Protocol for characterizing the inhibition of SARS-CoV-2 infection by a protein of interest in cultured cells.

Here, we present a protocol to characterize the antiviral ability of a protein of interest to SARS-CoV-2 infection in cultured cells, using MUC1 as an example. We use SARS-CoV-2 ΔN trVLP system, which utilizes transcription and replication-competent SARS-CoV-2 virus-like particles lacking nucleocapsid gene. We describe the optimized procedure to analyze protein interference of viral attachment and entry into cells, and qRT-PCR-based quantification of viral infection. The protocol can be applied to characterize more antiviral candidates and clarify their functioning stage. For complete details on the use and execution of this protocol, please refer to Lai et al. (2022).

Protocol for characterizing the inhibition of SARS-CoV-2 infection by a protein of interest in cultured cells

Here, we present a protocol to characterize the antiviral ability of a protein of interest to SARS-CoV-2 infection in cultured cells, using MUC1 as an example. We use SARS-CoV-2 ΔN trVLP system, which utilizes transcription and replication-competent SARS-CoV-2 virus-like particles lacking nucleocapsid gene. We describe the optimized procedure to analyze protein interference of viral attachment and entry into cells, and RT-qPCR-based quantification of viral infection. The protocol can be applied to characterize more antiviral candidates and clarify their functioning stage. Graphical Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Identified human breast milk compositions effectively inhibit SARS-CoV-2 and variants infection and replication.

The global pandemic of COVID-19 caused by the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection confers great threat to public health. Human breast milk is a complex nutritional composition to nourish infants and protect them from different kinds of infectious diseases including COVID-19. Here, we identified that lactoferrin (LF), mucin1 (MUC1), and α-lactalbumin (α-LA) from human breast milk inhibit SARS-CoV-2 infection using a SARS-CoV-2 pseudovirus system and transcription and replication-competent SARS-CoV-2 virus-like-particles (trVLP). In addition, LF and MUC1 inhibited multiple steps including viral attachment, entry, and postentry replication, whereas α-LA inhibited viral attachment and entry. Importantly, LF, MUC1, and α-LA possess potent antiviral activities toward variants such as B.1.1.7 (alpha), B.1.351 (beta), P.1 (gamma), and B.1.617.1 (kappa). Taken together, our study provides evidence that human breast milk components (LF, MUC1, and α-LA) are promising antiviral and potential therapeutic candidates warranting further development for treating COVID-19.

Identified human breastmilk compositions effectively inhibit SARS-CoV-2 and variants infection and replication

The global pandemic of COVID-19 caused by the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection confers great threat to the public health. Human breastmilk is a complex with nutritional composition to nourish infants and protect them from different kinds of infectious diseases including COVID-19. Here, we identified lactoferrin (LF), mucin1 (MUC1) and α-lactalbumin (α-LA) from human breastmilk inhibit SARS-CoV-2 infection using a SARS-CoV-2 pseudovirus system and transcription and replication-competent SARS-CoV-2 virus-like-particles (trVLP). Additionally, LF and MUC1 inhibited multiple steps including viral attachment, entry and post-entry replication, while α-LA inhibited viral attachment and entry. Importantly, LF, MUC1 and α-LA possessed potent antiviral activities towards variants such as B.1.1.7 (alpha), B.1.351 (beta), P.1 (gamma) and B.1.617.1 (kappa). Taken together, our study provides evidence that human breastmilk components (LF, MUC1 and α-LA) are promising antiviral and potential therapeutic candidates warranting further development or treating COVID-19. Graphical

Natural triterpenoids from licorice potently inhibit SARS-CoV-2 infection.

