Development of a Cell Culture Model for Inducible SARS-CoV-2 Replication.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) induces direct cytopathic effects, complicating the establishment of low-cytotoxicity cell culture models for studying its replication. We initially developed a DNA vector-based replicon system utilizing the CMV promoter to generate a recombinant viral genome bearing reporter genes. However, this system frequently resulted in drug resistance and cytotoxicity, impeding model establishment. Herein, we present a novel cell culture model with SARS-CoV-2 replication induced by Cre/LoxP-mediated DNA recombination. An engineered SARS-CoV-2 transcription unit was subcloned into a bacterial artificial chromosome (BAC) vector. To enhance biosafety, the viral spike protein gene was deleted, and the nucleocapsid gene was replaced with a reporter gene. An exogenous sequence was inserted within NSP1 as a modulatory cassette that is removable after Cre/LoxP-mediated DNA recombination and subsequent RNA splicing. Using the PiggyBac transposon strategy, the transcription unit was integrated into host cell chromatin, yielding a stable cell line capable of inducing recombinant SARS-CoV-2 RNA replication. The model exhibited sensitivity to the potential antivirals forsythoside A and verteporfin. An innovative inducible SARS-CoV-2 replicon cell model was introduced to further explore the replication and pathogenesis of the virus and facilitate screening and assessment of anti-SARS-CoV-2 therapeutics.

The Moderation Effect of Approach Motivation Between Schizotypy and Creative Ideational Behavior.

Introduction: The schizotypy-creativity link has been studied from different perspectives over the past few decades, yet the results of this relationship are inconsistent in the literature. Previous studies have suggested that two basic motivational systems-Behavioral Inhibition System (BIS, avoidance motivation) and Behavioral Activation System (BAS, approach motivation)-underlie the relationship between schizotypy and creativity. Few empirical studies, however, have examined how the relationship interacts with other variables. This study fills these gaps and explores the role of the approach and avoidance motivation assessed by trait behavioral activation and inhibition in the link between schizotypy as a dimensional personality trait and creative ideational behavior as a measure of creativity. Method: Undergraduate students (N = 388) completed questionnaires including the Schizotypal Personality Questionnaire (SPQ) measuring 3 dimensions of schizotypy, Runco Ideational Behavior Scale (RIBS) measuring creative ideational behavior, and BIS/BAS Scales measuring trait behavioral motivation. Bivariate Pearson correlation was computed, and hierarchical linear regression was performed to explore the effects of schizotypy, BIS/BAS, and their interaction on creative ideational behavior. The conditional effect of schizotypy based on different levels of the moderator was further tested. Results: The total score, positive dimension and disorganized dimension of SPQ were all positively correlated with RIBS, BAS, and BIS. Negative dimension of SPQ was not significantly correlated with the RIBS score but was positively correlated with BIS. Additionally, after controlling gender and age, BAS significantly moderated the relationship between the positive and disorganized dimensions of schizotypy and creative ideational behavior measured by RIBS. However, BIS was not a significant moderator. Discussion: The findings of this study regarding the relationships between different dimensions of schizotypy, two motivational systems, and creative ideational behavior were mostly consistent with previous findings. The significant moderated effect of BAS on the relationship between two dimensions of schizotypy and creative ideational behavior made significant contributions to the understanding of the relationship between schizotypy and creativity.

A wide drive frequency matching method for harmonic suppression and voltage stabilization of ultrasonic motors.

To reduce harmonic components, balance system impedance, and stabilize driving voltage, an additional matching circuit is required for ultrasonic motors (USMs) driver. However, the performance of inductor or capacitor matching can be seriously weakened with changes in driving frequency. Therefore, this paper presents a simple and effective LC matching method against driving frequency adjustment for USMs. First, the driving scheme of the USM is proposed and the electromechanical coupling model is analyzed. Subsequently, the output characteristics of the full-bridge inverter are derived theoretically when the driving frequency deviates from the mechanical resonant frequency. Then, the impedance circular transform method is proposed, which can intuitively analyze the effect of matching parameters on the voltage amplitude. A matching objective function is established that can consider both the voltage stabilization and harmonic suppression. The matching parameters are solved using random weight particle swarm optimization. Simulations and experiments demonstrate that within the operating frequency of the USM, the proposed matching method can effectively prevent overvoltage and suppress harmonic components. Furthermore, compared with the existing resonant matching method, the proposed matching method can realize more stable driving capability at different frequencies. The proposed method could be useful for USMs' variable-frequency driver design.

