Correction: Enterovirus 71 Protease 2Apro Targets MAVS to Inhibit Anti-Viral Type I Interferon Responses.

[This corrects the article DOI: 10.1371/journal.ppat.1003231.].

Regulating electronic structure of Fe single-atom site by S/N dual-coordination for efficient Fenton-like catalysis.

The activity of single-atom catalysts in peroxymonosulfate activation process is bound up with the local electronic state of metal center. However, the large electronegativity of N atoms in Metal-N4 restricts the electron transfer between center metal atom and peroxymonosulfate. Herein, we constructed Fe-SN-C catalyst by incorporating S atom in the first coordination sphere of Fe single-atom site (Fe-S1N3) for Fenton-like catalysis. The Fe-SN-C with a low valent Fe is found to exhibit excellent catalytic activity for bisphenol A degradation, and the corresponding rate constant reaches 0.405 min-1, 11.9-fold higher than the original Fe-N-C. Besides, the Fe-SN-C/PMS system exhibits ideal catalytic stability under the effect of wide pH range and background substrates by the fast generation of high-valent Fe species. Experimental results and theoretical calculations reveal that the dual coordination of S and N atoms notably increases the local electron density of Fe atoms and electron filling in eg orbital, causing a d band center shifting close to the fermi level and thereby optimizes the activation energy for peroxymonosulfate decomposition via Fe 3d-O 2p orbital interaction. This work provides further development of promising SACs for the efficient activation of peroxymonosulfate based on direct regulation of the coordination environment of active center metal atoms.

Generalized heterophily graph data augmentation for node classification.

Graph data augmentations have demonstrated remarkable performance on homophilic graph neural networks (GNNs). Nevertheless, when transferred to a heterophilic graph, these augmentations are less effective for GNN models and lead to reduced performance. To address this issue, we propose a unified augmentation approach called GePHo, a regularization technique for heterophilic graph neural networks based on self-supervised learning, leveraging graph data augmentation to acquire extra information to guide model learning. Specifically, we propose to generate a pseudo-homophily graph that is type-agnostic, enabling us to apply GePHo to both homophilic and heterophilic graphs. Then, we regularize the neighbors with a sharpening technique for data augmentation and generate the auxiliary pseudo-labels to classify the original GNN's output, whose operations are to constrain the local and global node representation, respectively. Extensive experiments on three homophilic graph and six heterophilic graph datasets demonstrate the competitive effectiveness of GePHo in node classification task, and the ablation experiments verify the efficacy of our GePHo in graph data augmentation.

DNA self-assembly nanoflower reverse P-glycoprotein mediated drug resistance in chronic myelogenous leukemia therapy.

Introduction: Chronic myelogenous leukemia (CML) is a clonal myeloproliferative disorder caused by the BCR-ABL chimeric tyrosine kinase. Vincristine (VCR) is widely used in leukemia therapy but is hindered by multidrug resistance (MDR). Methods: We prepared DNA nanoflower via self-assembly for the delivery of VCR and P-glycoprotein small interfering RNA (P-gp siRNA). Results and Discussion: The as-prepared nanoflower had a floriform shape with high loading efficiency of VCR (80%). Furthermore, the nanoflower could deliver VCR and P-gp siRNA into MDR CML cells and induce potent cytotoxicity both in vitro and in vivo, thus overcoming MDR of CML. Overall, this nanoflower is a promising tool for resistant CML therapy.

Expression, Purification, and Crystallization of the Vγ9Vδ2 T-cell Receptor Recognizing Protein/Peptide Antigens.

γδ T cells, especially Vγ9Vδ2 T cells, play an important role in mycobacterial infection. We have identified some Vγ9Vδ2 T cells that recognize protein/peptide antigens derived from mycobacteria, which may induce protective immune responses to mycobacterial infection. To clarify the structural basis of the molecular recognition mechanism, we tried many methods to express the Vγ9Vδ2 T-cell receptor (TCR). The Vγ9Vδ2 TCR was not expressed well in a prokaryotic expression system or a baculovirus expression system, even after extensive optimization. In a mammalian cell expression system, the Vγ9Vδ2 TCR was expressed in the form of a soluble heterodimer, which was suitable for crystal screening. Reduced-temperature cultivation (cold shock) increased the yield of the recombinant TCR. The recombinant purified TCR was used for crystal trials, and crystals that could be used for X-ray diffraction were obtained. Although we have not yet determined the crystal structure of the Vγ9Vδ2 TCR, we have established a procedure for Vγ9Vδ2 TCR expression and purification, which is useful for basic research and potentially for clinical application.

