Sputum production and salivary microbiome in COVID-19 patients reveals oral-lung axis (preprint)

Objective: SARS-CoV-2 is a severe respiratory disease that primarily targets the lungs and was the leading cause of death worldwide during the pandemic. Investigating the intricate interplay between the oral microbiome and inflammatory cytokines during the acute phase of infection is crucial for understanding host immune responses. This study aimed to explore the relationship between the oral microbiome and cytokines in COVID-19 patients, specifically examining those with and without sputum production. Methods: Saliva and blood samples from 50 COVID-19 patients were subjected to 16S ribosomal RNA gene sequencing to analyze the oral microbiome. Additionally, 65 saliva and serum cytokines were assessed using Luminex multiplex analysis. The Mann-Whitney test compared cytokine levels between individuals with and without sputum production. Results: Our study revealed significant differences in the membership (Jaccard dissimilarity: p=0.016) and abundance (PhILR dissimilarity: p=0.048; metagenomeSeq) of salivary microbial communities between COVID-19 patients with and without sputum production. Seven bacterial genera, including Prevotella, Streptococcus, Actinomyces, Atopobium, Filifactor, Leptotrichia, and Selenomonas, were present in statistically higher proportions of patients with sputum production (p<0.05, Fisher's exact test). Eight bacterial genera, including Prevotella, Megasphaera, Stomatobaculum, Leptotrichia, Veillonella, Actinomyces, Atopobium, and Corynebacteria were significantly more abundant in the sputum-producing group, while Lachnoacaerobaculum was notably more prevalent in the non-sputum-producing group (p<0.05, ANCOM-BC).We observed a significant positive correlation between salivary IFN-gamma (Interferon-gamma) and Eotaxin2/CCL24 (chemokine ligand 24) with sputum production. Conversely, negative correlations were noted in serum MCP3/CCL7 (monocyte-chemotactic protein 3/Chemokine ligand 7), MIG/CXCL9 (Monokine induced by gamma/Chemokine ligand 9), IL1 beta (interleukin 1 beta), and SCF (stem cell factor) with sputum production (p<0.05, Mann-Whitney test). Conclusion: Substantial distinctions in salivary microbial communities were evident between COVID-19 patients with and without sputum production, emphasizing the notable impact of sputum production on the oral microbiome and cytokine levels during the acute phase of infection.

[Traditional Chinese medicine treatment of asymptomatic infection of SARS-CoV-2].

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been infected rapidly and is generally susceptible to population. Moreover, it has become the most serious public health problem in the world. In the process of coronavirus disease 2019 (COVID-19) treatment, the traditional Chinese medicine (TCM) intervention has achieved positive efficacy and which is widely recognized. However, the COVID-19 epidemic is still very serious, especially due to the characteristics of asymptomatic infection such as concealment, limitations and subjective symptoms, which has increased the difficulty of prevention and control. In view of asymptomatic infection, isolation is the main management. There is little mention of specific treatment options in each version of COVID-19 treatment plan. Through systematic study of TCM theory, we explored the way of diagnosis and treatment of asymptomatic infections. Based on the theory of latent evil, clearing away latent pathogens to truncate the course of disease and reduce the incidence. Based on the theory of prevention of disease, strengthen the lung and stomach to protect the place from pathogen and prevent the transmission of disease evil. It is supplemented by accurate treatment according to individual, timing and local conditions, in order to provide reference for the treatment of asymptomatic infection.

Exosomes-based particles as inhalable COVID-19 vaccines

Coronavirus disease 2019 (COVID-19), a severely spreading pandemic, has dramatically brought physiological and economical burdens to people. Although the injectable vaccines have some achievements for coronavirus defense, they still generate accompanied pain, untoward reaction and cannot take part in mucosal immunity. Inhalable vaccines, as a safe, facile and efficient strategy, have been presented to protect body from virus by inducing robust mucosal immunity. Here, we give a perspective of an inhalable COVID-19 vaccine composed of lung-derived exosomes (a type of virus-like particle) conjugated with viral receptor-binding domain. The lung-derived exosomes act as carriers, such inhalable particles successfully reach at lung and reveal wider distribution and longer retention on respiratory mucosa. In addition, such vaccines induce the high production of specific antibodies and T cells in lung, significantly protecting host against coronavirus invasion. It is conceived that inhalable virus-like particles with long-term stability wound open a new avenue for vaccines delivery and further achieve vaccine popularization to against with COVID-19 pandemic.

