Proceedings of the 27th Annual Congress of the International Liver Transplantation Society.

After a virtual congress in 2021 and a previous absence in 2020 because of the coronavirus disease 2019 pandemic, the 27th Annual Congress of the International Liver Transplantation Society was held from May 4 to 7, 2022, in a hybrid format in Istanbul, with 1123 (58% on-site) liver transplant professionals from 61 countries attending the meeting. The hybrid format successfully achieved a balance of much yearned-for "in-person interaction" and global online participation. Almost 500 scientific abstracts were presented. In this report, the Vanguard Committee aims to present a summary of key invited lectures and selected abstracts for the liver transplant community.

Proteomic and phosphoproteomic characteristics of the cortex, hippocampus, thalamus, lung, and kidney in COVID-19-infected female K18-hACE2 mice

Background Neurological damage caused by coronavirus disease 2019 (COVID-19) has attracted increasing attention. Recently, through autopsies of patients with COVID-19, the direct identification of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in their central nervous system (CNS) has been reported, indicating that SARS-CoV-2 might directly attack the CNS. The need to prevent COVID-19-induced severe injuries and potential sequelae is urgent, requiring the elucidation of large-scale molecular mechanisms in vivo. Methods In this study, we performed liquid chromatography-mass spectrometry-based proteomic and phosphoproteomic analyses of the cortex, hippocampus, thalamus, lungs, and kidneys of SARS-CoV-2-infected K18-hACE2 female mice. We then performed comprehensive bioinformatic analyses, including differential analyses, functional enrichment, and kinase prediction, to identify key molecules involved in COVID-19. Findings We found that the cortex had higher viral loads than did the lungs, and the kidneys did not have SARS-COV-2. After SARS-CoV-2 infection, RIG-I-associated virus recognition, antigen processing and presentation, and complement and coagulation cascades were activated to different degrees in all five organs, especially the lungs. The infected cortex exhibited disorders of multiple organelles and biological processes, including dysregulated spliceosome, ribosome, peroxisome, proteasome, endosome, and mitochondrial oxidative respiratory chain. The hippocampus and thalamus had fewer disorders than did the cortex;however, hyperphosphorylation of Mapt/Tau, which may contribute to neurodegenerative diseases, such as Alzheimer's disease, was found in all three brain regions. Moreover, SARS-CoV-2-induced elevation of human angiotensin-converting enzyme 2 (hACE2) was observed in the lungs and kidneys, but not in the three brain regions. Although the virus was not detected, the kidneys expressed high levels of hACE2 and exhibited obvious functional dysregulation after infection. This indicates that SARS-CoV-2 can cause tissue infections or damage via complicated routes. Thus, the treatment of COVID-19 requires a multipronged approach. Interpretation This study provides observations and in vivo datasets for COVID-19-associated proteomic and phosphoproteomic alterations in multiple organs, especially cerebral tissues, of K18-hACE2 mice. In mature drug databases, the differentially expressed proteins and predicted kinases in this study can be used as baits to identify candidate therapeutic drugs for COVID-19. This study can serve as a solid resource for the scientific community. The data in this manuscript will serve as a starting point for future research on COVID-19-associated encephalopathy. Funding This study was supported by grants from the 10.13039/501100005150Chinese Academy of Medical Sciences Innovation Fund for Medical Sciences, the 10.13039/501100001809National Natural Science Foundation of China, and the 10.13039/501100004826Natural Science Foundation of Beijing.

Emerging trends and hot spots in subacute thyroiditis research from 2001 to 2022: A bibliometric analysis.

