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Abstract Introduction Artificial cervical disc replacement (ACDR) is a widely accepted surgical procedure in the treatment of cervical radiculopathy and myelopathy. However, some research suggests that ACDR may redistribute more load onto the facet joints, potentially leading to postoperative axial pain in certain patients. Earlier studies have indicated that facet tropism is prevalent in the lower cervical spine and can significantly increase facet joint pressure. The present study aims to investigate the changes in the biomechanical environment of the cervical spine after ACDR using different prosthese when facet tropism is present. Methods A C2-C7 cervical spine finite element model was created. Symmetrical, moderate asymmetrical (7 degrees tropism), and severe asymmetrical (14 degrees tropism) models were created at the C5/C6 level by adjusting the left-side facet. C5/C6 ACDR with Prestige LP, Prodisc-C vivo, and Mobi-C were simulated in all models. A 75 N follower load and 1 N⋅m moment was applied to initiate flexion, extension, lateral bending, and axial rotation, and the range of motions (ROMs), facet contact forces(FCFs), and facet capsule stress were recorded. Results In the presence of facet tropism, all ACDR models exhibited significantly higher FCFs and facet capsule stress compared to the intact model. In the asymmetric model, FCFs on the right side were significantly increased in neutral position, extension, left bending and right rotation, and on both sides in right bending and left rotation compared to the symmetric model. All ACDR model in the presence of facet tropism, exhibited significantly higher facet capsule stresses at all positions compared to the symmetric model. The stress distribution on the facet surface and the capsule ligament in the asymmetrical models was different from that in the symmetrical model. Conclusions The existence of facet tropism could considerably increase FCFs and facet capsule stress after ACDR with Prestige-LP, Prodisc-C Vivo, and Mobi-C. None of the three different designs of implants were able to effectively protect the facet joints in the presence of facet tropism. Research into designing new implants may be needed to improve this situation. Clinical trials are needed to validate the impact of facet tropism.

Abstract Background Artificial Cervical Disc Replacement (ACDR) is an effective treatment for cervical degenerative disc diseases. However, clinical information regarding the facet joint alterations after ACDR was limited. Facet tropism is common in the sub-axial cervical spine. Our previous research indicated that facet tropism could lead to increased pressure on the cervical facet joints. This study aimed to assess the impact of facet tropism on the facet contact force and facet capsule stress after ACDR. Methods A C2–T1 cervical finite element model was constructed from computed tomography (CT) scans of a 28-year-old male volunteer. Symmetrical, moderate asymmetrical (7 degrees tropism), and severe asymmetrical (14 degrees tropism) models were created at the C5/C6 level by altering the facet orientation at the C5-C6 level. The C5/C6 ACDR was simulated in the intact, moderate asymmetrical and severe asymmetrical models. A 75-N follower load with 1.0-Nm moments was applied to the top of C2 vertebra in the models to simulate flexion, extension, lateral bending, and axial rotation with the T1 vertebra fixed. The range of motions (ROMs) under all moments, facet contact forces (FCFs) and facet capsule strains were tested. Results In the asymmetrical model, the right FCFs considerably increased under flexion, extension, right bending, left rotation, especially under right bending the right sided FCF of the severe asymmetrical model was about 5.44 times of the neutral position, and 3.14 times of the symmetrical model. and concentrated on the cephalad part of the facets. The facet capsule stresses on both sides remarkably increased under extension, lateral bending and right rotation. In the moderate and severe asymmetrical models, the capsule strain was greater on both sides of each position than in the symmetric model. Conclusions The face tropism increased facet contact force and facet capsule strain after ACDR, especially under extension, lateral bending, and rotation, and also could result in abnormal stress distribution on the facet joint surface and facet joint capsule. The results suggest that face tropism might be a risk factor for post-operative facet joint degeneration progression after ACDR. Facet tropism may be noteworthy when ACDR is considered as a surgical option.

Abstract Hemorrhagic fever with renal syndrome (HFRS) occurs throughout Eurasia with considerable morbidity and mortality. Currently, the absence of specific treatments or effective antiviral drugs for hantavirus infection makes developing safe and effective vaccines a high priority. Here, we report the development of three novel nucleic acid vaccine candidates, mRNA, naked DNA, and DNA encapsulated in lipid nanoparticles, encoding the glycoproteins of the Hantaan virus (HTNV). To comprehensively evaluate the potential of candidate HTNV nucleic acid vaccines in preventing HFRS, we focus on evaluating their immunogenicity and efficacy in mice and comparing them with an inactivated vaccine as the benchmark. Our findings reveal that all candidate vaccines activated instant and sustained immune responses, offering comparable in vivo protective efficacy to the inactivated vaccines. Notably, compared to the inactivated vaccine, mRNA vaccine induced stronger virus-specific T-helper 1 cell immune response, while DNA-LNP elicited higher levels of neutralizing antibodies in mice. These results mark a significant step in developing nucleic acid vaccines for HTNV, suggesting that sequential immunization with DNA and mRNA vaccines could further amplify the advantages of nucleic acid vaccines.

