Mental health status of adolescents after family confinement during the COVID-19 outbreak in the general population: a longitudinal survey.

Few studies have examined the psychological impact on adolescents of family confinement and infection exposure during the COVID-19 pandemic. However, these surveys lacked follow-up data to determine how the family confinement affects participants' depression and anxiety. The purpose of this study was to evaluate the psychological status and related risk and protective factors of adolescents after two months of family confinement for preventing COVID-19 in China, and compare them with baseline data. We surveyed teenagers in January 2020 before the COVID-19 outbreak (T1) and after home confinement (T2). We used the Patient Health Questionnaire (PHQ), the Generalized Anxiety Disorder (GAD) Scale and the Childhood Trauma Questionnaire (CTQ). 13,637 valid questionnaires were collected at T1, of which 22.34% reported depressive symptoms (PHQ-9 ≥ 10) and 14.42% reported anxiety symptoms (GAD-7 ≥ 10). At T2, the rates decreased to 14.86 and 7.44%, respectively (all P < 0.0001). Of the adolescents, 223 reported potential risk of exposure to COVID-19. We then compared them to the 9639 non-risk adolescents using a propensity score matching analysis. The adolescents with potential exposure risk had higher rates of depression (26.91 vs 15.32%, P = 0.0035) and anxiety (14.80 vs 7.21%, P = 0.01) than risk-free adolescents. Among adolescents with an exposure risk, psychological resilience was protective in preventing depression and anxiety symptoms, while emotional abuse, a poor parent-child relationship were risk factors. Long-term home confinement had minimal psychological impact on adolescents, but COVID-19 infection rates accounted for 50% of the variance in depression and anxiety among adolescents even with low community rates.

Trafficked Malayan pangolins contain viral pathogens of humans.

Pangolins are the most trafficked wild animal in the world according to the World Wildlife Fund. The discovery of SARS-CoV-2-related coronaviruses in Malayan pangolins has piqued interest in the viromes of these wild, scaly-skinned mammals. We sequenced the viromes of 161 pangolins that were smuggled into China and assembled 28 vertebrate-associated viruses, 21 of which have not been previously reported in vertebrates. We named 16 members of Hunnivirus, Pestivirus and Copiparvovirus pangolin-associated viruses. We report that the L-protein has been lost from all hunniviruses identified in pangolins. Sequences of four human-associated viruses were detected in pangolin viromes, including respiratory syncytial virus, Orthopneumovirus, Rotavirus A and Mammalian orthoreovirus. The genomic sequences of five mammal-associated and three tick-associated viruses were also present. Notably, a coronavirus related to HKU4-CoV, which was originally found in bats, was identified. The presence of these viruses in smuggled pangolins identifies these mammals as a potential source of emergent pathogenic viruses.

Behavioral problems of pediatric patients recovered from COVID-19 in Wuhan, China.

BACKGROUND: Coronavirus disease 2019 (COVID-19) is profoundly affecting lives around the globe. Previous studies on COVID-19 mainly focused on epidemiological, clinical, and radiological features of patients with confirmed infection. Little attention has been paid to the follow-up of recovered patients. As a vulnerable population to adverse events, the health status of the COVID-19 recovered pediatric patients is of great concern. We aimed to investigate the prevalence of behavioral problems among pediatric patients recovered from the COVID-19 in Wuhan, China. METHODS: A total of 122 children who were suspected or confirmed COVID-19 cases and hospitalized for treatment were enrolled in the study between April 2020 and May 2020 in Wuhan, China. We collected related information about hospitalization and discharge of the children and emotional symptoms of their parents through electronic medical records and questionnaire. The behavioral problems of children were examined by applying the parent-reported the Strengths and Difficulties Questionnaire (SDQ). RESULTS: The participant children were discharged from hospital after about two months. Among them, 76 (62%) were boys, and the mean age was 6.71 years old. The highest prevalence of behavioral problems among pediatric children with COVID-19 was for prosocial behavior (15%), followed by total difficulties (13%), emotional symptoms (11%), hyperactivity (10%), conduct problems (9%), and peer problems (1%). With regarding to their parents, 26% reported having symptoms of anxiety and 23% as having symptoms of depression. The scores of SDQ were higher in those children whose parents have emotional problems compared to parents without. CONCLUSION: Long-term follow up studies on the psychological and behavioral problems of COVID-19 recovered children and their parents are warranted.