SARS-CoV-2 Spike Protein Reduces Burst Activities in Neurons Measured by Micro-Electrode Arrays (preprint)

SARS-CoV-2 caused a large-scale global pandemic between 2020 and 2022. Despite efforts to understand its biology and mechanisms of pathogenicity, the viral impact on the neurological systems remains unclear. The main goal of this study was to quantify the neurological phenotypes induced by SARS-CoV-2 spike protein in neurons, measured by in-vitro multi-well micro-electrode arrays (MEAs). We extracted the whole-brain neurons from the newborn P1 mice and plated them on multi-well micro-electrode arrays and administered purified recombinant spike proteins (S1 and S2 subunits respectively) from the SARS-CoV-2 virus. The signals from the MEAs were transmitted from an amplifier to a high-performance computer for recording and analysis. We used an in-house developed algorithm to quantify neuronal phenotypes. Among all the phenotypic features analyzed, we discovered that the S1 protein of SARS-CoV-2 decreased the mean burst numbers observed on each electrode; This effect was not observed for the spike 2 protein (S2) and could be rescued by an anti-S1 antibody. Finally, our data strongly suggest that the receptor binding domain (RBD) of S1 is responsible for the reduction of burst activities in neurons. Overall, our results strongly indicate that spike proteins may play an important role in altering neuronal phenotypes, specifically the burst patterns, when neurons are exposed during early development.

Effect of COVID-19 vaccination on semen parameters: A systematic review and meta-analysis.

The aim of this study was to investigate the effect of coronavirus disease 2019 (COVID-19) vaccination on semen parameters through systematic review and meta-analysis. PubMed, EMBASE, Web of Science, and Cochrane Library were comprehensively searched by June 2022. Studies were considered eligible if they compared semen parameters before and after COVID-19 vaccination or between vaccinated and unvaccinated men, with no restrictions on vaccine types or doses. The effect size was calculated as mean difference (MD) with 95% confidence interval (CI) using a random-effects model. Subgroup and sensitivity analyses were conducted to assess the sources of heterogeneity measured by the I2 statistic, with publication bias evaluated by Egger's test. Twelve cohort studies involving 914 participants fulfilled the inclusion criteria. In a comparison of vaccinated versus unvaccinated group, the pooled data revealed no significant differences in semen volume (MD = 0.18 ml, 95% CI -0.02 to 0.38), sperm concentration (MD = 1.16 million/ml, 95% CI -1.34 to 3.66), total sperm motility (MD = -0.14%, 95% CI -2.84 to 2.56), progressive sperm motility (MD = -1.06%, 95% CI -2.88 to 0.77), total sperm count (MD = 5.92 million, 95% CI -10.22 to 22.05), total motile sperm count (MD = 2.18 million, 95% CI -1.28 to 5.63), total progressively motile sperm count (MD = -3.87 million, 95% CI -13.16 to 5.43), and sperm morphology (MD = 0.07%, 95% CI -0.84 to 0.97). The results also remained similar across messenger ribonucleic acid, viral-vector, and inactivated COVID-19 vaccines. Sensitivity analysis identified two individual studies that contributed to heterogeneity, while the effect size was not materially altered. No obvious publication bias was detected among included studies. Our finding suggested that COVID-19 vaccination had no detrimental impact on semen quality, which could be potentially helpful to reduce male vaccine hesitancy and increase vaccination coverage.

A Structural Determinants Framework for Financial Well-Being.

This study develops a conceptual framework that provides a broad understanding of financial well-being. Using the 2018 National Financial Capability Study and structural equation modeling methods, this study provides empirical evidence for the proposed framework by identifying significant direct and indirect determinants of financial well-being. Previous personal financial wellness and financial satisfaction-related research provides a theoretical rationale for the construction of the conceptual framework in the current study. The results reported the relationships among these determinants, including financial perceptions and knowledge factors, financial stress, short- and long-term positive financial behavior, and financial satisfaction. The findings indicate that financial satisfaction, short-term financial behavior, perceived financial capability showed positive and direct associations with financial well-being, whereas financial stress and long-term financial behavior were negatively and directly associated with financial well-being. Financial perception and knowledge factors, financial stress, and short-term financial behavior also showed significant indirect relationships with financial well-being. The findings of this study contribute to the literature on financial well-being and provide significant policy and practical implications. Implications for financial practitioners and policy makers are discussed.

