A rapid and sensitive fluorescent chromatography with cloud system for MPXV point-of-care diagnosis.

Monkeypox (mpox) is spreading around the world, and its rapid diagnosis is of great significance. In the present study, a rapid and sensitive fluorescent chromatography assisted with cloud system was developed for point-of-care diagnosis of mpox. To screen high affinity antibodies, nanoparticle antigen AaLS-A29 was generated by conjugating A29 onto scaffold AaLS. Immunization with AaLS-A29 induced significantly higher antibody titers and monoclonal antibodies were generated with the immunized mice. A pair of monoclonal antibodies, MXV 14 and MXV 15, were selected for fluorescence chromatography development. The Time-Resolved Fluorescence Immunoassay (TRFIA) was used to develop the chromatography assay. After optimization of the label and concentration of antibodies, a sensitive TRFIA assay with detection limit of 20 pg/mL and good repeatability was developed. The detection of the surrogate Vaccinia virus (VACA) strain Tian Tan showed that the TRFIA assay was more sensitive than the SYBR green I based quantitative PCR. In real samples, the detection result of this assay were highly consistent with the judgement of Quantitative Real-Time PCR (Concordance Rate = 90.48%) as well as the clinical diagnosis (Kappa Value = 0.844, P < 0.001). By combining the portable detection and online cloud system, the detection results could be uploaded and shared, making this detection system an ideal system for point-of-care diagnosis of mpox both in field laboratory and outbreak investigation.

A Divalent Chikungunya and Zika Nanovaccine with Thermostable Self-Assembly Multivalent Scaffold LS-SUMO.

The convergence strategies of antigenic subunits and synthetic nanoparticle scaffold platform improve the vaccine production efficiency and enhance vaccine-induced immunogenicity. Selecting the appropriate nanoparticle scaffold is crucial to controlling target antigens immunologically. Lumazine synthase (LS) is an attractive candidate for a vaccine display system due to its thermostability, modification tolerance, and morphological plasticity. Here, the first development of a multivalent thermostable scaffold, LS-SUMO (SUMO, small ubiquitin-likemodifier), and a divalent nanovaccine covalently conjugated with Chikungunya virus E2 and Zika virus EDIII antigens, is reported. Compared with antigen monomers, LS-SUMO nanoparticle vaccines elicit a higher humoral response and neutralizing antibodies against both antigen targets in mouse sera. Mice immunized with LS-SUMO conjugates produce CD4+ T cell-mediated Th2-biased responses and promote humoral immunity. Importantly, LS-SUMO conjugates possess equivalent humoral immunogenicity after heat treatment. Taken together, LS-SUMO is a powerful biotargeting nanoplatform with high-yield production, thermal stability and opens a new avenue for multivalent presentation of various antigens.

Metabolomic landscape of macrophage discloses an anabolic signature of dengue virus infection and antibody-dependent enhancement of viral infection.

Dengue virus (DENV) infection causes dengue fever, the most prevalent arthropod-transmitted viral disease worldwide. Viruses are acellular parasites and obligately rely on host cell machinery for reproduction. Previous studies have indicated metabolomic changes in endothelial cell models and sera of animal models and patients with dengue fever. To probe the immunometabolic mechanism of DENV infection, here, we report the metabolomic landscape of a human macrophage cell model of DENV infection and its antibody-dependent enhancement. DENV infection of THP-1-derived macrophages caused 202 metabolic variants, of which amino acids occupied 23.7%, fatty acids 21.78%, carbohydrates 10.4%, organic acids 13.37%, and carnitines 10.4%. These metabolomic changes indicated an overall anabolic signature, which was characterized by the global exhaustion of amino acids, increases of cellular fatty acids, carbohydrates and pentoses, but decreases of acylcarnitine. Significant activation of metabolic pathways of glycolysis, pentose phosphate, amino acid metabolism, and tricarboxylic acid cycle collectively support the overall anabolism to meet metabolic demands of DENV replication and immune activation by viral infection. Totally 88 of 202 metabolic variants were significantly changed by DENV infection, 36 of which met the statistical standard (P<0.05, VIP>1.5) of differentially expressed metabolites, which were the predominantly decreased variants of acylcarnitine and the increased variants of fatty acids and carbohydrates. Remarkably, 11 differentially expressed metabolites were significantly distinct between DENV only infection and antibody-dependent enhancement of viral infection. Our data suggested that the anabolic activation by DENV infection integrates the viral replication and anti-viral immune activation.