Background: Subacute thyroiditis (SAT) is the most prevalent self-limiting thyroid disease that causes pain, accounting for about 5% of all clinical thyroid disorders. Numerous clinically noteworthy results have been published in this area over the last 20 years. However, no article has comprehensively assessed the relevant literature yet. We conducted a bibliometric analysis of SAT to provide light on the dynamic nature of scientific advancement and aid researchers in gaining a global perspective while examining research core themes and hotspots. Methods: SAT-related articles and reviews from 2001 to 2022 were retrieved from the Science Citation Index-Expanded of Web of Science Core Collection (WoSCC). We analyzed current research trends and hotspots in this area using CiteSpace and Vosviewer. Results: A total of 568 studies associated with SAT research were published in 282 academic journals by 2,473 authors in 900 institutions from 61 countries/regions. The United States was a crucial link in inter-country/region collaboration and was the most frequently involved country in international cooperation. The University of Missouri System was the top organization, and Braley-Mullen H. was the most productive researcher. Thyroid published the most papers, with 36 publications. The most co-cited article was "Clinical features and outcome of subacute thyroiditis in an incidence cohort: Olmsted County, Minnesota, study" (by Fatourechi V., 2003). The clustered network and timeline view of keywords showed that the prevalence, diagnosis, and treatment of SAT were the research core themes during the past 20 years. Analysis of keyword bursts indicated that the clinical characteristic and the influence of COVID-19 on SAT appeared to be the current research hotspots. Conclusion: This bibliometric analysis conducted a thorough review of the SAT research. The clinical characteristics and the genetic background of SAT under the influence of COVID-19 are current research hotspots. However, there is still a need for further study and global collaboration. Our findings can aid researchers in understanding the current status of SAT research and immediately pinpoint new directions for further investigation.

Proteomic and phosphoproteomic characteristics of the cortex, hippocampus, thalamus, lung, and kidney in COVID-19-infected female K18-hACE2 mice.

BACKGROUND: Neurological damage caused by coronavirus disease 2019 (COVID-19) has attracted increasing attention. Recently, through autopsies of patients with COVID-19, the direct identification of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in their central nervous system (CNS) has been reported, indicating that SARS-CoV-2 might directly attack the CNS. The need to prevent COVID-19-induced severe injuries and potential sequelae is urgent, requiring the elucidation of large-scale molecular mechanisms in vivo. METHODS: In this study, we performed liquid chromatography-mass spectrometry-based proteomic and phosphoproteomic analyses of the cortex, hippocampus, thalamus, lungs, and kidneys of SARS-CoV-2-infected K18-hACE2 female mice. We then performed comprehensive bioinformatic analyses, including differential analyses, functional enrichment, and kinase prediction, to identify key molecules involved in COVID-19. FINDINGS: We found that the cortex had higher viral loads than did the lungs, and the kidneys did not have SARS-COV-2. After SARS-CoV-2 infection, RIG-I-associated virus recognition, antigen processing and presentation, and complement and coagulation cascades were activated to different degrees in all five organs, especially the lungs. The infected cortex exhibited disorders of multiple organelles and biological processes, including dysregulated spliceosome, ribosome, peroxisome, proteasome, endosome, and mitochondrial oxidative respiratory chain. The hippocampus and thalamus had fewer disorders than did the cortex; however, hyperphosphorylation of Mapt/Tau, which may contribute to neurodegenerative diseases, such as Alzheimer's disease, was found in all three brain regions. Moreover, SARS-CoV-2-induced elevation of human angiotensin-converting enzyme 2 (hACE2) was observed in the lungs and kidneys, but not in the three brain regions. Although the virus was not detected, the kidneys expressed high levels of hACE2 and exhibited obvious functional dysregulation after infection. This indicates that SARS-CoV-2 can cause tissue infections or damage via complicated routes. Thus, the treatment of COVID-19 requires a multipronged approach. INTERPRETATION: This study provides observations and in vivo datasets for COVID-19-associated proteomic and phosphoproteomic alterations in multiple organs, especially cerebral tissues, of K18-hACE2 mice. In mature drug databases, the differentially expressed proteins and predicted kinases in this study can be used as baits to identify candidate therapeutic drugs for COVID-19. This study can serve as a solid resource for the scientific community. The data in this manuscript will serve as a starting point for future research on COVID-19-associated encephalopathy. FUNDING: This study was supported by grants from the Chinese Academy of Medical Sciences Innovation Fund for Medical Sciences, the National Natural Science Foundation of China, and the Natural Science Foundation of Beijing.

Research progress of electrochemical oxidation and self-action of electric field for medical wastewater treatment.

A large number of pathogenic microorganisms exist in medical wastewater, which could invade the human body through the water and cause harm to human health. With the global pandemic coronavirus (COVID-19), public health safety become particularly important, and medical wastewater treatment is an important part of it. In particular, electrochemical disinfection technology has been widely studied in medical wastewater treatment due to its greenness, high efficiency, convenient operation, and other advantages. In this paper, the development status of electrochemical disinfection technology in the treatment of medical wastewater is reviewed, and an electrochemical three-stage disinfection system is proposed for the treatment of medical wastewater. Moreover, prospects for the electrochemical treatment of medical wastewater will be presented. It is hoped that this review could provide insight and guidance for the research and application of electrochemical disinfection technology to treat medical wastewater.GRAPHICAL ABSTRACT.

