Network pharmacological mechanism of Runfei Ningshen Decoction in the treatment of insomnia caused by COVID-19

Objective: To study the mechanism of Runfei Ningshen Decoction in the treatment of insomnia caused by corona virus disease 2019(COVID-19) by using network pharmacology and molecular docking analysis. Methods: The chemical components and targets of Chinese medicinal materials of Runfei Ningshen Decoction in TCMSP, Batman, and CTD databases were searched. The relevant targets of novel coronavirus pneumonia and insomnia in Disgenet, GeneCards, CTD, and Malacards databases were searched. The component-target-disease network was established by using Cytoscape 3.2.1 software;The protein-protein intereation(PPI) network was constructed in string database. The common targets were enriched by using Cluster Profiler software package in R language software platform. The molecular docking of core targets related to insomnia caused by COVID-19 was carried out by using Discovery Studio 4.0 software. Results: 349 medicinal ingredients in Runfei Ningshen Decoction, 1 904 targets, 1 505 new coronavirus pneumonia-related targets, and 1 337 insomnia-related targets were collected. When the intersection of Venn diagrams were used, 404 common targets were obtained for the 2 diseases. 250 targets were intersected with the 2 diseases, and 33 core targets were screened out by the analysis of the interaction network between targets. Pathway enrichment analysis showed that Runfei Ningshen Decoction mainly acts on AKT1, INS, TP53, IL-6, key targets such as AKT1, INS, TP53, IL-6, JUN, CASP3, TNF, CAT, PTGS2 and CXCL8, which are involved in the important pathway processes such as human cytomegalovirus infection, fluid shear stress, and AGE-RAGE signaling pathways in complications of atherosclerosis and diabetes. The results of molecular docking showed that the core target has a high affinity with beta-sitosterol, 1-methoxy phaseolin, 3'-hydroxy-4'-O-methylglycyrrhizin, and anhydroicariin. The prescription treatment of insomnia caused by COVID-19 may be through the targets such as PTGS2, AR, PPARG, NOS2, HSP90 AA1 and so on. Conclusion: Runfei Ningshen Decoction can treat insomnia caused by COVID-19 by inhibiting IL-6 and TNF-a.

Study on the effect of Xuanfei Jiere Granules in the treatment of COVID-19 by network pharmacology and molecular docking

Objective: To explore the effect and mechanism of Xuanfei Jiere Granules in the treatment of corona virus disease 2019 (COVID-19) complicated with fever. Methods: The effective components of Xuanfei Jiere Granules were screened by Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP), and the potential target genes were predicted. The disease targets of COVID-19 complicated with fever were searched by GeneCards and CTD databases, and the common targets were obtained, and then introduced into Cytoscape to construct the "component-target-disease" network. The above 2 common targets were input into the online database of STRING protein interaction, and the results were imported into Cytoscape software to obtain Protein-Protein Interaction(PPI) network;R language was used to analyze the common targets with Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment analysis. Results: 84 active components and 82 targets were obtained. The pathway enrichment analysis showed that Xuanfei Jiere Granules mainly acted on protein kinase B(AKT1), tumor protein P53(TP53), interleukin-6(IL6) and other key targets in the treatment of COVID-19 complicated with fever. KEGG pathway enrichment analysis showed that the action was mainly related to the influence of fluid shear stress and atherosclerosis, human cytomegalovirus infection, and AGE-RAGE in diabetic complications and other signal pathways. The results of molecular docking showed that the core target had strong affinity with beta-sitosterol, formononetin, N-trans ferulyl tyramine and so on. Conclusion: This study preliminarily verified that Xuanfei Jiere Granules can play a role in the treatment of COVID-19 complicated with fever through multi-components, multi-targets and multi-pathways.

RAMIHM generates fully human monoclonal antibodies by rapid mRNA immunization of humanized mice and BCR-seq.

As a clinical vaccine, lipid nanoparticle (LNP) mRNA has demonstrated potent and broad antibody responses, leading to speculation about its potential for antibody discovery. Here, we developed RAMIHM, a highly efficient strategy for developing fully human monoclonal antibodies that employs rapid mRNA immunization of humanized mice followed by single B cell sequencing (scBCR-seq). We immunized humanized transgenic mice with RAMIHM and generated 15 top-ranked clones from peripheral blood, plasma B, and memory B cell populations, demonstrating a high rate of antigen-specificity (93.3%). Two Omicron-specific neutralizing antibodies with high potency and one broad-spectrum neutralizing antibody were discovered. Furthermore, we extended the application of RAMIHM to cancer immunotherapy targets, including a single transmembrane protein CD22 and a multi-transmembrane G protein-coupled receptor target, GPRC5D, which is difficult for traditional protein immunization methods. RAMIHM-scBCR-seq is a broadly applicable platform for the rapid and efficient development of fully human monoclonal antibodies against an assortment of targets.

