Screening of Efficient Adjuvants for the RBD-Based Subunit Vaccine of SARS-CoV-2.

The variants of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are more transmissible, with a reduced sensitivity to vaccines targeting the original virus strain. Therefore, developing an effective vaccine against both the original SARS-CoV-2 strain and its variants is an urgent need. It is known that the receptor-binding domain (RBD) in the S protein of SARS-CoV-2 is an important vaccine target, but subunit vaccines usually have lower immunogenicity and efficacy. Thus, selecting appropriate adjuvants to enhance the immunogenicity of protein-based subunit vaccine antigens is necessary. Here, an RBD-Fc subunit vaccine of SARS-CoV-2 has been generated, followed by vaccination in B6 mice, and four adjuvant regimens were investigated, including aluminum salts (Alum) + 3-O-desacyl-4'-monophosphoryl lipid A (MPL), AddaVax, QS21 + MPL, and Imiquimod. The adjuvant potency was evaluated by comparing the elicited polyclonal antibodies titers with measuring binding to RBD and S protein in ELISA and Western blot analysis, and also the cross-neutralizing antibodies titers using a pseudovirus infection assay of hACE2-expressing 293T cells, with pseudoviruses expressing the S protein of the SARS-CoV-2 original strain and Delta strain. The presence of QS21 + MPL adjuvant induced stronger polyclonal antibody response and neutralization potency blocking the original strain and Delta strain, as compared with the non-adjuvant RBD-Fc group and other adjuvant groups. Meanwhile, Imiquimod even had a negative effect in inducing specific antibodies and cross-neutralizing antibody production as an adjuvant.

Establishing an integrated prevention and control system to ensure zero nosocomial infections – an analysis of the logistics support in controlling nosocomial infections during the hospital's fight against the COVID-19 epidemic Aufbau eines integrierten Präventions- und Kontrollsystems zur Gewährleistung der Null-Strategie zur Prävention nosokomialer Infektionen – eine Analyse der logistischen Unterstützung zur Prävention nosokomialer Infektionen während des Kampfes des Krankenhauses gegen die COVID-19-Epidemie

Aim: To analyze the role of the logistics support services in nosocomial infection control during emergency periods, with a focus on job responsibilities including the organization of vehicle parking, supply of hospital meals, washing of medical bedding and clothing, disposal and management of medical sewage and waste, elevator services, disinfection of air conditioning systems, disinfection and cleaning of ambulances, management of hospital buildings, storage of sterilization supplies, reception and delivery of oxygen cylinders and protection of staff health as examples. Methods: The adjustment and optimization of the emergency support system and working mode as part of hospitals' response to major public emergencies were summarized, and the vital supporting role of the logistics support services in nosocomial infection control was analyzed. Results: The logistics support services played a crucial role in ensuring the high-performance operations of the hospitals and control of nosocomial infections, resulting in the excellent outcome of "zero infection” among hospital staff. Conclusion: Establishing a safe, flexible and efficient system for the logistics support services is important in ensuring an effective response by hospitals to health emergencies.

Identification of critical genes and molecular pathways in COVID-19 myocarditis and constructing gene regulatory networks by bioinformatic analysis.

BACKGROUND: There is growing evidence of a strong relationship between COVID-19 and myocarditis. However, there are few bioinformatics-based analyses of critical genes and the mechanisms related to COVID-19 Myocarditis. This study aimed to identify critical genes related to COVID-19 Myocarditis by bioinformatic methods, explore the biological mechanisms and gene regulatory networks, and probe related drugs. METHODS: The gene expression data of GSE150392 and GSE167028 were obtained from the Gene Expression Omnibus (GEO), including cardiomyocytes derived from human induced pluripotent stem cells infected with SARS-CoV-2 in vitro and GSE150392 from patients with myocarditis infected with SARS-CoV-2 and the GSE167028 gene expression dataset. Differentially expressed genes (DEGs) (adjusted P-Value <0.01 and |Log2 Fold Change| ≥2) in GSE150392 were assessed by NetworkAnalyst 3.0. Meanwhile, significant modular genes in GSE167028 were identified by weighted gene correlation network analysis (WGCNA) and overlapped with DEGs to obtain common genes. Functional enrichment analyses were performed by using the "clusterProfiler" package in the R software, and protein-protein interaction (PPI) networks were constructed on the STRING website (https://cn.string-db.org/). Critical genes were identified by the CytoHubba plugin of Cytoscape by 5 algorithms. Transcription factor-gene (TF-gene) and Transcription factor-microRibonucleic acid (TF-miRNA) coregulatory networks construction were performed by NetworkAnalyst 3.0 and displayed in Cytoscape. Finally, Drug Signatures Database (DSigDB) was used to probe drugs associated with COVID-19 Myocarditis. RESULTS: Totally 850 DEGs (including 449 up-regulated and 401 down-regulated genes) and 159 significant genes in turquoise modules were identified from GSE150392 and GSE167028, respectively. Functional enrichment analysis indicated that common genes were mainly enriched in biological processes such as cell cycle and ubiquitin-protein hydrolysis. 6 genes (CDK1, KIF20A, PBK, KIF2C, CDC20, UBE2C) were identified as critical genes. TF-gene interactions and TF-miRNA coregulatory network were constructed successfully. A total of 10 drugs, (such as Etoposide, Methotrexate, Troglitazone, etc) were considered as target drugs for COVID-19 Myocarditis. CONCLUSIONS: Through bioinformatics method analysis, this study provides a new perspective to explore the pathogenesis, gene regulatory networks and provide drug compounds as a reference for COVID-19 Myocarditis. It is worth highlighting that critical genes (CDK1, KIF20A, PBK, KIF2C, CDC20, UBE2C) may be potential biomarkers and treatment targets of COVID-19 Myocarditis for future study.

