Recent developments and trends of automatic nucleic acid detection systems.

Nucleic acid detection, widely used in clinical diagnosis, biological analysis, and environmental monitoring, is of great significance for disease diagnosis and basic research. With the outbreak of COVID-19, the demand for fast and high-throughput nucleic acid detection from large numbers of samples has increased sharply. Automated nucleic acid detection systems can meet these needs, and also play important roles in disease screening and infectious disease prevention and control. In this review, we introduce and compare the current mainstream nucleic acid automatic detection instruments and equipment, then discuss the future demands of nucleic acid detection.

Investigation of sport-related habits in college students after covid-19 epidemic / Investigação sobre hábitos relacionados ao esporte em estudantes universitários após epidemia de covid-19 / Investigación de los hábitos relacionados con el deporte en estudiantes universitarios tras la epidemia de covid-19

ABSTRACT Introduction: China's covid-19 epidemic is under control. Universities and schools across the country have resumed physical education classes, but the mental health knowledge of college students in the aftermath of the covid-19 outbreak has not been investigated. Objective: To study college students' mental health and sports-related habits after the covid-19 outbreak. Methods: This study comprehensively used literature materials, expert interviews, mathematical statistics, and other methods to study the sports-related attitudes and health awareness of college students before and after the outbreak of the new epidemic, examining the health knowledge domain of college students. Results: In the survey dimensions related to students' sports knowledge, the highest mean score was for "emotional control" at 3.48, followed by "participation motivation" at 3.41 and "participation attitude" at 3.35. The scores of sports participation attitude, motivation, and sports emotion for males were equal and higher than in females, where the measures of emotional factors and motivation to participate reached significant levels. After the epidemic, the mean scores for students' participation attitude, sports emotion, and motivation to participate in sports exceeded the theoretical mean. Conclusion: After the health awareness and prevention work in the community and school network, university students' sports attitudes and awareness improved significantly compared to before the Covid-19 outbreak. Level of evidence II; Therapeutic studies - investigation of treatment outcomes.

RESUMO Introdução: A epidemia de covid-19 em China encontra-se sob controle. Faculdades, escolas primárias e secundárias em todo o país retomaram as aulas de educação física, porém o conhecimento sobre a saúde mental dos estudantes universitários no estágio posterior do surto da Covid-19 não foi investigado. Objetivo: Estudar a saúde mental e os hábitos relacionados ao esporte nos universitários depois da epidemia da Covid-19. Métodos: Este estudo utilizou de forma abrangente materiais de literatura, entrevistas de especialistas, estatísticas matemáticas e outros métodos para estudar especificamente as atitudes relacionadas ao esporte e a conscientização na saúde dos estudantes universitários antes e depois do surto da nova epidemia, examinando o domínio do conhecimento em saúde dos estudantes universitários. Resultados: Nas dimensões da pesquisa relacionadas ao conhecimento esportivo dos alunos, a maior pontuação média foi de "controle emocional" como 3,48, seguida por "motivação de participação" como 3,41 e "atitude participativa" como 3,35. As pontuações de atitude de participação esportiva, motivação e emoção esportiva para os homens foram iguais e maior do que nas mulheres, onde as medidas de fator emocional e motivação para participar atingiram níveis significativos. Após a epidemia, as médias de atitude de participação dos alunos, emoção esportiva e motivação para participar do esporte superaram a média teórica. Conclusão: Após o trabalho de conscientização em saúde e prevenção, na rede comunitária e escolar, a atitude esportiva e a conscientização dos universitários foram significativamente aprimoradas em relação aos antecedentes ao surto de Covid-19. Nível de evidência II; Estudos terapêuticos - investigação dos resultados do tratamento.