An Integrated Approach with CLFPR-Based ANP and Fuzzy TOPSIS for Evaluating Business Performance of Rural Homestays: Study from China

Given the high importance of the performance of rural homestays to the local economy, this study aims to fill the gap in homestay performance research and make rural homestays more competitive and sustainable after the coronavirus disease epidemic (COVID-19). Integrating a consistent linguistic fuzzy preference relations-based analytic network process (CLFPR-based ANP) and a fuzzy technique for order performance by similarity to ideal solution (TOPSIS), this study constructs a comprehensive evaluation model of the performance of rural homestays and empirically analyzes homestay performance in Zhejiang. The results show the following: (1) Among the criteria's weights by CLFPR-based ANP, homestay operation and management, service quality, and homestay geist and community co-prosperity should be given much more attention;the importance of factors regarding the environment and building of rural homestays is gradually weakening. (2) In light of ranking alternatives based on performance evaluation by fuzzy TOPSIS, the homestay with the optimal performance has been found, which practitioners can use as a benchmark. Therefore, the priorities of these criteria further deepen the understanding of the performance of rural homestays and underline the development direction for practitioners. Simultaneously, in terms of feasibility and reliability, the integrated approach comprises a beneficial attempt and becomes an effective evaluation tool for practitioners to improve effectiveness.

High-Sensitivity Troponin I is an Indicator of Poor Prognosis in Patients with Severe COVID-19 Related Pneumonia.

Objective: Critical covid-19 patients have complications with acute myocardial injury is still unclear. We observed a series of critically ill patients, paying particular attention to the impact of myocardial injury at admission on short-term outcome. Methods: We prospectively collected and analyzed data from a series of severe covid-19 patients confirmed by real-time RT-PCR. Data were obtained from electronic medical records including clinical charts, nursing records, laboratory findings, and chest x-rays were from Feb 8, 2020, to April 7, 2020. The Acute Physiology and Chronic Health Evaluation (APACHE II) score, CURB-65 Pneumonia Severity Score, Sequential Organ Failure Assessment (SOFA) Score and pneumonia severity index (PSI) score were made within 24 hours of admission. Cardiac injury was diagnosed as hs-cTnI were above >28 pg/mL. The short-term outcome was defined as mortality in hospital. Results: A total of 100 patients met the diagnostic criteria of severe patients with COVID-19 during 2020.02.08-2020.04.07. The CURB 65, APACH2, SOFA, and PSI score were significantly higher in Critical group than in Severe group. Univariate regression analysis showed that oxygen flow, PO2/FiO2, SOFA and hs-cTnI were closely related to short-term outcome. The corresponding ROC of hs-cTnI, oxygen flow and SOFA for patient death prediction were 0.949, 0.906 and 0.652. hs-cTnI at 47.8 ng/liter predicted death, sensitivity 92.8%, specificity 92.9%; Oxygen flow at 5.5 liter/minute predicted death sensitivity 100%, specificity 77.9%; SOFA score at 5 predicted death sensitivity 100%, specificity 73.8%. Conclusion: Our cohort study demonstrated that inhaled oxygen flow, SOFA score, and myocardial injury at admission in critically ill COVID-19 patients were important indicators for predicting short-term death of patients, the hs-cTnI can be as a risk stratification, which may provide a simple method for the physicians to identify high-risk patients and give reasonable treatment in time.

Multiplexed LNP-mRNA vaccination against pathogenic coronavirus species.

Although COVID-19 vaccines have been developed, multiple pathogenic coronavirus species exist, urging on development of multispecies coronavirus vaccines. Here we develop prototype lipid nanoparticle (LNP)-mRNA vaccine candidates against SARS-CoV-2 Delta, SARS-CoV, and MERS-CoV, and we test how multiplexing LNP-mRNAs can induce effective immune responses in animal models. Triplex and duplex LNP-mRNA vaccinations induce antigen-specific antibody responses against SARS-CoV-2, SARS-CoV, and MERS-CoV. Single-cell RNA sequencing profiles the global systemic immune repertoires and respective transcriptome signatures of vaccinated animals, revealing a systemic increase in activated B cells and differential gene expression across major adaptive immune cells. Sequential vaccination shows potent antibody responses against all three species, significantly stronger than simultaneous vaccination in mixture. These data demonstrate the feasibility, antibody responses, and single-cell immune profiles of multispecies coronavirus vaccination. The direct comparison between simultaneous and sequential vaccination offers insights into optimization of vaccination schedules to provide broad and potent antibody immunity against three major pathogenic coronavirus species.

