The CRISPR/Cas System: A Customizable Toolbox for Molecular Detection.

Clustered regularly interspaced short palindromic repeats (CRISPR) and their associated proteins (Cas) are promising molecular diagnostic tools for rapidly and precisely elucidating the structure and function of genomes due to their high specificity, programmability, and multi-system compatibility in nucleic acid recognition. Multiple parameters limit the ability of a CRISPR/Cas system to detect DNA or RNA. Consequently, it must be used in conjunction with other nucleic acid amplification techniques or signal detection techniques, and the reaction components and reaction conditions should be modified and optimized to maximize the detection performance of the CRISPR/Cas system against various targets. As the field continues to develop, CRISPR/Cas systems have the potential to become an ultra-sensitive, convenient, and accurate biosensing platform for the detection of specific target sequences. The design of a molecular detection platform employing the CRISPR/Cas system is asserted on three primary strategies: (1) Performance optimization of the CRISPR/Cas system; (2) enhancement of the detection signal and its interpretation; and (3) compatibility with multiple reaction systems. This article focuses on the molecular characteristics and application value of the CRISPR/Cas system and reviews recent research progress and development direction from the perspectives of principle, performance, and method development challenges to provide a theoretical foundation for the development and application of the CRISPR/CAS system in molecular detection technology.

Health-Risk Behaviors among Chinese Adults during the COVID-19 Pandemic.

This study analyzed archived data from a previous large-scale survey study on multiple health-risk behaviors among Chinese adults in Hong Kong between 21 June and 31 August 2021. In addition, this study examined participants' perceptions of the risks associated with their behaviors, their attitudes toward adopting healthy behaviors, and the impact of COVID-19 on their health-risk behaviors. A total of 4605 participants who had at least one health-risk behavior were included in the analysis. The results showed that about half of the participants were unaware that non-communicable diseases (NCDs) can be caused by health-risk behaviors such as tobacco use, harmful use of alcohol, physical inactivity, and an unhealthy diet. More than half of the participants did not have regular body checkups or monitor their physical health at home. Many participants paid more attention to their health due to the COVID-19 pandemic, but few made changes to their unhealthy habits or adopted a healthier lifestyle. Of the 704 smokers, 60.9% did not realize that smoking increases the risk of transmitting COVID-19 to others. Only 32.2% and 11.9% smokers had the intention to quit smoking and reduced their cigarette consumption, respectively. Additionally, 13.6% reported that their daily cigarette consumption had increased, and 78.8% changed their smoking behavior during the pandemic. Healthcare professionals must educate the public about the association between health-risk behaviors and NCDs and between COVID-19 and NCDs. The government should formulate a long-term plan to strengthen the primary healthcare system and address the challenges posed by the rising prevalence of NCDs.

A novel RBD-protein/peptide vaccine elicits broadly neutralizing antibodies and protects mice and macaques against SARS-CoV-2.

The development of safe and effective vaccines to respond to COVID-19 pandemic/endemic remains a priority. We developed a novel subunit protein-peptide COVID-19 vaccine candidate (UB-612) composed of: (i) receptor binding domain of SARS-CoV-2 spike protein fused to a modified single-chain human IgG1 Fc; (ii) five synthetic peptides incorporating conserved helper and cytotoxic T lymphocyte (Th/CTL) epitopes derived from SARS-CoV-2 structural proteins (three from S2 subunit, one from membrane and one from nucleocapsid), and one universal Th peptide; (iii) aluminum phosphate as adjuvant. The immunogenicity and protective immunity induced by UB-612 vaccine were evaluated in four animal models: Sprague-Dawley rats, AAV-hACE2 transduced BALB/c mice, rhesus and cynomolgus macaques. UB-612 vaccine induced high levels of neutralizing antibody and T-cell responses, in all animals. The immune sera from vaccinated animals neutralized the SARS-CoV-2 original wild-type strains and multiple variants of concern, including Delta and Omicron. The vaccination significantly reduced viral loads, lung pathology scores, and disease progression after intranasal and intratracheal challenge with SARS-CoV-2 in mice, rhesus and cynomolgus macaques. UB-612 has been tested in primary regimens in Phase 1 and Phase 2 clinical studies and is currently being evaluated in a global pivotal Phase 3 clinical study as a single dose heterologous booster.