Behavioral problems of pediatric patients recovered from COVID-19 in Wuhan, China

Background Coronavirus disease 2019 (COVID-19) is profoundly affecting lives around the globe. Previous studies on COVID-19 mainly focused on epidemiological, clinical, and radiological features of patients with confirmed infection. Little attention has been paid to the follow-up of recovered patients. As a vulnerable population to adverse events, the health status of the COVID-19 recovered pediatric patients is of great concern. We aimed to investigate the prevalence of behavioral problems among pediatric patients recovered from the COVID-19 in Wuhan, China. Methods A total of 122 children who were suspected or confirmed COVID-19 cases and hospitalized for treatment were enrolled in the study between April 2020 and May 2020 in Wuhan, China. We collected related information about hospitalization and discharge of the children and emotional symptoms of their parents through electronic medical records and questionnaire. The behavioral problems of children were examined by applying the parent-reported the Strengths and Difficulties Questionnaire (SDQ). Results The participant children were discharged from hospital after about two months. Among them, 76 (62%) were boys, and the mean age was 6.71 years old. The highest prevalence of behavioral problems among pediatric children with COVID-19 was for prosocial behavior (15%), followed by total difficulties (13%), emotional symptoms (11%), hyperactivity (10%), conduct problems (9%), and peer problems (1%). With regarding to their parents, 26% reported having symptoms of anxiety and 23% as having symptoms of depression. The scores of SDQ were higher in those children whose parents have emotional problems compared to parents without. Conclusion Long-term follow up studies on the psychological and behavioral problems of COVID-19 recovered children and their parents are warranted.

Neuropilin-1-Mediated SARS-CoV-2 Infection in Bone Marrow-Derived Macrophages Inhibits Osteoclast Differentiation.

In humans, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection can cause medical complications across various tissues and organs. Despite the advances to understanding the pathogenesis of SARS-CoV-2, its tissue tropism and interactions with host cells have not been fully understood. Existing clinical data have revealed disordered calcium and phosphorus metabolism in Coronavirus Disease 2019 (COVID-19) patients, suggesting possible infection or damage in the human skeleton system by SARS-CoV-2. Herein, SARS-CoV-2 infection in mouse models with wild-type and beta strain (B.1.351) viruses is investigated, and it is found that bone marrow-derived macrophages (BMMs) can be efficiently infected in vivo. Single-cell RNA sequencing (scRNA-Seq) analyses of infected BMMs identify distinct clusters of susceptible macrophages, including those related to osteoblast differentiation. Interestingly, SARS-CoV-2 entry on BMMs is dependent on the expression of neuropilin-1 (NRP1) rather than the widely recognized receptor angiotensin-converting enzyme 2 (ACE2). The loss of NRP1 expression during BMM-to-osteoclast differentiation or NRP1 neutralization and knockdown can significantly inhibit SARS-CoV-2 infection in BMMs. Importantly, it is found that authentic SARS-CoV-2 infection impedes BMM-to-osteoclast differentiation. Collectively, this study provides evidence for NRP1-mediated SARS-CoV-2 infection in BMMs and establishes a potential link between disturbed osteoclast differentiation and disordered skeleton metabolism in COVID-19 patients.

Mental Health in Children in the Context of COVID-19: Focus on Discharged Children.