Development of an efficient reproducible cell-cell transmission assay for rapid quantification of SARS-CoV-2 spike interaction with hACE2.

Efficient quantitative assays for measurement of viral replication and infectivity are indispensable for future endeavors to develop prophylactic or therapeutic antiviral drugs or vaccines against SARS-CoV-2. We developed a SARS-CoV-2 cell-cell transmission assay that provides a rapid and quantitative readout to assess SARS-CoV-2 spike hACE2 interaction in the absence of pseudotyped particles or live virus. We established two well-behaved stable cell lines, which demonstrated a remarkable correlation with standard cell-free viral pseudotyping for inhibition by convalescent sera, small-molecule drugs, and murine anti-spike monoclonal antibodies. The assay is rapid, reliable, and highly reproducible, without a requirement for any specialized research reagents or laboratory equipment and should be easy to adapt for use in most investigative and clinical settings. It can be effectively used or modified for high-throughput screening for compounds and biologics that interfere with virus-cell binding and entry to complement other neutralization assays currently in use.

Omicron-specific mRNA vaccination alone and as a heterologous booster against SARS-CoV-2.

The Omicron variant of SARS-CoV-2 recently swept the globe and showed high level of immune evasion. Here, we generate an Omicron-specific lipid nanoparticle (LNP) mRNA vaccine candidate, and test its activity in animals, both alone and as a heterologous booster to WT mRNA vaccine. Our Omicron-specific LNP-mRNA vaccine elicits strong antibody response in vaccination-naïve mice. Mice that received two-dose WT LNP-mRNA show a > 40-fold reduction in neutralization potency against Omicron than WT two weeks post boost, which further reduce to background level after 3 months. The WT or Omicron LNP-mRNA booster increases the waning antibody response of WT LNP-mRNA vaccinated mice against Omicron by 40 fold at two weeks post injection. Interestingly, the heterologous Omicron booster elicits neutralizing titers 10-20 fold higher than the homologous WT booster against Omicron variant, with comparable titers against Delta variant. All three types of vaccination, including Omicron alone, WT booster and Omicron booster, elicit broad binding antibody responses against SARS-CoV-2 WA-1, Beta, Delta variants and SARS-CoV. These data provide direct assessments of an Omicron-specific mRNA vaccination in vivo, both alone and as a heterologous booster to WT mRNA vaccine.

Student Experience and Satisfaction in Academic Libraries: A Comparative Study among Three Universities in Wuhan

In recent years, the spatial renovation of university libraries in various countries has focused on readers’ needs and followed the trend to develop learning spaces as a primary spatial form. In this study, we reviewed six spatial dimensions affecting student users’ learning experience. Specifically, we built a theory- and practice-based conceptual analysis framework to measure users’ satisfaction with recent spatial renovations at three university libraries in Wuhan, China. We used SPSS statistical software to conduct multiple linear regression analyses of spatial satisfaction. The findings show that five spatial dimensions significantly affect students’ satisfaction with library space, namely, service facility availability, quality of interior design, physical environment elements, spatial diversity, and learning space controllability. Service facility availability is the most critical factor affecting spatial satisfaction. In this study, we present empirical, evidence-based space elements that enhance user satisfaction with library spaces, and provide targeted design suggestions for future library space renovation and the optimization of space allocation and expansion of space services at university libraries in China.

Variant-specific vaccination induces systems immune responses and potent in vivo protection against SARS-CoV-2.

Lipid nanoparticle (LNP)-mRNA vaccines offer protection against COVID-19; however, multiple variant lineages caused widespread breakthrough infections. Here, we generate LNP-mRNAs specifically encoding wild-type (WT), B.1.351, and B.1.617 SARS-CoV-2 spikes, and systematically study their immune responses. All three LNP-mRNAs induced potent antibody and T cell responses in animal models; however, differences in neutralization activity have been observed between variants. All three vaccines offer potent protection against in vivo challenges of authentic viruses of WA-1, Beta, and Delta variants. Single-cell transcriptomics of WT- and variant-specific LNP-mRNA-vaccinated animals reveal a systematic landscape of immune cell populations and global gene expression. Variant-specific vaccination induces a systemic increase of reactive CD8 T cells and altered gene expression programs in B and T lymphocytes. BCR-seq and TCR-seq unveil repertoire diversity and clonal expansions in vaccinated animals. These data provide assessment of efficacy and direct systems immune profiling of variant-specific LNP-mRNA vaccination in vivo.