Mental health issues in parents of children with autism spectrum disorder: A multi-time-point study related to COVID-19 pandemic.

Given the unpredictability and challenges brought about by the 2019 novel coronavirus (COVID-19) pandemic, this study aimed to investigate the impact trend of the prolonged pandemic on the mental health of parents of children with autism spectrum disorder (ASD). The 8112 participants included parents of children with ASD and parents of typically developing (TD) children at two sites (Heilongjiang and Fujian province, China). The parents completed a set of self-report questionnaires covering demographic characteristics, influences related to COVID-19, COVID-19 concerns and perceived behaviors, as well as the Connor-Davidson resilience scale (CD-RISC), self-rating anxiety scale (SAS), and self-rating depression scale (SDS) by means of an online survey platform. Data were collected by three cross-sectional surveys carried out in April 2020 (Time 1), October 2020 (Time 2), and October 2021 (Time 3). The results of quantitative and qualitative comparisons showed that: (i) parents of children with ASD had lower levels of resilience, and more symptoms of anxiety and depression than parents of TD children at each time point (all P < 0.05); and (ii) there were significant time-cumulative changes in resilience, anxiety, and depression among all participants (all P < 0.05). The logistic regression analyzes after adjusting for demographic characteristics revealed that the following factors were significantly associated with poor resilience and a higher rate of anxiety and depression in parents of children with ASD: time-point, the effect of COVID-19 on children's emotions and parents' emotions, changes in relationships, changes in physical exercise, changes in daily diet during the COVID-19 pandemic, and COVID-19-related psychological distress. In conclusions, the parents did not report improvements in resilience, anxiety, or depression symptoms from Time 1 to Time 2 or 3, indicating that cumulative mental health issues increased when, surprisingly, the COVID-19 restrictions were eased. The psychological harm resulting from the COVID-19 pandemic is far-reaching, especially among parents of children with ASD.

Structural characteristics of China's COVID-19 patent cooperation network at the province, city, and patent applicant levels.

The Chinese Plan has provided an important model for the global fight against COVID-19 since its outbreak. The present study describes the structural characteristics of China's COVID-19 patent cooperation network at the province, city, and applicant levels by using social network analysis based on data from the Incopat global patent database since 2020, which helps to clarify the current technology accumulation in this field in China, and provide patent information support for the scientific efforts to fight against COVID-19. The findings are as follows: First, the inter-regional cooperation level in China's COVID-19 patent cooperation network shows a decreasing trend from eastern to central to western regions. At the inter-applicant cooperation level, kinship-based cooperation is the strongest, business-based cooperation has the widest scope, while proximity-based cooperation exists throughout these two main models of cooperation. Second, coastal provinces and cities occupy a core position in the network, and play an important role in utilizing structural holes and bridging. Patent applicants with high centrality are mostly firms. Research institutes and universities mainly play the role of bridges. Third and lastly, there is no large number of cliques at the province and city levels. However, there is a tendency for cliques to develop at the applicant level. Hence, actions are needed to prevent the development of information barriers.

Spatiotemporal evolution of online attention to vaccines since 2011: An empirical study in China.

Since the outbreak of Coronavirus Disease 2019 (COVID-19), the Chinese government has taken a number of measures to effectively control the pandemic. By the end of 2021, China achieved a full vaccination rate higher than 85%. The Chinese Plan provides an important model for the global fight against COVID-19. Internet search reflects the public's attention toward and potential demand for a particular thing. Research on the spatiotemporal characteristics of online attention to vaccines can determine the spatiotemporal distribution of vaccine demand in China and provides a basis for global public health policy making. This study analyzes the spatiotemporal characteristics of online attention to vaccines and their influencing factors in 31 provinces/municipalities in mainland China with Baidu Index as the data source by using geographic concentration index, coefficient of variation, GeoDetector, and other methods. The following findings are presented. First, online attention to vaccines showed an overall upward trend in China since 2011, especially after 2016. Significant seasonal differences and an unbalanced monthly distribution were observed. Second, there was an obvious geographical imbalance in online attention to vaccines among the provinces/municipalities, generally exhibiting a spatial pattern of "high in the east and low in the west." Low aggregation and obvious spatial dispersion among the provinces/municipalities were also observed. The geographic distribution of hot and cold spots of online attention to vaccines has clear boundaries. The hot spots are mainly distributed in the central-eastern provinces and the cold spots are in the western provinces. Third, the spatiotemporal differences in online attention to vaccines are the combined result of socioeconomic level, socio-demographic characteristics, and disease control level.