RESUMEN Introducción: La epidemia de covid-19 en China está bajo control. Los colegios y las escuelas primarias y secundarias de todo el país han reanudado las clases de educación física, pero no se ha investigado el conocimiento sobre la salud mental de los estudiantes universitarios en la última etapa del brote de covid-19. Objetivo: Estudiar la salud mental y los hábitos relacionados con el deporte en los estudiantes universitarios tras el brote de covid-19. Métodos: Este estudio utilizó de forma exhaustiva materiales bibliográficos, entrevistas a expertos, estadísticas matemáticas y otros métodos para estudiar específicamente las actitudes relacionadas con el deporte y el conocimiento de la salud de los estudiantes universitarios antes y después del estallido de la nueva epidemia, examinando el dominio del conocimiento de la salud de los estudiantes universitarios. Resultados: En las dimensiones de la encuesta relacionadas con los conocimientos deportivos de los estudiantes, la puntuación media más alta correspondió al "control emocional" con un 3,48, seguido de la "motivación para la participación" con un 3,41 y la "actitud para la participación" con un 3,35. Las puntuaciones de la actitud de participación deportiva, la motivación y la emoción deportiva de los varones fueron iguales y superiores a las de las mujeres, donde las medidas del factor emocional y la motivación para participar alcanzaron niveles significativos. Después de la epidemia, las puntuaciones medias de la actitud de participación, la emoción deportiva y la motivación para participar en el deporte de los alumnos superaron la media teórica. Conclusión: Tras el trabajo de concienciación y prevención sanitaria en la comunidad y la red escolar, la actitud y la conciencia deportiva de los estudiantes universitarios mejoraron significativamente en comparación con las anteriores al brote de Covid-19. Nivel de evidencia II; Estudios terapéuticos - investigación de los resultados del tratamiento.

The potential of biomass-derived bio-liquid to prevent the spread of SARS-CoV-2 from waste and its production-based life cycle assessment.

COVID-19 pandemic caused a significant increase in medical and infected domestic waste, greatly increasing risk of human infected with SARS-CoV-2. Therefore, it is critical to prevent the spread of SARS-CoV-2 from solid waste to humans. Current commercial disinfectants present a high carbon footprint issue. Hence, we prepared a renewable wheat straw-based bio-liquid that can damage SARS-CoV-2 RNA and protein. The wet thermochemical extraction (WTE) bio-liquid, with total organic carbon concentration exceeding 1892 mg/L, could effectively damage the virus. However, dry thermochemical extraction (DTE) samples were not efficient due to their low content of effective compounds. The life cycle assessment showed that WTE bio-liquid production implies lower energy and environmental negative impacts than DTE. Moreover, the process by-product, char, can simultaneously reduce 3.1 million tonnes of global CO2 emissions while used as coal substitute. Yield of bio-liquid extremely exceed commercial disinfectant with just 1 % wheat straw utilisation, which meet the demand of processing solid waste. Further, their costs are significantly lower than commercial disinfectants, which are suitable for developing countries. Therefore, the antiviral bio-liquid produced from agricultural straw can be a new way to meet the needs of preventing the spread of SARS-CoV-2 and resume the sustainable development of society.

Alpha-1 antitrypsin protects against phosgene-induced acute lung injury by activating the ID1-dependent anti-inflammatory response (preprint)

Phosgene, a highly dangerous chemical warfare agent, is widely used as an industrial chemical. Phosgene inhalation causes acute lung injury (ALI), which may further progress into pulmonary edema. Currently, there is no known antidote for phosgene poisoning. Alpha-1 antitrypsin (α1-AT) is a protease inhibitor that has been used to treat emphysema patients, who are deficient in α1-AT, for decades. Recent studies have shown that α1-AT has both anti-inflammatory and anti-SARS-CoV-2 effects. In this study, we aimed to investigate the role of α1-AT in phosgene-induced ALI. We observed a time-dependent increase in α1-AT expression and secretion in the lungs of rats exposed to phosgene. Interestingly, α1-AT was derived from neutrophils, but not from macrophages or alveolar type II cells, and α1-AT knockdown aggravated phosgene- and lipopolysaccharide (LPS)-induced inflammation and cell death in human bronchial epithelial cells (BEAS-2B). Conversely, α1-AT administration suppressed the inflammatory response and prevented death in LPS- and phosgene-exposed BEAS-2B cells. Furthermore, α1-AT treatment increased the expression of the inhibitor of DNA binding (ID1) gene, which suppressed NF-κB pathway activation, reduced inflammation, and inhibited cell death. These data demonstrate that neutrophil-derived α1-AT protects against phosgene-induced ALI by activating the ID1-dependent anti-inflammatory response. This study may provide novel strategies for the treatment of patients with phosgene-induced ALI.