Monospecific and bispecific monoclonal SARS-CoV-2 neutralizing antibodies that maintain potency against B.1.617.

COVID-19 pathogen SARS-CoV-2 has infected hundreds of millions and caused over 5 million deaths to date. Although multiple vaccines are available, breakthrough infections occur especially by emerging variants. Effective therapeutic options such as monoclonal antibodies (mAbs) are still critical. Here, we report the development, cryo-EM structures, and functional analyses of mAbs that potently neutralize SARS-CoV-2 variants of concern. By high-throughput single cell sequencing of B cells from spike receptor binding domain (RBD) immunized animals, we identify two highly potent SARS-CoV-2 neutralizing mAb clones that have single-digit nanomolar affinity and low-picomolar avidity, and generate a bispecific antibody. Lead antibodies show strong inhibitory activity against historical SARS-CoV-2 and several emerging variants of concern. We solve several cryo-EM structures at ~3 Å resolution of these neutralizing antibodies in complex with prefusion spike trimer ectodomain, and reveal distinct epitopes, binding patterns, and conformations. The lead clones also show potent efficacy in vivo against authentic SARS-CoV-2 in both prophylactic and therapeutic settings. We also generate and characterize a humanized antibody to facilitate translation and drug development. The humanized clone also has strong potency against both the original virus and the B.1.617.2 Delta variant. These mAbs expand the repertoire of therapeutics against SARS-CoV-2 and emerging variants.

High-affinity, neutralizing antibodies to SARS-CoV-2 can be made without T follicular helper cells.

T follicular helper (TFH) cells are the conventional drivers of protective, germinal center (GC)­based antiviral antibody responses. However, loss of TFH cells and GCs has been observed in patients with severe COVID-19. As T cell­B cell interactions and immunoglobulin class switching still occur in these patients, noncanonical pathways of antibody production may be operative during SARS-CoV-2 infection. We found that both TFH-dependent and -independent antibodies were induced against SARS-CoV-2 infection, SARS-CoV-2 vaccination, and influenza A virus infection. Although TFH-independent antibodies to SARS-CoV-2 had evidence of reduced somatic hypermutation, they were still high affinity, durable, and reactive against diverse spike-derived epitopes and were capable of neutralizing both homologous SARS-CoV-2 and the B.1.351 (beta) variant of concern. We found by epitope mapping and B cell receptor sequencing that TFH cells focused the B cell response, and therefore, in the absence of TFH cells, a more diverse clonal repertoire was maintained. These data support an alternative pathway for the induction of B cell responses during viral infection that enables effective, neutralizing antibody production to complement traditional GC-derived antibodies that might compensate for GCs damaged by viral inflammation.

A smartphone-based visual biosensor for CRISPR-Cas powered SARS-CoV-2 diagnostics.

The pandemic of coronavirus disease 2019 (COVID-19) resulted from novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has become a worldwide concern. It is imperative to develop rapid, sensitive, and specific biosensing methods. Herein, we developed a CRISPR-Cas12a powered visual biosensor with a smartphone readout for ultrasensitive and selective detection of SARS-CoV-2. Simply, the SARS-CoV-2 derived nucleic acids triggered CRISPR-Cas12a based indiscriminate degradation of a single-stranded DNA that was supposed to link two gold nanoparticles, inducing the dis-aggregation of gold nanoparticles and thus generating observable color changes. This change can be readily distinguished by naked eyes as well as a smartphone with a Color Picker App. The proposed biosensor was successfully applied to detect SARS-CoV-2 gene in synthetic vectors, transcribed RNA and SARS-CoV-2 pseudoviruses. It rendered "single copy resolution" as evidenced by the 1 copy/µL limit of detection of pseudoviruses with no cross-reactivity. When the developed biosensor was challenged with SARS-CoV-2 clinical bio-samples, it provided 100% agreement (both positive and negative) with qPCR results. The sample-to-result time was roughly 90 min. Our work provides a novel and robust technology for ultrasensitive detection of SARS-CoV-2 that could be used clinically.