IgM-specific linear epitopes on the E2 protein for serodiagnosis of Chikungunya.

Chikungunya virus (CHIKV) and Dengue virus (DENV) are vector-borne diseases transmitted by Aedes aegypti and Aedes albopictus that pose a significant threat to global public health. Cases of acute Chikungunya fever often present similar clinical symptoms to other vector-borne diseases, such as Dengue fever. In regions where multiple vector-borne diseases coexist, CHIKV is often overlooked or misdiagnosed as Dengue virus, West Nile virus, Zika virus or other viral infections, which delays its prevention and control. However, IgM antibodies directed against the E2 protein of CHIKV have not yet been generalized to clinical settings due to the low sensitivity and high cost in commercial kits. Indirect ELISA with peptides provides an effective supplementary tool for detecting CHIKV IgM antibodies. Our study aims at examining the potential of linear epitopes on the E2 glycoprotein that specifically bind to IgM antibodies as serodiagnostic tool for CHIKV. The sensitivity of the established peptide indirect ELISA method for detecting clinical samples is significantly better than that of commercial kits, realizing a beneficial supplement to the existing IgM antibody assay. It also established the groundwork for comprehending the biological mechanisms of the CHIKV E2 protein and the advancement of innovative epitope peptide vaccines.

Driving effect of multiplex factors on Mpox in global high-risk region, implication for Mpox based on one health concept.

Mpox is an ongoing viral zoonotic disease epidemic worldwide. Being different from conventional animal-to-human transmission, the present outbreak is mainly caused by human-to-human transmission of Mpox virus, putting forward the risk of worldwide epidemic. The current spatial distribution characteristics and risk area prediction are urgently needed for preparedness for prevention and control of the disease based on the One Health strategy. In the present study, the global outbreak point of Mpox virus were collected and used to predict potential global risk of Mpox virus with ecological niche model constructed with a combination of eco-geographical, anthropoid, meteorological, and host variables. The results showed that human factors are the key to the risk and prevalence of Mpox. The risk map indicated that Mpox may affect extensive areas worldwide. Europe and North America have the highest risk of Mpox. Although most areas have never recorded Mpox before, there are some high-risk areas in Asia. Our findings highlight population density is the most important contributing factor for high-risk area. Many large cities with dense populations, developed transportation, and high migration rate in the world, are in high risks. At present, the spread of Mpox is highly valued in the world and strict prevention and control measures have been taken. However, under the influence of human factors, Mpox has the potential of a global pandemic. The risk area prediction and main risk factors provide key information for targeted preparedness for prevention and control of Mpox outbreak and avoiding potential global epidemic through the One Health approach.

Evaluating the zoonotic potential of RNA viromes of rodents provides new insight into rodent-borne zoonotic pathogens in Guangdong, China.

Emerging and re-emerging infectious diseases have been on the rise, with a significant proportion being zoonotic. Rodents, as the natural reservoirs of numerous diverse zoonotic viruses, pose a substantial threat to human health. To investigate the diversity of known and unknown viruses harbored by rodents in Guangdong (southern province of China), we conducted a comprehensive analysis of viral genomes through metagenomic sequencing of organs from 194 rodents. Our analysis yielded 2163 viral contigs that were assigned to 25 families known to infect a wide range of hosts, including vertebrates, invertebrates, amoebas, and plants. The viral compositions vary considerably among different organs, but not in rodent species. We also assessed and prioritized zoonotic potential of those detected viruses. Ninety-two viral species that are either known to infect vertebrates and invertebrates or only vertebrates were identified, among which 21 are considered high-risk to humans. The high-risk viruses included members of the Hantavirus, Picobirnaviruses, Astroviruses and Pestivirus. The phylogenetic trees of four zoonotic viruses revealed features of novel viral genomes that seem to fit evolutionarily into a zone of viruses that potentially pose a risk of transmission to humans. Recognizing that zoonotic diseases are a One Health issue, we approached the problem of identifying the zoonotic risk from rodent-transmitted disease in the Guangdong province by performing next-generation sequencing to look for potentially zoonotic viruses in these animals.