How does AI use drive individual digital resilience? a conservation of resources (COR) theory perspective

Artificial intelligence (AI) serves as a useful resource for replacing, supporting, and augmenting individuals in responding to external difficulties and enhancing individual resilience. However, little is known about the underlying laws of how AI can heighten individual resilience. This research examines the formation of individual resilience based on the consequences of different AI usage behaviours. Study 1 uses text mining to detect individual resilience based on the experience with AI. We identify not only individual resilience but also family resilience. Study 2, based on the context of the COVID-19 pandemic, collects online survey data from personal intelligent assistant users to investigate the transformation mechanism of AI usage behaviours to individual resilience. Drawing upon the conservation of resources theory, routine and infusion use are considered two levels of resource investments to strengthen the different degrees of individual resilience by coping responses (task-focused, emotion-focused, and avoidance coping). The findings confirm the differences between routine and infusion use in the formation of individual resilience, mediated by both task-focused and emotion-focused coping, without the mediating role of avoidance coping. Our research provides enlightenment for researchers and practitioners on building resilience and improving performance. [ FROM AUTHOR]

Fully Automated CRISPR-LAMP Platform for SARS-CoV-2 Delta and Omicron Variants.

Integrated clustered regularly interspaced short palindromic repeat (CRISPR)-loop-mediated amplification (LAMP) technology is of great importance in CRISPR-based diagnostic systems, which urgently needs to be developed to improve diagnostic accuracy. A labor-free, contamination-free, and fully automated droplet manipulation platform for the CRISPR-LAMP technology has not been developed before. Herein, we propose a fully automated CRISPR-LAMP platform, which can precisely manipulate the CRISPR-LAMP droplet and perform combined reactions with high sensitivity and specificity. SARS-CoV-2 Spike T478K, D614G, P681R, and P681H mutations, typical point mutations of B.1.617.2 (Delta) and Omicron variants, are monitored with this platform with a detection limit of 102 copies/µL. Allele discrimination between the mutants and wild type is significant with the designed one/two-mismatch CRISPR RNA (crRNA) at a limit of 102 copies/µL. Chemically synthesized and modified crRNAs greatly increase the CRISPR-LAMP signal, which advance the wide application. Combined with the previously developed RdRp CRISPR-LAMP assay, clinical results showed that Spike T478K and P681H can discriminate the mutant type form the wild type with 70% (49.66-85.50%, 95% confidence interval) and 78% (57.27-90.62%, 95% confidence interval) sensitivity, respectively, and 100% specificity (51.68-100%, 95% confidence interval), and the RdRp target can detect SARS-CoV-2 strains with 85% sensitivity (65.39-95.14%, 95% confidence interval) and 100% specificity (51.68-100%, 95% confidence interval). We believe that this automatic digital microfluid (DMF) system can advance the integrated CRISPR-LAMP technology with higher stability, sensitivity, and practicability, also for other CRISPR-associated diagnostic platforms.

Mind the feline coronavirus: comparison with SARS-CoV-2

Both feline coronavirus (FCoV) and SARS-CoV-2 are coronaviruses that infect cats and humans, respectively. However, cats have been shown to be susceptible to SARS-CoV-2, and FCoV also had been shown to infect human. To elucidate the relationship between FCoV and SARS-CoV-2, we highlight the main characteristics of the genome, the receptor usage, and the correlation of the receptor-binding domain (RBD) of spike proteins in FCoV and SARS-CoV-2. It is demonstrated that FCoV and SARS-CoV-2 are closely related to the main characteristics of the genome, receptor usage, and RBD of spike proteins with similar furin cleavage sites. In particular, the affinity of the conserved feline angiotensin-converting enzyme 2 (fACE2) receptor to the RBD of SARS-CoV-2 suggests that cats are susceptible to SARS-CoV-2. In addition, cross-species of coronaviruses between cats and humans or other domesticated animals are also discussed. This review sheds light on cats as potential intermediate hosts for SARS-CoV-2 transmission, and cross-species transmission or zoonotic infection of FCoV and SARS-CoV-2 between cats and humans was identified.

Recent developments and trends of automatic nucleic acid detection systems

Nucleic acid detection, widely used in clinical diagnosis, biological analysis, and environmental monitoring, is of great significance for disease diagnosis and basic research. With the outbreak of COVID-19, the demand for fast and high-throughput nucleic acid detection from large numbers of samples has increased sharply. Automated nucleic acid detection systems can meet these needs, and also play important roles in disease screening and infectious disease prevention and control. In this review, we introduce and compare the current mainstream nucleic acid automatic detection instruments and equipment, then discuss the future demands of nucleic acid detection.