Slow stripe rusting in Chinese wheat Jimai 44 conferred by Yr29 in combination with a major QTL on chromosome arm 6AL.

KEY MESSAGE: YrJ44, a more effective slow rusting gene than Yr29, was localized to a 3.5-cM interval between AQP markers AX-109373479 and AX-109563479 on chromosome 6AL. "Slow rusting" (SR) is a type of adult plant resistance (APR) that can provide non-specific durable resistance to stripe rust in wheat. Chinese elite wheat cultivar Jimai 44 (JM44) has maintained SR to stripe rust in China since its release despite exposure to a changing and variable pathogen population. An F2:6 population comprising 295 recombinant inbred lines (RILs) derived from a cross between JM44 and susceptible cultivar Jimai 229 (JM229) was used in genetic analysis of the SR. The RILs and parental lines were evaluated for stripe rust response in five field environments and genotyped using the Affymetrix Wheat55K SNP array and 13 allele-specific quantitative PCR-based (AQP) markers. Two stable QTL on chromosome arms 1BL and 6AL were identified by inclusive composite interval mapping. The 1BL QTL was probably the pleiotropic gene Lr46/Yr29/Sr58. QYr.nwafu-6AL (hereafter named YrJ44), mapped in a 3.5-cM interval between AQP markers AX-109373479 and AX-109563479, was more effective than Yr29 in reducing disease severity and relative area under the disease progress curve (rAUDPC). RILs harboring both YrJ44 and Yr29 displayed levels of SR equal to the resistant parent JM44. The AQP markers linked with YrJ44 were polymorphic and significantly correlated with stripe rust resistance in a panel of 1,019 wheat cultivars and breeding lines. These results suggested that adequate SR resistance can be obtained by combining YrJ44 and Yr29 and the AQP markers can be used in breeding for durable stripe rust resistance.

Genotypic spectrum of α-thalassemia and ß-thalassemia in newborns of the Li minority in Hainan province, China.

Purpose: To explore the genotypes and allele frequencies of α, ß and α+ß thalassemias in Li minorities, which resided in Hainan Province of China for a long time. Methods: In the present study, 1,438 newborns of the Li minority were collected from January 2020 to April 2021. The genotypes of thalassemia were detected by fluorescence PCR and verified by flow-through hybridization PCR analyses. Rare genotypes were detected by restriction fragment length polymorphism electrophoresis and Sanger DNA sequencing. Results: Among 1,438 participants, 1,024 (71.2%) were diagnosed with any kind of thalassemia. Among all thalassemia carriers, 902 (88.09%) subjects were diagnosed with α-thalassemia, and 18 subtypes of α-thalassemia were detected, with the top three genotypes being -α4.2/αα (25.39%), -α3.7/αα (22.62%) and αWSα/αα (16.96%). Thirty-two (3.13%) patients were ß-thalassemia carriers, and 6 types of ß-thalassemia genotypes were detected. The top two genotypes were ßCD41-42/ßN (46.88%) and ß-28/ßN (18.75%). Additionally, 90 (8.79%) cases were α + ß-thalassemia, and the top two genotypes were -α3.7/αα, ßCD41-42/ßN (30.00%) and -α4.2/αα, ßCD41-42/ßN (26.67%). Furthermore, two genotypes (-α4.2/HKαα and ßCD76 GCT > CCT/ßN) were first identified in Hainan Province, and ßCD76 GCT > CCT/ßN was first identified in China. Conclusion: Newborns of Li have a higher prevalence of thalassemia for a long period, and further education on the impact of thalassemia, follow-up studies of the clinical manifestation and treatment and proper intervention methods should be designed to reduce the burden of thalassemia and enhance the quality of life in Li newborns.

Rapid determination and health risk assessment of neonicotinoids in source water and tap water of the tropical Hainan Island, China.