Predictive Value of Systemic Immune-Inflammation index and Neutrophil-to-Lymphocyte Ratio in Patients with Severe COVID-19.

Objective: It was initially reported that a novel coronavirus (COVID-19) had been identified in Wuhan, China, in December 2019.To date, COVID-19 is still threatening all humanity and has affected the public healthcare system and the world economic situation. Neutrophil-to-lymphocyte ratio (NLR) has also been demonstrated that associated with severity of COVID-19, but little is known about systemic immune-inflammation index (SII) relation with COVID-19. Methods: One hundred and twenty-five patients with diagnosed COVID-19 including non-severe cases (n = 77) and severe cases (n = 48) were enrolled in this study. Each patient of clinical characteristic information, blood routine parameters, and the haemogram-derived ratios were collected, calculated, and retrospectively analyzed. Receiver operating characteristics (ROC) was performed to investigate whether these parameters could be used to the predictive value of patients with severe COVID-19. Results: White blood cell count (WBC), neutrophil count (NEU), red cell volume distribution width (RDW), NLR, Platelet to lymphocyte ratio (PLR), neutrophil-to-platelet ratio (NPR), and SII were significantly higher in the severe groups than in the non-severe group (p < 0.01).Conversely, the severe group had a markedly decreased lymphocyte count, basophil (Baso#) count, red blood cell count (RBC), Hemoglobin (HGB), hematocrit (HCT), and lymphocyte-to-monocyte ratio (LMR) (P < 0.01).ROC curve analysis showed the AUC, optimal cut-off value, sensitivity, specificity of NLR and SII to early predict severe-patients with COVID-19 were 0.867, 7.25, 70.83%, 92.21% and 0.860, 887.20, 81.25%, 81.82%, respectively. Conclusion The results suggest that the SII and NLR is a potential new diagnosed biomarker in severe-patients with COVID-19.

Cold chain and severe acute respiratory syndrome coronavirus 2 transmission: a review for challenges and coping strategies.

Since June 2020, the re-emergence of coronavirus disease 2019 (COVID-19) epidemics in parts of China was linked to the cold chain, which attracted extensive attention and heated discussions from the public. According to the typical characteristics of these epidemics, we speculated a possible route of transmission from cold chain to human. A series of factors in the supply chain contributed to the epidemics if the cold chain were contaminated by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), such as temperature, humidity, personal hygiene/protection, and disinfection. The workers who worked in the cold chain at the receiving end faced a higher risk of being infected when they were not well protected. Facing the difficult situation, China put forward targeted and powerful countermeasures to block the cold chain-related risk. However, in the context of the unstable pandemic situation globally, the risk of the cold chain needs to be recognized and evaluated seriously. Hence, in this review, we reviewed the cold chain-related epidemics in China, analyzed the possible mechanisms, introduced the Chinese experience, and suggested coping strategies for the global epidemic prevention and control.

Inhibition of SARS-CoV-2 replication by zinc gluconate in combination with hinokitiol.

The Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) pandemic is currently the major challenge to global public health. Two proteases, papain-like protease (PLpro) and the 3-chymotrypsin-like protease (3CLpro or Mpro), are indispensable for SARS-CoV-2 replication, making them attractive targets for antiviral therapy development. Here we screened a panel of essential metal ions using a proteolytic assay and identified that zinc gluconate, a widely-used zinc supplement, strongly inhibited the proteolytic activities of the two proteases in vitro. Biochemical and crystallographic data reveal that zinc gluconate exhibited the inhibitory function via binding to the protease catalytic site residues. We further show that treatment of zinc gluconate in combination with a small molecule ionophore hinokitiol, could lead to elevated intracellular Zn2+ level and thereby significantly impaired the two protease activities in cellulo. Particularly, this approach could also be applied to rescue SARS-CoV-2 infected mammalian cells, indicative of potential application to combat coronavirus infections. Our studies provide the direct experimental evidence that elevated intracellular zinc concentration directly inhibits SARS-CoV-2 replication and suggest the potential benefits to use the zinc supplements for coronavirus disease 2019 (COVID-19) treatment.