Abstract The CRISPR-Cas12a system has revolutionized nucleic acid testing (NAT) with its rapid and precise capabilities, yet it traditionally required RNA pre-amplification. Here we develop rapid fluorescence and lateral flow NAT assays utilizing a split Cas12a system (SCas12a), consisting of a Cas12a enzyme and a split crRNA. The SCas12a assay enables highly sensitive, amplification-free, and multiplexed detection of miRNAs and long RNAs without complex secondary structures. It can differentiate between mature miRNA and its precursor (pre-miRNA), a critical distinction for precise biomarker identification and cancer progression monitoring. The system’s specificity is further highlighted by its ability to detect DNA and miRNA point mutations. Notably, the SCas12a system can quantify the miR-21 biomarker in plasma from cervical cancer patients and, when combined with RPA, detect HPV at attomole levels in clinical samples. Together, our work presents a simple and cost-effective SCas12a-based NAT platform for various diagnostic settings.

Abstract To investigate how cell elongation impacts extracellular electron transfer (EET) of electroactive microorganisms (EAMs), the division of model EAM Shewanella oneidensis (S. oneidensis) MR‐1 is engineered by reducing the formation of cell divisome. Specially, by blocking the translation of division proteins via anti‐sense RNAs or expressing division inhibitors, the cellular length and output power density are all increased. Electrophysiological and transcriptomic results synergistically reveal that the programmed cell elongation reinforces EET by enhancing NADH oxidation, inner‐membrane quinone pool, and abundance of c‐type cytochromes. Moreover, cell elongation enhances hydrophobicity due to decreased cell‐surface polysaccharide, thus facilitates the initial surface adhesion stage during biofilm formation. The output current and power density all increase in positive correction with cellular length. However, inhibition of cell division reduces cell growth, which is then restored by quorum sensing‐based dynamic regulation of cell growth and elongation phases. The QS‐regulated elongated strain thus enables a cell length of 143.6 ± 40.3 µm (72.6‐fold of that of S. oneidensis MR‐1), which results in an output power density of 248.0 ± 10.6 mW m−2 (3.41‐fold of that of S. oneidensis MR‐1) and exhibits superior potential for pollutant treatment. Engineering cellular length paves an innovate avenue for enhancing the EET of EAMs.

Abstract Interfacial electron transfer between electroactive microorganisms (EAMs) and electrodes underlies a wide range of bio‐electrochemical systems with diverse applications. However, the electron transfer rate at the biotic‐electrode interface remains low due to high transmembrane and cell‐electrode interfacial electron transfer resistance. Herein, a modular engineering strategy is adopted to construct a Shewanella oneidensis‐carbon felt biohybrid electrode decorated with bacterial cellulose aerogel‐electropolymerized anthraquinone to boost cell‐electrode interfacial electron transfer. First, a heterologous riboflavin synthesis and secretion pathway is constructed to increase flavin‐mediated transmembrane electron transfer. Second, outer membrane c‐Cyts OmcF is screened and optimized via protein engineering strategy to accelerate contacted‐based transmembrane electron transfer. Third, a S. oneidensis‐carbon felt biohybrid electrode decorated with bacterial cellulose aerogel and electropolymerized anthraquinone is constructed to boost the interfacial electron transfer. As a result, the internal resistance decreased to 42 Ω, 480.8‐fold lower than that of the wild‐type (WT) S. oneidensis MR‐1. The maximum power density reached 4286.6 ± 202.1 mW m−2, 72.8‐fold higher than that of WT. Lastly, the engineered biohybrid electrode exhibited superior abilities for bioelectricity harvest, Cr6+ reduction, and CO2 reduction. This study showed that enhancing transmembrane and cell‐electrode interfacial electron transfer is a promising way to increase the extracellular electron transfer of EAMs.

Abstract Proteins containing a ubiquitin regulatory X (UBX) domain are cofactors of Cell Division Cycle 48 (CDC48) and function in protein quality control. However, whether and how UBX-containing proteins participate in host–microbe interactions remain unclear. Here we show that MoNLE1, an effector from the fungal pathogen Magnaporthe oryzae, is a core virulence factor that suppresses rice immunity by specifically interfering with OsPUX8B.2. The UBX domain of OsPUX8B.2 is required for its binding to OsATG8 and OsCDC48-6 and controls its 26 S proteasome–dependent stability. OsPUX8B.2 and OsCDC48-6 positively regulate plant immunity against blast fungus, while the high-temperature tolerance heat-shock protein OsBHT, a putative cytoplasmic substrate of OsPUX8B.2–OsCDC48-6, negatively regulates defense against blast infection. MoNLE1 promotes the nuclear migration and degradation of OsPUX8B.2 and disturbs its association with OsBHT. Given the high conservation of MoNLE1 among fungal isolates, plants with broad and durable blast resistance might be generated by engineering intracellular proteins resistant to MoNLE1.