Construction and efficacy testing of DNA vaccines containing HLA-A*02:01-restricted SARS-CoV-2 T-cell epitopes predicted by immunoinformatics.

Vaccines play essential roles in the fight against the COVID-19 pandemic. The development and assessment of COVID-19 vaccines have generally focused on the induction and boosting of neutralizing antibodies targeting the SARS-CoV-2 spike (S) protein. Due to rapid and continuous variation in the S protein, such vaccines need to be regularly updated to match newly emerged dominant variants. T-cell vaccines that target MHC I- or II-restricted epitopes in both structural and non-structural viral proteins have the potential to induce broadly cross-protective and long-lasting responses. In this work, the entire proteome encoded by SARS-CoV-2 (Wuhan-hu-1) is subjected to immunoinformatics-based prediction of HLA-A*02:01-restricted epitopes. The immunogenicity of the predicted epitopes is evaluated using peripheral blood mononuclear cells from convalescent Wuhan-hu-1-infected patients. Furthermore, predicted epitopes that are conserved across major SARS-CoV-2 lineages and variants are used to construct DNA vaccines expressing multi-epitope polypeptides. Most importantly, two DNA vaccine constructs induce epitope-specific CD8 + T-cell responses in a mouse model of HLA-A*02:01 restriction and protect immunized mice from challenge with Wuhan-hu-1 virus after hACE2 transduction. These data provide candidate T-cell epitopes useful for the development of T-cell vaccines against SARS-CoV-2 and demonstrate a strategy for quick T-cell vaccine candidate development applicable to other emerging pathogens.

Hepatitis B virus core protein as a Rab-GAP suppressor driving liver disease progression.

Chronic hepatitis B virus (HBV) infection can lead to advanced liver pathology. Here, we establish a transgenic murine model expressing a basic core promoter (BCP)-mutated HBV genome. Unlike previous studies on the wild-type virus, the BCP-mutated HBV transgenic mice manifest chronic liver injury that culminates in cirrhosis and tumor development with age. Notably, agonistic anti-Fas treatment induces fulminant hepatitis in these mice even at a negligible dose. As the BCP mutant exhibits a striking increase in HBV core protein (HBc) expression, we posit that HBc is actively involved in hepatocellular injury. Accordingly, HBc interferes with Fis1-stimulated mitochondrial recruitment of Tre-2/Bub2/Cdc16 domain family member 15 (TBC1D15). HBc may also inhibit multiple Rab GTPase-activating proteins, including Rab7-specific TBC1D15 and TBC1D5, by binding to their conserved catalytic domain. In cells under mitochondrial stress, HBc thus perturbs mitochondrial dynamics and prevents the recycling of damaged mitochondria. Moreover, sustained HBc expression causes lysosomal consumption via Rab7 hyperactivation, which further hampers late-stage autophagy and substantially increases apoptotic cell death. Finally, we show that adenovirally expressed HBc in a mouse model is directly cytopathic and causes profound liver injury, independent of antigen-specific immune clearance. These findings reveal an unexpected cytopathic role of HBc, making it a pivotal target for HBV-associated liver disease treatment. The BCP-mutated HBV transgenic mice also provide a valuable model for understanding chronic hepatitis B progression and for the assessment of therapeutic strategies.

TGF-ß1/SMAD3-driven GLI2 isoform expression contributes to aggressive phenotypes of hepatocellular carcinoma.