The microbiome in post-acute infection syndrome (PAIS).

Post-Acute Infection Syndrome (PAIS) is a relatively new medical terminology that represents prolonged sequelae symptoms after acute infection by numerous pathogenic agents. Imposing a substantial public health burden worldwide, PASC (post-acute sequelae of COVID-19 infection) and ME/CFS (myalgic encephalomyelitis/chronic fatigue syndrome) are two of the most recognized and prevalent PAIS conditions. The presences of prior infections and similar symptom profiles in PAIS reflect a plausible common etiopathogenesis. The human microbiome is known to play an essential role in health and disease. In this review, we reviewed and summarized available research on oral and gut microbiota alterations in patients with different infections or PAIS conditions. We discussed key theories about the associations between microbiome dysbiosis and PAIS disease development, aiming to explore the mechanistic roles and potential functions the microbiome may have in the process. Additionally, we discuss the areas of knowledge gaps and propose the potential clinical applications of the microbiome for prevention and treatment of PAIS conditions.

T Cell Activating Thermostable Self-Assembly Nanoscaffold Tailored for Cellular Immunity Antigen Delivery.

Antigen delivery based on non-virus-like particle self-associating protein nanoscffolds, such as Aquifex aeolicus lumazine synthase (AaLS), is limited due to the immunotoxicity and/or premature clearance of antigen-scaffold complex resulted from triggering unregulated innate immune responses. Here, using rational immunoinformatics prediction and computational modeling, we screen the T epitope peptides from thermophilic nanoproteins with the same spatial structure as hyperthermophilic icosahedral AaLS, and reassemble them into a novel thermostable self-assembling nanoscaffold RPT that can specifically activate T cell-mediated immunity. Tumor model antigen ovalbumin T epitopes and the severe acute respiratory syndrome coronavirus 2 receptor-binding domain are loaded onto the scaffold surface through the SpyCather/SpyTag system to construct nanovaccines. Compared to AaLS, RPT -constructed nanovaccines elicit more potent cytotoxic T cell and CD4+ T helper 1 (Th1)-biased immune responses, and generate less anti-scaffold antibody. Moreover, RPT significantly upregulate the expression of transcription factors and cytokines related to the differentiation of type-1 conventional dendritic cells, promoting the cross-presentation of antigens to CD8+ T cells and Th1 polarization of CD4+ T cells. RPT confers antigens with increased stability against heating, freeze-thawing, and lyophilization with almost no antigenicity loss. This novel nanoscaffold offers a simple, safe, and robust strategy for boosting T-cell immunity-dependent vaccine development.

A meta-transcriptomic study of mosquito virome and blood feeding patterns at the human-animal-environment interface in Guangdong Province, China.

