A Study of the Detection of SARS-CoV-2 by the Use of Electrochemiluminescent Biosensor Based on Asymmetric Polymerase Chain Reaction Amplification Strategy.

A new and reliable method has been constructed for detecting severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2) open reading frames 1ab (ORF1ab) gene via highly sensitive electrochemiluminescence (ECL) biosensor technology based on highly efficient asymmetric polymerase chain reaction (asymmetric PCR) amplification strategy. This method uses magnetic particles coupled with biotin-labeled one complementary nucleic acid sequence of the SARS-CoV-2 ORF1ab gene as the magnetic capture probes, and [Formula: see text]-labeled amino-modified another complementary nucleic acid sequence as the luminescent probes, and then a detection model of magnetic capture probes-asymmetric PCR amplification nucleic acid products-[Formula: see text]-labeled luminescent probes is formed, which combines the advantages of highly efficient asymmetric PCR amplification strategy and highly sensitive ECL biosensor technology, enhancing the method sensitivity of detecting the SARS-CoV-2 ORF1ab gene. The method enables the rapid and sensitive detection of the ORF1ab gene and has a linear range of 1-[Formula: see text] copies/[Formula: see text], a regression equation of [Formula: see text] = [Formula: see text] + 2919.301 ([Formula: see text] = 0.9983, [Formula: see text] = 7), and a limit of detection (LOD) of 1 copy/[Formula: see text]. In summary, it can meet the analytical requirements for simulated saliva and urine samples and has the benefits of easy operation, reasonable reproducibility, high sensitivity, and anti-interference abilities, which can provide a reference for developing efficient field detection methods for SARS-CoV-2.

Identify the Virus-like Models for COVID-19 as Bio-Threats: Combining Phage Display, Spectral Detection and Algorithms Analysis.

The rapid identification and recognition of COVID-19 have been challenging since its outbreak. Multiple methods were developed to realize fast monitoring early to prevent and control the pandemic. In addition, it is difficult and unrealistic to apply the actual virus to study and research because of the highly infectious and pathogenic SARS-CoV-2. In this study, the virus-like models were designed and produced to replace the original virus as bio-threats. Three-dimensional excitation-emission matrix fluorescence and Raman spectroscopy were employed for differentiation and recognition among the produced bio-threats and other viruses, proteins, and bacteria. Combined with PCA and LDA analysis, the identification of the models for SARS-CoV-2 was achieved, reaching a correction of 88.9% and 96.3% after cross-validation, respectively. This idea might provide a possible pattern for detecting and controlling SARS-CoV-2 from the perspective of combining optics and algorithms, which could be applied in the early-warning system against COVID-19 or other bio-threats in the future.

Utility of Epstein-Barr Virus DNA in Nasopharynx Swabs as a Reflex Test to Triage Seropositive Individuals in Nasopharyngeal Carcinoma Screening Programs.

BACKGROUND: Epstein-Barr virus (EBV) DNA detection in the nasopharynx is considered a biomarker for nasopharyngeal carcinoma (NPC). We evaluated its performance as a reflex test to triage EBV seropositives within an NPC screening program in China. METHODS: The study population was embedded within an ongoing NPC screening trial and included 1111 participants who screened positive for anti-EBV VCA (antibodies against EBV capsid antigens)/EBNA1 (EBV nuclear antigen1)-IgA antibodies (of 18 237 screened). Nasopharynx swabs were collected/tested for EBNA1 gene EBV DNA load. We evaluated performance of EBV DNA in the nasopharynx swab as a reflex test to triage EBV serological high-risk (those referred to endoscopy/MRI) and medium-risk (those referred to accelerated screening) individuals. RESULTS: By the end of 2019, we detected 20 NPC cases from 317 serological high-risk individuals and 4 NPC cases from 794 medium-risk individuals. When used to triage serological high-risk individuals, nasopharynx swab EBV DNA was detected in 19/20 cases (positivity rate among cases: 95.0%; 95% CI, 75.1%-99.9%), with a referral rate of 63.4% (201/317, 95% CI, 57.8%-68.7%) and NPC detection rate among positives of 9.5% (19/201, 95% CI, 5.8%-14.4%). The performance of an algorithm that combined serology with triage of serology high-risk individuals using EBV DNA testing yielded a sensitivity of 72.4% (95% CI, 3.0%-81.4%) and specificity of 97.6% (95% CI, 97.2%-97.9%). When used to triage EBV serological medium-risk individuals, the positivity rate among cases was 75.0% (95% CI, 19.4%-99.4%), with a referral rate of 61.8% (95% CI, 58.4%-65.2%) and NPC detection rate among positives of 0.6% (95% CI, 0.1%-1.8%). CONCLUSIONS: Nasopharynx swab EBV DNA showed promise as a reflex test to triage serology high-risk individuals, reducing referral by ca. 40% with little reduction in sensitivity compared to a serology-only screening program.

