Screening and characterization of potential anti-gout components from Polygonum cuspidatum by integration off-line two-dimensional liquid chromatography-mass spectrometry with affinity ultrafiltration and on-line HPLC-ABTS.

Polygonum cuspidatum (P. cuspidatum) is a traditional herbal medicine with a long history and proven efficacy in treating gout. However, due to the complexity of composition and extensive content distribution, the substance basis of its anti-gout effectiveness is still unclear. A strategy was proposed via integrating off-line two-dimensional liquid chromatography (2D-LC) and targeted rapid screening technology based on ultrafiltration-liquid chromatography-mass spectrometry (UF-LC/MS) and on-line high-performance liquid chromatography-2, 2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (HPLC-ABTS) to accomplish high coverage and high throughput screening of anti-gout components from P. cuspidatum. As a result, twenty components were screened from P. cuspidatum extract with both xanthine oxidase (XOD) inhibitory activity and free radical scavenging activity, then were preliminarily identified by high-resolution electrospray ionization-quadrupole-time-of-flight mass spectrometer (ESI-Q-TOF/MS). The screened results were verified by the in vitro assays. Meanwhile, molecular docking further elucidated that the screened bioactive ingredients had favourable binding capabilities with XOD. The performance of this study can achieve high efficiency and high coverage screening of the anti-gout components from P. cuspidatum, which provides methodology and strategy support for the rapid screening of bioactive ingredients from complex medicinal plants.

Caregivers' understanding of childhood influenza vaccination during the epidemic in China. A mixed-methods study.

Introduction: Influenza vaccination uptake among young children has been poor in China, but it is unclear how it changed during the COVID-19. This study aimed to investigate the uptake status and reasons of childhood influenza vaccination during the pandemic in China. Methods: A mixed-methods study combining a questionnaire survey and semi-structured interviews was conducted in Anhui, Shaanxi, and Guangdong provinces between September and November 2021. 2081 caregivers completed the valid questionnaire. 38 caregivers participated in interviews, and data were analyzed thematically, using deductive and inductive coding. Results: A total of 2081 caregivers completed the valid questionnaire, and 38 caregivers participated in interviews. Among the caregivers, a total of 1796 were in the age group for high-risk groups in the 2019-2020 flu season, and 46.10% reported that their children received influenza vaccination in the 2019-2020 flu season; 43.63% said that they vaccinated their children against influenza in the 2020-2021 flu season. Many caregivers indicated that the adoption of nonpharmacologic interventions (NPIs) during COVID-19 reduced the risk of influenza infection for children. Most caregivers consider the severity of influenza to be low, and some confused the common cold with influenza. Meanwhile, some caregivers lack confidence in the vaccine's effectiveness and importance. They thought that vaccines are not effective in preventing the constantly mutating virus. Despite clear perceptions about the severity of influenza and the effectiveness of the vaccine, we found that most caregivers did not receive any relevant medical information, and the communication about vaccines between caregivers and professional information sources, such as healthcare workers, is inadequate. Hence, caregivers have no scientific evidence to back up their perceptions. In terms of access to vaccination service, caregivers reported conflicts between time of vaccination service and their schedule, and the need for vaccine prices to be reduced. Discussion: Targeted interventions are needed to address caregivers' lack of risk perception on influenza during COVID-19 and promote communication between caregivers and professional information sources. Extending vaccination service hours and increasing the number of vaccine clinics close to residential areas and expansion of financing sources for self-paid vaccination could facilitate the access to influenza vaccination service.

Hyperprogression of cutaneous T cell lymphoma after anti-PD-1 treatment.