Comparison of Prevalence and Risk Factors of PTSS Between Chinese Patients With Depression and Non-depressed Controls During COVID-19 Outbreak.

Background: COVID-19 pandemic is a traumatic event all over the world, and may lead to post-traumatic stress symptom (PTSS) in different population who are under the threat of novel corona virus. Therefore, the aim of our study was to compare the prevalence and risk factors of PTSS between Chinese patients with depression and non-depressed controls during the COVID-19 outbreak. Methods: 437 depressed patients and 2,940 non-depressed controls were enrolled in this cross-sectional study between February 14 and May 9, 2020.The Impact of Events Scale-Revised (IES-R), Zung Self-Rating Depression Scale (SDS), Zung Self-Rating Anxiety Scale (SAS) and Pittsburgh Sleep Quality Index (PSQI) were used to evaluate the psychological status of all the participants. Results: The prevalence of PTSS (IES-R ≥ 33) in depressed patients (45.08%) was higher than that in non-depressed controls (5.31%). Patients with depression were 16 times more likely to suffer from PTSS than those without depression. Correlation analyses showed that the IES-R total score was positively correlated with SDS, SAS, and PSQI scores in both depressed and non-depressed groups (Bonferroni corrected all p < 0.001). Multiple linear regression analysis showed that SAS score, and PSQI score were independently associated with IES-R total score in both depression and non-depression groups. In depressed patients, education level and duration of media exposure to COVID-19 were positively associated with PTSS, while in the non-depressed group, subjects who were married, in the 31-50 year group or with higher SDS score were more likely to develop PTSS. Conclusions: These results indicate that the prevalence rate of PTSS in patients with depression is very higher than that in subjects without depression. PTSS are associated with a number of socio-demographic and clinical variables.

Exploring the Potential Targets and Mechanisms of Huang Lian Jie Du Decoction in the Treatment of Coronavirus Disease 2019 Based on Network Pharmacology.

BACKGROUND: In December 2019, coronavirus disease 2019 (COVID-19) caused by a novel coronavirus (severe acute respiratory syndrome coronavirus 2, SARS-CoV-2; previously known as 2019-nCoV) emerged in Wuhan, China, and caused many infections and deaths. At present, there are no specific drugs for the etiology and treatment of COVID-19. A combination of traditional Chinese and western medicine is proposed to treat COVID-19, in which Huang Lian Jie Du decoction (HLJDD) is recommended for the treatment of COVID-19 in many provinces in China and has been widely used in the clinic. This study explored the potential targets of HLJDD in the treatment of COVID-19 based on network pharmacology. METHODS: First, the chemical composition and targets of HLJDD and COVID-19-related targets were obtained through the TCMSP, UniProt, GeneCards and OMIM databases. Second, HLJDD target and HLJDD-COVID-19 target networks were constructed via the STRING database and Cytoscape software. Finally, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis of the HLJDD-COVID-19 targets was applied via the DAVID database. RESULTS: Our study identified a total of 67 active ingredients of HLJDD and 204 targets of HLJDD. A total of 502 COVID-19-related targets were obtained, of which 47 were intersecting targets of HLJDD and COVID-19. A total of 179 GO terms and 77 KEGG terms, including the TNF signaling pathway, NF-κB signaling pathway and HIF-1 signaling pathway, were identified. CONCLUSION: The present study explored the potential targets and signaling pathways of HLJDD during the treatment of COVID-19, which may provide a basis for the research and development of drugs for the treatment of COVID-19.