Construction, Characterization, and Application of a Nonpathogenic Virus-like Model for SARS-CoV-2 Nucleocapsid Protein by Phage Display.

With the outbreak and spread of COVID-19, a deep investigation of SARS-CoV-2 is urgent. Direct usage of this virus for scientific research could provide reliable results and authenticity. However, it is strictly constrained and unrealistic due to its high pathogenicity and infectiousness. Considering its biosafety, different systems and technologies have been employed in immunology and biomedical studies. In this study, phage display technology was used to construct a nonpathogenic model for COVID-19 research. The nucleocapsid protein of SARS-CoV-2 was fused with the M13 phage capsid p3 protein and expressed on the M13 phages. After validation of its successful expression, its potential as the standard for qPCR quantification and affinity with antibodies were confirmed, which may show the possibility of using this nonpathogenic bacteriophage to replace the pathogenic virus in scientific research concerning SARS-CoV-2. In addition, the model was used to develop a system for the classification and identification of different samples using ATR-FTIR, which may provide an idea for the development and evaluation of virus monitoring equipment in the future.

Should we support prophylactic intervention for asymptomatic kidney stones? A retrospective cohort study with long-term follow-up.

The purpose of this study was to evaluate the long-term outcomes of patients undergoing prophylactic intervention or active surveillance for asymptomatic kidney stones and identify factors influencing the outcomes. In this retrospective cohort study, we reviewed the medical records of patients with asymptomatic kidney stones in two institutes between November 2014 and November 2019. Standardized questions were asked via phone calls to supplement the outcomes. Pain, hydronephrosis, stone growth, serious infection, gross hematuria, and spontaneous passage were defined as stone-related events. Future intervention was also recorded to evaluate management. A total of 101 patients with 120 kidney units were enrolled in this study. The median follow-up time was 63 months. The patients were classified into the control group (79 cases) or exposure group (41 cases) according to whether they underwent prophylactic intervention before any stone-related events. Generally, the rates of stone-related events and future intervention were significantly different between the two groups (57.0 vs. 12.2%, p < 0.001; and 31.6 vs. 4.9%, p = 0.002, respectively). After applying stabilized inverse probability of treatment weighted, Cox regression suggested that patients who underwent prophylactic intervention were less likely to experience stone-related events and future intervention (HR = 0.175, and HR = 0.028, respectively). In conclusion, patients who underwent prophylactic intervention had a lower risk of stone-related events and future intervention, although they had some slight complications.

A Study of the Detection of SARS-CoV-2 ORF1ab Gene by the Use of Electrochemiluminescent Biosensor Based on Dual-Probe Hybridization.

To satisfy the need to develop highly sensitive methods for detecting the severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2) and further enhance detection efficiency and capability, a new method was created for detecting SARS-CoV-2 of the open reading frames 1ab (ORF1ab) target gene by a electrochemiluminescence (ECL) biosensor based on dual-probe hybridization through the use of a detection model of "magnetic capture probes-targeted nucleic acids-Ru(bpy)32+ labeled signal probes". The detection model used magnetic particles coupled with a biotin-labeled complementary nucleic acid sequence of the SARS-CoV-2 ORF1ab target gene as the magnetic capture probes and Ru(bpy)32+ labeled amino modified another complementary nucleic acid sequence as the signal probes, which combined the advantages of the highly specific dual-probe hybridization and highly sensitive ECL biosensor technology. In the range of 0.1 fM~10 µM, the method made possible rapid and sensitive detection of the ORF1ab gene of the SARS-CoV-2 within 30 min, and the limit of detection (LOD) was 0.1 fM. The method can also meet the analytical requirements for simulated samples such as saliva and urine with the definite advantages of a simple operation without nucleic acid amplification, high sensitivity, reasonable reproducibility, and anti-interference solid abilities, expounding a new way for efficient and sensitive detection of SARS-CoV-2.

Research Progress and Model Construction for Online Health Information Seeking Behavior.

Under the background of coronavirus disease 2019 (COVID-19), online health information seeking has become one of the most important information needs of the public and even the only channel for health information seeking in this special period. A review of the research on online health information-seeking behavior will help give full play to the previous academic research, further emphasize the necessity of online health information-seeking research, and promote the development of research in this field. This study firstly presents the research overview of online health information-seeking behavior by using the informetric method. Secondly, an overview is carried out from the perspective of online health information platforms, groups, quality, satisfaction, etc., to explore the influencing factors and their relationships in the process of online health information seeking. On this basis, the existing behavioral models are integrated and sorted out to build a new behavioral theoretical model in line with the current online health information seeking.

The Association of Maternal Emotional Status With Child Over-Use of Electronic Devices During the COVID-19 Pandemic.

