ABSTRACT
Piglet diarrhea caused by the porcine epidemic diarrhea virus (PEDV) is a common problem on pig farms in China associated with high morbidity and mortality rates. In this study, three PEDV isolates were successfully detected after the fourth blind passage in Vero cells. The samples were obtained from infected piglet farms in Jilin (Changchun), and Shandong (Qingdao) Provinces of China and were designated as CH/CC-1/2018, CH/CC-2/2018, and CH/QD/2018. According to the analysis of the complete S protein gene sequence, the CH/CC-1/2018 and CH/CC-2/2018 were allocated to the G2b branch, while CH/QD/2018 was located in the G1a interval and was closer to the vaccine strain CV777. Successful detection and identification of the isolated strains were carried out using electron microscopy and indirect immunofluorescence. Meanwhile, animal challenge experiments and viral RNA copies determination were used to compare the pathogenicity. The results showed that CH/CC-1/2018 in Changchun was more pathogenic than CH/QD/2018 in Qingdao. In conclusion, the discovery of these new strains is conducive to the development of vaccines to prevent the pandemic of PEDV, especially that the CH/CC-1/2018, and CH/CC-2/2018 were not related to the classical vaccine strain CV777.
ABSTRACT
High infection caused by mutations of SARS-CoV-2 calls for new prevention strategy. Ganoderma lucidum known as a superior immunoenhancer exhibits various antiviral effects, whether it can resist SARS-CoV-2 remains unclear. Herein, virtual screening combined with in vitro hACE2 inhibition assays were used to investigate its anti SARS-CoV-2 effect. Potential 54 active components, 80 core targets and 20 crucial pathways were identified by the component-target-pathway network. The binding characters of these components to hACE2 and its complexes with spike protein including omicron variant was analyzed by molecular docking. Lucidenic acid A was selected as the top molecule with high affinity to all receptors by forming hydrogen bonds. Molecular dynamics simulation showed it had good binding stability with the receptor proteins. Finally, in vitro FRET test demonstrated it inhibited the hACE2 activity with IC50 2 µmol/mL. Therefore, lucidenic acid A can prevent the virus invasion by blocking hACE2 binding with SARS-CoV-2.
Subject(s)
Angiotensin-Converting Enzyme 2 , Antiviral Agents , COVID-19 , Cholic Acids , SARS-CoV-2 , Spike Glycoprotein, Coronavirus , Virus Internalization , Humans , Angiotensin-Converting Enzyme 2/chemistry , Angiotensin-Converting Enzyme 2/metabolism , Antiviral Agents/pharmacology , Cholic Acids/pharmacology , COVID-19/prevention & control , Molecular Docking Simulation , Protein Binding , SARS-CoV-2/drug effects , Spike Glycoprotein, Coronavirus/chemistry , Spike Glycoprotein, Coronavirus/metabolism , Virus Internalization/drug effects , Reishi/chemistryABSTRACT
Objective: To establish a rapid, high-throughput detection assay of serum peptidome profiling for the diagnosis of SARS-CoV-2 infection based on matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS).
ABSTRACT
CT image of COVID-19 is disturbed by impulse noise during transmission and acquisition. Aiming at the problem that the early lesions of COVID-19 are not obvious and the density is low, which is easy to confuse with noise. A median filtering algorithm based on adaptive two-stage threshold is proposed to improve the accuracy for noise detection. In the advanced stage of ground-glass lesion, the density is uneven and the boundary is unclear. It has similar gray value to the CT images of suspected COVID-19 cases such as adenovirus pneumonia and mycoplasma pneumonia (reticular shadow and strip shadow). Aiming at the problem that the traditional weighted median filter has low contrast and fuzzy boundary, an adaptive weighted median filter image denoising method based on hybrid genetic algorithm is proposed. The weighted denoising parameters can adaptively change according to the detailed information of lung lobes and ground-glass lesions, and it can adaptively match the cross and mutation probability of genetic combined with the steady-state regional population density, so as to obtain a more accurate COVID-19 denoised image with relatively few iterations. The simulation results show that the improved algorithm under different density of impulse noise is significantly better than other algorithms in peak signal-to-noise ratio (PSNR), image enhancement factor (IEF) and mean absolute error (MSE). While protecting the details of lesions, it enhances the ability of image denoising.