Hedgehog signaling is activated in response to liver injury, and modulates organogenesis. However, the role of non-canonical hedgehog activation via TGF-ß1/SMAD3 in hepatic carcinogenesis is poorly understood. TGF-ß1/SMAD3-mediated non-canonical activation was found in approximately half of GLI2-positive hepatocellular carcinoma (HCC), and two new GLI2 isoforms with transactivating activity were identified. Phospho-SMAD3 interacted with active GLI2 isoforms to transactivate downstream genes in modulation of stemness, epithelial-mesenchymal transition, chemo-resistance and metastasis in poorly-differentiated hepatoma cells. Non-canonical activation of hedgehog signaling was confirmed in a transgenic HBV-associated HCC mouse model. Inhibition of TGF-ß/SMAD3 signaling reduced lung metastasis in a mouse in situ hepatic xenograft model. In another cohort of 55 HCC patients, subjects with high GLI2 expression had a shorter disease-free survival than those with low expression. Moreover, co-positivity of GLI2 with SMAD3 was observed in 87.5% of relapsed HCC patients with high GLI2 expression, indicating an increased risk of post-resection recurrence of HCC. The findings underscore that suppressing the non-canonical hedgehog signaling pathway may confer a potential strategy in the treatment of HCC.

Push-Pull Inverter Using Amplitude Control and Frequency Tracking for Piezoelectric Transducers.

Frequency tracking and amplitude control are essential for piezoelectric transducers. Frequency tracking ensures the piezoelectric transducer operates at the resonant frequency for maximum power output, and amplitude control regulates the mechanical motion of the output. This paper presents a novel driver based on a push-pull inverter for piezoelectric transducers. The proposed driver realizes the frequency tracking and amplitude control scheme by a voltage sensing bridge in the case of transformer secondary matching, guaranteeing automatic frequency tracking and precise mechanical functions regardless of environmental and load variations. The proposed scheme is verified by the ultrasonic scalpel and the ultrasonic motor (USM). The experimental results show that this scheme reduces the build-up time from 10 ms to 3 ms and loaded frequency variations from 250 Hz to 200 Hz. In addition, the amplitude control performance was further observed on USM for various loads. The overshoot is less than 5.4% under different load torques. Therefore, the proposed scheme improves the load adaptability and stability of piezoelectric transducers and promotes the application of piezoelectric transducers under various conditions.

Canocapavir Is a Novel Capsid Assembly Modulator Inducing a Conformational Change of the Linker Region of HBV Core Protein.

Canocapavir is a novel antiviral agent with characteristics of core protein allosteric modulators (CpAMs) that is currently in a phase II clinical trial for treatment of hepatitis B virus (HBV) infection. Herein, we show that Canocapavir prevented the encapsidation of HBV pregenomic RNA and increased the accumulation of cytoplasmic empty capsids, presumably by targeting the hydrophobic pocket at the dimer-dimer interface of HBV core protein (HBc). Canocapavir treatment markedly reduced the egress of naked capsids, which could be reversed by Alix overexpression through a mechanism other than direct association of Alix with HBc. Moreover, Canocapavir interfered with the interaction between HBc and HBV large surface protein, resulting in diminished production of empty virions. Of particular note, Canocapavir induced a conformational change of capsids, with the C-terminus of HBc linker region fully exposed on the exterior of capsids. We posit that the allosteric effect may have great importance in the anti-HBV activity of Canocapavir, given the emerging virological significance of HBc linker region. In support of this notion, the mutation at HBc V124W typically recapitulated the conformational change of the empty capsid with aberrant cytoplasmic accumulation. Collectively, our results indicate Canocapavir as a mechanistically distinct type of CpAMs against HBV infection.

Suppression of Vanadium Oxide Dissolution via Cation Metathesis within a Coordination Supramolecular Network for Durable Aqueous Zn-V2 O5 Batteries.

Aqueous zinc metal batteries (ZMBs) are a promising sustainable technology for large-scale energy storage applications. However, the water is often associated with problematic parasitic reactions on both anode and cathode, leading to the low durability and reliability of ZMBs. Here, a multifunctional separator for the Zn-V2 O5 batteries by growing the coordination supramolecular network (CSN:Zn-MBA, MBA = 2-mercaptobenzoic acid) on the conventional non-woven fabrics (NWF) is developed. CSN tends to form a stronger coordination bond as a softer cation, enabling a thermodynamically preferred Zn2+ to VO2 + substitution in the network, leading to the formation of VO2 -MBA interface, that strongly obstructs the VO2 (OH)2 - penetration but simultaneously allows Zn2+ transfer. Moreover, Zn-MBA molecules can adsorb the OTF- and distribute the interfacial Zn2+ homogeneous, which facilitate a dendrite-free Zn deposition. The Zn-V2 O5 cells with Zn-MBA@NWF separator realize high capacity of 567 mAh g-1 at 0.2 A g-1 , and excellent cyclability over 2000 cycles with capacity retention of 82.2% at 5 A g-1 . This work combines the original advantages of the template and new function of metals via cation metathesis within a CSN, provides a new strategy for inhibiting vanadium oxide dissolution.