Introduction: To date, the mental health consequences of children hospitalized with COVID-19 remain unclear. We aimed to assess mental health status in children in the context of COVID-19, with a focus on discharged children. Methods: We recruited discharged children who recovered from COVID-19 and healthy controls between July and September 2020 in Wuhan Children's Hospital. Post-traumatic stress disorder (PTSD), anxiety, depression, and sleep problems were assessed in these children using questionnaires. Univariable and multivariable logistic and linear regressions were conducted to identify risk factors. Results: Totally, there were 152 children (61 discharged children and 91 healthy controls) aged 7-18 years old in our study. An increasing trend in the prevalence of PTSD, anxiety, and depression was observed in the discharged children compared with healthy controls (PTSD: 8.20 vs. 2.20%, anxiety: 22.95 vs. 13.19%; depression: 47.54 vs. 32.97%). Discharged children tended to report more depressive symptoms (ß = 0.39) and less sleep problems (ß = -0.37). Discharged children who lived in nuclear families and had longer hospital stays were more likely to report depression [odds ratio (OR) = 3.68 and 1.14, respectively]. Anxiety symptoms and the severity of sleep problems of discharged children were positively associated with caregivers' depression and PTSD symptoms (OR = 21.88 and 31.09, respectively). Conclusion: In conclusion, PTSD, anxiety, and depression symptoms were common among recovered children 4 months after COVID-19 hospitalization. Children from nuclear family and those had longer hospital stays need special attention. In addition, parental mental health had a significant impact on their children's mental resilience and recovery.

Depression, anxiety, and stress symptoms in pregnant women before and during the COVID-19 pandemic.

OBJECTIVE: This study evaluates depression, anxiety, and stress symptoms in pregnant women before and during COVID-19 pandemic and analyzes their risk factors. METHODS: This was a cross-sectional analyses included pregnant women with depression, anxiety, and stress levels evaluated both in the Novel Coronavirus-Pregnancy Cohort study (NCP) and the Healthy Baby Cohort study (HBC). NCP was conducted during COVID-19 pandemic, while HBC was performed before the pandemic. Multiple logistic regressions were employed to evaluate the associations between COVID-19 pandemic and other co-variables and maternal mental health. RESULTS: NCP and HBC studies respectively included 531 and 2352 participants. Depression rates differed significantly between the two studies (p < 0.05). The mild and moderate-to-severe depression rates in NCP study were 25.8% and 10.36%, respectively, and 19.94% and 0.55% in HBC study. The stress rate of participants was higher in HBC study (69.39%) than in NCP study (60.45%) (p < 0.05). COVID-19 pandemic was correlated with higher depression but lower stress risks (p < 0.05) in pregnant women, with OR and 95% CI as 1.68 (1.16, 2.44) and 0.42 (0.29, 0.61), respectively. Pregnant women with pre-pregnancy obesity and high educational levels might have lower risks for depression, anxiety, and stress than those with normal weight and low educational levels. CONCLUSIONS: Depression among pregnant women was impacted by the pandemic. Apart from COVID-19 pandemic impact, pre-pregnant weight status and educational level might also influence depression, anxiety and stress statuses in pregnant women.

Clinical features and follow-up of pediatric patients hospitalized for COVID-19.

OBJECTIVE: This report summarizes the clinical features and 1-month follow-up observations for pediatric patients who were hospitalized with coronavirus disease 2019 (COVID-19) in Wuhan Women and Children's Hospital. METHODS: The 1-month follow-up data included clinical manifestations and results from serum severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) IgG and IgM tests, reverse-transcription polymerase chain reaction (RT-PCR) for SARS-CoV-2, lung computed tomography (CT) scans, and laboratory tests. RESULTS: Between January 20 and March 15, 2020, 127 patients aged 0-15 years were hospitalized for COVID-19 treatment, including 3 severe cases and 124 mild or moderate cases. The main therapies included inhalation of aerosolized interferon-α (122/127) and additional antiviral drugs (28/127). Among the 81 patients who had pneumonia at admission, 35 with right lobe pneumonia had the longest hospital stay (mean 14.5 ± 7 days); 17 with left lobe pneumonia had the highest creatine kinase (154 ± 106 U/L) and creatine kinase myocardial band  (CK-MB, 43 ± 48 U/L) levels; and 29 with bilateral pneumonia had the highest white blood cell counts (8.3 ± 4 × 109 /L). Among the 46 patients who were successfully followed up 1 month after discharge, two notable findings were right lobe pneumonia in 22% (95% confidence interval [CI]: 11%-37%) of patients and persistently elevated serum creatine kinase and CK-MB levels. The median duration of elevated CK-MB was 45 days. The mean concentrations of serum SARS-CoV-2 IgG and IgM in 41 patients were 8.0 ± 7.5 and 98 ± 40 ng/ml, respectively. At follow-up, four patients retested positive for SARS-CoV-2. CONCLUSIONS: The involvement of different lung lobes in patients with COVID-19 was associated with variations in the persistence of pneumonia and elevation of CK-MB levels and body temperature.

