Engineering antiviral immune-like systems for autonomous virus detection and inhibition in mice.

The ongoing COVID-19 pandemic has demonstrated that viral diseases represent an enormous public health and economic threat to mankind and that individuals with compromised immune systems are at greater risk of complications and death from viral diseases. The development of broad-spectrum antivirals is an important part of pandemic preparedness. Here, we have engineer a series of designer cells which we term autonomous, intelligent, virus-inducible immune-like (ALICE) cells as sense-and-destroy antiviral system. After developing a destabilized STING-based sensor to detect viruses from seven different genera, we have used a synthetic signal transduction system to link viral detection to the expression of multiple antiviral effector molecules, including antiviral cytokines, a CRISPR-Cas9 module for viral degradation and the secretion of a neutralizing antibody. We perform a proof-of-concept study using multiple iterations of our ALICE system in vitro, followed by in vivo functionality testing in mice. We show that dual output ALICESaCas9+Ab system delivered by an AAV-vector inhibited viral infection in herpetic simplex keratitis (HSK) mouse model. Our work demonstrates that viral detection and antiviral countermeasures can be paired for intelligent sense-and-destroy applications as a flexible and innovative method against virus infection.

An Empirical Study on the Incentive Mechanism for Public Active Involvement in Grass-Roots Social Governance Based on Stimulus-Organism-Response Theory

In the period of the normalization of COVID-19 prevention and control, Chinese grass-roots social governance, under the guidance of the dynamic zero-COVID policy, was unique, and the experience of actively mobilizing the public to be actively involved in grass-roots social governance, represented by epidemic prevention and control, has a profound internal logic. The Chinese government has long been committed to building a social governance community, and further empirical studies are needed to motivate the public to participate in grass-roots social governance in a sustainable manner. This study uses 428 members of the public who have experience in grass-roots social governance in 20 street offices in 11 cities, including Nanjing, Wuhan and Chengdu, as a valid sample to empirically test the incentive mechanism for the public's active involvement in grass-roots social governance, from the perspective of Stimulus-Organism-Response Theory. The empirical results show that exogenously driven organizational institutional factors will eventually positively influence the incentive effect on the public's active involvement in grass-roots social governance through the mediating effect of the individual's endogenous drive. By adjusting organizational institutional factors to meet the public's inner drive for acquisition, bond, comprehension, and defense, public motivation can be mobilized and the public can be motivated to be involved in grass-roots social governance in a sustainable manner. The results of the study reveal the incentive mechanism for the public's active involvement in grass-roots social governance, analyze the internal logic of Chinese characteristics in motivating the public's active involvement in grass-roots governance, and expand the scope of the application of Stimulus-Organism-Response Theory in studying the incentives for the public's active involvement in grass-roots social governance, which is important for revealing the characteristic laws in a Chinese context with empirical research.

Microfluidic Chip with Two-Stage Isothermal Amplification Method for Highly Sensitive Parallel Detection of SARS-CoV-2 and Measles Virus.

A two-stage isothermal amplification method, which consists of a first-stage basic recombinase polymerase amplification (RPA) and a second-stage fluorescence loop-mediated isothermal amplification (LAMP), as well as a microfluidic-chip-based portable system, were developed in this study; these enabled parallel detection of multiplex targets in real time in around one hour, with high sensitivity and specificity, without cross-contamination. The consumption of the sample and the reagent was 2.1 µL and 10.6 µL per reaction for RPA and LAMP, respectively. The lowest detection limit (LOD) was about 10 copies. The clinical amplification of about 40 nasopharyngeal swab samples, containing 17 SARS-CoV-2 (severe acute respiratory syndrome coronavirus) and 23 measles viruses (MV), were parallel tested by using the microfluidic chip. Both clinical specificity and sensitivity were 100% for MV, and the clinical specificity and sensitivity were 94.12% and 95.83% for SARS-CoV-2, respectively. This two-stage isothermal amplification method based on the microfluidic chip format offers a convenient, clinically parallel molecular diagnostic method, which can identify different nucleic acid samples simultaneously and in a timely manner, and with a low cost of the reaction reagent. It is especially suitable for resource-limited areas and point-of-care testing (POCT).