Modeling and Analyzing Transmission of Infectious Diseases Using Generalized Stochastic Petri Nets

Some infectious diseases such as COVID-19 have the characteristics of long incubation period, high infectivity during the incubation period, and carriers with mild or no symptoms which are more likely to cause negligence. Global researchers are working to find out more about the transmission of infectious diseases. Modeling plays a crucial role in understanding the transmission of the new virus and helps show the evolution of the epidemic in stages. In this paper, we propose a new general transmission model of infectious diseases based on the generalized stochastic Petri net (GSPN). First, we qualitatively analyze the transmission mode of each stage of infectious diseases such as COVID-19 and explain the factors that affect the spread of the epidemic. Second, the GSPN model is built to simulate the evolution of the epidemic. Based on this model’s isomorphic Markov chain, the equilibrium state of the system and its changing laws under different influencing factors are analyzed. Our paper demonstrates that the proposed GSPN model is a compelling tool for representing and analyzing the transmission of infectious diseases from system-level understanding, and thus contributes to providing decision support for effective surveillance and response to epidemic development.

Nonstructural Protein 1 of SARS-CoV-2 Is a Potent Pathogenicity Factor Redirecting Host Protein Synthesis Machinery toward Viral RNA.

The causative virus of the COVID-19 pandemic, SARS-CoV-2, uses its nonstructural protein 1 (Nsp1) to suppress cellular, but not viral, protein synthesis through yet unknown mechanisms. We show here that among all viral proteins, Nsp1 has the largest impact on host viability in the cells of human lung origin. Differential expression analysis of mRNA-seq data revealed that Nsp1 broadly alters the cellular transcriptome. Our cryo-EM structure of the Nsp1-40S ribosome complex shows that Nsp1 inhibits translation by plugging the mRNA entry channel of the 40S. We also determined the structure of the 48S preinitiation complex formed by Nsp1, 40S, and the cricket paralysis virus internal ribosome entry site (IRES) RNA, which shows that it is nonfunctional because of the incorrect position of the mRNA 3' region. Our results elucidate the mechanism of host translation inhibition by SARS-CoV-2 and advance understanding of the impacts from a major pathogenicity factor of SARS-CoV-2.

Pulmonary contusion mimicking COVID-19: A case report.

BACKGROUND: Coronavirus disease 2019 (COVID-19) is a major public health emergency with obvious characteristics of human-to-human transmission, and there are infective asymptomatic carriers. Early identification and proper management of patients with COVID-19 are important. Features in chest computed tomography (CT) can facilitate identifying newly infected individuals. However, CT findings of some lung contusions are similar to those of COVID-19, as shown in the present case. CASE SUMMARY: A 46-year-old woman was admitted to hospital for backache and foot pain caused by a fall injury 1 d before hospitalization. She was suspected of having COVID-19, since there was a confirmed COVID-19 case near her residence. But she had no fever, cough, chest tightness, difficult breathing, nausea, vomiting, or diarrhea, etc. On physical examination, the lower posterior chest of both sides showed dullness on percussion and moist rales at the end of inspiration on auscultation. The white blood cell count and lymphocyte count were 10.88 × 109/L and 1.04 × 109/L, respectively. CT performed on February 7, 2020 revealed that both lungs were scattered with patchy ground-glass opacity. The patient was diagnosed with pulmonary contusion with thoracic spinal fracture (T12), calcaneal fracture, and pelvic fracture. On day 9 after conservative treatment, her condition was alleviated. On review of the chest CT, the previous shadows were significantly reduced. CONCLUSION: Differential diagnosis of lung contusion and COVID-19 must be emphasized. Both conditions require effective prompt actions, especially COVID-19.

Prognostic value of NT-proBNP in patients with severe COVID-19 (preprint)

The outbreak of coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in China has been declared a public health emergency of international concern. The cardiac injury was dominate in the process. However, whether N terminal pro B type natriuretic peptide (NT-proBNP) predicted outcome of COVID-19 patients was unknown. The study initially enrolled 102 patients with severe COVID-19 pneumonia from a continuous sample. After screening out the ineligible cases, 54 patients were analyzed in this study. Results found that patients with higher NT-proBNP (above 88.64 pg/mL) level had more risks of in-hospital death. After adjusting for potential cofounders in separate modes, NT-proBNP presented as an independent risk factor of in-hospital death in patients with severe COVID-19.