Abstract Background Mecapegfilgrastim, a long‐acting granulocyte‐colony stimulating factor has been approved for reducing the incidence of infection, particularly febrile neutropenia (FN), in China. Objective We conducted a multicenter prospective observational study to examine the safety and effectiveness of mecapegfilgrastim in preventing neutropenia in gastrointestinal patients receiving the chemotherapy, including S‐1/capecitabine‐based regimens or the fluorouracil, leucovorin, oxaliplatin, and irinotecan (FOLFOXIRI)/fluorouracil, leucovorin, and oxaliplatin (FOLFOX)/fluorouracil, leucovorin, oxaliplatin, and irinotecan (FOLFIRINOX) regimens. Method Five hundred and sixty‐one gastrointestinal patients from 40 sites across China, between May 2019 and November 2021, were included. The administration of mecapegfilgrastim was prescribed at the discretion of local physicians. Results The most common adverse drug reactions (ADRs) of any grade for all patients was increased white blood cells (2.9%). Grade 3/4 ADRs were observed for anemia (0.2%), decreased white blood cells (0.2%), and decreased neutrophil count (0.2%). Among the 116 patients who received S‐1/capecitabine‐based chemotherapy throughout all cycles, ADRs of any grade included anemia (1.7%), myalgia (0.9%), and increased alanine aminotransferase (0.9%). No grade 3/4 ADRs were observed. In 414 cycles of patients who underwent S‐1/capecitabine‐based regimens, only one (0.2%) cycle experienced grade 4 neutropenia. In the FOLFIRINOX, FOLFOXIRI, and FOLFOX chemotherapy regimens, grade 4 neutropenia occurred in one (2.7%) of 37 cycles, four (4.7%) of 85 cycles, and two (1.2%) of 167 cycles, respectively. Conclusion In a real‐world setting, mecapegfilgrastim has proven effective in preventing severe neutropenia in gastrointestinal patients following chemotherapy. This includes commonly used moderate or high‐risk FN regimens or regimens containing S1/capecitabine, all of which have demonstrated favorable efficacy and safety profiles.

Marburg hemorrhagic fever (MHF) is a fatal infectious disease caused by Marburg virus (MARV) infection, and MARV has been identified as a priority pathogen for vaccine development by the WHO. The glycoprotein (GP) of MARV mediates viral adhesion and invasion of host cells and therefore can be used as an effective target for vaccine development. Moreover, DNA vaccines have unique advantages, such as simple construction processes, low production costs, and few adverse reactions, but their immunogenicity may decrease due to the poor absorption rate of plasmids. Lysosome-associated membrane protein 1 (LAMP1) can direct antigens to lysosomes and endosomes and has great potential for improving the immunogenicity of nucleic acid vaccines. Therefore, we constructed a DNA vaccine based on a codon-optimized MARV GP (ID MF939097.1) fused with LAMP1 and explored the effect of a LAMP targeting strategy on improving the immunogenicity of the MARV DNA vaccine. ELISA, ELISpot, and flow cytometry revealed that the introduction of LAMP1 into the MARV DNA candidate vaccine improved the humoral and cellular immune response, enhanced the secretion of cytokines, and established long-term immune protection. Transcriptome analysis revealed that the LAMP targeting strategy significantly enriched antigen processing and presentation-related pathways, especially the MHC class II-related pathway, in the candidate vaccine. Our study broadens the strategic vision for enhanced DNA vaccine design and provides a promising candidate vaccine for MHF prevention.

Vaccines has long been the focus of antiviral immunotherapy research. Viral epitopes are thought to be useful biomarkers for immunotherapy (both antibody-based and cellular). In this study, we designed a novel vaccine molecule, the Hantaan virus (HTNV) glycoprotein (GP) tandem Th epitope molecule (named the Gnc molecule), in silico. Subsequently, computer analysis was used to conduct a comprehensive and in-depth study of the various properties of the molecule and its effects as a vaccine molecule in the body. The Gnc molecule was designed for DNA vaccines and optimized with a lysosomal-targeting membrane protein (LAMP) strategy. The effects of GP-derived Th epitopes and multiepitope vaccines were initially verified in animals. Our research has resulted in the design of two vaccines based on effective antiviral immune targets. The effectiveness of molecular therapies has also been preliminarily demonstrated in silico and in laboratory animals, which lays a foundation for the application of a vaccines strategy in the field of antivirals.

The maize anthracnose stalk rot and leaf blight diseases caused by the fungal pathogen Colletotrichum graminicola is emerging as an important threat to corn production worldwide. In this work, we provide an improved genome assembly of a C. graminicola strain (TZ-3) by using the PacBio Sequel II and Illumina high-throughput sequencing technologies. The genome of TZ-3 consists of 36 contigs with a length of 59.3 Mb. After correction and evaluation with the Illumina sequencing data and BUSCO, this genome showed a high assembly quality and integrity. Gene annotation of this genome predicted 11,911 protein-coding genes, among which 983 secreted protein-coding genes and 332 effector genes were predicted. Compared with previous genomes of C. graminicola strains, TZ-3 genome is superior in nearly all parameters. The genome assembly and annotation will enhance our knowledge of the genetic makeup of the pathogen and molecular mechanisms underlying its pathogenicity and will provide valuable insights into genome variation across different regions. [Graphic: see text] Copyright © 2023 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Abstract Background The temporomandibular joint (TMJ) is a complex joint consisting of the condyle, the temporal articular surface, and the articular disc. Functions such as mastication, swallowing and articulation are accomplished by the movements of the TMJ. To date, the TMJ has been studied more extensively, but the types of TMJ cells, their differentiation, and their interrelationship during growth and development are still unclear and the study of the TMJ is limited. The aim of this study was to establish a molecular cellular atlas of the human embryonic temporomandibular joint condyle (TMJC) by single-cell RNA sequencing, which will contribute to understanding and solving clinical problems. Results Human embryos at 3 and 4 months of age are an important stage of TMJC development. We performed a comprehensive transcriptome analysis of TMJC tissue from human embryos at 3 and 4 months of age using single-cell RNA sequencing. A total of 16,624 cells were captured and the gene expression profiles of 15 cell clusters in human embryonic TMJC were determined, including 14 known cell types and one previously unknown cell type, "transition state cells (TSCs)". Immunofluorescence assays confirmed that TSCs are not the same cell cluster as mesenchymal stem cells (MSCs). Pseudotime trajectory and RNA velocity analysis revealed that MSCs transformed into TSCs, which further differentiated into osteoblasts, hypertrophic chondrocytes and tenocytes. In addition, chondrocytes (CYTL1high + THBS1high) from secondary cartilage were detected only in 4-month-old human embryonic TMJC. Conclusions Our study provides an atlas of differentiation stages of human embryonic TMJC tissue cells, which will contribute to an in-depth understanding of the pathophysiology of the TMJC tissue repair process and ultimately help to solve clinical problems.