The predictive role of lymphocyte subsets and laboratory measurements in COVID-19 disease: a retrospective study.

AIM: The aim of this study was to investigate the predictive role of lymphocyte subsets and other laboratory measurements in patients with COVID-19. METHODS: Electronic medical records of adult patients with confirmed diagnosis of COVID-19 from the Shanghai Public Health Clinical Center were reviewed retrospectively to obtain relevant data. RESULTS: The mean age of patients was 40.98 ± 15.95 years, with 58% of the patients being males. The cutoff values at the intensive care unit (ICU) admission, mechanical ventilation, and mortality were CD4+ cells (267, 198, and 405), CD8+ cells (263, 203, and 182), and CD4+ /CD8+ cells (1.4, 1.8, and 1.4). The cutoffs below these values indicate the higher chances of disease progression. Higher CD4+ cell count led to lesser chances for ICU admission [odds ratio (OR) (95% confidence interval (CI): 0.994 (0.991, 0.997); p = 0.0002] and mortality [OR (95% CI): 0.988 (0.979, 0.99); p = 0.001], higher CD8+ count was an independent risk factor for ICU admission. T-cell count positively correlated with total lymphocyte count and platelets, while negatively correlated with D-dimer and lactate dehydrogenase (LDH). Among patients with non-severe COVID-19, median CD8+ T cell, CD4+ T cell, total lymphocyte count, and platelets were 570, 362, 1.45, and 211, respectively, while median values decreased to 149, 106, 0.64, and 172, respectively, in patients with severe COVID-19. CONCLUSION: Lower T lymphocyte subsets were significantly associated with higher admission to ICU, mechanical ventilation, and mortality among patients with COVID-19. A cutoff value of ICU admission, mechanical ventilation, and mortality below CD4+ cells (267, 198, and 405), CD8+ cells (263, 203, 182), and CD4+/CD8+ cells (1.4, 1.8, 1.4) may help identify patients at high risk of disease progression. The continuous evaluation of laboratory indices may help with dismal prognosis and prompt intervention to improve outcomes.

Point-of-care training program on COVID-19 infection prevention and control for pediatric healthcare workers: a multicenter, cross-sectional questionnaire survey in Shanghai, China.

BACKGROUND: To evaluate the effectiveness of training on knowledge and practice of infection prevention and control (IPC) among pediatric health care workers (HCW) in Shanghai, China, in the context of COVID-19 pandemic. METHODS: An online training program was designed by the Shanghai Pediatric Clinical Quality Control Center (SPQCC) during the early phase of COVID-19 pandemic on disease knowledge and practice of IPC. Training took place in the 81 partner hospitals affiliated with SPQCC. A multicenter, cross-sectional questionnaire survey was designed with a 25-item self-administered questionnaire to evaluate the knowledge gained from the training. Stratified-random sampling was used to select HCW according to three professionals (i.e., pediatricians, nurses and administrators) within each partner hospital. Awareness and knowledge of COVID-19 and its related infection control and practice were assessed by comparing survey results between different types of hospitals, professionals and professional ranks. A higher survey score meant that the respondent was more prepared and knowledgeable about COVID-19 and its infection control measures. RESULTS: Completed questionnaires were returned from 1,062 subjects (385 pediatricians, 410 nurses, and 267 administrators), giving a response rate of 96.5%. Overall, awareness of clinical information related to COVID-19, importance of personal hygiene and isolation policy was high among the respondents. No statistical difference of scores on knowledge of COVID-19, IPC and relevant practice between the tertiary and peripheral hospitals. Among all respondents, middle-ranked health care personnel were most knowledgeable and achieved the highest score. CONCLUSIONS: Majority of pediatric HCW showed good recognition and practice in infection protection and control measures. The online training was able to achieve its aim to enhance knowledge and awareness and could have contributed to the zero infection rate among HCW caring for confirmed COVID-19 cases in Shanghai.

COVID-19 infection in patients with connective tissue disease: A multicity study in Hubei province, China.