Universally Stable and Precise CRISPR-LAMP Detection Platform for Precise Multiple Respiratory Tract Virus Diagnosis Including Mutant SARS-CoV-2 Spike N501Y.

Nowadays, rapid and accurate diagnosis of respiratory tract viruses is an urgent need to prevent another epidemic outbreak. To overcome this problem, we have developed a clustered, regularly interspaced short palindromic repeats (CRISPR) loop mediated amplification (LAMP) technology to detect influenza A virus, influenza B virus, respiratory syncytial A virus, respiratory syncytial B virus, and severe acute respiratory syndrome coronavirus 2, including variants of concern (B.1.1.7), which utilized CRISPR-associated protein 12a (Cas12a) to advance LAMP technology with the sensitivity increased 10 times. To reduce aerosol contamination in CRISPR-LAMP technology, an uracil-DNA-glycosylase-reverse transcription-LAMP system was also developed which can effectively remove dUTP-incorporated LAMP amplicons. In vitro Cas12a cleavage reaction with 28 crRNAs showed that there were no position constraints for Cas12a/CRISPR RNA (crRNA) recognition and cleavage in LAMP amplicons, and even the looped position of LAMP amplicons could be effectively recognized and cleaved. Wild-type or spike N501Y can be detected with a limit of detection of 10 copies/µL (wild-type) even at a 1% ratio level on the background (spike N501Y). Combining UDG-RT-LAMP technology, CRISPR-LAMP design, and mutation detection design, we developed a CRISPR-LAMP detection platform that can precisely diagnose pathogens with better stability and significantly improved point mutation detection efficiency.

Characteristics and Prognosis of Antibody Non-Responders with COVID-19 (preprint)

Background：The coronavirus disease 2019 (COVID-19) caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been spreading globally. The information regarding the characteristics and prognosis of antibody non-responders with COVID-19 is scarce.Method: In this retrospective, single-center study, we included all the patients with confirmed COVID-19 using real-time reverse transcriptase-polymerase chain reaction (RT-PCR) admitted to the Fire God Mountain hospital from February 3, 2020, to April 14, 2020. A total of 1921 patients were divided into the antibody-negative group (n=94) and antibody-positive group (n=1827), and the 1:1 propensity score matching (PSM) was used to match two groups.Results: In the antibody negative group, 40 patients (42.6%) were male, 54 patients (57.4%) were female, and 49 patients (52.1%) were older than 65 years old. Cough was the most common symptoms in the antibody negative group. White blood cell counts (WBC) 6.6×109/L [5.0, 9.1], Neutrophils 4.3×109/L [3.1, 6.6], C-reactive protein 7.3 mg/L [1.3, 49.0], Procalcitonin (PCT) 0.1 ng/mL [0.0, 0.2], Interleukin-6 (IL-6) 64.2 [1.5, 28.7], Lactate dehydrogenase (LDH) 193.8 U/L [154.9,260.6], Creatine kinase 60.5 U/L [40.5, 103.7], Creatine kinase isoenzyme 10.3 ng/mL [8.2, 14.5], Urea nitrogen 5.3 mmol/L [4.0, 8.7] and Creatinine 77.7 μmol/L [60.6, 98.7] were significantly higher in antibody negative patients than in antibody positive group (P<0.005). The days of nucleic acid negative conversion in the antibody negative group was shorter than that in the antibody positive group (P < 0.001). Meanwhile, the hospitalization time of antibody negative patients was shorter than that of antibody positive patients (8.0 [6.0, 10.0] VS 13.0 [8.2, 23.0], P < 0.001).Conclusion: Some COVID-19 patients without specific antibodies had mild symptoms, but the inflammatory reaction caused by innate clinical immunity was more intense than those with antibodies, and the virus was cleared faster. The production of specific antibodies was unnecessary for SARS-CoV-2 clearance, and non-specific immune responses played an essential role in virus clearance.

Risk Factors for Anxiety and Depressive Symptoms in Doctors During the Coronavirus Disease 2019 Pandemic.