Receptor-Binding Domain Proteins of SARS-CoV-2 Variants Elicited Robust Antibody Responses Cross-Reacting with Wild-Type and Mutant Viruses in Mice.

Multiple variants of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) have spread around the world, but the neutralizing effects of antibodies induced by the existing vaccines have declined, which highlights the importance of developing vaccines against mutant virus strains. In this study, nine receptor-binding domain (RBD) proteins of the SARS-CoV-2 variants (B.1.1.7, B.1.351 and P.1 lineages) were constructed and fused with the Fc fragment of human IgG (RBD-Fc). These RBD-Fc proteins contained single or multiple amino acid substitutions at prevalent mutation points of spike protein, which enabled them to bind strongly to the polyclonal antibodies specific for wild-type RBD and to the recombinant human ACE2 protein. In the BALB/c, mice were immunized with the wild-type RBD-Fc protein first and boosted twice with the indicated mutant RBD-Fc proteins later. All mutant RBD-Fc proteins elicited high-level IgG antibodies and cross-neutralizing antibodies. The RBD-Fc proteins with multiple substitutions tended to induce higher antibody titers and neutralizing-antibody titers than the single-mutant RBD-Fc proteins. Meanwhile, both wild-type RBD-Fc protein and mutant RBD-Fc proteins induced significantly decreased neutralization capacity to the pseudovirus of B.1.351 and P.1 lineages than to the wild-type one. These data will facilitate the design and development of RBD-based subunit vaccines against SARS-COV-2 and its variants.

The high diversity of SARS-CoV-2-related coronaviruses in pangolins alerts potential ecological risks.

Understanding the zoonotic origin and evolution history of SARS-CoV-2 will provide critical insights for alerting and preventing future outbreaks. A significant gap remains for the possible role of pangolins as a reservoir of SARS-CoV-2 related coronaviruses (SC2r-CoVs). Here, we screened SC2r-CoVs in 172 samples from 163 pangolin individuals of four species, and detected positive signals in muscles of four Manis javanica and, for the first time, one M. pentadactyla. Phylogeographic analysis of pangolin mitochondrial DNA traced their origins from Southeast Asia. Using in-solution hybridization capture sequencing, we assembled a partial pangolin SC2r-CoV (pangolin-CoV) genome sequence of 22 895 bp (MP20) from the M. pentadactyla sample. Phylogenetic analyses revealed MP20 was very closely related to pangolin-CoVs that were identified in M. javanica seized by Guangxi Customs. A genetic contribution of bat coronavirus to pangolin-CoVs via recombination was indicated. Our analysis revealed that the genetic diversity of pangolin-CoVs is substantially higher than previously anticipated. Given the potential infectivity of pangolin-CoVs, the high genetic diversity of pangolin-CoVs alerts the ecological risk of zoonotic evolution and transmission of pathogenic SC2r-CoVs.

Maternal and Neonatal Outcomes of Pregnant Women With Coronavirus Disease 2019 (COVID-19) Pneumonia: A Case-Control Study.

BACKGROUND: The ongoing pandemic of coronavirus disease 2019 (COVID-19) has caused serious concerns about its potential adverse effects on pregnancy. There are limited data on maternal and neonatal outcomes of pregnant women with COVID-19 pneumonia. METHODS: We conducted a case-control study to compare clinical characteristics and maternal and neonatal outcomes of pregnant women with and without COVID-19 pneumonia. RESULTS: During the period 24 January-29 February 2020, there were 16 pregnant women with confirmed COVID-19 pneumonia and 18 suspected cases who were admitted to labor in the third trimester. Two had vaginal delivery and the rest were cesarean delivery. Few patients presented respiratory symptoms (fever and cough) on admission, but most had typical chest computed tomographic images of COVID-19 pneumonia. Compared to the controls, patients with COVID-19 pneumonia had lower counts of white blood cells (WBCs), neutrophils, C-reactive protein (CRP), and alanine aminotransferase on admission. Increased levels of WBCs, neutrophils, eosinophils, and CRP were found in postpartum blood tests of pneumonia patients. Three (18.8%) of the mothers with confirmed COVID-19 pneumonia and 3 (16.7%) with suspected COVID-19 pneumonia had preterm delivery due to maternal complications, which were significantly higher than in the control group. None experienced respiratory failure during their hospital stay. COVID-19 infection was not found in the newborns, and none developed severe neonatal complications. CONCLUSIONS: Severe maternal and neonatal complications were not observed in pregnant women with COVID-19 pneumonia who had vaginal or cesarean delivery. Mild respiratory symptoms of pregnant women with COVID-19 pneumonia highlight the need of effective screening on admission.