Mosquitoes are a formidable reservoir of viruses and important vectors of zoonotic pathogens. Blood-fed mosquitoes have been utilized to determine host infection status, overcoming the difficulties associated with sampling from human and animal populations. Comprehensive surveillance of potential pathogens at the interface of humans, animals, and the environment is currently an accredited method to provide an early warning of emerging or re-emerging infectious diseases and to proactively respond to them. Herein we performed comprehensive sampling of mosquitoes from seven habitats (residential areas, hospital, airplane, harbor, zoo, domestic sheds, and forest park) across five cities in Guangdong Province, China. Our aim was to characterize the viral communities and blood feeding patterns at the human-animal-environment interface and analyze the potential risk of cross-species transmission using meta-transcriptomic sequencing. 1898 female adult mosquitoes were collected, including 1062 Aedes and 836 Culex mosquitoes, of which approximately 12% (n = 226) were satiated with blood. Consequently, 101 putative viruses were identified, which included DNA and RNA viruses, and positive-stranded RNA viruses (+ssRNA) were the most abundant. According to viral diversity analysis, the composition of the viral structure was highly dependent on host species, and Culex mosquitoes showed richer viral diversity than Aedes mosquitoes. Although the virome of mosquitoes from different sampling habitats showed an overlap of 39.6%, multiple viruses were specific to certain habitats, particularly at the human-animal interface. Blood meal analysis found four mammals and one bird bloodmeal source, including humans, dogs, cats, poultry, and rats. Further, the blood feeding patterns of mosquitoes were found to be habitat dependent, and mosquitoes at the human-animal interface and from forests had a wider choice of hosts, including humans, domesticated animals, and wildlife, which in turn considerably increases the risk of spillover of potential zoonotic pathogens. To summarize, we are the first to investigate the virome of mosquitoes from multiple interfaces based on the One Health concept. The characteristics of viral community and blood feeding patterns of mosquitoes at the human-animal-environment interface were determined. Our findings should support surveillance activities to identify known and potential pathogens that are pathogenic to vertebrates.

A Multivalent and Thermostable Nanobody Neutralizing SARS-CoV-2 Omicron (B.1.1.529).

Background: The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) Omicron variants have risen to dominance, which contains far more mutations in the spike protein in comparison to previously reported variants, compromising the efficacy of most existing vaccines or therapeutic monoclonal antibodies. Nanobody screened from high-throughput naïve libraries is a potential candidate for developing preventive and therapeutic antibodies. Methods: Four nanobodies specific to the SARS-CoV-2 wild-type receptor-binding domain (RBD) were screened from a naïve phage display library. Their affinity and neutralizing activity were evaluated by surface plasmon resonance assays, surrogate virus neutralization tests, and pseudovirus neutralization assays. Preliminary identification of the binding epitopes of nanobodies by peptide-based ELISA and competition assay. Then four multivalent nanobodies were engineered by attaching the monovalent nanobodies to an antibody-binding nanoplatform constructed based on the lumazine synthase protein cage nanoparticles isolated from the Aquifex aeolicus (AaLS). Finally, the differences in potency between the monovalent and multivalent nanobodies were compared using the same methods. Results: Three of the four specific nanobodies could maintain substantial inhibitory activity against the Omicron (B.1.1.529), of them, B-B2 had the best neutralizing activity against the Omicron (B.1.1.529) pseudovirus (IC50 = 1.658 µg/mL). The antiviral ability of multivalent nanobody LS-B-B2 was improved in the Omicron (B.1.1.529) pseudovirus assays (IC50 = 0.653 µg/mL). The results of peptide-based ELISA indicated that LS-B-B2 might react with the linear epitopes in the SARS-CoV-2 RBD conserved regions, which would clarify the mechanisms for the maintenance of potent neutralization of Omicron (B.1.1.529) preliminary. Conclusion: Our study indicated that the AaLS could be used as an antibody-binding nanoplatform to present nanobodies on its surface and improve the potency of nanobodies. The multivalent nanobody LS-B-B2 may serve as a potential agent for the neutralization of SARS-CoV-2 variants.

Effect of dietary consumption on the survival of esophageal squamous cell carcinoma: a prospective cohort study.