Introduction: Small-scaled robotic walkers play an increasingly important role in Activity of Daily Living (ADL) assistance in the face of ever-increasing rehab requirements and existing equipment drawbacks. This paper proposes a Rehabilitation Robotic Walker (RRW) for walking assistance and body weight support (BWS) during gait rehabilitation.Methods: The walker provides the patients with weight offloading and guiding force to mimic a series of the physiotherapist’s (PT’s) movements, and creates a natural, comfortable, and safe environment. This system consists of an omnidirectional mobile platform, a BWS mechanism, and a pelvic brace to smooth the motions of the pelvis. To recognize the human intentions, four force sensors, two joysticks, and one depth-sensing camera were used to monitor the human-machine information, and a multimodal fusion algorithm for intention recognition was proposed to improve the accuracy. Then the system obtained the heading angle E, the pelvic pose F, and the motion vector H via the camera, the force sensors, and the joysticks respectively, classified the intentions with feature extraction and information fusion, and finally outputted the motor speed control through the robot’s kinematics.Results: To validate the validity of the algorithm above, a preliminary test with three volunteers was conducted to study the motion control. The results showed that the average error of the integral square error (ISE) was 2.90 and the minimum error was 1.96.Discussion: The results demonstrated the efficiency of the proposed method, and that the system is capable of providing walking assistance.

ProblemsPlant Disease diagnosis based on deep learning mechanisms has been extensively studied and applied. However, the complex and dynamic agricultural growth environment results in significant variations in the distribution of state samples, and the lack of sufficient real disease databases weakens the information carried by the samples, posing challenges for accurately training models.AimThis paper aims to test the feasibility and effectiveness of Denoising Diffusion Probabilistic Models (DDPM), Swin Transformer model, and Transfer Learning in diagnosing citrus diseases with a small sample.MethodsTwo training methods are proposed: The Method 1 employs the DDPM to generate synthetic images for data augmentation. The Swin Transformer model is then used for pre-training on the synthetic dataset produced by DDPM, followed by fine-tuning on the original citrus leaf images for disease classification through transfer learning. The Method 2 utilizes the pre-trained Swin Transformer model on the ImageNet dataset and fine-tunes it on the augmented dataset composed of the original and DDPM synthetic images.Results and conclusionThe test results indicate that Method 1 achieved a validation accuracy of 96.3%, while Method 2 achieved a validation accuracy of 99.8%. Both methods effectively addressed the issue of model overfitting when dealing with a small dataset. Additionally, when compared with VGG16, EfficientNet, ShuffleNet, MobileNetV2, and DenseNet121 in citrus disease classification, the experimental results demonstrate the superiority of the proposed methods over existing approaches to a certain extent.

Abstract Traffic data prediction offers a significant way to evaluate the future traffic congestion status; many deep learning based approaches have been widely applied in this field. Most current methods only consider short‐term traffic data forecasting; however, long‐term prediction, which supports the optimized distribution of traffic resources, is not well studied. Besides, multiple traffic parameters enable stronger constraints for the data estimation, but the correlation between them in both spatial and temporal domains has not been efficiently learned. Geometric algebra, as a generalization of linear algebra, provides a framework to encode multidimensional data and analyze the correlation. By combining the advantages of the deep neural network and geometric algebra, a multi‐channel geometric algebra residual network (MGAResNet) is proposed to address the problem of long‐term traffic data prediction. Traffic data obtained from two urban expressways are employed and experimental results demonstrate that the approach outperforms the state‐of‐the‐art work.