Background: The purpose of our study was to investigate the prevalence of anxiety and depressive symptoms and their risk factors among doctors during the coronavirus disease 2019 (COVID-19) pandemic. Methods: A hospital-based survey study was conducted. Anxiety symptoms were assessed using the Self-Rating Anxiety Scale (SAS), and depressive symptoms were assessed using the Self-Rating Depression Scale (SDS). Multivariable logistic regression was used to analyze anxiety and depressive symptoms across independent variables. Results: A total of 1,521 doctors were included; 11.11% (169/1,521) of the doctors had anxiety symptoms, and 16.90% (257/1,521) had depressive symptoms. Female sex [adjusted odds ratio (aOR), 1.69; 95% confidence interval (CI), 1.21-2.34; P = 0.002] and having a minor child (aOR, 2.31; 95% CI, 1.50-3.56; P < 0.001) were associated with an increased risk of anxiety symptoms. Female sex (aOR, 1.56; 95% CI, 1.18-2.06; P = 0.002) and having a minor child (aOR, 1.48; 95% CI, 1.06-2.01; P = 0.022) were associated with an increased risk of depressive symptoms. Older age (aOR, 0.97; 95% CI, 0.98-0.99; P = 0.008) was associated with a decreased risk of depressive symptoms. Conclusions: Anxiety and depressive symptoms have been common mental health problems in doctors during the COVID-19 pandemic. We found that female sex, having a minor child, and younger age were major risk factors for the development of anxiety and depressive symptoms among doctors during the COVID-19 pandemic.

Immune responses to SARS-CoV-2 infection in Humans and ACE2 humanized mice

The coronavirus disease 2019 (COVID-19) pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) represents a major public health threat worldwide. Insight into protective and pathogenic aspects of SARS-CoV-2 immune responses is critical to work out effective therapeutics and develop vaccines for controlling the disease. Here, we review the present literature describing the innate and adaptive immune responses including innate immune cells, cytokine responses, antibody responses and T cell responses against SARS-CoV-2 in human infection, as well as in AEC2-humanized mouse infection. We also summarize the now known and unknown about the role of the SARS-CoV-2 immune responses. By better understanding the mechanisms that drive the immune responses, we can tailor treatment strategies at specific disease stages and improve our response to this worldwide public health threat.

Contrastive Cross-site Learning with Redesigned Net for COVID-19 CT Classification (preprint)

The pandemic of coronavirus disease 2019 (COVID-19) has lead to a global public health crisis spreading hundreds of countries. With the continuous growth of new infections, developing automated tools for COVID-19 identification with CT image is highly desired to assist the clinical diagnosis and reduce the tedious workload of image interpretation. To enlarge the datasets for developing machine learning methods, it is essentially helpful to aggregate the cases from different medical systems for learning robust and generalizable models. This paper proposes a novel joint learning framework to perform accurate COVID-19 identification by effectively learning with heterogeneous datasets with distribution discrepancy. We build a powerful backbone by redesigning the recently proposed COVID-Net in aspects of network architecture and learning strategy to improve the prediction accuracy and learning efficiency. On top of our improved backbone, we further explicitly tackle the cross-site domain shift by conducting separate feature normalization in latent space. Moreover, we propose to use a contrastive training objective to enhance the domain invariance of semantic embeddings for boosting the classification performance on each dataset. We develop and evaluate our method with two public large-scale COVID-19 diagnosis datasets made up of CT images. Extensive experiments show that our approach consistently improves the performances on both datasets, outperforming the original COVID-Net trained on each dataset by 12.16% and 14.23% in AUC respectively, also exceeding existing state-of-the-art multi-site learning methods.

On-site analysis of COVID-19 on the surfaces in wards.

SARS-Cov-2 has erupted across the globe, and confirmed cases of COVID-19 pose a high infection risk. Infected patients typically receive their treatment in specific isolation wards, where they are confined for at least 14 days. The virus may contaminate any surface of the room, especially frequently touched surfaces. Therefore, surface contamination in wards should be monitored for disease control and hygiene purposes. Herein, surface contamination in the ward was detected on-site using an RNA extraction-free rapid method. The whole detection process, from surface sample collection to readout of the detection results, was finished within 45 min. The nucleic acid extraction-free method requires minimal labor. More importantly, the tests were performed on-site and the results were obtained almost in real-time. The test confirmed that 31 patients contaminated seven individual sites. Among the sampled surfaces, the electrocardiogram fingertip presented a 72.7% positive rate, indicating that this surface is an important hygiene site. Meanwhile, the bedrails showed the highest correlation with other surfaces, so should be detected daily. Another surface with high contamination risk was the door handle in the bathroom. To our knowledge, we present the first on-site analysis of COVID-19 surface contamination in wards. The results and applied technique provide a potential further reference for disease control and hygiene suggestions.