Exploring the Natural Origins of SARS-CoV-2 in the Light of Recombination.

The lack of an identifiable intermediate host species for the proximal animal ancestor of SARS-CoV-2, and the large geographical distance between Wuhan and where the closest evolutionary related coronaviruses circulating in horseshoe bats (members of the Sarbecovirus subgenus) have been identified, is fueling speculation on the natural origins of SARS-CoV-2. We performed a comprehensive phylogenetic study on SARS-CoV-2 and all the related bat and pangolin sarbecoviruses sampled so far. Determining the likely recombination events reveals a highly reticulate evolutionary history within this group of coronaviruses. Distribution of the inferred recombination events is nonrandom with evidence that Spike, the main target for humoral immunity, is beside a recombination hotspot likely driving antigenic shift events in the ancestry of bat sarbecoviruses. Coupled with the geographic ranges of their hosts and the sampling locations, across southern China, and into Southeast Asia, we confirm that horseshoe bats, Rhinolophus, are the likely reservoir species for the SARS-CoV-2 progenitor. By tracing the recombinant sequence patterns, we conclude that there has been relatively recent geographic movement and cocirculation of these viruses' ancestors, extending across their bat host ranges in China and Southeast Asia over the last 100 years. We confirm that a direct proximal ancestor to SARS-CoV-2 has not yet been sampled, since the closest known relatives collected in Yunnan shared a common ancestor with SARS-CoV-2 approximately 40 years ago. Our analysis highlights the need for dramatically more wildlife sampling to: 1) pinpoint the exact origins of SARS-CoV-2's animal progenitor, 2) the intermediate species that facilitated transmission from bats to humans (if there is one), and 3) survey the extent of the diversity in the related sarbecoviruses' phylogeny that present high risk for future spillovers.

Inhibitory Effect of Baicalin on 2019-NCOV Invasion (preprint)

Objective To study the the inhibitory effect of baicalin on novel Coronavirus (2019-NCOV) entry process in vitro. Methods The pseudovirus system of SARS-CoV-2 S protein constructed with luciferase reporter gene was used in the study. A luciferase kit was used to detect the changes of luciferase expression in Huh-7 cells, and then the virus inhibition curve was plotted. Results Baicalin can significantly inhibited the infection rate of pseudovirus. There was no significant difference in the virus inhibition curve between the baicalin&virus pre-incubation group and co-incubation at different concentrations, indicating that baicalin could not directly bind to virus, but inhibited the virus S protein mediated cell fusion process. We futher found that, the inhibition rate of baicalin to virus decreased significantly in the 4h group, but had no significant difference in the 0h and 2h groups at the concentration of 0.125mg/ mL, indicating that baicalin may have an inhibitory effect on virus invasion stage rather than adsorption stage, and the mediated inhibition stage occurred within 4h. Conclusion Baicalin may mediate the fusion of SARS-CoV-2 S protein with cell surface receptor and exert anti-novel coronavirus activity by playing a role in inhibitting virus invasion in non-adsorption stage.

Psychological experience of patients with confirmed COVID-19 at the initial stage of pandemic in Wuhan, China: a qualitative study.