BACKGROUNDImmune checkpoint blockade is an emerging treatment for T cell non-Hodgkin's lymphoma (T-NHL), but some patients with T-NHL have experienced hyperprogression with undetermined mechanisms upon anti-PD-1 therapy.METHODSSingle-cell RNA-Seq, whole-genome sequencing, whole-exome sequencing, and functional assays were performed on primary malignant T cells from a patient with advanced cutaneous T cell lymphoma who experienced hyperprogression upon anti-PD-1 treatment.RESULTSThe patient was enrolled in a clinical trial of anti-PD-1 therapy and experienced disease hyperprogression. Single-cell RNA-Seq revealed that PD-1 blockade elicited a remarkable activation and proliferation of the CD4+ malignant T cells, which showed functional PD-1 expression and an exhausted status. Further analyses identified somatic amplification of PRKCQ in the malignant T cells. PRKCQ encodes PKCθ; PKCθ is a key player in the T cell activation/NF-κB pathway. PRKCQ amplification led to high expressions of PKCθ and p-PKCθ (T538) on the malignant T cells, resulting in an oncogenic activation of the T cell receptor (TCR) signaling pathway. PD-1 blockade in this patient released this signaling, derepressed the proliferation of malignant T cells, and resulted in disease hyperprogression.CONCLUSIONOur study provides real-world clinical evidence that PD-1 acts as a tumor suppressor for malignant T cells with oncogenic TCR activation.TRIAL REGISTRATIONClinicalTrials.gov (NCT03809767).FUNDINGThe National Natural Science Foundation of China (81922058), the National Science Fund for Distinguished Young Scholars (T2125002), the National Science and Technology Major Project (2019YFC1315702), the National Youth Top-Notch Talent Support Program (283812), and the Peking University Clinical Medicine plus X Youth Project (PKU2019LCXQ012) supported this work.

The Prevalence, Features, Influencing Factors, and Solutions for COVID-19 Vaccine Misinformation: Systematic Review.

BACKGROUND: During the COVID-19 pandemic, infodemic spread even more rapidly than the pandemic itself. The COVID-19 vaccine hesitancy has been prevalent worldwide and hindered pandemic exiting strategies. Misinformation around COVID-19 vaccines is a vital contributor to vaccine hesitancy. However, no evidence systematically summarized COVID-19 vaccine misinformation. OBJECTIVE: This review aims to synthesize the global evidence on misinformation related to COVID-19 vaccines, including its prevalence, features, influencing factors, impacts, and solutions for combating misinformation. METHODS: We performed a systematic review by searching 5 peer-reviewed databases (PubMed, Embase, Web of Science, Scopus, and EBSCO). We included original articles that investigated misinformation related to COVID-19 vaccines and were published in English from January 1, 2020, to August 18, 2022. We excluded publications that did not cover or focus on COVID-19 vaccine misinformation. The Appraisal tool for Cross-Sectional Studies, version 2 of the Cochrane risk-of-bias tool for randomized trials (RoB 2), and Critical Appraisal Skills Programme Checklist were used to assess the study quality. The review was guided by PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) and registered with PROSPERO (CRD42021288929). RESULTS: Of the 8864 studies identified, 91 observational studies and 11 interventional studies met the inclusion criteria. Misinformation around COVID-19 vaccines covered conspiracy, concerns on vaccine safety and efficacy, no need for vaccines, morality, liberty, and humor. Conspiracy and safety concerns were the most prevalent misinformation. There was a great variation in misinformation prevalence, noted among 2.5%-55.4% in the general population and 6.0%-96.7% in the antivaccine/vaccine hesitant groups from survey-based studies, and in 0.1%-41.3% on general online data and 0.5%-56% on antivaccine/vaccine hesitant data from internet-based studies. Younger age, lower education and economic status, right-wing and conservative ideology, and having psychological problems enhanced beliefs in misinformation. The content, format, and source of misinformation influenced its spread. A 5-step framework was proposed to address vaccine-related misinformation, including identifying misinformation, regulating producers and distributors, cutting production and distribution, supporting target audiences, and disseminating trustworthy information. The debunking messages/videos were found to be effective in several experimental studies. CONCLUSIONS: Our review provides comprehensive and up-to-date evidence on COVID-19 vaccine misinformation and helps responses to vaccine infodemic in future pandemics. TRIAL REGISTRATION: PROSPERO CRD42021288929; https://tinyurl.com/2prejtfa.

Childhood Influenza Vaccination and Its Determinants during 2020-2021 Flu Seasons in China: A Cross-Sectional Survey.