Perinatal Depression of Exposed Maternal Women in the COVID-19 Pandemic in Wuhan, China.

Objective: This study aims to investigate perinatal depression in women who gave birth during the COVID-19 pandemic in Wuhan, and to evaluate the effect of the pandemic on perinatal depression prevalence. Methods: A cross-sectional investigation was conducted into women hospitalized for delivery in Hubei Maternity and Child Healthcare Hospital from December 31, 2019 to March 22, 2020, a period which encompasses the entire time frame of the COVID-19 pandemic in Wuhan. The Edinburgh Postnatal Depression Scale (EPDS) was adopted to evaluate perinatal depression status. A Chi-square test and logistic regression model were utilized for data analysis. Results: A total of 2,883 participants were included, 33.71% of whom were found to suffer from depressive symptoms. In detail, 27.02%, 5.24%, and 1.46% were designated as having mild, moderate, and severe depressive symptoms, respectively. The perinatal depression prevalence increased as the COVID-19 pandemic worsened. Compared to the period from December 31, 2019 to January 12, 2020, perinatal depression risk significantly decreased within the 3 weeks of March 2-22, 2020 (1st week: OR = 0.39, 95% CI: 0.20, 0.78; 2nd week: OR = 0.35, 95% CI: 0.17, 0.73; and 3rd week: OR = 0.48, 95% CI: 0.25, 0.94); and the postnatal depression risk significantly rose within the four weeks of January 27-February 23, 2020 (1st week: OR = 1.78, 95% CI: 1.18, 2.68; 2nd week: OR = 2.03, 95% CI: 1.35, 3.04; 3rd week: OR = 1.48, 95% CI: 1.02, 2.14; and 4th week: OR = 1.73, 95% CI: 1.20, 2.48). Conclusion: The dynamic change of perinatal depression was associated with the progression of the COVID-19 pandemic among new mothers who were exposed to the pandemic. An elevated risk of postnatal depression was also observed during the COVID-19 pandemic.

Vaginal delivery in women with COVID-19: report of two cases.

BACKGROUND: During the ongoing global outbreak of COVID-19, pregnant women who are susceptible to COVID-19 should be highly concerned. The issue of vertical transmission and the possibility of neonatal infection is a major concern. CASE PRESENTATION: Case 1: A 35-year-old pregnant woman with a gestational age of 37 weeks and 6 days was admitted to our hospital at the point of giving birth. Except for the abnormalities in her chest CT image, she was asymptomatic. She had an uncomplicated spontaneous vaginal delivery, and her infant was discharged home for isolation. Because of the positive result of the maternal swabs for SARS-CoV-2 obtained on the 2nd day after sampling, we transferred the mother to the designated hospital and followed up with her by telephone interviews. Luckily, it was confirmed on February 23 that the newborn did not develop any COVID-19 symptoms after observation for 14 days after birth. Case 2: Another pregnant woman, with a gestational age of 38 weeks and 2 days, was also admitted to our hospital because of spontaneous labor with cervical dilation of 5 cm. Since she had the typical manifestations of COVID-19, including cough, lymphopenia, and abnormal chest CT images, she was highly suspected of having COVID-19. Based on the experience from case 1, we helped the mother deliver a healthy baby by vaginal delivery. On the 2nd day after delivery, the maternal nasopharyngeal swab result was positive, while the infant's result was negative. CONCLUSION: There is still insufficient evidence supporting maternal-fetal vertical transmission for COVID-19-infected mothers in late pregnancy, and vaginal delivery may not increase the possibility of neonatal infection.

Prevalence, risk factors and clinical correlates of depression in quarantined population during the COVID-19 outbreak.