A Spike-destructing human antibody effectively neutralizes Omicron-included SARS-CoV-2 variants with therapeutic efficacy.

Neutralizing antibodies (nAbs) are important assets to fight COVID-19, but most existing nAbs lose the activities against Omicron subvariants. Here, we report a human monoclonal antibody (Ab08) isolated from a convalescent patient infected with the prototype strain (Wuhan-Hu-1). Ab08 binds to the receptor-binding domain (RBD) with pico-molar affinity (230 pM), effectively neutralizes SARS-CoV-2 and variants of concern (VOCs) including Alpha, Beta, Gamma, Mu, Omicron BA.1 and BA.2, and to a lesser extent for Delta and Omicron BA.4/BA.5 which bear the L452R mutation. Of medical importance, Ab08 shows therapeutic efficacy in SARS-CoV-2-infected hACE2 mice. X-ray crystallography of the Ab08-RBD complex reveals an antibody footprint largely in the ß-strand core and away from the ACE2-binding motif. Negative staining electron-microscopy suggests a neutralizing mechanism through which Ab08 destructs the Spike trimer. Together, our work identifies a nAb with therapeutic potential for COVID-19.

Anchoring I3- via Charge-Transfer Interaction by a Coordination Supramolecular Network Cathode for a High-Performance Aqueous Dual-Ion Battery.

Iodine is considered to have broad application prospects in the field of electrochemical energy storage. However, the high solubility of I3- severely hampers its practical application, and the lack of research on the anchoring mechanism of I3- has seriously hindered the development of advanced cathode materials for iodine batteries. Herein, based on the molecular orbital theory, we studied the charge-transfer interaction between the acceptor of I3- with a σ* empty antibonding orbital and the donor of pyrimidine nitrogen with lone-pair electrons, which is proved by the results of UV-vis absorption spectroscopy, Raman spectroscopy, and density functional theory (DFT) calculations. The prepared dual-ion battery (DIB) exhibits a high voltage platform of 1.2 V, a remarkable discharge-specific capacity of up to 207 mAh g-1, and an energy density of 233 Wh kg-1 at a current density of 5 A g-1, as well as outstanding cycle stability (operating stably for 5000 cycles) with a high Coulombic efficiency of 97%, demonstrating excellent electrochemical performance and a promising prospect in stationary energy storage.

The positive role of parental attachment and communication in Chinese adolescents' health behavior and mental health during COVID-19.

INTRODUCTION: Given that coronavirus disease 2019 (COVID-19) has largely influenced adolescents' physical and mental health around the globe, it is important to identify protective factors that may promote adolescents' positive adjustment during the pandemic. This study aimed to examine the role of parental attachment and COVID-19 communication in adolescents' health behavior and mental health during COVID-19. METHODS: A total of 442 Chinese parent-adolescent dyads (mean age of adolescents = 13.35 years; 50% girls) completed two-wave longitudinal surveys over the span of 2 months during the pandemic (Wave 1: July 2020; Wave 2: September 2020). At each wave, adolescents reported on their COVID-19-related health behavior, general health behavior, depressive symptoms, and anxiety symptoms. At Wave 1, parent-adolescent attachment security and COVID-19 communication were also assessed. RESULTS: Adolescents' attachment security to parents was associated with their increased COVID-19-related and general health behavior as well as decreased depression and anxiety over 2 months during COVID-19. Moreover, more frequent parent-adolescent COVID-19 communication was associated with adolescents' increased COVID-19-related and general health behavior over time. Notably, attachment security's and COVID-19 communication's associations with health behavior largely remained the same after taking into account both factors simultaneously. In addition, results from exploratory analyses suggest that more frequent COVID-19 communication mediates the link between attachment security and increased health behavior. CONCLUSIONS: These findings highlight the importance of promoting attachment security and COVID-19 communication between parents and adolescents during the pandemic, which may play a positive role in adolescents' health behavior and mental health.