Neonicotinoids (NEOs) pesticides are widely used around the world, especially in the tropics with greater frequency and intensity. However, little is known about NEOs residue in drinking water of tropics. In this study, a highly efficient method using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was established for determining eight NEOs in source water and tap water of Hainan Island, China. The method adopted a high-throughput direct aqueous injection without sample concentration steps, with a rapid analyzing period of 5.0 min, method detection limits (MDLs) in the range of 0.84-1.82 ng/L and the average recoveries ranged from 83% to 116%. NEOs were detected in all source water samples and at an upper level as compared with other parts of China. The most frequently detected NEO was imidacloprid with a detection frequency of 94%, followed by clothianidin (88%) and thiamethoxam (78%), with maximum concentrations of 86.4, 164, and 188 ng/L, respectively. Moreover, seasonal and spatial variations had remarkable impacts on NEO contamination in source water. Drinking water treatment processes removed approximately 20% of NEOs from surface water. However, 90% of tap water samples contained at least one NEO, With 3 samples' concentration of single NEO exceeding the acceptable value recommended by the European Union (100 ng/L). Therefore, the risk of human exposure through drinking water was evaluated for 4 age group and 2 genders. Young children aged 9 months to 3 years old were found to have the highest risk, with the median exposure up to 4 times greater than teenagers and adults. Next, water intake is likely only a small part of the daily intake of these individuals, thus the potential health problems caused by NEOs present in the tap water of Hainan should not be ignored.

Improved Electronic Structure from Spin-State Reconstruction of a Heteronuclear Fe-Co Diatomic Pair to Boost the Fenton-like Reaction.

Dual-atom catalysts (DACs) are promising candidates for various catalytic reactions, including electrocatalysis, chemical synthesis, and environmental remediation. However, the high-activity origin and mechanism underlying intrinsic activity enhancement remain elusive, especially for the Fenton-like reaction. Herein, we systematically compared the catalytic performance of dual-atom FeCo-N/C with its single-atom counterparts by activating peroxymonosulfate (PMS) for pollutant abatement. The unusual spin-state reconstruction on FeCo-N/C is demonstrated to effectively improve the electronic structure of Fe and Co in the d orbital and enhance the PMS activation efficiency. Accordingly, the dual-atom FeCo-N/C with an intermediate-spin state remarkably boosts the Fenton-like reaction by almost 1 order of magnitude compared with low-spin Co-N/C and high-spin Fe-N/C. Moreover, the established dual-atom-activated PMS system also exhibits excellent stability and robust resistance against harsh conditions. Combined theoretical calculations reveal that unlike unitary Co atom or Fe atom transferring electrons to the PMS molecule, the Fe atom of FeCo-N/C provides extra electrons to the neighboring Co atom and positively shifts the d band of the Co center, thereby optimizing the PMS adsorption and decomposition into a unique high-valent FeIV-O-CoIV species via a low-energy barrier pathway. This work advances a conceptually novel mechanistic understanding of the enhanced catalytic activity of DACs in Fenton-like reactions and helps to expand the application of DACs in various catalytic reactions.

Performance optimization and mechanism analysis of applied Enteromorpha-based composite dust suppressant.

In order to solve the problem of environmental pollution caused by the escape of coal dust in open-pit coal mines, a composite dust suppressant was prepared from Enteromorpha, and the preparation factors (water-soluble polymer, temperature, solid content and surfactant) were optimized. The mechanism of dust suppression and the possibility of large-scale field application were discussed. The research results on the related properties of dust suppressants showed that the performance of Enteromorpha-based dust suppressants prepared by this method was excellent compared with similar studies. Among them, polyacrylamide (PAM) Enteromorpha-based dust suppressant had the best performance, with viscosity of 25.1 mPa s and surface tension of 27.05 mN/m. Moreover, PAM Enteromorpha-based dust suppressant had the best effect, with the mass loss of 2.94% under the wind speed of 10 m/s and the coal dust loss rate of 4.6% after rain erosion, and it had strong water retention performance. Through the discussion of dust suppression mechanism, it was found that the mechanical entangled network structure with hydrogen bonds as nodes was formed after the graft copolymerization of PAM and Enteromorpha. It had high permeability and good adhesion. After quickly wetting coal dust, it formed a dense package for coal dust. The field experiment also showed that the use of Enteromorpha-based dust suppressant can effectively inhibit the escape of coal dust. From the point of view of economy and efficiency, Enteromorpha can save 30% of the material cost and the dust suppression efficiency can reach 89-94%. Therefore, the Enteromorpha-based dust suppressant may stably suppress coal dust on the basis of reducing the cost.