Development of a Sensitive Immunochromatographic Method Using Lanthanide Fluorescent Microsphere for Rapid Serodiagnosis of COVID-19.

The SARS-CoV-2 infection that caused the COVID-19 pandemic quickly spread worldwide within two months. Rapid diagnosis of the disease and isolation of patients are effective ways to prevent and control the spread of COVID-19. Therefore, a sensitive immunofluorescent assay method was developed for rapid detection of special IgM and IgG of COVID-19 in human serum within 10 min. The recombinant nucleocapsid protein of 2019 novel coronavirus was used as capture antigen. Lanthanide, Eu(III) fluorescent microsphere, was used to qualitatively/semiquantitatively determine the solid phase immunochromatographic assay. A total of 28 clinical positive and 77 negative serum or plasma samples were included in the test. Based on the analysis of serum or plasma from COVID-19 patients and healthy people, the sensitivity and specificity of the immunochromatographic assay were calculated as 98.72% and 100% (IgG), and 98.68% and 93.10% (IgM), respectively. The results demonstrated that rapid immunoassay has high sensitivity and specificity and was useful for rapid serodiagnosis of COVID-19.

Safety, Tolerability and Immunogenicity of a Recombinant Adenovirus Type 5 Vectored COVID-19 Vaccine in Healthy Adults in China: Preliminary Report of a First-In Human Single-Center, Open-Label, Dose-Escalating Clinical Trial (preprint)

Background: We aimed to assess the safety, tolerability and immunogenicity of a recombinant adenovirus type 5 (Ad5) vectored COVID-19 vaccine expressing the spike glycoprotein of a SARS-Cov-2 strain. This is the first-in-human study of a candidate vaccine against COVID-19.Methods: We conducted a single-center, open-label, dose-escalating clinical trial of Ad5 vectored COVID-19 vaccine. Healthy adults aged between 18-60 years were sequentially enrolled and allocated to receive a single intramuscular injection in one of three dose groups: 5 × 10^10, 1×10^11, and 1·5×10^11 viral particles. Safety was assessed over the next 28 days. Specific antibodies were measured on enzyme-linked immunosorbent assay (ELISA), and the neutralizing antibody responses induced by vaccination were detected by using SARS-CoV-2 virus neutralization and pseudovirus neutralization tests. T-cell responses were accessed by enzyme-linked immunospot (ELISpot) and flow-cytometry assays. Results: A total of 108 participants were recruited and received low dose, middle dose, or high dose vaccine, with 36 in each dose group. 30(83·3%), 30(83·3%), and 27(75·0%) recipients in the low dose, middle dose, and high dose groups reported at least one adverse reaction within the first 7 days after the vaccination. The most common injection-site adverse reaction was pain, the most commonly reported systematic adverse reactions were fever, fatigue, headache, and muscle pain. A majority of the adverse reactions that were reported in all dose groups were mild or moderate in severity. No serious adverse event was noted within 28 days post-vaccination. Both ELISA antibodies and neutralizing antibodies increased significantly at day 14, and peaked 28 days post-vaccination. Specific T cell response peaked at day 14 post-vaccination.Conclusions: The Ad5 vectored COVID-19 vaccine is tolerable and immunogenic. Humoral responses against SARS-CoV-2 peaked at day 28 post-vaccination in health adults, and rapid specific T cell responses were noted since day 14.Trial Registration: The study is registered with ClinicalTrials.gov, number NCT04313127. Funding Statement: National Key R&D Program of China (2020YFC10841400), National Science and Technology Major Project (2016ZX10004001, 2018ZX09201005), and CanSino Biotechnology Inc.Declaration of Interests: Mr. Gou report being employees of Tianjin CanSino Biotechnology Inc, No other potential conflict of interest relevant to this article was reported.Ethics Approval Statement: The protocol and informed consent were approved by the Institutional Review Board of the Jiangsu Provincial Center of Disease Control and Prevention. Written informed consents from all participants were obtained before screening. This study was undertaken by Jiangsu Provincial Center of Disease Control and Prevention, Hubei Provincial Center for Disease Control and Prevention and Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology in accordance with the Declaration of Helsinki and Good Clinical Practice.