Novel Coronavirus disease 2019 (COVID-19) has spread rapidly around the world. Individuals with immune dysregulation and/or on immunosuppressive therapy, such as rheumatic patients, are considered at greater risk for infections. However, the risks of patients with each subcategory of rheumatic diseases have not been reported. Here, we identified 100 rheumatic patients from 18,786 COVID-19 patients hospitalized in 23 centers affiliated to Hubei COVID-19 Rheumatology Alliance between January 1 and April 1, 2020. Demographic information, medical history, length of hospital stay, classification of disease severity, symptoms and signs, laboratory tests, disease outcome, computed tomography, and treatments information were collected. Compared to gout and ankylosing spondylitis (AS) patients, patients with connective tissue disease (CTD) tend to be more severe after COVID-19 infection (p = 0.081). CTD patients also had lower lymphocyte counts, hemoglobin, and platelet counts (p values were 0.033, < 0.001, and 0.071, respectively). Hydroxychloroquine therapy and low- to medium-dose glucocorticoids before COVID-19 diagnosis reduced the progression of COVID-19 to severe/critical conditions (p = 0.001 for hydroxychloroquine; p = 0.006 for glucocorticoids). Our data suggests that COVID-19 in CTD patients may be more severe compared to patients with AS or gout.

The Important Role of Volatile Components From a Traditional Chinese Medicine Dayuan-Yin Against the COVID-19 Pandemic.

Aromatic Chinese herbs have been used to prevent plagues since ancient times. Traditional Chinese medicine has unique advantages in the prevention and treatment of epidemic diseases. According to the traditional Chinese medicine treatment plan in the National COVID-19 Diagnosis and Treatment Plan (Trial Seventh Edition) of the National Health Commission, Chinese patent medicines or prescriptions rich in aromatic Chinese herbs are selected for prevention and treatment during the period of medical observation, clinical treatment, and recovery of confirmed COVID-19 patients. Some local health committees or traditional Chinese medicine administrations recommend a variety of other ways of using traditional aromatic Chinese herbs to prevent and cure COVID-19. These involve external fumigation, use of moxibustion, and wearing of sachet. The efficacy of aromatic Chinese herbs plays a decisive role in the prevention and treatment of COVID-19. The unique properties, chemical composition, and mechanism of action of aromatic Chinese herbs are worthy of extensive and in-depth experimental and clinical research. The findings are expected to provide a reference for follow-up treatment of novel coronavirus and the development of corresponding drugs. In 2003, Dayuan-Yin produced excellent results in the treatment of the SARS virus. Individually, 112 confirmed cases were administered this drug between January and April 2003, and more than 93.7% of the patients showed noticeable mitigation of the symptoms, as well as recovery. Dayuan-Yin also was selected as one of the nationally recommended prescriptions for the COVID-19. Based on the national recommendation of Dayuan-Yin prescription, this review discusses the role of volatile components in the prevention and treatment of COVID-19, and speculates the possible mechanism of action, so as to provide a basis for the prevention and treatment of COVID-19.

The mental health of neurological doctors and nurses in Hunan Province, China during the initial stages of the COVID-19 outbreak.

BACKGROUND: Neurological symptoms are increasingly being noted among COVID-19 patients. Currently, there is little data on the mental health of neurological healthcare workers. The aim of this study was to identify the prevalence and influencing factors on anxiety and depression in neurological healthcare workers in Hunan Province, China during the early stage of the Coronavirus Disease 2019 (COVID-19) outbreak. METHODS: An online cross-sectional study was conducted among neurological doctors and nurses in early February 2020 in Hunan Province. Symptoms of anxiety and depression were assessed by the Chinese version of the Self-Rating Anxiety Scale (SAS) (defined as a total score ≥ 50) and Self-Rating Depression Scale (SDS) (defined as a total score ≥ 53). The prevalences of probable anxiety and depression were compared between different groups, and multivariate logistic regression analysis was used to understand the independent influencing factors on anxiety and depression. RESULTS: The prevalence of probable anxiety and depression in neurological nurses (20.3 and 30.2%, respectively) was higher than that in doctors (12.6 and 20.2%, respectively). Female healthcare workers (18.4%) had a higher proportion of anxiety than males (10.8%). Probable anxiety and depression were more prevalent among nurses, younger workers (≤ 40 years), and medical staff with junior titles. Logistic regression analysis showed that a shortage of protective equipment was independently associated with probable anxiety (OR = 1.980, 95% CI: 1.241-3.160, P = 0.004), while young age was a risk factor for probable depression (OR = 2.293, 95% CI: 1.137-4.623, P = 0.020) among neurological healthcare workers. CONCLUSIONS: Probable anxiety and depression were more prevalent among neurological nurses than doctors in Hunan Province. The shortage of protective equipment led to probable anxiety, and young age led to probable depression in healthcare workers in neurology departments, which merits attention during the battle against COVID-19.