Young children aged 6−59 months are recommended as one of the priority groups for seasonal influenza vaccination in China. This study assessed influenza vaccination coverage and the factors associated with vaccination uptake among children in three Chinese provinces. In September 2021, 2081 caregivers with children <5 years completed self-administered questionnaires as part of a cross-sectional survey. Logistic regression was used to assess determinants of childhood influenza vaccination. A total of 43.63% of respondents reported vaccinating their children against influenza during the 2020−2021 flu season. Caregivers who lived in Anhui province, had a bachelor degree or above, and an annual household income <20,000 RMB were more likely to vaccinate their children against influenza. Confidence in the importance (OR: 2.50; 95%CI: 1.77−3.54), safety (OR: 1.60; 95%CI: 1.29−1.99), and effectiveness (OR: 1.54; 95%CI: 1.23−1.93) of influenza vaccine was significantly associated with childhood vaccine acceptance. Respondents who saw that other caregivers were vaccinating their children had significantly higher odds of vaccinating their own children. Caregivers' receiving positive influence from healthcare workers (OR: 1.33; 95%CI: 1.00−1.77), family members, or friends (OR: 1.30; 95%CI: 1.14−1.49) were also significantly associated with childhood influenza vaccination. Poor access, including conflicts between caregivers' availability and vaccination service schedules and inconvenient transportation to the vaccination site were negatively associated with childhood flu vaccination. To promote childhood influenza vaccination, public health information campaigns need to target wealthier and less educated caregivers to enhance caregivers' confidence in influenza vaccination. Targeted interventions are also needed to optimize access to vaccination services, including extending vaccination service hours and increasing the number of vaccination sites close to residential areas. Interventions are also needed to encourage primary care providers to play a greater role in promoting vaccination. Finally, the dissemination of related information and the public response need to be monitored for the timely understanding of public perceptions.

Erratum: Author Correction: Mapping global acceptance and uptake of COVID-19 vaccination: a systematic review and meta-analysis.

[This corrects the article DOI: 10.1038/s43856-022-00177-6.].

Mapping global acceptance and uptake of COVID-19 vaccination: A systematic review and meta-analysis.

Background: The COVID-19 pandemic exit strategies depend on widespread acceptance of COVID-19 vaccines. We aim to estimate the global acceptance and uptake of COVID-19 vaccination, and their variations across populations, countries, time, and sociodemographic subgroups. Methods: We searched four peer-reviewed databases (PubMed, EMBASE, Web of Science, and EBSCO) for papers published in English from December 1, 2019 to February 27, 2022. This review included original survey studies which investigated acceptance or uptake of COVID-19 vaccination, and study quality was assessed using the Appraisal tool for Cross-Sectional Studies. We reported the pooled acceptance or uptake rates and 95% confidence interval (CI) using meta-analysis with a random-effects model. Results: Among 15690 identified studies, 519 articles with 7,990,117 participants are eligible for meta-analysis. The global acceptance and uptake rate of COVID-19 vaccination are 67.8% (95% CI: 67.1-68.6) and 42.3% (95% CI: 38.2-46.5), respectively. Among all population groups, pregnant/breastfeeding women have the lowest acceptance (54.0%, 46.3-61.7) and uptake rates (7.3%, 1.7-12.8). The acceptance rate varies across countries, ranging from 35.9% (34.3-37.5) to 86.9% (81.4-92.5) for adults, and the lowest acceptance is found in Russia, Ghana, Jordan, Lebanon, and Syria (below 50%). The acceptance rate declines globally in 2020, then recovers from December 2020 to June 2021, and further drops in late 2021. Females, those aged < 60 years old, Black individuals, those with lower education or income have the lower acceptance than their counterparts. There are large gaps (around 20%) between acceptance and uptake rates for populations with low education or income. Conclusion: COVID-19 vaccine acceptance needs to be improved globally. Continuous vaccine acceptance monitoring is necessary to inform public health decision making.

TDO2+ myofibroblasts mediate immune suppression in malignant transformation of squamous cell carcinoma.