Skint: Retirement? Financial Hardship and Retirement Planning Behaviors.

This study used data from the 2018 National Financial Capability Study to investigate the association between financial hardship and retirement planning behaviors. Results from logistic regressions showed that respondents with high difficulty making ends meet were more likely to calculate retirement needs and more likely to own a non-employer sponsored retirement plan. The perceived over-indebtedness was positively associated with owning an employer-sponsored account while negatively associated with owning a non-employer-sponsored account. Financial fragility was associated with a lower likelihood of calculating retirement needs and having a retirement account. The results of additional generational analyses revealed that the difficulty making ends meet and the perceived over-indebtedness showed different patterns with retirement planning behavior across three generations. In contrast, financial fragility showed consistent and negative associations with the retirement planning behaviors across generations.

Community Innovation and Regional Sustainability: A Case Study on an Elder Social Survey Conducted by National Cheng Kung University

National Cheng Kung University (NCKU) promotes university social responsibility and strives to ensure that all of its teaching staff and students engage in academics in a manner that incorporates both social awareness and social practice. The NCKU team has launched a series of social practice and research projects focused on accompanying and caring for older adults who live in the community, looking to establish a social practice model that bridges university teaching and research to practical community needs. The objective of this initiative is to develop an innovative model of social support and a new model for younger generations to work with older adults living in the community. The results of this survey project show better well-being in older adults to be associated with the following sets of traits: exercising regularly and lacking dental problems;engaging in daily laughter and holding positive aging perceptions;and having a higher subjective social status, more family support, and a sense of community attachment. The study results imply that multiple factors affect well-being in the context of caring for community-dwelling older adults and social development, especially during the current COVID-19 pandemic.

CLEC4s as Potential Therapeutic Targets in Hepatocellular Carcinoma Microenvironment.

Immunosuppressive tumor microenvironment in hepatocellular carcinoma (HCC) is critical in tumor development. C-type (Ca2+ -dependent) lectin (CLEC) receptors, essential in innate pattern recognition, have potential regulatory effects on immune cell trafficking and modulatory effects on cancer cell activity. However, information on the expression and prognostic value of CLECs in HCC is scanty. Herein, we explored the potential role of CLECs in HCC based on TCGA, ONCOMINE, GEPIA, UALCAN, cBioPortal, Metascape, TRRUST, and TIMER databases. Results demonstrated a significantly higher mRNA level of CLEC4A and CLEC4L in HCC tissues than normal liver tissues. Contrarily, we found significantly low CLEC4G/H1/H2/M expression in HCC tissues. The IHC analysis revealed the following: Absence of CLEC4A/J/K/M in normal and liver cancer tissues; high CLEC4C expression in HCC tissues; low expression and zero detection of CLEC4D/E/H1/H2/L in HCC tissues and normal tissues, respectively. And the HepG2 and LX-2 were used to verify the expression level of CLEC4s via qRT-PCR in vitro. Furthermore, the expression of CLEC4H1 (ASGR1) and CLEC4H2 (ASGR2) exhibited a significant relation to clinical stages. However, the expression of CLEC4A, CLEC4D, CLEC4E, CLEC4J (FCER2), CLEC4K (CD207), CLEC4G, CLEC4H1, CLEC4M, and CLEC4H2 decreased with tumor progression. Patients expressing higher CLEC4H1/H2 levels had longer overall survival than patients exhibiting lower expression. Moreover, CLEC4A/D/E/J/K/G/H1/M/H2 had significant down-regulated levels of promoter methylation. The expression level of CLEC4s was correlated with the infiltration of B cells, CD8 + T cells, CD4 + T cells, macrophage cells, neutrophil cells, and dendritic cells. Functional analysis revealed the potential role of CLECL4s in virus infection, including COVID-19. Also, hsa-miR-4278 and hsa-miR-324-5p, two potential miRNA targets of CLEC4s, were uncovered. This article demonstrates that CLEC4 is crucial for the development of HCC and is associated with infiltration of various immune cells, providing evidence for new immunotherapy targets in HCC.

Evaluation of serum hepatic enzyme activities in different COVID-19 phenotypes.