Neuropilin-1 Mediates SARS-CoV-2 Infection in Bone Marrow-derived Macrophages (preprint)

SARS-CoV-2 infection in human can cause medical complications across various tissues and organs. Despite of the advances to understanding the pathogenesis of SARS-CoV-2, its tissue tropism and interactions with host cells have not been fully understood. Existing clinical data have suggested possible SARS-CoV-2 infection in human skeleton system. In the present study, we found that authentic SARS-CoV-2 could efficiently infect human and mouse bone marrow-derived macrophages (BMMs) and alter the expression of macrophage chemotaxis and osteoclast-related genes. Importantly, in a mouse SARS-CoV-2 infection model that was enabled by the intranasal adenoviral (AdV) delivery of human angiotensin converting enzyme 2 (hACE2), SARS-CoV-2 was found to be present in femoral BMMs as determined by in situ immunofluorescence analysis. Using single-cell RNA sequencing (scRNA-Seq), we characterized SARS-CoV-2 infection in BMMs. Importantly, SARS-CoV-2 entry on BMMs appeared to be dependent on the expression of neuropilin-1 (NRP1) rather than the widely recognized receptor ACE2. It was also noted that unlike brain macrophages which displayed aging-dependent NRP1 expression, BMMs from neonatal and aged mice had constant NRP1 expression, making BMMs constantly vulnerable target cells for SARS-CoV-2. Furthermore, it was found that the abolished SARS-CoV-2 entry in BMM-derived osteoclasts was associated with the loss of NRP1 expression during BMM-to-osteoclast differentiation. Collectively, our study has suggested that NRP1 can mediate SARS-CoV-2 infection in BMMs, which precautions the potential impact of SARS-CoV-2 infection on human skeleton system.

Development of a Broadly Applicable Cas12a-Linked Beam Unlocking Reaction for Sensitive and Specific Detection of Respiratory Pathogens Including SARS-CoV-2.

The outbreak of novel coronavirus SARS-CoV-2 has caused a worldwide threat to public health. COVID-19 patients with SARS-CoV-2 infection can develop clinical symptoms that are often confused with the infections of other respiratory pathogens. Sensitive and specific detection of SARS-CoV-2 with the ability to discriminate from other viruses is urgently needed for COVID-19 diagnosis. Herein, we streamlined a highly efficient CRISPR-Cas12a-based nucleic acid detection platform, termed Cas12a-linked beam unlocking reaction (CALIBURN). We show that CALIBURN could detect SARS-CoV-2 and other coronaviruses and influenza viruses with little cross-reactivity. Importantly, CALIBURN allowed accurate diagnosis of clinical samples with extremely low viral loads, which is a major obstacle for the clinical applications of existing CRISPR diagnostic platforms. When tested on the specimens from SARS-CoV-2-positive and negative donors, CALIBURN exhibited 73.0% positive and 19.0% presumptive positive rates and 100% specificity. Moreover, unlike existing CRISPR detection methods that were mainly restricted to respiratory specimens, CALIBURN displayed consistent performance across both respiratory and nonrespiratory specimens, suggesting its broad specimen compatibility. Finally, using a mouse model of SARS-CoV-2 infection, we demonstrated that CALIBURN allowed detection of coexisting pathogens without cross-reactivity from a single tissue specimen. Our results suggest that CALIBURN can serve as a versatile platform for the diagnosis of COVID-19 and other respiratory infectious diseases.