Depressive symptoms and physical function among the elderly in nursing homes during the COVID-19 pandemic in China: A cross-sectional study

The coronavirus disease 2019 (COVID-19) pandemic has placed a heavy burden on global healthcare. Depressive symptoms and physical function impairment are 2 common health problems among the elderly, but the association between depressive symptoms and physical function in nursing homes have not been extensively investigated during the COVID-19 pandemic. The purpose of this study was to investigate the current status of depressive symptoms and physical function and analyze the prevalence and related factors of depression among elderly people in nursing homes during the COVID-19 pandemic in China. A cross-sectional study was conducted. 381 elderly people were included in 4 nursing homes who were 60 to 100 years old with more than 3 months’ residential in Weifang City, Shandong Province using convenience cluster sampling. The Patient Health Questionnaire (PHQ-9) was performed to evaluate geriatric depression, the Barthel Index (BI) was administered to assess the activities of daily living, and a self-designed demographic data questionnaire was used to collect the demographic data. Multiple logistic regression analysis was conducted. 103 (27.0%) old residents reported depression according to PHQ-9. 279 (73.2%) old residents reported impaired self-care ability according to BI. The mean score of PHQ-9 and BI in the elderly was 3.56 ± 3.76 and 5.76 ± 7.05. The total PHQ-9 score of the elderly in nursing homes was positively correlated with the total activities of daily living score (R = 0.503, P < .01). Regression analysis showed that gender, self-care ability, more chronic diseases and medicines, especially Alzheimer’s disease and cataract were risk factors for depression among elderly people in nursing homes (P < .05). Our study showed 27.0% depression rate among old residents in nursing homes in China in the context of the COVID-19 pandemic. Depression is relatively prevalent among the elderly in China, and we should pay attention to those with poor self-care ability and more chronic diseases and medicines.

Function of Perceived Corporate Social Responsibility in Safety of Sports Activities and Home Aerobic Equipment in the Late Period of COVID-19.

The pandemic has impacted various industries, including the sports industry. However, corporate social responsibility (CSR) can mitigate the adverse effects of the crisis and promote the sports industry. To analyze the effect of CSR, the study examined the impact of perceived corporate social responsibility on injury prevention expectation, injury risk perception, and health up-gradation with the mediation of sports safety measures. There are 259 sportsmen of local sports bodies provided the data through a self-administered survey. Data analysis was conducted through Smart-PLS and SEM techniques. The outcome of the analysis showed that perceived corporate social responsibility leads to injury prevention expectation, injury risk perception, and health up-gradation. Also, the study found that sports safety measure mediates the relationship between perceived corporate social responsibility and injury prevention expectation, between perceived corporate social responsibility and injury risk perception, and between perceived corporate social responsibility and health up-gradation among sportsmen of local sports bodies. The theoretical implications were presented related to the significance of CSR and sports safety measure and their impact on sportsmen injury prevention expectation, health, and risk perception. The practical implications were related to the management of local sports bodies and how they can induce CSR initiatives and programs. Some limitations related to sample size, incorporating other variables, examining the model in other contexts, and using different study designs, have also been mentioned in the study.

Proceedings of the 26th Annual Virtual Congress of the International Liver Transplantation Society.

After a 1-y absence due to the coronavirus disease 2019 pandemic, the 26th Annual Congress of the International Liver Transplantation Society was held from May 15 to 18, 2021, in a virtual format. Clinicians and researchers from all over the world came together to share their knowledge on all the aspects of liver transplantation (LT). Apart from a focus on LT in times of coronavirus disease 2019, featured topics of this year's conference included infectious diseases in LT, living donation, machine perfusion, oncology, predictive scoring systems and updates in anesthesia/critical care, immunology, radiology, pathology, and pediatrics. This report presents highlights from invited lectures and a review of the select abstracts. The aim of this report, generated by the Vanguard Committee of International Liver Transplantation Society, is to provide a summary of the most recent developments in clinical practice and research in LT.

Imatinib and methazolamide ameliorate COVID-19-induced metabolic complications via elevating ACE2 enzymatic activity and inhibiting viral entry.