Characterization of the dynamic change in the immunological landscape during malignant transformation from precancerous lesions to cancerous lesions in squamous cell carcinoma (SCC) is critical for the application of immunotherapy. Here, we performed single-cell RNA-Seq (scRNA-Seq) of 131,702 cells from 13 cancerous tissues of oral squamous cell carcinoma (OSCC), 3 samples of precancerous oral leukoplakia, and 8 adjacent normal samples. We found that tumor-infiltrating CD4+ and CD8+ T cells were functionally inhibited by immunosuppressive ligands expressed on various types of myeloid cells or neutrophils in the process of oral carcinogenesis. Notably, we identified a subset of myofibroblasts that exclusively expressed tryptophan 2,3-dioxygenase (TDO2). These TDO2+ myofibroblasts were located distally from tumor nests, and both CD4+ and CD8+ T cells were enriched around them. Functional experiments revealed that TDO2+ myofibroblasts were more likely to possess the ability for chemotaxis toward T cells but induced the transformation of CD4+ T cells into Tregs and caused CD8+ T cell dysfunction. We further showed that use of the TDO2 inhibitor LM10 attenuated the inhibitory states of T cells, restored the T cell antitumor response, and prevented the progression of OSCC malignant transformation in murine models. Our study reveals a multistep transcriptomic landscape of OSCC and demonstrates that TDO2+ myofibroblasts are potential targets for immunotherapy.

Influenza vaccination hesitancy and its determinants among elderly in China: A national cross-sectional study.

BACKGROUND: The Chinese elderly face a significant threat from seasonal influenza, owing to the consistently low vaccination coverage. This study investigated the prevalence and determinants of influenza vaccination hesitancy among the Chinese elderly. METHODS: In 2019, 3849 elderly individuals from 10 provinces in China were recruited in a cross-sectional survey. Multinomial logistic regression was applied to investigate the determinants of influenza vaccination hesitancy. RESULTS: Among the elderly respondents, 37.18% expressed some degree of hesitancy towards influenza vaccination: 19.28% were hesitant, and 17.90% refused influenza vaccination, including 19.28% acceptors with doubts and 17.90% refusers. Only 39.10% of the respondents considered themselves as the priority group for influenza vaccination, and 13.93% reported receiving a recommendation for vaccination from healthcare workers. Respondents with higher education levels and from urban areas had significantly higher odds of vaccine hesitancy than their counterparts. Confidence in the safety of vaccines was negatively associated with vaccine hesitancy, but confidence in vaccine efficacy had no such association. Respondents who perceived themselves as highly susceptible to influenza (AOR = 0.85; 95 %CI = 0.77-0.93) and those aware of the elderly as a priority group for influenza vaccination (AOR = 0.51; 95 %CI = 0.41-0.64) had a significantly lower odds of being refusers. CONCLUSION: This study found a high prevalence of hesitancy towards influenza vaccination among the Chinese elderly, especially well-educated and urban-dwelling respondents. The government should address vaccine hesitancy through culturally appropriate communication, subsidies for vaccination, and actively promoting vaccines through primary care professionals.

Single-cell transcriptomics links malignant T cells to the tumor immune landscape in cutaneous T cell lymphoma.

Cutaneous T cell lymphoma (CTCL) represents a heterogeneous group of non-Hodgkin lymphoma distinguished by the presence of clonal malignant T cells. The heterogeneity of malignant T cells and the complex tumor microenvironment remain poorly characterized. With single-cell RNA analysis and bulk whole-exome sequencing on 19 skin lesions from 15 CTCL patients, we decipher the intra-tumor and inter-lesion diversity of CTCL patients and propose a multi-step tumor evolution model. We further establish a subtyping scheme based on the molecular features of malignant T cells and their pro-tumorigenic microenvironments: the TCyEM group, demonstrating a cytotoxic effector memory T cell phenotype, shows more M2 macrophages infiltration, while the TCM group, featured by a central memory T cell phenotype and adverse patient outcome, is infiltrated by highly exhausted CD8+ reactive T cells, B cells and Tregs with suppressive activities. Our results establish a solid basis for understanding the nature of CTCL and pave the way for future precision medicine for CTCL patients.