MeCas12a, a Highly Sensitive and Specific System for COVID-19 Detection.

Cas12a-based systems, which detect specific nucleic acids via collateral cleavage of reporter DNA, display huge potentials for rapid diagnosis of infectious diseases. Here, the Manganese-enhanced Cas12a (MeCas12a) system is described, where manganese is used to increase the detection sensitivity up to 13-fold, enabling the detection of target RNAs as low as five copies. MeCas12a is also highly specific, and is able to distinguish between single nucleotide polymorphisms (SNPs) differing by a single nucleotide. MeCas12a can detect severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in clinical samples and distinguish between SARS-CoV-2 and Middle East respiratory syndrome coronavirus (MERS-CoV) RNA in simulated samples, thus offering an attractive alternative to other methods for the diagnosis of infectious diseases including COVID-19 and MERS.

Structure and Drug Binding of the SARS-CoV-2 Envelope Protein in Phospholipid Bilayers (preprint)

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent of the ongoing COVID-19 pandemic. Successful development of vaccines and antivirals against SARS-CoV-2 requires a comprehensive understanding of the essential proteins of the virus. The envelope (E) protein of SARS-CoV-2 assembles into a cation-selective channel that mediates virus budding, release, and host inflammation response. E blockage reduces virus pathogenicity while E deletion attenuates the virus. Here we report the 2.4 Å structure and drug-binding site of E’s transmembrane (TM) domain, determined using solid-state nuclear magnetic resonance (NMR) spectroscopy. In lipid bilayers that mimic the endoplasmic reticulum Golgi intermediate compartment (ERGIC) membrane, ETM forms a five-helix bundle surrounding a narrow central pore. The middle of the TM segment is distorted from the ideal a-helical geometry due to three regularly spaced phenylalanine residues, which stack within each helix and between neighboring helices. These aromatic interactions, together with interhelical Val and Leu interdigitation, cause a dehydrated pore compared to the viroporins of influenza and HIV viruses. Hexamethylene amiloride and amantadine bind shallowly to polar residues at the N-terminal lumen, while acidic pH affects the C-terminal conformation. These results indicate that SARS-CoV-2 E forms a structurally robust but bipartite channel whose N- and C-terminal halves can interact with drugs, ions and other viral and host proteins semi-independently. This structure establishes the atomic basis for designing E inhibitors as antiviral drugs against SARS-CoV-2.

Clinical efficacy and safety of hydroxychloroquine sulfate combined with azithromycin in treatment of patients with refractory common coronavirus disease 2019

Objective To evaluate the clinical efficacy and safety of hydroxychloroquine sulfate combined with azithromycin in the treatment of refractory common coronavirus disease 2019 (COVID-19) patients. Methods The clinical data of 11 refractory common COVID-19 patients, who were admitted to Guanggu Branch of Maternity and Child Healthcare Hospital of Hubei Province from Mar. 22 to 25, 2020, were retrospectively collected. The patients all received combined treatment regimens: hydroxychloroquine sulfate orally 200 mg three times daily for 7 days;and azithromycin orally 500 mg once daily on day 1 and then 250 mg once daily from day 2 to day 4. The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) nucleic acid test of throat swab was performed once daily from day 4 to day 10 after combined administration, and the blood routine and other laboratory indicators were tested within 3 days before administration and on the 8th days after administration. Results All the 11 patients had common COVID-19, seven of them were consistently positive for SARSCoV- 2 nucleic acid test, and four were positive again after negative results. The average course of disease of 11 patients before combined administration was 50.2 days. The treatment process was uneventful. Zero case of SARS-CoV-2 nucleic acid test result turned negative on day 4 after administration, two cases on day 5, two cases on day 6, two cases on day 7, one case on day 8 and one case on day 9. No patients progressed to severe or critical illness, and no severe side effects were found. Conclusion Hydroxychloroquine sulfate combined with azithromycin is safe and effective in the treatment of refractory common COVID-19 patients who have ailed in other treatments and are consistently positive for SARS-CoV-2 nucleic acid.