Neutralization mechanism of a human antibody with pan-coronavirus reactivity including SARS-CoV-2.

Frequent outbreaks of coronaviruses underscore the need for antivirals and vaccines that can counter a broad range of coronavirus types. We isolated a human antibody named 76E1 from a COVID-19 convalescent patient, and report that it has broad-range neutralizing activity against multiple α- and ß-coronaviruses, including the SARS-CoV-2 variants. 76E1 also binds its epitope in peptides from γ- and δ-coronaviruses. 76E1 cross-protects against SARS-CoV-2 and HCoV-OC43 infection in both prophylactic and therapeutic murine animal models. Structural and functional studies revealed that 76E1 targets a unique epitope within the spike protein that comprises the highly conserved S2' site and the fusion peptide. The epitope that 76E1 binds is partially buried in the structure of the SARS-CoV-2 spike trimer in the prefusion state, but is exposed when the spike protein binds to ACE2. This observation suggests that 76E1 binds to the epitope at an intermediate state of the spike trimer during the transition from the prefusion to the postfusion state, thereby blocking membrane fusion and viral entry. We hope that the identification of this crucial epitope, which can be recognized by 76E1, will guide epitope-based design of next-generation pan-coronavirus vaccines and antivirals.

Constructing Hydrophobic Interface with Close-Packed Coordination Supramolecular Network for Long-Cycling and Dendrite-Free Zn-Metal Batteries.

Commercialization of aqueous zinc-metal batteries remains unrealistic due to the substantial dendrite growth and side reaction issues on the zinc anodes. It is highly demanded to develop easy-to-handle approaches for constructing stable, dense, as well as homogeneous solid anode/electrolyte interfaces. Herein, the authors construct the zinc anode interface with a close-packed Zn-TSA (TSA = thiosalicylate) coordination supramolecular network through the facile and up-scalable wet-chemical method. The hydrophobic Zn-TSA network can block solvated water and establish a solid-state diffusion barrier to well-distribute the interfacial Zn2+ , thus inhibiting hydrogen evolution and zinc dendrite growth on the anode. Meanwhile, the Zn-TSA network induces the formation of a uniform and stable solid electrolyte interphase composed of multiple inorganic-organic compounds. This denser structure can accommodate and self-heal the crack/degradation of the anode interphase associated with the repeated volume changes, and suppress the generation of detrimental by-product, Znx (OTF- )y (OH)2x-y ·nH2 O. Such a rationally fabricated anode/electrolyte interface further endows the assembled symmetric cells with superior plating/stripping stability for over 2000 h without dendrite formation (at 1 mA cm-2 and 1 mAh cm-2 ). Furthermore, this zinc anode has practical application in the Zn-MoS2 and Zn-V2 O5 full cells. This study provides a new train of thought for constructing the dense interface of zinc-metal anode.

Broad neutralization of SARS-CoV-2 variants by an inhalable bispecific single-domain antibody.

The effectiveness of SARS-CoV-2 vaccines and therapeutic antibodies have been limited by the continuous emergence of viral variants and by the restricted diffusion of antibodies from circulation into the sites of respiratory virus infection. Here, we report the identification of two highly conserved regions on the Omicron variant receptor-binding domain recognized by broadly neutralizing antibodies. Furthermore, we generated a bispecific single-domain antibody that was able to simultaneously and synergistically bind these two regions on a single Omicron variant receptor-binding domain as revealed by cryo-EM structures. We demonstrated that this bispecific antibody can be effectively delivered to lung via inhalation administration and exhibits exquisite neutralization breadth and therapeutic efficacy in mouse models of SARS-CoV-2 infections. Importantly, this study also deciphered an uncommon and highly conserved cryptic epitope within the spike trimeric interface that may have implications for the design of broadly protective SARS-CoV-2 vaccines and therapeutics.

The Role of Empathy in Chinese Adolescents' Preventive Health Behavior During COVID-19.