BACKGROUND: The COVID-19 outbreak has brought tremendous psychological pressure to the general population, which may lead to depression. Therefore, this study aim to evaluate the prevalence and clinical correlates of depressive symptoms in the general population quarantined during the COVID-19 outbreak in Shenzhen. METHODS: 2237 quarantined general individuals participated in this cross-sectional study from February 14 to March 4, 2020, during their 14 days quarantine. They completed the Zung's Self-Rating Depression Scale (SDS) for depression, Zung's self-rating anxiety scale (SAS) for anxiety, the Pittsburgh Sleep Quality Index (PSQI) for sleep quality, and the Impact of Events Scale-Revised (IES-R) for post-traumatic stress symptoms (PTSS). RESULTS: The prevalence of depressive symptom was 6.21% in quarantined individuals. The depressed group were younger, less married and educated, and had higher SAS, PSQI, IES-R total scores (all p<0.05), as well as more avoidance, intrusion and hyperarousal symptoms than the non-depressed group. Correlation analysis showed significant correlations between SDS score and the following parameters: age, marriage, education, SAS, PSQI, IES-R total and its three subscale scores (Bonferroni corrected all p<0.05). Further multiple regression indicated that age, marriage, education, SAS, PSQI, IES-R total score, Avoidance and Hyperarousal factor were independent predictors of depressive symptom. LIMITATIONS: This study adopted a cross-sectional design and used self-report questionnaires. CONCLUSIONS: Our results suggest an elevated prevalence of depressive symptom in quarantined general individuals in Shenzhen. Some demographic and clinical variables were associated with depressive symptoms.

Clinical analysis of ten pregnant women with COVID-19 in Wuhan, China: A retrospective study.

BACKGROUND: COVID-19 is spreading globally. This study aims to evaluate the clinical characteristics and outcomes of pregnant women confirmed with COVID-19 to provide reference for clinical work. METHODS: The clinical features and outcomes of 10 pregnant women confirmed with COVID-19 at Maternal and Child Health Hospital of Hubei Province, Tongji Medical College, Huazhong University of Science and Technology, a tertiary- care teaching hospital in Hubei province, Wuhan, China from January 23 to February 23, 2020 were retrospectively analyzed. RESULTS: All the 10 observed pregnant women including 9 singletons and 1 twin were native people in Wuhan. All of them were diagnosed mild COVID-19, and none of the patients developed severe COVID-19 or died. Among the 10 patients, two patients underwent vaginal delivery, two patients underwent intrapartum cesarean section, and the remaining six patients underwent elective cesarean section. All of 10 patients showed lung abnormalities by pulmonary CT images after delivery. Their eleven newborns were recorded and no neonatal asphyxia was observed. CONCLUSIONS: Pulmonary CT screening on admission may be necessary to reduce the risk of nosocomial transmission of COVID-19 during the outbreak period. And COVID-19 is not an indication of cesarean section.

Clinical features and outcomes of pregnant women suspected of coronavirus disease 2019.

BACKGROUND: 2019 novel coronavirus disease (COVID-19) has become a worldwide pandemic. Under such circumstance pregnant women are also affected significantly. OBJECTIVE: This study aims to observe the clinical features and outcomes of pregnant women who have been confirmed with COVID-19. METHODS: The research objects were 55 cases of suspected COVID-19 pregnant women who gave a birth from Jan 20th 2020 to Mar 5th 2020 in our hospital-a big birth center delivering about 30,000 babies in the last 3 years. These cases were subjected to pulmonary CT scan and routine blood test, manifested symptoms of fever, cough, chest tightness or gastrointestinal symptoms. They were admitted to an isolated suite, with clinical features and newborn babies being carefully observed. Among the 55 cases, 13 patients were assigned into the confirmed COVID-19 group for being tested positive sever acute respiratory syndrome coronavirus 2(SARS-CoV-2) via maternal throat swab test, and the other 42 patients were assigned into the control group for being ruled out COVID-19 pneumonia based on new coronavirus pneumonia prevention and control program(the 7th edition). RESULTS: There were 2 fever patients during the prenatal period and 8 fever patients during the postpartum period in the confirmed COVID-19 group. In contrast, there were 11 prenatal fever patients and 20 postpartum fever patients in the control group (p>0.05). Among 55 cases, only 2 case had cough in the confirmed group. The imaging of pulmonary CT scan showed ground- glass opacity (46.2%, 6/13), patch-like shadows(38.5%, 5/13), fiber shadow(23.1%, 3/13), pleural effusion (38.5%, 5/13)and pleural thickening(7.7%, 1/13), and there was no statistical difference between the confirmed COVID-19 group and the control group (p>0.05). During the prenatal and postpartum period, there was no difference in the count of WBC, Neutrophils and Lymphocyte, the radio of Neutrophils and Lymphocyte and the level of CRP between the confirmed COVID-19 group and the control group(p<0.05). 20 babies (from confirmed mother and from normal mother) were subjected to SARS-CoV-2 examination by throat swab samples in 24 h after birth and no case was tested positive. CONCLUSION: The clinical symptoms and laboratory indicators are not obvious for asymptomatic and mild COVID-19 pregnant women. Pulmonary CT scan plus blood routine examination are more suitable for finding pregnancy women with asymptomatic or mild COVID-19 infection, and can be used screening COVID-19 pregnant women in the outbreak area of COVID-19 infection.