BACKGROUND/OBJECTIVES: This prospective cohort study was to assess the association of pre-diagnostic dietary intake and dietary pattern with the survival of esophageal squamous cell carcinoma (ESCC) patients. SUBJECTS/METHODS: 855 patients were recruited and successfully followed. Information on diet over past five years before diagnosis was collected using a food frequency questionnaire, and dietary patterns were extracted using principal component analysis. Hazard ratio (HR) with 95% confidence interval (95% CI) was calculated using the Cox proportional hazard model. RESULTS: 164 (19.18%) ESCC patients survived during the follow-up. Every 25-g increment intake of pickled vegetables was associated with a 6.0% (HR: 1.060, 95% CI: 1.003-1.121) increased risk of death after adjustment for covariates. When comparing the highest with lowest tertiles of energy-adjusted intake, pickled vegetables intake was associated with a 21.9% elevated risk of death (HR: 1.219, 95% CI: 1.014-1.465), while fish and shrimp intake was associated with a 19.4% (HR: 0.816, 95% CI: 0.675-0.986) reduced risk of death. Three dietary patterns were defined and labeled as patterns I, II, and III. Every 10-score increment of dietary pattern II, characterized with a higher loading of preserved vegetables, pickled vegetables, and salted meat, was associated with a 1.7% (HR: 1.017, 95% CI: 1.003-1.032) increased risk of death. CONCLUSIONS: A diet characterized with higher loading of preserved vegetables, pickled vegetables, and salted meat, was negatively associated with death risk among ESCC patients. Prospective studies concerning the role of post-diagnosis dietary intake in ESCC prognosis are needed.

Effect of physical activity on the risk of frailty: A systematic review and meta-analysis.

OBJECTIVE: The relationship between physical activity (PA) and the risk of frailty has not reached a conclusive result. This systematic review with meta-analysis aimed to evaluate the effect of PA on the onset of frailty in the community-dwelling middle and older age adults by pooling data from cohort studies. METHODS: A systematic literature search was performed via PubMed, Embase, and Web of Science up to June 01, 2021. Pooled adjusted effect estimates (ES) with 95% confidence interval (CI) were calculated by using the random-effect model and by comparing the highest with lowest levels of PA. Heterogeneity was tested using the I2 statistic and Q-test. The quality of evidence was evaluated by using the Grading of Recommendations Assessment, Development and Evaluation (GRADE) approach. RESULTS: A total of ten cohort studies with 14 records were selected, and the GRADE approach classified the quality of evidence as low. In comparison with the lowest level of PA, the highest level of PA was associated with 41% decreased odds of frailty (ES: 0.59, 95% CI: 0.51-0.67; I2 = 70.0%, P-heterogeneity < 0.001) after pooling results from included studies. In stratified analysis by frailty assessment approach, the highest level of PA was significantly associated with 37% (ES 0.63, 95% CI: 0.52-0.77, 49% (ES: 0.51, 95% CI: 0.41-0.63), and 30% (ES: 0.70, 95% CI: 0.65-0.75) reduced odds of frailty when pooling studies using criteria of physical frailty, multidimensional model, and accumulation of disability, respectively. Stratified analyses further by PA indicators and PA assessment tools yielded similar protective effects in any subgroups. CONCLUSIONS: This study with moderate-certainty evidence shows that a higher level of PA was associated with lower odds of frailty, and the benefits of PA for frailty prevention were independent of frailty assessment tools, PA indicators, and PA assessment methods. Findings from this study may help implement active exercise strategies to prevent frailty.

Meta-analysis of 16S rRNA microbial data identified alterations of the gut microbiota in COVID-19 patients during the acute and recovery phases.

BACKGROUND: Dozens of studies have demonstrated gut dysbiosis in COVID-19 patients during the acute and recovery phases. However, a consensus on the specific COVID-19 associated bacteria is missing. In this study, we performed a meta-analysis to explore whether robust and reproducible alterations in the gut microbiota of COVID-19 patients exist across different populations. METHODS: A systematic review was conducted for studies published prior to May 2022 in electronic databases. After review, we included 16 studies that comparing the gut microbiota in COVID-19 patients to those of controls. The 16S rRNA sequence data of these studies were then re-analyzed using a standardized workflow and synthesized by meta-analysis. RESULTS: We found that gut bacterial diversity of COVID-19 patients in both the acute and recovery phases was consistently lower than non-COVID-19 individuals. Microbial differential abundance analysis showed depletion of anti-inflammatory butyrate-producing bacteria and enrichment of taxa with pro-inflammatory properties in COVID-19 patients during the acute phase compared to non-COVID-19 individuals. Analysis of microbial communities showed that the gut microbiota of COVID-19 recovered patients were still in unhealthy ecostates. CONCLUSIONS: Our results provided a comprehensive synthesis to better understand gut microbial perturbations associated with COVID-19 and identified underlying biomarkers for microbiome-based diagnostics and therapeutics.