Electroactive microorganisms (EAMs) play important roles in biogeochemical redox processes and have been of great interest in the fields of energy recovery, waste treatment, and environmental remediation. However, the currently identified EAMs are difficult to be widely used in complex and diverse environments, due to the existence of poor electron transfer capability, weak environmental adaptability, and difficulty with engineering modifications, etc. Therefore, rapid and efficient screening of high performance EAMs from environments is an effective strategy to facilitate applications of microbial fuel cells (MFCs). In this study, to achieve efficient degradation of methyl orange (MO) by MFC and electricity harvest, a more efficient exoelectrogen Shewanella carassii-D5 that belongs to Shewanella spp. was first isolated from activated sludge by WO3 nanocluster probe technique. Physiological properties experiments confirmed that S. carassii-D5 is a Gram-negative strain with rounded colonies and smooth, slightly reddish surface, which could survive in media containing lactate at 30 °C. Moreover, we found that S. carassii-D5 exhibited remarkable MO degradation ability, which could degrade 66% of MO within 72 h, 1.7 times higher than that of Shewanella oneidensis MR-1. Electrochemical measurements showed that MFCs inoculated with S. carassii-D5 could generate a maximum power density of 704.6 mW/m2, which was 5.6 times higher than that of S. oneidensis MR-1. Further investigation of the extracellular electron transfer (EET) mechanism found that S. carassii-D5 strain had high level of c-type cytochromes and strong biofilm formation ability compared with S. oneidensis MR-1, thus facilitating direct EET. Therefore, to enhance indirect electron transfer and MO degradation capacity, a synthetic gene cluster ribADEHC encoding riboflavin synthesis pathway from Bacillus subtilis was heterologously expressed in S. carassii-D5, increasing riboflavin yield from 1.9 to 9.0 mg/g DCW with 1286.3 mW/m2 power density output in lactate fed-MFCs. Furthermore, results showed that the high EET rate endowed a faster degradation efficient of MO from 66% to 86% with a maximum power density of 192.3 mW/m2, which was 1.3 and 1.6 times higher than that of S. carassii-D5, respectively. Our research suggests that screening and engineering high-efficient EAMs from sludge is a feasible strategy in treating organic pollutants.

(1) Background and Purpose: Ebola virus (EBOV) is the causative agent of Ebola virus disease (EVD), which causes extremely high mortality and widespread epidemics. The only glycoprotein (GP) on the surface of EBOV particles is the key to mediating viral invasion into host cells. DNA vaccines for EBOV are in development, but their effectiveness is unclear. The lack of immune characteristics resides in antigenic MHC class II reactivity. (2) Methods: We selected MHC-II molecules from four human leukocyte antigen II (HLA-II) superfamilies with 98% population coverage and eight mouse H2-I alleles. IEDB, NetMHCIIpan, SYFPEITHI, and Rankpep were used to screen MHC-II-restricted epitopes with high affinity for EBOV GP. Further immunogenicity and conservation analyses were performed using VaxiJen and BLASTp, respectively. EpiDock was used to simulate molecular docking. Cluster analysis and binding affinity analysis of EBOV GP epitopes and selected MHC-II molecules were performed using data from NetMHCIIpan. The selective GP epitopes were verified by the enzyme-linked immunospot (ELISpot) assay using splenocytes of BALB/c (H2d), C3H, and C57 mice after DNA vaccine pVAX-GPEBO immunization. Subsequently, BALB/c mice were immunized with Protein-GPEBO, plasmid pVAX-GPEBO, and pVAX-LAMP/GPEBO, which encoded EBOV GP. The dominant epitopes of BALB/c (H-2-I-AdEd genotype) mice were verified by the enzyme-linked immunospot (ELISpot) assay. It is also used to evaluate and explore the advantages of pVAX-LAMP/GPEBO and the reasons behind them. (3) Results: Thirty-one HLA-II-restricted and 68 H2-I-restricted selective epitopes were confirmed to have high affinity, immunogenicity, and conservation. Nineteen selective epitopes have cross-species reactivity with good performance in MHC-II molecular docking. The ELISpot results showed that pVAX-GPEBO could induce a cellular immune response to the synthesized selective peptides. The better immunoprotection of the DNA vaccines pVAX-LAMP/GPEBO coincides with the enhancement of the MHC class II response. (4) Conclusions: Promising MHC-II-restricted candidate epitopes of EBOV GP were identified in humans and mice, which is of great significance for the development and evaluation of Ebola vaccines.

Electroactive biofilm plays a crucial rule in the electron transfer efficiency of microbial electrochemical systems (MES). However, the low ability to form biofilm and the low conductivity of the formed biofilm substantially limit the extracellular electron transfer rate of microbial cells to the electrode surfaces in MES. To promote biofilm formation and enhance biofilm conductivity, we develop synthetic biology approach to systematically engineer Shewanella oneidensis, a model exoelectrogen, via modular manipulation of the full-cycle different stages of biofilm formation, namely, from initial contact, cell adhesion, and biofilm growth stable maturity to cell dispersion. Consequently, the maximum output power density of the engineered biofilm reaches 3.62 ± 0.06 W m−2, 39.3-fold higher than that of the wild-type strain of S. oneidensis, which, to the best our knowledge, is the highest output power density that has ever been reported for the biofilms of the genetically engineered Shewanella strains.

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 β-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.

BackgroundThe purpose of this study was to evaluate the efficacy and safety of BAL in treating MPP.MethodsFrom January 2013 to January 2019, 1,689 pediatric patients with MPP were analyzed retrospectively. Patients were subdivided into BAL group and non-BAL group according to whether they received BAL treatment within seven days after admission. The propensity score matching method matched patients' baseline characteristics (1:1). The primary outcomes were hospital stays and the cure rate. Secondary outcomes included mortality, co-infection, repeat hospitalization within 30 days, and total cost of treatment.ResultsAfter matching, 524 patients (BAL: 262; control: 262) were recorded. The BAL group had significantly shorter hospital stays (OR: 0.5, 95% CI: 0.4–0.7). Meanwhile, BAL did not significantly modify the cost, co-infection rate, and mortality. In subgroup analyses, the group with BAL intervention within three days had a significantly shorter hospital stay (OR: 0.4, 95% CI: 0.3–0.5) compared with the group with BAL intervention three days after admission.ConclusionsEarly BAL intervention is a better treatment than conventional drug therapy alone, and no significant complications were seen in this study. BAL intervention has an excellent clinical benefit. The earlier the intervention, the better the effect.