Coronavirus disease 2019 (COVID-19) is a newly emerging infectious disease. Our understanding of the clinical characteristics of liver damage and the relationship with disease severity in COVID-19 is still limited. To investigate the serum hepatic enzyme activities in different phenotypes of COVID-19 patients, evaluate their relationship with the illness severity and analyze the correlation of glycyrrhizin treatment and abnormal liver enzyme activities, one hundred and forty-seven patients with COVID-19 were enrolled in a retrospective study that investigated hepatic dysfunction. Liver alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactic dehydrogenase (LDH), Y-glutamyl transferase (GGT), superoxide dismutase (SOD), and alkaline phosphatase (ALP) were analyzed in these patients. Patients with diammonium glycyrrhizinate (DG) treatment were further investigated. Of the 147 patients, 56 (38.1%) had abnormal ALT activity and 80 (54.4%) had abnormal AST activity. The peak of abnormal hepatic enzyme activities occurred at 3 to 6 days after on admission. Serum AST and LDH levels were elevated, while the SOD level was decreased in severe and critical patients, compared with mild cases. DG treatment may alleviate the abnormal liver enzyme activities in non-critical COVID-19 patients. Abnormal liver functions may be observed in patients with COVID-19, and were associated with SARS-CoV-2-induced acute liver damage. Glycyrrhizin treatment may be an effective therapeutic approach for the outcome of abnormal hepatic enzyme activities in severe COVID-19 cases. Serum hepatic enzyme tests may reflect the illness severity and should be monitored.

Correlation of Krebs von den Lungen-6 and fibronectin with pulmonary fibrosis in coronavirus disease 2019.

OBJECTIVE: Coronavirus Disease 2019 (COVID-19) caused by a novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is still spreading worldwide, which may progress to pulmonary fibrosis (PF), leading to the worsen outcome. As the markers of lung injury, the correlation of Krebs von den Lungen-6 (KL-6) and fibronectin (Fn) with pulmonary fibrosis in COVID-19 was still unclear. METHODS: 113 patients diagnosed as COVID-19 were enrolled in this retrospective study, and divided into three categories as mild, moderate and severe cases. The concentrations of serum KL-6 and Fn at hospital admission were tested using the method of latex agglutination assay and immunoturbidimetic assay, respectively. RESULTS: Compared with that in the non-severe COVID-19 cases and normal control subjects, serum KL-6 concentration on admission was significantly higher in the severe group, which was positively correlated with C-reactive protein, and negatively correlated with lymphocytes count. Whereas, no obvious elevation in serum Fn concentration was investigated in COVID-19 patients with the different phenotypes. The severe cases displayed the higher incident rate of pulmonary fibrosis at hospital discharge. Compared with non-PF patients, the COVID-19 cases with PF had the higher serum KL-6 values. CONCLUSION: Serum KL-6 concentration was significantly elevated in severe COVID-19 patients, which may be useful for evaluating the disease severity. For early prevention of the development of pulmonary fibrosis, high concentrations of serum KL-6 in the early stage of COVID-19 should be paid close attention.

COVID-19 Quarantine Reveals Grade-specific Behavioral Modification of Myopia: One-Million Chinese Schoolchildren Study (preprint)