ABSTRACT
Interferon (IFN)-λ plays an essential role in mucosal cells which exhibit strong antiviral activity. Lactobacillus plantarum (L. plantarum) has substantial application potential in the food and medical industries because of its probiotic properties. Alphacoronaviruses, especially porcine epidemic diarrhea virus (PEDV) and transmissible gastroenteritis virus (TGEV), cause high morbidity and mortality in piglets resulting in economic loss. Co-infection by these two viruses is becoming increasingly frequent. Therefore, it is particularly important to develop a new drug to prevent diarrhea infected with mixed viruses in piglets. In this study, we first constructed an anchored expression vector with CWA (C-terminal cell wall anchor) on L. plantarum. Second, we constructed two recombinant L. plantarum strains that anchored IFN-λ3 via pgsA (N-terminal transmembrane anchor) and CWA. Third, we demonstrated that both recombinant strains possess strong antiviral effects against coronavirus infection in the intestinal porcine epithelial cell line J2 (IPEC-J2). However, recombinant L. plantarum with the CWA anchor exhibited a more powerful antiviral effect than recombinant L. plantarum with pgsA. Consistent with this finding, Lb.plantarum-pSIP-409-IFN-λ3-CWA enhanced the expression levels of IFN-stimulated genes (ISGs) (ISG15, OASL, and Mx1) in IPEC-J2 cells more than did recombinant Lb.plantarum-pSIP-409-pgsA'-IFN-λ3. Our study verifies that recombinant L. plantarum inhibits PEDV and TGEV infection in IPEC-J2 cells, which may offer great potential for use as a novel oral antiviral agent in therapeutic applications for combating porcine epidemic diarrhea and transmissible gastroenteritis. This study is the first to show that recombinant L. plantarum suppresses PEDV and TGEV infection of IPEC-J2 cells.
Subject(s)
Coronavirus Infections/veterinary , Gastroenteritis, Transmissible, of Swine/prevention & control , Interferons/administration & dosage , Lactobacillus plantarum/genetics , Swine Diseases/prevention & control , Animals , Coronavirus Infections/immunology , Coronavirus Infections/prevention & control , Coronavirus Infections/virology , Epithelial Cells/immunology , Epithelial Cells/virology , Female , Gastroenteritis, Transmissible, of Swine/genetics , Gastroenteritis, Transmissible, of Swine/immunology , Gastroenteritis, Transmissible, of Swine/virology , Gene Expression , Interferons/genetics , Interferons/immunology , Lactobacillus plantarum/metabolism , Male , Porcine epidemic diarrhea virus/physiology , Swine , Swine Diseases/genetics , Swine Diseases/immunology , Swine Diseases/virology , Transmissible gastroenteritis virus/physiologyABSTRACT
Understanding the spatio-temporal characteristics or patterns of the 2019 novel coronavirus (2019-nCoV) epidemic is critical in effectively preventing and controlling this epidemic. However, no research analyzed the spatial dependency and temporal dynamics of 2019-nCoV. Consequently, this research aims to detect the spatio-temporal patterns of the 2019-nCoV epidemic using spatio-temporal analysis methods at the county level in Hubei province. The Mann-Kendall and Pettitt methods were used to identify the temporal trends and abrupt changes in the time series of daily new confirmed cases, respectively. The local Moran's I index was applied to uncover the spatial patterns of the incidence rate, including spatial clusters and outliers. On the basis of the data from January 26 to February 11, 2020, we found that there were 11 areas with different types of temporal patterns of daily new confirmed cases. The pattern characterized by an increasing trend and abrupt change is mainly attributed to the improvement in the ability to diagnose the disease. Spatial clusters with high incidence rates during the period were concentrated in Wuhan Metropolitan Area due to the high intensity of spatial interaction of the population. Therefore, enhancing the ability to diagnose the disease and controlling the movement of the population can be confirmed as effective measures to prevent and control the regional outbreak of the epidemic.