The quarantine during the COVID-19 pandemic may generate high levels of maternal depression/anxiety, and maternal emotional status may affect child behavioral development. Online education during the pandemic may induce child over-use of electronic-devices. However, child electronic-device over-use (especially among children under 12 who are immature in physical and mental development) during the pandemic has not attracted sufficient attention, and the association of child over-use with maternal emotional status remains unknown. Therefore, this study aims to assess the characteristics of child electronic-device over-use and the association between maternal emotional status and child over-use among 1,300 children from nurseries (<3 years), kindergartens (3-6 years), and primary schools (6-12 years) in Shanghai and Wuhan during COVID-19. Mothers completed an online questionnaire (including the Self-Rating-Depression/Anxiety-Scales and Family-Environment-Scale). The use of electronic devices (mobile-phones, iPads, computers, and televisions) and online courses taken by the children were investigated. Associations of maternal emotional status with electronic-device-use by child age were analyzed. The proportions of children in nurseries, kindergartens and primary schools were 8.5, 44.5, and 47.0%, their percentages following online-courses were 24.5, 48.4, and 99.0%, and their rates of electronic-device over-use were 34.2, 62.2, and 93.4%, respectively. Significant associations were observed between higher maternal anxiety/depression levels and higher risks of mobile-phone/iPad over-use among preschoolers and primary-school students. Lower family intimacy and higher conflict levels were associated with higher maternal depression/anxiety levels and higher risks of electronic-device over-use. Our findings suggested that over-use of electronic-devices among children under 12 was common during COVID-19, especially among children ≥6 years, and online-teaching may exacerbate over-use. Maternal anxiety/depression levels were associated with over-use of portable internet-devices (mobile-phone/iPad), especially among preschoolers and school-aged students, and family environment may mediate the association. These findings may contribute to a better understanding of factors leading to over-use of electronic-device and developing strategies to decrease over-use during COVID-19.

A Vimentin-Targeting Oral Compound with Host-Directed Antiviral and Anti-Inflammatory Actions Addresses Multiple Features of COVID-19 and Related Diseases.

Damage in COVID-19 results from both the SARS-CoV-2 virus and its triggered overactive host immune responses. Therapeutic agents that focus solely on reducing viral load or hyperinflammation fail to provide satisfying outcomes in all cases. Although viral and cellular factors have been extensively profiled to identify potential anti-COVID-19 targets, new drugs with significant efficacy remain to be developed. Here, we report the potent preclinical efficacy of ALD-R491, a vimentin-targeting small molecule compound, in treating COVID-19 through its host-directed antiviral and anti-inflammatory actions. We found that by altering the physical properties of vimentin filaments, ALD-491 affected general cellular processes as well as specific cellular functions relevant to SARS-CoV-2 infection. Specifically, ALD-R491 reduced endocytosis, endosomal trafficking, and exosomal release, thus impeding the entry and egress of the virus; increased the microcidal capacity of macrophages, thus facilitating the pathogen clearance; and enhanced the activity of regulatory T cells, therefore suppressing the overactive immune responses. In cultured cells, ALD-R491 potently inhibited the SARS-CoV-2 spike protein and human ACE2-mediated pseudoviral infection. In aged mice with ongoing, productive SARS-CoV-2 infection, ALD-R491 reduced disease symptoms as well as lung damage. In rats, ALD-R491 also reduced bleomycin-induced lung injury and fibrosis. Our results indicate a unique mechanism and significant therapeutic potential for ALD-R491 against COVID-19. We anticipate that ALD-R491, an oral, fast-acting, and non-cytotoxic agent targeting the cellular protein with multipart actions, will be convenient, safe, and broadly effective, regardless of viral mutations, for patients with early- or late-stage disease, post-COVID-19 complications, and other related diseases. IMPORTANCE With the Delta variant currently fueling a resurgence of new infections in the fully vaccinated population, developing an effective therapeutic drug is especially critical and urgent in fighting COVID-19. In contrast to the many efforts to repurpose existing drugs or address only one aspect of COVID-19, we are developing a novel agent with first-in-class mechanisms of action that address both the viral infection and the overactive immune system in the pathogenesis of the disease. Unlike virus-directed therapeutics that may lose efficacy due to viral mutations, and immunosuppressants that require ideal timing to be effective, this agent, with its unique host-directed antiviral and anti-inflammatory actions, can work against all variants of the virus, be effective during all stages of the disease, and even resolve post-disease damage and complications. Further development of the compound will provide an important tool in the fight against COVID-19 and its complications, as well as future outbreaks of new viruses.

Ethanol Production from Colpomenia sinuosa by an Alginate Fermentation Strain Meyerozyma guilliermondii.