BackgroundHigh prevalence of myopia of adolescent has been a global public health concern. Their risk factors and effective prevention methods for myopia across schoolchildren developmental stages are critically needed but remain uncertain due to the difficulty in implementing intervention measurements under normal life situation. We aimed to study the impact of the COVID-19 quarantine on myopia development among over one-million schoolchildren. MethodsWe designed the ongoing longitudinal project of Myopic Epidemiology and Intervention Study (MEIS) to biannually examine myopia among millions of schoolchildren for ten years in Wenzhou City, Zhejiang Province, China. In the present study, we performed three examinations of myopia in 1,305 elementary and high schools for schoolchildren in June 2019, December 2019 and June 2020. We used the normal period (June-December 2019) and COVID-19 quarantine period (January-June 2020) for comparisons. Myopia was defined as an uncorrected visual acuity of 20/25 or less and a spherical equivalent refraction (SER) of -0.5 diopters (D) or less. High myopia was defined as an SER of -6.0 D or less. FindingsIn June 2019, 1,001,749 students aged 7-18 were eligible for examinations. In the 6-month and 12-month follow-up studies, there were 813,755 eligible students (81.2%) and 768,492 eligible students (76.7%), respectively. Among all students, we found that half-year myopia progression increased approximate 1.5 times from -0.263 D (95% CI, -0.262 to -0.264) during normal period to -0.39 D (95% CI, -0.389 to -0.391) during COVID-19 quarantine (P < 0.001). Multivariate Cox regression analysis identified grade rather than age was significantly associated with myopia (Hazard ratio [HR]: 1.10, 95% CI, 1.08 to 1.13; P < 0.001) and high myopia (HR: 1.40, 95% CI, 1.35 to 1.46; P < 0.001) after adjustment for other factors. The prevalence, progression, and incidence of myopia and high myopia could be categorized into two grade groups: I (grades 1-6) and II (grades 7-12). Specifically, COVID-19 quarantine for 6 months sufficiently increased risk of developing myopia (OR: 1.36, 95% CI, 1.33 to 1.40) or high myopia (OR: 1.30, 95% CI, 1.22 to 1.39) in Grade Group I, but decreased risk of developing myopia (OR: 0.45, 95% CI, 0.43 to 0.48) or high myopia (OR: 0.57, 95% CI, 0.54 to 0.59) in Grade Group II. InterpretationThe finding that behavioral modifications for six months during COVID-19 quarantine sufficiently and grade-specifically modify myopia development offers the largest human behavioral intervention data at the one million scale to identify the grade-specific causal factors and effective prevention methods for guiding the formulation of myopia prevention and control policies. FundingKey Program of National Natural Science Foundation of China; the National Natural Science Foundation of China; Scientific Research Foundation for Talents of Wenzhou Medical University; Key Research and Development Program of Zhejiang Province. Research in contextO_ST_ABSEvidence before this studyC_ST_ABSMyopia is the most-common refractive error worldwide. Myopia with younger onset may result in developing high myopia, which is associated with sight-threatening ocular diseases such as maculopathy, retinal detachment, opticneuropathy, glaucoma, retinal atrophy, choroidal neovascularization. In light of the increasing prevalence of myopia and high myopia has been a global public health concern, the impact of COVID-19 lockdown on myopia development has gained substantial attention. We searched PubMed, Google Scholar, and MEDLINE databases for original articles reported between database inception and November 10, 2020, using the following search terms: (coronavirus OR COVID* OR SARS-COV-2 OR lockdown OR quarantine) AND (myopia OR short-sightedness OR refractive error). To date, there was no original study reported to uncover the influence of COVID-19 quarantine on myopia progression. Added value of this studyThis study provides the largest longitudinal intervention data on myopia progression in Chinese schoolchildren covering all grades of schoolchildren at one-million scale. COVID-19 quarantine model uncovers that behavioral modifications for six months may lead to significant increase of overall prevalence of myopia associated with their increased screen times and decreased outdoor activity times. Importantly, their effects on developing myopia or high myopia of students are grade-dependent, which were risk factors for elementary schools period but protective factors for high schools period partly due to reduced school education burden. Implications of all the available evidenceThis one-million schoolchildren myopia survey offers evidence that six months behavioral modifications sufficiently and grade-specifically change the progression of myopia and high myopia. In view of the increased use of electronic devices is an unavoidable trend, effective myopia prevention strategy according to grade among students is urgently needed. Since COVID-19 outbreak is still ongoing and spreading, international collaborate efforts are warranted to uncover the influence of COVID-19 on myopia progression to further substantiate these findings.

Highland of COVID-19 outside Hubei: epidemic characteristics, control and projections of Wenzhou, China (preprint)

In late December 2019, Chinese authorities reported a cluster of pneumonia cases of unknown aetiology in Wuhan, China. Wenzhou, as a southeast coastal city with the most cases outside Hubei Province, its policy control and epidemic projections have certain references for national and worldwide epidemic prevention and control. The maximum number of illness onset in Wenzhou appeared on January 26 (38 cases). Since January 27, the incidence per day in Wenzhou has continued to decline is likely to be due to isolation of people return to Wenzhou, implementation of region quarantine and suspension of public transportation system. Under the current control efforts, we estimate that the daily incidence by March 3-9, 2020 will drop to 0 in Wenzhou using SEIR model. The total number of affected people is 538.