The role of dietary carotenoids in preventing the development of esophageal squamous cell carcinoma.

Background and aims: Experimental studies showed that carotenoids had anti-carcinogenesis properties, but epidemiological studies on the association between dietary carotenoids and esophageal squamous cell carcinoma (ESCC) risk were limited, and the findings were inconsistent. This study aimed to investigate the roles of intake of dietary carotenoids in the development of ESCC among a rural Chinese population. Methods: A population-based case-control study was conducted in Southwest China. A total of 915 incident ESCC cases and 925 community-based controls were included. A validated food frequency questionnaire with 76-item was adopted to collect information about dietary consumption. Intake of dietary calories and each carotenoid was calculated according to the China food composition tables. Odds ratios (OR) with 95% confidence intervals (CI) were calculated by using logistic regression model, with adjustments for age, gender, body mass index, family cancer history, cigarette smoking, alcohol drinking, education, marital status, prudent pattern score, and total calories. Results: In comparison of the highest with lowest intake quartiles, intake of total carotene (OR: 0.70, 95% CI: 0.52-0.96, Ptrend: 0.024), α-carotene (OR: 0.62, 95% CI: 0.46-0.83, Ptrend: 0.014), ß-carotene (OR: 0.63, 95% CI: 0.46-0.86, P-trend: 0.005), and the sum of lutein and zeaxanthin (OR: 0.41, 95% CI: 0.29-0.56, Ptrend<0.001) was significantly associated with a decreased risk of ESCC after adjustment for confounders. Conclusions: The results indicated that a higher intake of total carotene, α-carotene, ß-carotene, and the sum of lutein and zeaxanthin was associated with a lower risk of ESCC.

Inhibition of Chikungunya virus early replication by intracellular nanoantibodies targeting nsP2 Epitope Rich Region.

Chikungunya fever, caused by Chikungunya virus (CHIKV), is an Aedes mosquito-borne disease present worldwide, and millions of CHIKV infections have been reported. Treatment for CHIKV includes supportive care and anti-inflammatory medications, but there are currently no antiviral treatments or vaccines. Nonstructural protein 2 (nsP2) of CHIKV is the most important functional protein mediating virus replication and amplification, making it an ideal antiviral target for CHIKV. In this study, we determined the CHIKV nsP2 Epitope Rich Region, expressed recombinant nsP2 protein, and isolated 5 nsP2-specific nanobodies (Nb-A2, Nb-A9, Nb-D7, Nb-D12 and Nb-E12) from a phage display library comprising variable domains of Camellidae heavy chain-only antibodies (VHH). We subsequently established a stable Nbs-expressing HEK293T cell line to explore antiviral function. The results showed that Nb-A9 inhibited CHIKV replication at the early stage of CHIKV infection in HEK293T cells, and protected cells against CHIKV-induced cytopathic effect (CPE). This is possibly the first report of an Nbs-based strategy against CHIKV nsP2, Nb-A9 has great potential for developing a novel antiviral drug to treat CHIKV infection. The acquisition of antibodies has laid a foundation for further research on the function of CHIKV nsP2 and the development of therapeutic drugs.

Collaboration between meteorology and public health: Predicting the dengue epidemic in Guangzhou, China, by meteorological parameters.