Coronavirus disease 2019 (COVID-19) represents a systemic disease that may cause severe metabolic complications in multiple tissues including liver, kidney, and cardiovascular system. However, the underlying mechanisms and optimal treatment remain elusive. Our study shows that impairment of ACE2 pathway is a key factor linking virus infection to its secondary metabolic sequelae. By using structure-based high-throughput virtual screening and connectivity map database, followed with experimental validations, we identify imatinib, methazolamide, and harpagoside as direct enzymatic activators of ACE2. Imatinib and methazolamide remarkably improve metabolic perturbations in vivo in an ACE2-dependent manner under the insulin-resistant state and SARS-CoV-2-infected state. Moreover, viral entry is directly inhibited by these three compounds due to allosteric inhibition of ACE2 binding to spike protein on SARS-CoV-2. Taken together, our study shows that enzymatic activation of ACE2 via imatinib, methazolamide, or harpagoside may be a conceptually new strategy to treat metabolic sequelae of COVID-19.

Automatic Sequence-Based Network for Lung Diseases Detection in Chest CT.

OBJECTIVE: To develop an accurate and rapid computed tomography (CT)-based interpretable AI system for the diagnosis of lung diseases. BACKGROUND: Most existing AI systems only focus on viral pneumonia (e.g., COVID-19), specifically, ignoring other similar lung diseases: e.g., bacterial pneumonia (BP), which should also be detected during CT screening. In this paper, we propose a unified sequence-based pneumonia classification network, called SLP-Net, which utilizes consecutiveness information for the differential diagnosis of viral pneumonia (VP), BP, and normal control cases from chest CT volumes. METHODS: Considering consecutive images of a CT volume as a time sequence input, compared with previous 2D slice-based or 3D volume-based methods, our SLP-Net can effectively use the spatial information and does not need a large amount of training data to avoid overfitting. Specifically, sequential convolutional neural networks (CNNs) with multi-scale receptive fields are first utilized to extract a set of higher-level representations, which are then fed into a convolutional long short-term memory (ConvLSTM) module to construct axial dimensional feature maps. A novel adaptive-weighted cross-entropy loss (ACE) is introduced to optimize the output of the SLP-Net with a view to ensuring that as many valid features from the previous images as possible are encoded into the later CT image. In addition, we employ sequence attention maps for auxiliary classification to enhance the confidence level of the results and produce a case-level prediction. RESULTS: For evaluation, we constructed a dataset of 258 chest CT volumes with 153 VP, 42 BP, and 63 normal control cases, for a total of 43,421 slices. We implemented a comprehensive comparison between our SLP-Net and several state-of-the-art methods across the dataset. Our proposed method obtained significant performance without a large amount of data, outperformed other slice-based and volume-based approaches. The superior evaluation performance achieved in the classification experiments demonstrated the ability of our model in the differential diagnosis of VP, BP and normal cases.

The Influences of COVID-19 on Patients With Chronic Kidney Disease: A Multicenter Cross-Sectional Study

Background: The outbreak of coronavirus disease 2019 (COVID-19) has attracted global attention. During the lockdown period of COVID-19, follow-up of many patients with chronic disease had been interrupted, which brought severe challenges to better management of their disease. This study aimed at exploring the change of illness, daily life, and psychological responses during the COVID-19 pandemic among chronic kidney disease (CKD) patients. Methods: A total of 612 patients were enrolled in this study;282 patients were categorized into the CKD stage 1–2 group and 330 patients were categorized into the CKD stage 3–5 group. Among two groups, 168 (27.5%) and 177 (28.9%) patients were female with a median age of 42 and 45, respectively. The study was conducted by collecting the questionnaires in five nephrology centers. The questionnaire consisted of assessment of anxiety by using the Self-Rating Anxiety Scale and the influences of COVID-19, which included basic demographic data, the influences of COVID-19 on illness and daily life, as well as the patients' psychological responses during the epidemic. Results: A total of 612 patients were included and divided into two groups according to eGFR. Ninety-six patients (34%) in the CKD stage 1–2 group and 141 patients (42.7%) in the CKD stage 3–5 group had reduced their follow-up frequency (p = 0.031). More patients with CKD stages 1–2 consulted online (25.9%), p = 0.005. Besides, patients in the CKD stage 3–5 group tended to be more anxious about follow-up (p = 0.002), fearful of being infected with COVID-19 (p = 0.009), and more likely to feel symptoms getting worse (p = 0.006). The standard scores of SAS were 48.58 ± 7.082 and 51.19 ± 5.944 in the CKD stage 1–2 group and the CKD stage 3–5 group, respectively (p < 0.001). There were significant differences in the severity of anxiety (p = 0.004). Conclusion: COVID-19 had a greater impact on patients with CKD stages 3–5 than those with stages 1–2 in terms of illness, daily life, and psychological disorder. Patients with CKD stages 3–5 were more anxious during the COVID-19 pandemic.