Abstract Background The crosstalk between the ubiquitin‐proteasome and the immune system plays an important role in the health and pathogenesis of viral infection. However, there have been few studies of ubiquitin activation in severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) infection. Methods We investigated the effect of ubiquitination on SARS‐CoV‐2 infection and patient prognosis by integrating published coronavirus disease 2019 (COVID‐19) multi‐transcriptome data and bioinformatics methods. Results The differential expression of COVID‐19 samples revealed changed ubiquitination in most solid and hollow organs, and it was activated in lymphatic and other immune tissues. In addition, in the respiratory system of COVID‐19 patients, the immune response was mainly focused on the alveoli, and the expression of ubiquitination reflected increasing immune infiltration. Ubiquitination stratification could significantly differentiate patients' prognosis and inflammation levels through the general transcriptional analysis of the peripheral blood of patients with COVID‐19. Moreover, high ubiquitination levels were associated with a favourable prognosis, low inflammatory response, and reduced mechanical ventilation and intensive care unit. Moreover, high ubiquitination promoted a beneficial immune response while inhibiting immune damage. Finally, prognostic stratification and biomarker screening based on ubiquitination traits played an important role in clinical management and drug development. Conclusion Ubiquitination characteristics provides new ideas for clinical intervention and prognostic guidance for COVID‐19 patients.

This study explored the safety of COVID-19 vaccine (Aikewei) and the role of clinical pharmacists in the implementation of COVID-19 vaccination. A total of 2305 hospital employees in Children’s Hospital of Fudan University in Shanghai, China received the COVID-19 vaccine. The whole process of vaccination was monitored by clinical pharmacists, and the occurrence, types, severity of adverse reactions were recorded in detail. Through the investigation and analysis on the safety of COVID-19 vaccination of the 2305 people, the important role and value of clinical pharmacists in the vaccination process was elaborated. Common adverse reactions included local pain, dizziness and fatigue, with the incidence rates of 2.09%, 0.67% and 0.49%, respectively. Others such as headache, nausea, skin itching, cough, palpitation, dry mouth, hand anesthesia, local induration, muscle soreness, local rash, and chill had incidence rates of less than 0.30%. Three cases of serious adverse events that occurred in this vaccination returned to normal after treatment, with no subsequent discomfort. Clinical pharmacists played an important role in the safety monitoring of COVID-19 vaccination. The safety of the inactivated COVID-19 vaccine is good. Most of the common adverse reactions were mild and tolerable, with generally low incidence. The work of clinical pharmacists is important and can be expanded in the future to ensure the safety of vaccination and to provide better health care service.

This paper aims to disclose how the induction coil parameters affects the temperature and flow fields of the inductively coupled plasma (ICP) torch. For this purpose, a numerical model was established for the balanced discharge of the ICP torch based on COMSOL Multiphysics, and used to simulate the effects of coil spacing and induction wire diameter on the distribution of temperature and flow fields. The results show that a ring-shaped high-temperature area will form inside the ICP torch, regardless of the values of coil parameters; the smaller the coil spacing, the faster for the working medium in the torch to generate the high-temperature, high-velocity area; the smaller the induction wire diameter, the slower for the working medium in the torch to generate the high-temperature, high-velocity area. The research results provide a reference for the optimization of the ICP torch.

Electroactive bacteria could perform bi-directional extracellular electron transfer (EET) to exchange electrons and energy with extracellular environments, thus playing a central role in microbial electro-fermentation (EF) process. Unbalanced fermentation and microbial electrosynthesis are the main pathways to produce value-added chemicals and biofuels. However, the low efficiency of the bi-directional EET is a dominating bottleneck in these processes. In this review, we firstly demonstrate the main bi-directional EET mechanisms during EF, including the direct EET and the shuttle-mediated EET. Then, we review representative milestones and progresses in unbalanced fermentation via anode outward EET and microbial electrosynthesis via inward EET based on these two EET mechanisms in detail. Furthermore, we summarize the main synthetic biology strategies in improving the bi-directional EET and target products synthesis, thus to enhance the efficiencies in unbalanced fermentation and microbial electrosynthesis. Lastly, a perspective on the applications of microbial electro-fermentation is provided.