Succession of endophytic bacterial community and its contribution to cinnamon oil production during cinnamon shade-drying process.

Cinnamon oil is a blend of secondary metabolites and is widely used as spice. Endophytic bacteria are always related to the secondary metabolites production. However, the potential of endophytic bacteria communities for cinnamon oil production during cinnamon shade-drying process is still not clear. In this study, we investigated the composition and metabolic function of endophytic bacterial community during 80-day shade-drying process. The temporal dynamics of essential oil content and its dominant constituents were analyzed. The succession of endophytic bacterial community from d0 to d80 was identified. The influence of endophytic bacterial community evolution on cinnamon oil is significant positive. Predictive functional analysis indicated that shade-drying process was rich in Saccharopolyspora that produce enzymes for the conversion of phenylalanine to cinnamaldehyde. These findings enhance our understanding of the functional bacterial genera and functional genes involved in the production of cinnamon oil during cinnamon shade-drying process.

Synthesis, Herbicidal Activity and Toxicity Evaluation of Secondary Ammonium Salts from Turpentine Oil for Sustainable Weed Control.

Three series of secondary ammonium chloride from turpentine were synthesized and evaluated as botanical herbicides. The preemergence herbicidal activities against ryegrass (Loliun multiflorum) and barnyard grass (Echinochloa crus-galli) were investigated using water as the only solvent. Their toxicity was evaluated by cytotoxicity assays. Preliminary results demonstrated that the herbicidal performance of the prepared salts was similar or much higher than that of corresponding secondary amines and even commercial herbicide glyphosate. Promisingly, compound 14e containing a cyclohexyl-substituted p-menthene skeleton with an IC50 value of 0.0014 mM against root growth of ryegrass showed 39-fold higher herbicidal activity than glyphosate. Besides, this compound was found to be nontoxic to human and animal cells, indicating the potential application as a water-soluble herbicide for ryegrass control.

Establishing a Thermal Imaging Technology (IRT) Based System for Evaluating Rat Erectile Function.

INTRODUCTION: Measurement of intra-cavernous pressure (ICP) is an internationally recognized method to evaluate erectile function of animals, however, this process is invasive, destructive, and cannot be repeated, leading to a daunting challenge for monitoring the changes in erectile function throughout the whole treatment duration. AIM: To verify whether infrared ray thermography technology based system could be a good substitution of ICP for evaluating rat erectile function. METHODS: A novel thermal image-based method, infrared ray thermography technology (IRT) was employed to monitor erectile function in erectile dysfunction (ED) rats. To detect the sensitivity and specificity of this new technology, 4 ED rat models (Diabetic, nerve-injury, vascular-injury and aged ED models) were established and subjected to both ICP and IRT test. OUTCOMES: Statistical comparisons were done to test the effectiveness of this new way for detecting and dynamically monitoring erectile function. RESULTS: Based on the data curves obtained from ICP and IRT, the IRT showed a similar trend (including peak value, climbing speed) as that of ICP. IRT is considered as a precise way to monitor the real-time changes of erectile function in all ED rat models. The AUC of peak temperature detected by IRT in DMED, aged ED, vascular-injury ED, the nerve-injury ED and total ED rat models were 0.9811,0.9836,0.9893,0.9989 and 0.9882, respectively. Meanwhile, the AUC of temperature climbing rate were 0.6486,0.8357,0.9184,0.8675and 0.8168.Also,it is a non-invasive process of dynamically monitoring erectile function of a same rat at different time points (before and after drug intervention). The data showed that the real-time recovery by tadalafil was obtained by IRT methods even after treatment for only 2 weeks in the diabetic ED (DMED) rat model. CONCLUSION: A novel noninvasive method for monitoring erectile function in rat ED models was established, and can replace or supplement ICP test. Liu S, Zhao Z, Wang Z et al. Establishing a Thermal Imaging Technology (IRT) Based System for Evaluating Rat Erectile Function. Sex Med 2022;10:100475.

ID2 inhibits innate antiviral immunity by blocking TBK1- and IKKε-induced activation of IRF3.