With the consumption of energy and the spread of COVID-19, the demand for ethanol production is increasing in the world. The industrial ethanol fermentation microbes cannot metabolize the alginate component of macro algae, which affects the ethanol yield. In this research, the ethanol production process from macro algae by an alginate fermentation yeast Meyerozyma guilliermondii, especially the pretreatment process of Colpomenia sinuosa, was studied. At the same time, the experimental design of Box-Behnken was carried out to achieve the optimum fermentation performance. The concentration of KH2PO4 (A: 2-6 g.L-1), pH (B: 4-7), reaction time (C: 60-120 h) and temperature (D: 24-34 °C) were variable input parameters. During the ethanol production process, the algae powder was firstly mixed with water at 90 °C for 0.5 h. Later the fermentation culture medium was prepared and then it was fermented by the yeast Meyerozyma guilliermondii to produce ethanol. And the optimal fermentation parameters were as follows: fermentation temperature of 28 °C, KH2PO4 dosage of 4.7 g.L-1, initial pH of 6, and fermentation time of 99 h. The ethanol yield reached 0.268 g.g-1 (ethanol to algae), close to the predicted value of model. The generation of alginate lyase during the fermentation of algae was also examined. The highest alginate lyase activity reached 46.42 U.mL-1.

Associations of childhood unintentional injuries with maternal emotional status during COVID-19.

BACKGROUND: To explore the characteristics of unintentional childhood-injury during the COVID-19 pandemic and assess the association of unintentional-injury with maternal emotional status. METHODS: A cross-sectional survey was conducted with a convenience sample of 1300 children under 12-years-old from 21 schools (including nurseries/ kindergartens/ primary schools) in Wuhan and Shanghai during March to April 2020, and the mothers completed questionnaires online. Self-rating Depression/Anxiety Scales were used to evaluate maternal emotional status, questions on child unintentional-injury were based on the International-Statistical-Classification-of-Diseases-and-Related-Health-Problems-version-10 (ICD-10), and a total of 11 kinds of unintentional injuries were inquired. Information on socio-demographic and family-background factors was also collected. RESULTS: The children of 0-4, 5-9, and 10-12 years accounted for 29.2, 55.2 and 15.6%, respectively, the unintentional-injury rates were 10.29, 4.18 and 3.45%, respectively (P < 0.001), and boys had higher rates than girls. The three leading causes included "being struck by/against", falls and animal bites (traffic-injury accounted for a small proportion). Lower maternal educational, living in suburban/rural (vs. urban) areas, grandparents (vs. mothers) being main caregivers, more child exposure to secondhand smoke, close relatives being suspected/ confirmed COVID-19 cases were associated with a higher risk of child unintentional-injury. After adjusting for related confounders, higher maternal depression levels were associated with a higher risk of unintentional injury. CONCLUSIONS: The characteristics of unintentional childhood injury were different from those in non-pandemic periods. The main causes, risk factors and the association of unintentional injury with maternal depression deserve attention for development of effective measures for preventing children from unintentional injury during COVID-19 pandemic.

Prevalence of Sleep Disturbances and Sleep Quality in Chinese Healthcare Workers During the COVID-19 Pandemic: A Systematic Review and Meta-Analysis.

Objectives: Healthcare workers (HWs) experienced high levels of work stress during the COVID-19 pandemic, leading to a high risk of sleep disturbances. This meta-analysis aimed to explore the prevalence of sleep disturbances and sleep quality in Chinese HWs during the COVID-19 pandemic. Methods: English (PubMed, EMBASE, PsycINFO, Web of Science, and the Cochrane Library) and Chinese databases (WanFang, Chinese National Knowledge Infrastructure, and SinoMed) were systematically and independently searched for relevant studies published from December 1, 2019, to May 20, 2020. The pooled prevalence of sleep disturbances and sleep quality were calculated using a random-effects model. Results: A total of 17 studies involving 12,682 Chinese HWs were included in the meta-analysis. The pooled prevalence of sleep disturbances in Chinese HWs was 45.1% (95% CI: 37.2-53.1%). We found that the prevalence of sleep disturbances varied among frontline, infected, and non-frontline HWs (Q = 96.96, p < 0.001); females and males (Q = 9.10, p = 0.003); studies using different assessment instruments (Q = 96.05, p < 0.001); and studies with different sample sizes (Q = 5.77, p = 0.016) and cut-off values (Q = 62.28, p < 0.001). The pooled mean total score of the Pittsburgh Sleep Quality Index (PSQI) was 9.83 (95% CI: 8.61-11.04). HWs in Wuhan had a higher total PSQI score than those in other regions (Q = 9.21, p = 0.002). Conclusion: Sleep disturbances were common in Chinese HWs during the COVID-19 pandemic, particularly in frontline and infected HWs. Our results indicate the heavy mental health burden on HWs during the COVID-19 pandemic in China and can provide other countries with valuable information to assist HWs during the crisis.