Introduction: The COVID-19 global epidemic caused by severe acute respiratory syndrome coronavirus (SARS-CoV-2) is a great public health emergency. Discovering antiviral drug candidates is urgent for the prevention and treatment of COVID-19. Objectives: This work aims to discover natural SARS-CoV-2 inhibitors from the traditional Chinese herbal medicine licorice. Methods: We screened 125 small molecules from Glycyrrhiza uralensis Fisch. (licorice, Gan-Cao) by virtual ligand screening targeting the receptor-binding domain (RBD) of SARS-CoV-2 spike protein. Potential hit compounds were further evaluated by ELISA, SPR, luciferase assay, antiviral assay and pharmacokinetic study. Results: The triterpenoids licorice-saponin A3 (A3) and glycyrrhetinic acid (GA) could potently inhibit SARS-CoV-2 infection, with EC50 of 75 nM and 3.17 µM, respectively. Moreover, we reveal that A3 mainly targets the nsp7 protein, and GA binds to the spike protein RBD of SARS-CoV-2. Conclusion: In this work, we found GA and A3 from licorice potently inhibit SARS-CoV-2 infection by affecting entry and replication of the virus. Our findings indicate that these triterpenoids may contribute to the clinical efficacy of licorice for COVID-19 and could be promising candidates for antiviral drug development.

Collection calendar: the diversity and local knowledge of wild edible plants used by Chenthang Sherpa people to treat seasonal food shortages in Tibet, China.

BACKGROUND: Wild edible plants (WEPs) are non-cultivated and non-domesticated plants used for food. WEPs provided food, nutrition, herbs and other plant products for people in underdeveloped areas, such as the Everest region, to maintain their daily lives. Chenthang Town is the only Sherpa ethnic township in Tibet, China. The core purpose of this research is to investigate, collect and record the WEPs and related local knowledge and functions within the Sherpa community. The ultimate goal is to answer the question of why Sherpa people choose these particular plants. MATERIALS AND METHODS: The field study was carried out in the six Sherpa communities of Chenthang Township from September 2019 to August 2020. The WEPs and related local knowledge were collected through semistructured interviews and direct observations. The field work was performed with the assistance of local guides. During the field survey, we collected plant specimens based on the principle of one plant with one vernacular name. In this study, we utilised a use report (UR) and cultural importance index (CI) to evaluate the comprehensive utilization value of WEPs in the daily diet of Sherpa people. RESULTS: We interviewed 78 people individually who provided us with 1199 use reports. In total, we collected 84 WEPs belonging to 65 genera in 41 families. These species were identified as 78 distinct ethno-species by local people, and the vernacular name of each ethno-species was recorded. Then, these use reports were classified into six use categories. All these plants were native wild plants. In these plants, Arisaema utile, Sorbus cuspidata and Elaeagnus umbellata have been introduced into home gardens by local people. Following the description of the Sherpa people, we articulated a collection calendar for WEPs. The Sherpa collect WEPs throughout nearly the entire year, January and February being the exceptions. CONCLUSION: The collection calendar of wild edible plants reflects the wisdom of the Sherpa in terms of survival. The Sherpa cleverly survive the food shortage periods by harnessing the phenology of different species. In general, WEPs can provide the Sherpa with seasonal carbohydrates, nutrition, healthcare supplements and other products and services necessary for survival, which is likely why the Sherpa choose these plants.

Public Awareness, Individual Prevention Practice, and Psychological Effect at the Beginning of the COVID-19 Outbreak in China.

BACKGROUND: The COVID-19 has spread to more than 200 countries and territories. But less is known about the knowledge, protection behavior and anxiety regarding the outbreak among the general population. METHODS: A cross-sectional, population-based online survey was conducted in China and abroad from January 28 to February 1, 2020. Socio-demographic information was collected and knowledge scores, practice scores, anxiety scores and perceived risk were calculated. General linear model and binary logistic regression were used to identify possible associations. RESULTS: We included 9,764 individuals in this study, and 156 (1.6%) were from Hubei Province. The average knowledge score was 4.7 (standard deviation, 1.0) (scored on a 6-point scale); 96.1% maintained hand hygiene, and 90.3% of participants had varying levels of anxiety. People in Hubei Province were the most anxious, followed by those in Beijing and Shanghai. People who had experienced risk behaviors did not pay more attention to wearing masks and hand hygiene. CONCLUSIONS: The public had high awareness on knowledge of COVID-19 outbreak, and a high proportion of people practiced good hand hygiene behavior. Many people claimed anxiety, especially in heavily affected areas during pandemic, suggesting the importance of closing the gap between risk awareness and good practice and conduct psychological counseling to public and patients.