Model-Informed Drug Development Approaches to Assist New Drug Development in the COVID-19 Pandemic.

Leveraging limited clinical and nonclinical data through modeling approaches facilitates new drug development and regulatory decision making amid the coronavirus disease 2019 (COVID-19) pandemic. Model-informed drug development (MIDD) is an essential tool to integrate those data and generate evidence to (i) provide support for effectiveness in repurposed or new compounds to combat COVID-19 and dose selection when clinical data are lacking; (ii) assess efficacy under practical situations such as dose reduction to overcome supply issues or emergence of resistant variant strains; (iii) demonstrate applicability of MIDD for full extrapolation to adolescents and sometimes to young pediatric patients; and (iv) evaluate the appropriateness for prolonging a dosing interval to reduce the frequency of hospital visits during the pandemic. Ongoing research activities of MIDD reflect our continuous effort and commitment in bridging knowledge gaps that leads to the availability of effective treatments through innovation. Case examples are presented to illustrate how MIDD has been used in various stages of drug development and has the potential to inform regulatory decision making.

The development and improvement of ribonucleic acid therapy strategies.

The biological understanding of RNA has evolved since the discovery of catalytic RNAs in the early 1980s and the establishment of RNA interference (RNAi) in the 1990s. RNA is no longer seen as the simple mid-product between transcription and translation but as potential molecules to be developed as RNA therapeutic drugs. RNA-based therapeutic drugs have gained recognition because of their ability to regulate gene expression and perform cellular functions. Various nucleobase, backbone, and sugar-modified oligonucleotides have been synthesized, as natural oligonucleotides have some limitations such as poor low nuclease resistance, binding affinity, poor cellular uptake, and toxicity, which affect their use as RNA therapeutic drugs. In this review, we briefly discuss different RNA therapeutic drugs and their internal connections, including antisense oligonucleotides, small interfering RNAs (siRNAs) and microRNAs (miRNAs), aptamers, small activating RNAs (saRNAs), and RNA vaccines. We also discuss the important roles of RNA vaccines and their use in the fight against COVID-19. In addition, various chemical modifications and delivery systems used to improve the performance of RNA therapeutic drugs and overcome their limitations are discussed.

Anti-SARS-CoV-2 Repurposing Drug Database: Clinical Pharmacology Considerations.

A critical step to evaluate the potential in vivo antiviral activity of a drug is to connect the in vivo exposure to its in vitro antiviral activity. The Anti-SARS-CoV-2 Repurposing Drug Database is a database that includes both in vitro anti-SARS-CoV-2 activity and in vivo pharmacokinetic data to facilitate the extrapolation from in vitro antiviral activity to potential in vivo antiviral activity for a large set of drugs/compounds. In addition to serving as a data source for in vitro anti-SARS-CoV-2 activity and in vivo pharmacokinetic information, the database is also a calculation tool that can be used to compare the in vitro antiviral activity with in vivo drug exposure to identify potential anti-SARS-CoV-2 drugs. Continuous development and expansion are feasible with the public availability of this database.

Human ACE2 receptor polymorphisms and altered susceptibility to SARS-CoV-2.

COVID-19 is a respiratory illness caused by a novel coronavirus called SARS-CoV-2. The viral spike (S) protein engages the human angiotensin-converting enzyme 2 (ACE2) receptor to invade host cells with ~10-15-fold higher affinity compared to SARS-CoV S-protein, making it highly infectious. Here, we assessed if ACE2 polymorphisms can alter host susceptibility to SARS-CoV-2 by affecting this interaction. We analyzed over 290,000 samples representing >400 population groups from public genomic datasets and identified multiple ACE2 protein-altering variants. Using reported structural data, we identified natural ACE2 variants that could potentially affect virus-host interaction and thereby alter host susceptibility. These include variants S19P, I21V, E23K, K26R, T27A, N64K, T92I, Q102P and H378R that were predicted to increase susceptibility, while variants K31R, N33I, H34R, E35K, E37K, D38V, Y50F, N51S, M62V, K68E, F72V, Y83H, G326E, G352V, D355N, Q388L and D509Y were predicted to be protective variants that show decreased binding to S-protein. Using biochemical assays, we confirmed that K31R and E37K had decreased affinity, and K26R and T92I variants showed increased affinity for S-protein when compared to wildtype ACE2. Consistent with this, soluble ACE2 K26R and T92I were more effective in blocking entry of S-protein pseudotyped virus suggesting that ACE2 variants can modulate susceptibility to SARS-CoV-2.