Background: Dengue has become an increasing public health threat around the world, and climate conditions have been identified as important factors affecting the transmission of dengue, so this study was aimed to establish a prediction model of dengue epidemic by meteorological methods. Methods: The dengue case information and meteorological data were collected from Guangdong Provincial Center for Disease Prevention and Control and Guangdong Meteorological Bureau, respectively. We used spatio-temporal analysis to characterize dengue epidemics. Spearman correlation analysis was used to analyze the correlation between lagged meteorological factors and dengue fever cases and determine the maximum lagged correlation coefficient of different meteorological factors. Then, Generalized Additive Models were used to analyze the non-linear influence of lagged meteorological factors on local dengue cases and to predict the number of local dengue cases under different weather conditions. Results: We described the temporal and spatial distribution characteristics of dengue fever cases and found that sporadic single or a small number of imported cases had a very slight influence on the dengue epidemic around. We further created a forecast model based on the comprehensive consideration of influence of lagged 42-day meteorological factors on local dengue cases, and the results showed that the forecast model has a forecast effect of 98.8%, which was verified by the actual incidence of dengue from 2005 to 2016 in Guangzhou. Conclusion: A forecast model for dengue epidemic was established with good forecast effects and may have a potential application in global dengue endemic areas after modification according to local meteorological conditions. High attention should be paid on sites with concentrated patients for the control of a dengue epidemic.

Effects of COVID-19 Non-Pharmacological Interventions on Dengue Infection: A Systematic Review and Meta-Analysis.

Non-pharmacological interventions (NPIs) implemented during the coronavirus disease 2019 (COVID-19) pandemic have demonstrated significant positive effects on other communicable diseases. Nevertheless, the response for dengue fever has been mixed. To illustrate the real implications of NPIs on dengue transmission and to determine the effective measures for preventing and controlling dengue, we performed a systematic review and meta-analysis of the available global data to summarize the effects comprehensively. We searched Embase, PubMed, and Web of Science in line with PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) guidelines from December 31, 2019, to March 30, 2022, for studies of NPI efficacy on dengue infection. We obtained the annual reported dengue cases from highly dengue-endemic countries in 2015-2021 from the European Centre for Disease Prevention and Control to determine the actual change in dengue cases in 2020 and 2021, respectively. A random-effects estimate of the pooled odds was generated with the Mantel-Haenszel method. Between-study heterogeneity was assessed using the inconsistency index (I2 ) and subgroup analysis according to country (dengue-endemic or non-endemic) was conducted. This review was registered with PROSPERO (CRD42021291487). A total of 17 articles covering 32 countries or regions were included in the review. Meta-analysis estimated a pooled relative risk of 0.39 (95% CI: 0.28-0.55), and subgroup revealed 0.06 (95% CI: 0.02-0.25) and 0.55 (95% CI: 0.44-0.68) in dengue non-endemic areas and dengue-endemic countries, respectively, in 2020. The majority of highly dengue-endemic countries in Asia and Americas reported 0-100% reductions in dengue cases in 2020 compared to previous years, while some countries (4/20) reported a dramatic increase, resulting in an overall increase of 11%. In contrast, there was an obvious reduction in dengue cases in 2021 in almost all countries (18/20) studied, with an overall 40% reduction rate. The overall effectiveness of NPIs on dengue varied with region and time due to multiple factors, but most countries reported significant reductions. Travel-related interventions demonstrated great effectiveness for reducing imported cases of dengue fever. Internal movement restrictions of constantly varying intensity and range are more likely to mitigate the entire level of dengue transmission by reducing the spread of dengue fever between regions within a country, which is useful for developing a more comprehensive and sustainable strategy for preventing and controlling dengue fever in the future.

Effect of Integrated Care on Patients With Atrial Fibrillation: A Systematic Review of Randomized Controlled Trials.

Aims: The integrated management was evidenced to improve the hospitalization and its associated complications in patients with atrial fibrillation (AF), but the strategies of integrated care varied and results were inconsistent. This systematic review and meta-analysis aimed to evaluate the effect of integrated care on AF-related outcomes with comparison with usual care. Methods: PubMed, Embase, and Web of Science were searched for articles published until 10th January 2022. Eligible studies were randomized controlled trials to study the effect of integrated care on AF-related outcomes. Meta-analysis with a random-effect model was used to calculate risk ratio (RR) and 95% confidence interval (CI) by comparing the integrated care with usual care. Results: A total of five studies with 6,486 AF patients were selected. By synthesizing available data, integrated care effectively reduced the risk of all-cause mortality (RR = 0.54, 95% CI = 0.42-0.69), cardiovascular hospitalization (RR = 0.72, 95% CI = 0.55-0.94), and cardiovascular mortality (RR = 0.52, 95% CI = 0.36-0.78) when compared with usual care; however, there was no superior effect on preventing AF-related hospitalization (RR = 0.86, 95% CI = 0.72-1.02), cerebrovascular events (RR = 1.13, 95% CI = 0.75-1.70), and major bleeding (RR = 1.29, 95% CI = 0.86-1.94) when comparing integrated care with usual care. Conclusion: Integrated care can reduce the risk of all-cause mortality, cardiovascular mortality, and cardiovascular hospitalizations in AF patients compared with usual care, while the benefit was not observed in other outcomes.

