclusions: The hospitalized CAP incidence in China was higher than previously known. CAP etiology showed that differences in age, seasons, regions, and respiratory viruses were detected at a higher rate than bacterial infection overall. Air pollutants and temperature have an influence on the detection of pathogens.

Molecular characterization of enterovirus 71 sibling strains for thermal adaption in Vero cells with adaptive laboratory evolution.

Enterovirus 71 is the main pathogen that causes severe and fatal hand-foot-mouth-disease (HFMD) cases. As the enterovirus virus mutation has implications for pathogenesis, vaccine development, antiviral therapy, and epidemiological disease management of the virus. In this study, we investigated the variations of enterovirus 71 in thermal adaption, using the method of adaptive laboratory evolution. The sibling virus strains were isolated from a 2-year-old severe case of HFMD (#100) and her symptomless close contact (#101). Both strains were cultured in Vero cells by serial passage of 36 generations at the temperatures of 28.0 °C, 33.0 °C and 39.5 °C to construct adaptive lineages. According to the comparative analysis of phenotypes between adapted strains and parental strains, differences in growth rate were observed in the sibling lineages and a larger plaque was found mainly in the hot adapted strains for lineage #101. Two sets of adaptive strains from six time points (parental, 12th 17th, 31st, 35th passage and endpoint) were sequenced and analyzed by both Sanger sequencing and Next Generation Sequencing. Several variations in most coding genes and one reverse mutation in 5'UTR was observed, along with the identity of 99.8% for complete genome for both lineages. Notably, thermal specific non-synonymous mutations were found in the gene of VP1\VP3\3A\2C\3C. Moreover, the concurrent mutations A292G, A434G and A355C/T of sibling lineages in VP1 showed quantificational trace with distinguishing patterns for different temperatures, which were suspected to be the thermo-sensitive mutation hotspots. These results highlight the possible rules of thermal adaption in enterovirus 71, produce a novel picture of genome evolution of the virus, and shed light on viral variation and evolution.

Prevalence, Molecular Characterization, and Antimicrobial Susceptibility of Methicillin-Resistant Staphylococcus aureus from Different Origins in Sichuan Province, China, 2007-2015.

Methicillin-resistant Staphylococcus aureus (MRSA) is a major multidrug-resistant (MDR) opportunistic pathogen that is responsible for nosocomial infections worldwide. The emergence of MRSA in food-producing animals has heightened concerns regarding the presence of MRSA in foods having an animal origin. The objectives of this study were to determine the prevalence of MRSA in different sources, including retail food, food-producing animals, and food handlers in Sichuan province. During 2007-2015, S. aureus was isolated from samples having different origins. Susceptibilities of MRSA to a panel of 12 antimicrobial agents were determined according to Clinical and Laboratory Standards Institute (CSLI) procedures. Pulsed-field gel electrophoresis, spa typing, staphylococcal chromosomal cassette (SCC) mec typing, and multilocus sequence typing (MLST) were used to characterize the relationship between these isolates. A total of 756 S. aureus were isolated from 11,067 samples. Of these, 52 isolates were classified as MRSA based on the presence of mecA genes. An antimicrobial susceptibility assay indicated that CHL-CLI-ERY-FOX-OXA-PEN-TET (n = 10) and CHL-CIP-CLI-ERY-FOX-OXA-PEN-TET (n = 8) were the two predominant MDR profiles in the isolates. Using 60% genetic similarity as a cutoff, the 52 MRSA isolates were grouped into 6 clusters having 34 pulsotypes. MLST typing showed seven multilocus sequence types (STs) with ST59 and ST9 being the most common. Six SCCmec types were identified in all MRSA isolates. The MRSA isolates had relatively low prevalence in Sichuan province. Clonal expansion is not involved in the dissemination of MRSA from food origins. Considering the threat of MRSA to public health, further surveillance is required to monitor the prevalence of food-related MRSA.