Mental Health Status of Adolescents after Family Confinement During the COVID-19 Outbreak (preprint)

Background: With the global spread of Coronavirus Disease 2019 (COVID-19), governments around the world have implemented strategies for closing schools and teaching online from home. However, the psychological impact of family confinement on adolescents has not been examined. Methods: From November 2019 to January 2020, just before the COVID-19 outbreak, 14,241 Chinese teenagers aged 12-17 years from 5 middle schools across North China were surveyed in the first round of the national mental health investigation of adolescents. They were assessed with paper questionnaires, including the Chinese version of the 9-item Patient Health Questionnaire, the 7-item Generalized Anxiety Disorder Scale, the short form of the Childhood Trauma Questionnaire, and the Connor-Davidson Resilience Scale. After two months of home confinement, by March 2020, 10,768 of them were re-evaluated online using the same scales. Propensity score matching analysis was performed to analyze the psychological status of adolescents who reported potential risk of exposure to COVID-19. Findings: The first survey round yielded 13,637 valid questionnaires, on which 51.51% reported depressive symptoms and 38.53% reported anxiety symptoms. The second round yielded 10,216 valid questionnaires, on which the rates decreased to 38.29% and 23.73% respectively (all P<0.0001). Of the 10,216 adolescents, 223 reported potential risk of exposure to COVID-19. Propensity score matching analysis showed that adolescents with potential exposure risks had significantly higher rates of depression (60.54% vs 45.95% P=0.002) and anxiety (41.26% vs 28.83% P=0.007) symptoms than risk-free adolescents. Multivariate logistic regression analysis showed that potential exposure risk to COVID-19 was an independent risk factor for depression symptoms (OR, 2.241; 95%CI, 1.659-3.027; PInterpretations: Long-term home confinement had no adverse psychological impact on adolescents from regions with a low incidence of COVID-19. However, several adolescent groups had serious psychological complications and needed interventions in time. These adolescents included those at potential risk of exposure to COVID-19. Therefore, the strategy of closing schools should be implemented at early stage of pandemic to minimize the adverse psychological impact on adolescents.Funding Statement: This work was supported by the National Natural Science Foundation of China (Grant No.81573905) and program of National Clinical/Scientific Excellence in Chinese Medicine.Declaration of Interests: There are no conflicts of interest.Ethics Approval Statement: Our data collection protocol followed the American Association for Public Opinion Research (AAPOR) reporting guidelines, and was approved by the Ethics in Human Research Committee of the Third Affiliated Hospital of Beijing University of Chinese Medicine (No. BZYSY-2019KYKTPJ-21). We obtained informed consents from guardians before including participants in the study. The survey was anonymous and the information from the participants was confidential.

Clinical symptoms and psychological changes of patients with COVID-19 in Jiangxi Province (preprint)

Objective The purpose of this study was to determine the prevalence and differences in etiology, clinical manifestations, and psychological activity of coronavirus disease-19 (COVID-19) among patients. Results We recruited 90 subjects, 30 were healthy controls, 30 were patients with moderate infection, and 30 were patients with severe/critical infections. No significant differences were noted in the sex ratio, mean age, body mass index, or blood type; however, the history of exposure of the patients with COVID-19 compared with healthy controls was noteworthy. The erythrocyte sedimentation rate, as well as the levels of C-reactive protein and serum amyloid A (SAA) were all increased. In terms of mental health, there were significant differences in the worry scores between severely and moderately infected patients and healthy controls. There was a significant difference in depression scores between patients with moderate infection and healthy hypertension, and there was also a significant difference in dream worry scores. Analysis of the Mini-Mental State Examination scores showed that for patients with moderate infection, the depression score was moderately and positively correlated with the dream anxiety score. For patients with severe infection, the anxiety score was positively correlated with the dream anxiety score, and the depression score was moderately and positively correlated with the dream anxiety score. Conclusion Patients with severe infection showed increased pain and sputum in the pharyngeal area compared with patients with moderate infection. Patients with blood type A may be more susceptible to COVID-19, and lymphopenia may indicate worsening of COVID-19.