A spatial and cellular distribution of rabies virus infection in the mouse brain revealed by fMOST and single-cell RNA sequencing.

BACKGROUND: Neurotropic virus infection can cause serious damage to the central nervous system (CNS) in both humans and animals. The complexity of the CNS poses unique challenges to investigate the infection of these viruses in the brain using traditional techniques. METHODS: In this study, we explore the use of fluorescence micro-optical sectioning tomography (fMOST) and single-cell RNA sequencing (scRNA-seq) to map the spatial and cellular distribution of a representative neurotropic virus, rabies virus (RABV), in the whole brain. Mice were inoculated with a lethal dose of a recombinant RABV encoding enhanced green fluorescent protein (EGFP) under different infection routes, and a three-dimensional (3D) view of RABV distribution in the whole mouse brain was obtained using fMOST. Meanwhile, we pinpointed the cellular distribution of RABV by utilizing scRNA-seq. RESULTS: Our fMOST data provided the 3D view of a neurotropic virus in the whole mouse brain, which indicated that the spatial distribution of RABV in the brain was influenced by the infection route. Interestingly, we provided evidence that RABV could infect multiple nuclei related to fear independent of different infection routes. More surprisingly, our scRNA-seq data revealed that besides neurons RABV could infect macrophages and the infiltrating macrophages played at least three different antiviral roles during RABV infection. CONCLUSION: This study draws a comprehensively spatial and cellular map of typical neurotropic virus infection in the mouse brain, providing a novel and insightful strategy to investigate the pathogenesis of RABV and other neurotropic viruses.

Hospitalization Costs and Financial Burden on Families with Children with Depression: A Cross-Section Study in Shandong Province, China.

BACKGROUND: Depression, one of the most frequent mental disorders, affects more than 350 million people of all ages worldwide, with China facing an increased prevalence of depression. Childhood depression is on the rise; globally, and in China. This study estimates the hospitalization costs and the financial burden on families with children suffering from depression and recommends strategies both to improve the health care of children with depression and to reduce their families' financial burden. METHODS: The data were obtained from the hospitalization information system of 297 general hospitals in six regions of Shandong Province, China. We identified 488 children with depression. The information on demographics, comorbidities, medical insurance, hospitalization costs and insurance reimbursements were extracted from the hospital's information systems. Descriptive statistics were presented, and regression analyses were conducted to explore the factors associated with hospitalization costs. STATA14 software was used for analysis. RESULTS: The mean age of children with depression was 13.46 ± 0.13 years old. The availability of medical insurance directly affected the hospitalization costs of children with depression. The children with medical insurance had average total hospitalization expenses of RMB14528.05RMB (US$2111.91) and length of stay in hospital of 38.87 days compared with the children without medical insurance of hospital with expenses of RMB10825.55 (US$1573.69) and hospital stays of 26.54 days. Insured children's mean out-of-pocket expenses (6517.38RMB) was lower than the those of uninsured children (RMB10825.55 or US$1573.69), significant at 0.01 level. Insured children incurred higher treatment costs, drug costs, bed fees, check-up fees, test costs and nursing fees than uninsured patients (p < 0.01). CONCLUSIONS: Children suffering from depression with medical insurance had higher hospitalization costs and longer hospitalization stays than children without medical insurance. While uninsured inpatients experienced larger out-of-pocket costs than insured patients, out-of-pocket hospital expenses strained all family budgets, pushing many, especially low-income, families into poverty-insured or uninsured. The different hospital cost structures for drugs, treatment, bed fees, nursing and other costs, between insured and uninsured children with depression, suggest the need for further investigations of treatment regimes, including over-demand by parents for treatment of their children, over-supply of treatment by medical staff and under-treatment of uninsured patients. We recommend more careful attention paid to diagnosing depression in girls and further reform to China's health insurance schemes-especially to allow migrant families to gain basic medical insurance.

EWASdb: epigenome-wide association study database.