COVID-19 in patients with rheumatic disease in Hubei province, China: a multicentre retrospective observational study.

BACKGROUND: In the ongoing COVID-19 pandemic, the susceptibility of patients with rheumatic diseases to COVID-19 remains unclear. We aimed to investigate susceptibility to COVID-19 in patients with autoimmune rheumatic diseases during the ongoing COVID-19 pandemic. METHODS: We did a multicentre retrospective study of patients with autoimmune rheumatic diseases in Hubei province, the epicentre of the COVID-19 outbreak in China. Patients with rheumatic diseases were contacted through an automated telephone-based survey to investigate their susceptibility to COVID-19. Data about COVID-19 exposure or diagnosis were collected. Families with a documented history of COVID-19 exposure, as defined by having at least one family member diagnosed with COVID-19, were followed up by medical professionals to obtain detailed information, including sex, age, smoking history, past medical history, use of medications, and information related to COVID-19. FINDINGS: Between March 20 and March 30, 2020, 6228 patients with autoimmune rheumatic diseases were included in the study. The overall rate of COVID-19 in patients with an autoimmune rheumatic disease in our study population was 0·43% (27 of 6228 patients). We identified 42 families in which COVID-19 was diagnosed between Dec 20, 2019, and March 20, 2020, in either patients with a rheumatic disease or in a family member residing at the same physical address during the outbreak. Within these 42 families, COVID-19 was diagnosed in 27 (63%) of 43 patients with a rheumatic disease and in 28 (34%) of 83 of their family members with no rheumatic disease (adjusted odds ratio [OR] 2·68 [95% CI 1·14-6·27]; p=0·023). Patients with rheumatic disease who were taking hydroxychloroquine had a lower risk of COVID-19 infection than patients taking other disease-modifying anti-rheumatic drugs (OR 0·09 [95% CI 0·01-0·94]; p=0·044). Additionally, the risk of COVID-19 was increased with age (adjusted OR 1·04 [95%CI 1·01-1·06]; p=0·0081). INTERPRETATION: Patients with autoimmune rheumatic disease might be more susceptible to COVID-19 infection than the general population. FUNDING: National Natural Science Foundation of China and the Tongji Hospital Clinical Research Flagship Program.

Epidemiology and clinical course of COVID-19 in Shanghai, China.

Background: Novel coronavirus pneumonia (COVID-19) is prevalent around the world. We aimed to describe epidemiological features and clinical course in Shanghai. Methods: We retrospectively analysed 325 cases admitted at Shanghai Public Health Clinical Center, between January 20 and February 29, 2020. Results: 47.4% (154/325) had visited Wuhan within 2 weeks of illness onset. 57.2% occurred in 67 clusters; 40% were situated within 53 family clusters. 83.7% developed fever during the disease course. Median times from onset to first medical care, hospitalization and negative detection of nucleic acid by nasopharyngeal swab were 1, 4 and 8 days. Patients with mild disease using glucocorticoid tended to have longer viral shedding in blood and feces. At admission, 69.8% presented with lymphopenia and 38.8% had elevated D-dimers. Pneumonia was identified in 97.5% (314/322) of cases by chest CT scan. Severe-critical patients were 8% with a median time from onset to critical disease of 10.5 days. Half required oxygen therapy and 7.1% high-flow nasal oxygen. The case fatality rate was 0.92% with median time from onset to death of 16 days. Conclusion: COVID-19 cases in Shanghai were imported. Rapid identification, and effective control measures helped to contain the outbreak and prevent community transmission.