BACKGROUND: Coronavirus disease 2019 (COVID-19) quickly developed into a global pandemic and affected patients' mental health. However, little is known about psychological experience of patients with COVID-19. The aim was to elucidate the psychological experience of patients with confirmed COVID-19 in Wuhan, at the initial stage of the pandemic. METHODS: This study was conducted using a phenomenological approach in a qualitative study. Thirteen patients with confirmed COVID-19 from a COVID-19-designated hospital in Wuhan, were recruited between March 15th and April 20th, 2020 via purposive sampling. Semi-structured in-depth interviews were conducted face-to-face. The interview data were analyzed using inductive thematic analysis. RESULTS: The psychological experience of patients was summarized into three themes: mental distress related to COVID-19, expectations of life scenarios after discharge, and making sense of the experience. These themes were classified into 10 sub-themes. Patients experienced confusion, uncertainty, worry, guilt and concern. Both positive and negative expectations of life scenarios after discharge were reported, manifested as expectations about making up for lost time with family, anxiety about social discrimination and feelings of helplessness about poor financial security. Moreover, patients perceived strength of abundant social support and awareness of social responsibility from their unique experience to cope with their condition. CONCLUSIONS: This study demonstrated that patients with confirmed COVID-19 in Wuhan underwent complex psychological experience, both positive and negative at the initial stage of the pandemic. These findings will contribute to the delivery of effective mental health care to safeguard patients' wellbeing.

Oxidative stress transforms 3CLpro into an insoluble and more active form to promote SARS-CoV-2 replication.

3CLpro is a key proteinase for SARS-CoV-2 replication and serves as an important target for antiviral drug development. However, how its activity is regulated intracellularly is still obscure. In this study, we developed a 3CLpro protease activity reporter system to examine the impact of various factors, including nutrient supplements, ions, pHs, or oxidative stress inducers, on 3CLpro protease activity. We found that oxidative stress could increase the overall activity of 3CLpro. Not altering the expression, oxidative stress decreased the solubility of 3CLpro in the lysis buffer containing 1% Triton-X-100. The Triton-X-100-insoluble 3CLpro was correlated with aggregates' formation and responsible for the increased enzymatic activity. The disulfide bonds formed between Cys85 sites of 3CLpro protomers account for the insolubility and the aggregation of 3CLpro. Besides being regulated by oxidative stress, 3CLpro impaired the cellular antioxidant capacity by regulating the cleavage of GPx1 at its N-terminus. This cleavage could further elevate the 3CLpro-proximate oxidative activity, favor aggregation and activation of 3CLpro, and thus lead to a positive feedback loop. In summary, we reported that oxidative stress transforms 3CLpro into a detergent-insoluble form that is more enzymatically active, leading to increased viral replication/transcription. Our study provided mechanistic evidence that suggests the therapeutic potential of antioxidants in the clinical treatment of COVID-19 patients.

The Physical Fitness Level of College Students Before and After Web-Based Physical Education During the COVID-19 Pandemic.

Background: The COVID-19 pandemic has been an emergency worldwide. Web-based physical education is a choice for college students to keep on their study. The aim of this study was to compare the data of physical fitness of college students before and after web-based physical education. Methods: All the students of 2018 and 2019 in Wuhan University of Technology who had taken the web-based physical education class in 2020 were included in this study. The records of annual physical fitness tests of all the subjects in 2019 and 2020 which were carried out in September were reviewed, including weight, height, body mass index (BMI), vital capacity (VC), 50-m dash, sit-and-reach, standing long jump, male-specific pull-ups and 1,000-m race, and female-specific sit-ups and 800-m race. Results: There were 24,112 male and 9,690 female records of physical fitness tests included in our study. The results of 11,219 male and 4,651 female students who completed both physical fitness tests in 2019 and 2020 were employed for Wilcoxon signed-rank test. Declined performance was observed on male 50-m dash by 0.1 s, male 1,000-m race by 14 s, and female 800-m race by 11 s. Notably, the percentage of male obesity, based on BMI, rose from 10.6 to 15.2% and 17.1 to 21.8% for male overweight; correspondingly, the percentage of male normal weight declined from 55.9 to 51.9% and 16.4 to 11.1% for male thinness. The trend of increasing BMI in males should be paid attention to. Improved results on vital capacity, sit-and-reach, standing long jump for both males and females, female 50-m dash, female sit-ups, and male pull-ups were observed in 2020. All the results of physical fitness tests were significantly different between 2019 and 2020 (p < 0.01) by Wilcoxon signed-rank test. Conclusions: The changes of physical fitness tests before and after web-based physical education suggested that the focus should be placed on improvement for running tests through appropriate alternatives, such as fast running in place and shuttle run. In addition, the simple, convenient, and practical sport that require available equipment and little field should be considered for web-based physical education.