DNA methylation, the most intensively studied epigenetic modification, plays an important role in understanding the molecular basis of diseases. Furthermore, epigenome-wide association study (EWAS) provides a systematic approach to identify epigenetic variants underlying common diseases/phenotypes. However, there is no comprehensive database to archive the results of EWASs. To fill this gap, we developed the EWASdb, which is a part of 'The EWAS Project', to store the epigenetic association results of DNA methylation from EWASs. In its current version (v 1.0, up to July 2018), the EWASdb has curated 1319 EWASs associated with 302 diseases/phenotypes. There are three types of EWAS results curated in this database: (i) EWAS for single marker; (ii) EWAS for KEGG pathway and (iii) EWAS for GO (Gene Ontology) category. As the first comprehensive EWAS database, EWASdb has been searched or downloaded by researchers from 43 countries to date. We believe that EWASdb will become a valuable resource and significantly contribute to the epigenetic research of diseases/phenotypes and have potential clinical applications. EWASdb is freely available at http://www.ewas.org.cn/ewasdb or http://www.bioapp.org/ewasdb.

EWAS: epigenome-wide association study software 2.0.

Motivation: With the development of biotechnology, DNA methylation data showed exponential growth. Epigenome-wide association study (EWAS) provide a systematic approach to uncovering epigenetic variants underlying common diseases/phenotypes. But the EWAS software has lagged behind compared with genome-wide association study (GWAS). To meet the requirements of users, we developed a convenient and useful software, EWAS2.0. Results: EWAS2.0 can analyze EWAS data and identify the association between epigenetic variations and disease/phenotype. On the basis of EWAS1.0, we have added more distinctive features. EWAS2.0 software was developed based on our 'population epigenetic framework' and can perform: (i) epigenome-wide single marker association study; (ii) epigenome-wide methylation haplotype (meplotype) association study and (iii) epigenome-wide association meta-analysis. Users can use EWAS2.0 to execute chi-square test, t-test, linear regression analysis, logistic regression analysis, identify the association between epi-alleles, identify the methylation disequilibrium (MD) blocks, calculate the MD coefficient, the frequency of meplotype and Pearson's correlation coefficients and carry out meta-analysis and so on. Finally, we expect EWAS2.0 to become a popular software and be widely used in epigenome-wide associated studies in the future. Availability and implementation: The EWAS software is freely available at http://www.ewas.org.cn or http://www.bioapp.org/ewas.

A Comparison of Co-methylation Relationships Between Rheumatoid Arthritis and Parkinson's Disease.

Rheumatoid arthritis (RA) is a complex autoimmune disease. Recent studies have identified the DNA methylation loci associated with RA and found that DNA methylation was a potential mediator of genetic risk. Parkinson's disease (PD) is a common neurodegenerative disease. Several studies have indicated that DNA methylation levels are linked to PD, and genes related to the immune system are significantly enriched in PD-related methylation modules. Although recent studies have provided profound insights into the DNA methylation of both RA and PD, no shared co-methylation relationships have been identified to date. Therefore, we sought to identify shared co-methylation relationships linked to RA and PD. Here, we calculated the Pearson's correlation coefficient (PCC) of 225,239,700 gene pairs and determined the differences and similarities between the two diseases. The global co-methylation change between in PD cases and controls was larger than that between RA cases and controls. We found 337 gene pairs with large changes that were shared between RA and PD. This co-methylation relationship study represents a new area of study for both RA and PD and provides new ideas for further study of the shared biological mechanisms of RA and PD.

Comparison of the general co-expression landscapes between human and mouse.

The murine model serves as an important experimental system in biomedical science because of its high degree of similarities at the sequence level with human. Recent studies have compared the transcriptional landscapes between human and mouse, but the general co-expression landscapes have not been characterized. Here, we calculated the general co-expression coefficients and constructed the general co-expression maps for human and mouse. The differences and similarities of the general co-expression maps between the two species were compared in detail. The results showed low similarities in the human and mouse, with only about 36.54% of the co-expression relationships conserved between the two species. These results indicate that researchers should pay attention to these differences when performing research using the expression data of human and mouse. To facilitate use of this information, we also developed the human-mouse general co-expression difference database (coexpressMAP) to search differences in co-expression between human and mouse. This database is freely available at http://www.bioapp.org/coexpressMAP.