Aquaporin-9 (AQP9) is an aquaglyceroporin strongly expressed in the basolateral membrane of hepatocytes facing the sinusoids. AQP9 is permeable to hydrogen peroxide (H2O2) and glycerol as well as to water. Here, we report impaired liver regeneration in AQP9−/− mice which involves altered steady-state H2O2 concentration and glucose metabolism in hepatocytes. AQP9−/− mice showed remarkably delayed liver regeneration and increased mortality following 70% or 90% partial hepatectomy. Compared to AQP9+/+ littermates, AQP9−/− mice showed significantly greater hepatic H2O2 concentration and more severe liver injury. Fluorescence measurements indicated impaired H2O2 transport across plasma membrane of primary cultured hepatocytes from AQP9−/− mice, supporting the hypothesis that AQP9 deficiency results in H2O2 accumulation and oxidative injury in regenerating liver because of reduced export of intracellular H2O2 from hepatocytes. The H2O2 overload in AQP9−/− hepatocytes reduced PI3K-Akt and insulin signaling, inhibited autophagy and promoted apoptosis, resulting in impaired proliferation and increased cell death. In addition, hepatocytes from AQP9−/− mice had low liver glycerol and high blood glycerol levels, suggesting decreased glycerol uptake and gluconeogenesis in AQP9−/− hepatocytes. Adeno-associated virus (AAV)-mediated expression of hepatic expression of aquaglyceroporins AQP9 and AQP3 in AQP9−/− mice, but not water-selective channel AQP4, fully rescued the impaired liver regeneration phenotype as well as the oxidative injury and abnormal glucose metabolism. Our data revealed a pivotal role of AQP9 in liver regeneration by regulating hepatocyte H2O2 homeostasis and glucose metabolism, suggesting AQP9 as a novel target to enhance liver regeneration following injury, surgical resection or transplantation.

Background: Microbial fuel cells (MFCs) are a novel bioelectrochemical devices that can use exoelectrogens as biocatalyst to convert various organic wastes into electricity. Among them, acetate, a major component of industrial biological wastewater and by-product of lignocellulose degradation, could release eight electrons per mole when completely degraded into CO2 and H2O, which has been identified as a promising carbon source and electron donor. However, Shewanella oneidensis MR-1, a famous facultative anaerobic exoelectrogens, only preferentially uses lactate as carbon source and electron donor and could hardly metabolize acetate in MFCs, which greatly limited Coulombic efficiency of MFCs and the capacity of bio-catalysis.Results: Here, to enable acetate as the sole carbon source and electron donor for electricity production in S. oneidensis, we successfully constructed three engineered S. oneidensis (named AceU1, AceU2, and AceU3) by assembling the succinyl-CoA:acetate CoA-transferase (SCACT) metabolism pathways, including acetate coenzyme A transferase encoded by ato1 and ato2 gene from G. sulfurreducens and citrate synthase encoded by the gltA gene from S. oneidensis, which could successfully utilize acetate as carbon source under anaerobic and aerobic conditions. Then, biochemical characterizations showed the engineered strain AceU3 generated a maximum power density of 8.3 ± 1.2 mW/m2 with acetate as the sole electron donor in MFCs. In addition, when further using lactate as the electron donor, the maximum power density obtained by AceU3 was 51.1 ± 3.1 mW/m2, which approximately 2.4-fold higher than that of wild type (WT). Besides, the Coulombic efficiency of AceU3 strain could reach 12.4% increased by 2.0-fold compared that of WT, which demonstrated that the engineered strain AceU3 can further utilize acetate as an electron donor to continuously generate electricity.Conclusion: In the present study, we first rationally designed S. oneidensis for enhancing the electron generation by using acetate as sole carbon source and electron donor. Based on synthetic biology strategies, modular assembly of acetate metabolic pathways could be further extended to other exoelectrogens to improve the Coulombic efficiency and broaden the spectrum of available carbon sources in MFCs for bioelectricity production.

One-third of patients with epilepsy suffer from drug-resistant epilepsy (DRE). Valproic acid (VPA) is a classic anticonvulsant drug, and its resistance is a crucial predictor of DRE, but the pathogenesis remain unknown. Most patients with VPA-resistant epilepsy appear distinct inflammatory response and local hypoxia. Hypoxia-inducible factor (HIF)-1α is an essential effector molecule of hypoxia and inflammation, and may exert therefore a significant effect on the development of VPA-resistant epilepsy. We systematically assess the significance of HIF-1α on children and mice with VPA-resistant epilepsy, and investigated the micro (mi) RNAs that regulate HIF-1α expression. We established models of VPA-sensitive epilepsy and VPA-resistant epilepsy in mice, and confirmed that they had significant differences in epileptic behavior and electroencephalography data. Through proteomics analysis, we identified that HIF-1α was overexpressed in mice with VPA-resistant epilepsy, and regulated the expression of interleukin-1β and tumor necrosis factor-α. Increased expression of HIF-1α led to the increase of microglia and induced their polarization from the M2 phenotype to M1 phenotype, which triggered the release of proinflammatory mediators. Bioinformatics analysis of public databases demonstrated that miR-221-3p was reduced in VPA-resistant epilepsy, and negatively regulated HIF-1α expression. Intervention using miR-221-3p mimics reduced HIF-1α expression markedly and suppressed the activation of microglia and the release of inflammatory mediators, which relieved epileptic seizures of VPA-resistant epilepsy. These observations reveal miR-221-3p/HIF-1α as essential component in pathogenesis of VPA-resistant epilepsy which represent therapeutic antiseizure targets.