Allosteric inhibition of SARS-CoV-2 3CL protease by colloidal bismuth subcitrate.

The SARS-CoV-2 3-chymotrypsin-like protease (3CLpro or Mpro) is a key cysteine protease for viral replication and transcription, making it an attractive target for antiviral therapies to combat the COVID-19 disease. Here, we demonstrate that bismuth drug colloidal bismuth subcitrate (CBS) is a potent inhibitor for 3CLpro in vitro and in cellulo. Rather than targeting the cysteine residue at the catalytic site, CBS binds to an allosteric site and results in dissociation of the 3CLpro dimer and proteolytic dysfunction. Our work provides direct evidence that CBS is an allosteric inhibitor of SARS-CoV-2 3CLpro.

Optimizing Safe Dental Practice During the COVID-19 Pandemic: Recommendations Based on a Guide Developed for Dental Practices in China.

The current global coronavirus disease 2019 (COVID-19) outbreak is still exerting severe global implications, and its development in various regions is complex and variable. The high risk of cross-infection poses a great challenge to the dental practice environment; it is therefore urgent to develop a set of pandemic prevention measures to ensure dental practice safety during the COVID-19 outbreak. Therefore, we combined the epidemiological characteristics of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), public emergency measures for COVID-19, characteristics of dental practice, and relevant literature reports to develop a set of dynamic practice measures for dental practices in high-, medium-, and low-risk areas affected by COVID-19. This will help dental practices to achieve standard prevention and ensure their safe and smooth operation during the pandemic. It is hoped that these measures will provide a reference basis for dental hospitals and dental clinics in their care and pandemic prevention work.

Review of the risk factors for SARS-CoV-2 transmission.

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic, which has lasted for nearly a year, has made people deeply aware of the strong transmissibility and pathogenicity of SARS-CoV-2 since its outbreak in December 2019. By December 2020, SARS-CoV-2 had infected over 65 million people globally, resulting in more than 1 million deaths. At present, the exact animal origin of SARS-CoV-2 remains unclear and antiviral vaccines are now undergoing clinical trials. Although the social order of human life is gradually returning to normal, new confirmed cases continue to appear worldwide, and the majority of cases are sporadic due to environmental factors and lax self-protective consciousness. This article provides the latest understanding of the epidemiology and risk factors of nosocomial and community transmission of SARS-CoV-2, as well as strategies to diminish the risk of transmission. We believe that our review will help the public correctly understand and cope with SARS-CoV-2.

The relationship between 2019-nCoV and psychological distress among parents of children with autism spectrum disorder.

OBJECTIVES: The psychological distress caused by COVID-19 may be pronounced among the parents of children with autism spectrum disorder (ASD). This study aimed to investigate psychological distress among parents of children with ASD during the COVID-19 pandemic. METHODS: A total of 1764 parents of children with ASD and 4962 parents of typically developing (TD) children were recruited. The participants completed an online survey which contained demographic information, the impact due to COVID-19 crisis, resilience, coping styles, anxiety and depression. Hierarchical linear regression was used to assess the contributions of these variables to anxiety and depression. RESULTS: After adjusting for demographic variables, the following factors were associated with parents' anxiety and depression symptoms: (i) Whether or not the participants had a child with ASD; (ii) resilience; (iii) coping strategies, and; (iv) the impact due to COVID-19. Among these, the psychological stress caused by COVID-19 played the most important role in parental anxiety (ß = 0.353) and depression (ß = 0.242) symptoms. Parents of children with ASD had lower levels of resilience and positive coping, and used more negative coping strategies than parents of TD children. Among all participants, 8.0 and 24.2% of parents had symptoms of anxiety and depression, respectively. Compared to parents of TD children, more parents of children with ASD exhibited symptoms of anxiety and depression (12.2% vs. 6.6%; 31.0% vs. 21.7%, respectively). CONCLUSIONS: During the COVID-19 pandemic, parents experienced varying levels of anxiety and depression, particularly, parents of children with ASD. More specific attention should be paid to parental mental health and long-term effective intervention programs, that are targeted towards parents of children with ASD, and such programs should be promoted around China in the wake of the COVID-19 crisis.