Association of Dietary Phytosterols Intake and Survival of Esophageal Squamous Cell Carcinoma: A Prospective Cohort Study.

BACKGROUND: The association of dietary phytosterols intake with survival of esophageal squamous cell carcinoma (ESCC) remains unclear. This study was to examine the effect of dietary phytosterols intake on ESCC survival in a Chinese rural population. METHODS: A total of 942 incident ESCC patients diagnosed between 2011 and 2013 in Yanting area were followed up until March 1st, 2020. Dietary intake five years before ESCC diagnosis was collected using a food frequency questionnaire. The outcome of interest was all-cause mortality. Cox proportional hazards regression model was used to estimate hazard ratio (HR) and 95% confidence intervals (CI). RESULTS: When comparing the highest with lowest intake quartiles, intake of five specific and total phytosterols was not significantly associated with risk of death after adjustment for covariates, the adjusted HR (95% CI) for ß-sitosterol, campesterol, stigmasterol, ß-sitostanol, campestanol and total phytosterols was 0.90 (95% CI: 0.70-1.16), 0.92 (95% CI: 0.71-1.19), 0.86 (95% CI: 0.66-1.12), 0.93 (95% CI: 0.73-1.20), 0.94 (95% CI: 0.72-1.21), 0.89 (95% CI: 0.69-1.15), respectively. CONCLUSION: This study does not find any association between pre-diagnostic phytosterols intake and risk of all-cause mortality among ESCC patients. Further research is required to determine the effect of post-diagnostic phytosterols intake on ESCC survival.

Highly potent multivalent VHH antibodies against Chikungunya isolated from an alpaca naïve phage display library.

BACKGROUND: Chikungunya virus (CHIKV) is a re-emerged mosquito-borne alphavirus that can cause musculoskeletal diseases, imposing a substantial threat to public health globally. High-affinity antibodies are need for diagnosis and treatment of CHIKV infections. As a potential diagnostic and therapeutic agent, the multivalent VHH antibodies is a promising tookit in nanomedicine. Here, we developed potent multivalent VHH antibodies from an alpaca naïve phage display library targeting the E2 glycoprotein of the CHIKV virus. RESULTS: In the present study, we generated 20 VHH antibodies using a naïve phage display library for binders to the CHIKV E2 glycoprotein. Of these, multivalent VHH antibodies Nb-2E8 and Nb-3C5 had specific high-affinity binding to E2 protein within the nanomolar range. The equilibrium dissociation constant (KD) was between 2.59-20.7 nM, which was 100-fold stronger than the monovalent antibodies' affinity. Moreover, epitope mapping showed that Nb-2E8 and Nb-3C5 recognized different linear epitopes located on the E2 glycoprotein domain C and A, respectively. A facile protocol of sandwich ELISA was established using BiNb-2E8 as a capture antibody and HRP-conjugated BiNb-3C5 as a detection antibody. A good linear correlation was achieved between the OD450 value and the E2 protein concentration in the 5-1000 ng/mL range (r = 0.9864, P < 0.0001), indicating its potential for quantitative detection of the E2 protein. CONCLUSIONS: Compared to monovalent antibodies, multivalent VHH antibodies Nb-2E8 and Nb-3C5 showed high affinity and are potential candidates for diagnostic applications to better detect CHIKV virions in sera.