The transcription regulator ID2 plays an essential role in the development and differentiation of immune cells. Here, we report that ID2 also negatively regulates antiviral innate immune responses. During viral infection of human epithelial cells, ID2 bound to TANK-binding kinase 1 (TBK1) and to inhibitor of nuclear factor κB kinase ε (IKKε). These interactions inhibited the recruitment and activation of interferon (IFN) regulatory factor 3 (IRF3) by TBK1 or IKKε, leading to a reduction in the expression of IFN-ß1 (IFNB1). IFN-ß induced the nuclear export of ID2 to form a negative feedback loop. Knocking out ID2 in human cells enhanced innate immune responses and suppressed infection by different viruses, including SARS-CoV-2. Mice with a myeloid-specific deficiency of ID2 produced more IFN-α in response to viral infection and were more resistant to viral infection than wild-type mice. Our findings not only establish ID2 as a modulator of IRF3 activation induced by TBK1 and/or IKKε but also introduce a mechanism for cross-talk between innate immunity and cell development and differentiation.

Effects of the different waxy proteins on starch biosynthesis, starch physicochemical properties and Chinese noodle quality in wheat.

Noodles are an important food in Asia. Wheat starch is the most important component in Chinese noodles. Loss of the waxy genes leads to lower activity of starch synthesis enzymes and decreased amylose content that further affects starch properties and noodle quality. To study the effects of different waxy (Wx) protein subunits on starch biosynthesis and processing quality, the high-yielding wheat cultivar Jimai 22 was treated with the mutagen ethyl methane sulfonate (EMS) to produce a population of Wx lines and chosen 7 Wx protein combinations. The amylose content increased but swelling power decreased as the number of Wx proteins increased. Both GBSS activity and gene expression were the lowest for the waxy mutant, followed by the mutants with 1 Wx protein. The combinations of these mutant alleles lead to reductions in both RNA expression and protein levels. Noodles made from materials with 2 Wx protein subunits had the highest score, which agreed with peak viscosity. The influence of the Wx-B1 protein on amylose synthesis and noodle quality was the highest, whereas the influence of Wx-A1 protein was the lowest. Mutants with lower amylose content caused by the absence of 1 subunit, especially the Wx-B1 subunit, had superior noodle quality. Additionally, the identified mutant lines can be used as intermediate materials to improve wheat quality. Supplementary Information: The online version contains supplementary material available at 10.1007/s11032-022-01292-x.

Pollutant removal from municipal sewage by a microaerobic up-flow oxidation ditch coupled with micro-electrolysis.

The development of efficient and low-cost wastewater treatment processes remains an important challenge. A microaerobic up-flow oxidation ditch (UOD) with micro-electrolysis by waterfall aeration was designed for treating real municipal wastewater. The effects of influential factors such as up-flow rate, waterfall height, reflux ratio, number of stages and iron dosing on pollutant removal were fully investigated, and the optimum conditions were obtained. The elimination efficiencies of chemical oxygen demand (COD), ammonia nitrogen (NH4 +-N), total nitrogen (TN) and total phosphorus (TP) reached up to 84.33 ± 2.48%, 99.91 ± 0.09%, 93.63 ± 0.60% and 89.27 ± 1.40%, respectively, while the effluent concentrations of COD, NH4 +-N, TN and TP were 20.67 ± 2.85, 0.02 ± 0.02, 1.39 ± 0.09 and 0.27 ± 0.02 mg l-1, respectively. Phosphorous removal was achieved by iron-carbon micro-electrolysis to form an insoluble ferric phosphate precipitate. The microbial community structure indicated that carbon and nitrogen were removed via multiple mechanisms, possibly including nitrification, partial nitrification, denitrification and anammox in the UOD.

Hepatitis C virus induces oxidation and degradation of apolipoprotein B to enhance lipid accumulation and promote viral production.