Vaccination willingness, vaccine hesitancy, and estimated coverage at the first round of COVID-19 vaccination in China: A national cross-sectional study.

BACKGROUND: Vaccination against coronavirus disease 2019 (COVID-19) has become an important public health solution. To date, there has been a lack of data on COVID-19 vaccination willingness, vaccine hesitancy, and vaccination coverage in China since the vaccine has become available. METHODS: We designed and implemented a cross-sectional, population-based online survey to evaluate the willingness, hesitancy, and coverage of the COVID-19 vaccine among the Chinese population. 8742 valid samples were recruited and classified as the vaccine-priority group (n = 3902; 44.6%) and the non-priority group (n = 4840; 55.4%). RESULTS: The proportion of people's trust in the vaccine, delivery system, and government were 69.0%, 78.0% and 81.3%, respectively. 67.1% of the participants were reportedly willing to accept the COVID-19 vaccination, while 9.0% refused it. 834 (35.5%) reported vaccine hesitancy, including acceptors with doubts (48.8%), refusers (39.4%), and delayers (11.8%). The current coverage was 34.4%, far from reaching the requirements of herd immunity. The predicted rate of COVID-19 vaccination was 64.9%, 68.9% and 81.1% based on the rates of vaccine hesitancy, willingness, and refusal, respectively. CONCLUSIONS: The COVID-19 vaccine rate is far from reaching the requirements of herd immunity, which will require more flexible and comprehensive efforts to improve the population's confidence and willingness to vaccinate. It should be highlighted that vaccination alone is insufficient to stop the pandemic; further efforts are needed not only to increase vaccination coverage but also to maintain non-specific prevention strategies.

Harnessing peak transmission around symptom onset for non-pharmaceutical intervention and containment of the COVID-19 pandemic.

Within a short period of time, COVID-19 grew into a world-wide pandemic. Transmission by pre-symptomatic and asymptomatic viral carriers rendered intervention and containment of the disease extremely challenging. Based on reported infection case studies, we construct an epidemiological model that focuses on transmission around the symptom onset. The model is calibrated against incubation period and pairwise transmission statistics during the initial outbreaks of the pandemic outside Wuhan with minimal non-pharmaceutical interventions. Mathematical treatment of the model yields explicit expressions for the size of latent and pre-symptomatic subpopulations during the exponential growth phase, with the local epidemic growth rate as input. We then explore reduction of the basic reproduction number R0 through specific transmission control measures such as contact tracing, testing, social distancing, wearing masks and sheltering in place. When these measures are implemented in combination, their effects on R0 multiply. We also compare our model behaviour to the first wave of the COVID-19 spreading in various affected regions and highlight generic and less generic features of the pandemic development.

Molecular epidemiology of SARS-CoV-2 clusters caused by asymptomatic cases in Anhui Province, China.

BACKGROUND: COVID-19 is a newly emerging disease caused by a novel coronavirus (SARS-CoV-2), which spread globally in early 2020. Asymptomatic carriers of the virus contribute to the propagation of this disease, and the existence of asymptomatic infection has caused widespread fear and concern in the control of this pandemic. METHODS: In this study, we investigated the origin and transmission route of SARS-CoV-2 in Anhui's two clusters, analyzed the role and infectiousness of asymptomatic patients in disease transmission, and characterized the complete spike gene sequences in the Anhui strains. RESULTS: We conducted an epidemiological investigation of two clusters caused by asymptomatic infections sequenced the spike gene of viruses isolated from 12 patients. All cases of the two clusters we investigated had clear contact histories, both from Wuhan, Hubei province. The viruses isolated from two outbreaks in Anhui were found to show a genetically close link to the virus from Wuhan. In addition, new single nucleotide variations were discovered in the spike gene. CONCLUSIONS: Both clusters may have resulted from close contact and droplet-spreading and asymptomatic infections were identified as the initial cause. We also analyzed the infectiousness of asymptomatic cases and the challenges to the current epidemic to provided information for the development of control strategies.