PURPOSE: Preventive health behavior during COVID-19 protects not only oneself but also the welfare of others. However, little attention has been paid to prosocial motivation in adolescents, who are often viewed as selfish and egocentric. Therefore, the current study aimed to explore the role of empathy in adolescents' preventive health behavior using longitudinal data. METHODS: A total of 442 Chinese adolescents (mean age of youth = 13.35 years; 49.5% girls and 50.5% boys) completed two-wave longitudinal surveys over the span of two months during the pandemic (Time 1: July 2020; Time 2: September 2020). At T1, participants reported on their empathic concern, perspective taking, and concern for personal health. At both T1 and T2, participants reported on their preventive health behavior and COVID-related worry. RESULTS: Adolescents who showed greater empathic concern tend to engage in more preventive health behavior over time (p < .01). However, greater empathic concern also predicted adolescents' greater worry about COVID-19 over time (p < .01). In comparison, adolescents' perspective-taking and concern for personal health did not predict their health behavior or worry over time. Notably, the longitudinal effect of empathic concern on preventive health behavior and COVID-related worry remained (ps < .05) after taking into account adolescents' perspective-taking and concern for personal health. CONCLUSIONS: These findings highlight adolescents' prosocial motivation in engaging in preventive health behavior during the pandemic and also point out the potential negative influence of empathic concern on adolescent mental health.

Numerical analysis of hemodynamic effect under different enhanced external counterpulsation (EECP) frequency for cerebrovascular disease: a simulation study.

Based on the changes in phase characteristics of blood flow and pressure, enhanced external counterpulsation (EECP) reduces cardiac load and improves cerebral perfusion in patients with cerebrovascular diseases. However, increased cerebral blood flow (CBF) is associated with the rise in blood pressure and its complications. Increased EECP frequency is a valuable solution when combined with the electrical equivalent impedance characteristics of the lumped parameter model (LPM) of the human blood circulation system. Herein, to investigate the effect of different EECP frequencies on CBF perfusion, an LPM was established with cardiopulmonary circulation and eight systemic blood flow units with cerebral autoregulation module of ischemic stroke patients. Then, using differential equations, we analyzed those parameters through hemodynamic simulations in four EECP modes. With related influencing parameters remaining constant, we adjusted the pressure frequency of EECP and found that when compared to the traditional sequential EECP mode, the relative increase rate of CBF was 16.68%, 18.95%, and 21.21% from 1 to 3 Hz, respectively. This study validates the effect of improving blood prefusion with increasing EECP frequency through numerical analysis.

Receptome profiling identifies KREMEN1 and ASGR1 as alternative functional receptors of SARS-CoV-2.

Host cellular receptors play key roles in the determination of virus tropism and pathogenesis. However, little is known about SARS-CoV-2 host receptors with the exception of ACE2. Furthermore, ACE2 alone cannot explain the multi-organ tropism of SARS-CoV-2 nor the clinical differences between SARS-CoV-2 and SARS-CoV, suggesting the involvement of other receptor(s). Here, we performed genomic receptor profiling to screen 5054 human membrane proteins individually for interaction with the SARS-CoV-2 capsid spike (S) protein. Twelve proteins, including ACE2, ASGR1, and KREMEN1, were identified with diverse S-binding affinities and patterns. ASGR1 or KREMEN1 is sufficient for the entry of SARS-CoV-2 but not SARS-CoV in vitro and in vivo. SARS-CoV-2 utilizes distinct ACE2/ASGR1/KREMEN1 (ASK) receptor combinations to enter different cell types, and the expression of ASK together displays a markedly stronger correlation with virus susceptibility than that of any individual receptor at both the cell and tissue levels. The cocktail of ASK-related neutralizing antibodies provides the most substantial blockage of SARS-CoV-2 infection in human lung organoids when compared to individual antibodies. Our study revealed an interacting host receptome of SARS-CoV-2, and identified ASGR1 and KREMEN1 as alternative functional receptors that play essential roles in ACE2-independent virus entry, providing insight into SARS-CoV-2 tropism and pathogenesis, as well as a community resource and potential therapeutic strategies for further COVID-19 investigations.