Hepatitis C virus (HCV) infection induces the degradation and decreases the secretion of apolipoprotein B (ApoB). Impaired production and secretion of ApoB-containing lipoprotein is associated with an increase in hepatic steatosis. Therefore, HCV infection-induced degradation of ApoB may contribute to hepatic steatosis and decreased lipoprotein secretion, but the mechanism of HCV infection-induced ApoB degradation has not been completely elucidated. In this study, we found that the ApoB level in HCV-infected cells was regulated by proteasome-associated degradation but not autophagic degradation. ApoB was degraded by the 20S proteasome in a ubiquitin-independent manner. HCV induced the oxidation of ApoB via oxidative stress, and oxidized ApoB was recognized by the PSMA5 and PSMA6 subunits of the 20S proteasome for degradation. Further study showed that ApoB was degraded at endoplasmic reticulum (ER)-associated lipid droplets (LDs) and that the retrotranslocation and degradation of ApoB required Derlin-1 but not gp78 or p97. Moreover, we found that knockdown of ApoB before infection increased the cellular lipid content and enhanced HCV assembly. Overexpression of ApoB-50 inhibited lipid accumulation and repressed viral assembly in HCV-infected cells. Our study reveals a novel mechanism of ApoB degradation and lipid accumulation during HCV infection and might suggest new therapeutic strategies for hepatic steatosis.

MARCH8 inhibits influenza A virus infection by targeting viral M2 protein for ubiquitination-dependent degradation in lysosomes.

The membrane-associated RING-CH (MARCH) proteins are E3 ligases that regulate the stability of various cellular membrane proteins. MARCH8 has been reported to inhibit the infection of HIV-1 and a few other viruses, thus plays an important role in host antiviral defense. However, the antiviral spectrum and the underlying mechanisms of MARCH8 are incompletely defined. Here, we demonstrate that MARCH8 profoundly inhibits influenza A virus (IAV) replication both in vitro and in mice. Mechanistically, MARCH8 suppresses IAV release through redirecting viral M2 protein from the plasma membrane to lysosomes for degradation. Specifically, MARCH8 catalyzes the K63-linked polyubiquitination of M2 at lysine residue 78 (K78). A recombinant A/Puerto Rico/8/34 virus carrying the K78R M2 protein shows greater replication and more severe pathogenicity in cells and mice. More importantly, we found that the M2 protein of the H1N1 IAV has evolved to acquire non-lysine amino acids at positions 78/79 to resist MARCH8-mediated ubiquitination and degradation. Together, our data support the important role of MARCH8 in host anti-IAV intrinsic immune defense by targeting M2, and suggest the inhibitory pressure of MARCH8 on H1N1 IAV transmission in the human population.

Enterovirus 71 Induces INF2 Cleavage via Activated Caspase-2 in Infected RD Cells.

Enterovirus 71 (EV71) is the major causative pathogen of hand, foot, and mouth disease. The lack of understanding of the virus's pathogenesis hinders the development of anti-virus drugs and the control of EV71 infection. Our previous studies have demonstrated that both mitochondria and endoplasmic reticulum (ER) were altered significantly in EV71 infected cells, but the mechanism is still unclear. In this study, we investigated the effects of EV71 infection on the expression of INF2, a key regulator factor in ER-Mitochondria communication and mitochondrial fission. We found that INF2 was cleaved in EV71 infected RD cells. The INF2 cleavage occurred at Aspartic 1,051 of INF2 and is mediated by activated caspases, predominantly by activated caspase-2. The subcellular localization of INF2 and caspase-2 was significantly altered in infected cells. We speculate that caspase-2-mediated INF2 cleavage is involved in forming viral replication organelles (ROs) and is a positive feedback regulatory mechanism of mitochondrial disorders caused by EV71 infection.

Construction of Non-infectious SARS-CoV-2 Replicons and Their Application in Drug Evaluation.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused a devastating pandemic worldwide. Vaccines and antiviral drugs are the most promising candidates for combating this global epidemic, and scientists all over the world have made great efforts to this end. However, manipulation of the SARS-CoV-2 should be performed in the biosafety level 3 laboratory. This makes experiments complicated and time-consuming. Therefore, a safer system for working with this virus is urgently needed. Here, we report the construction of plasmid-based, non-infectious SARS-CoV-2 replicons with turbo-green fluorescent protein and/or firefly luciferase reporters by reverse genetics using transformation-associated recombination cloning in Saccharomyces cerevisiae. Replication of these replicons was achieved simply by direct transfection of cells with the replicon plasmids as evident by the expression of reporter genes. Using SARS-CoV-2 replicons, the inhibitory effects of E64-D and remdesivir on SARS-CoV-2 replication were confirmed, and the half-maximal effective concentration (EC50) value of remdesivir and E64-D was estimated by different quantification methods respectively, indicating that these SARS-CoV-2 replicons are useful tools for antiviral drug evaluation.