Virus inactivator can inactivate cell-free virions without relying on their replication cycle, potentially reducing the impact of viral infection on cells. Previously, we successfully constructed a HIV-1 protein inactivator, 2DLT, by conjugating the D1D2 region of CD4 to the fusion inhibitor T1144 via a 35-amino acid linker. Therefore, it targets both the CD4 binding site in gp120 and NHR region in gp41. Considering that small-molecule agents have the advantages of fast production, low cost, good stability, and oral availability, we herein report the design of a new small-molecule HIV-1 inactivator, FD028, by conjugating FD016 (an analog of NBD-556, a gp120-CD4 binding inhibitor) with FD017 (an analog of 11d, an HIV-1 fusion inhibitor). The results showed that FD028 inactivated cell-free virions at a moderate nanomolar concentration by targeting both HIV-1 gp120 and gp41. Moreover, FD028 has broad-spectrum inhibition and inactivation activity against HIV-1 resistant strains and primary isolates of different subtypes without significant cytotoxicity. Therefore, FD028 has potential for further development as an HIV-1 inactivator-based therapeutic.

Clarifying the maize kernel movement during the crushing process is critical for improving the design and optimization of the impact mill. Rather than through experiments, maize kernel movement can be quantitatively analyzed through the discrete element method (DEM), and this could contribute more to the study of the crushing mechanism and equipment optimization. However, having an accurate particle model and contact parameters are prerequisites to ensure the accuracy of the DEM simulation. In this study, we proposed a maize kernel model construction method for the Rocky DEM simulation and a calibration method to calibrate contact parameters. The three-axis size, volume, and shape information of real maize kernels were obtained by 3D scanning, and then the maize kernel model was constructed by the section method. The particle–low-carbon-plate (p–w) and particle–particle (p–p) restitution coefficients were calibrated by using the improved inclined surface drop method. In addition, the angle of repose (AoR) and discharging time were considered together to calibrate the dynamical friction coefficients of p–w and p–p through the funnel method. Additionally, the maize kernel model and calibrated parameter values were used in a DEM simulation of the inclined surface drop test and the funnel test. The maximum relative errors between the simulation values and the measured values of the inclined surface drop test and the funnel test were 4.38% and 6.98%, respectively, which further verified that the proposed maize kernel model construction and contact parameter calibration methods are feasible and accurate. The research method used in this study is a novel idea that can be applied for the construction of the particle model and calibration of the contact parameters of granular materials with complex geometric structures, as well as the maize kernel model, and shows that calibrated contact parameters can provide a reference for the maize kernel crushing simulation to optimize the impact mill.

MHC-I antigen processes and presentation trigger host-specific anti-viral cellular responses during infection, in which epitope-recognizing cytotoxic T lymphocytes eliminate infected cells and contribute to viral clearance through a cytolytic killing effect. In this study, Hantaan virus (HTNV) GP-derived 9-mer dominant epitopes were obtained with high affinity to major HLA-I and H-2 superfamilies. Further immunogenicity and conservation analyses selected 11 promising candidates, and molecule docking (MD) was then simulated with the corresponding MHC-I alleles. Two-way hierarchical clustering revealed the interactions between GP peptides and MHC-I haplotypes. Briefly, epitope hotspots sharing good affinity to a wide spectrum of MHC-I molecules highlighted the biomedical practice for vaccination, and haplotype clusters represented the similarities among individuals during T-cell response establishment. Cross-validation proved the patterns observed through both MD simulation and public data integration. Lastly, 148 HTNV variants yielded six types of major amino acid residue replacements involving four in nine hotspots, which minimally influenced the general potential of MHC-I superfamily presentation. Altogether, our work comprehensively evaluates the pan-MHC-I immunoreactivity of HTNV GP through a state-of-the-art workflow in light of comparative immunology, acknowledges present discoveries, and offers guidance for ongoing HTNV vaccine pursuit.

Traffic-data recovery plays an important role in traffic prediction, congestion judgment, road network planning and other fields. Complete and accurate traffic data help to find the laws contained in the data more efficiently and effectively. However, existing methods still have problems to cope with the case when large amounts of traffic data are missed. As a generalization of vector algebra, geometric algebra has more powerful representation and processing capability for high-dimensional data. In this article, we are thus inspired to propose the geometric-algebra-based generative adversarial network to repair the missing traffic data by learning the correlation of multidimensional traffic parameters. The generator of the proposed model consists of a geometric algebra convolution module, an attention module and a deconvolution module. Global and local data mean squared errors are simultaneously applied to form the loss function of the generator. The discriminator is composed of a multichannel convolutional neural network which can continuously optimize the adversarial training process. Real traffic data from two elevated highways are used for experimental verification. Experimental results demonstrate that our method can effectively repair missing traffic data in a robust way and has better performance when compared with the state-of-the-art methods.

Antibiotic resistance is a serious global health concern that may have significant social and financial consequences. Methicillin-resistant Staphylococcus aureus (MRSA) infection is responsible for substantial morbidity and leads to the death of 21.8% of infected patients annually. A lack of novel antibiotics has prompted the exploration of therapies targeting bacterial virulence mechanisms. The two-component signal transduction system (TCS) enables microbial cells to regulate gene expression and the subsequent metabolic processes that occur due to environmental changes. The YycFG TCS in S. aureus is essential for bacterial viability, the regulation of cell membrane metabolism, cell wall synthesis and biofilm formation. However, the role of YycFG-associated biofilm organization in S. aureus antimicrobial drug resistance and gene regulation has not been discussed in detail. We reviewed the main molecules involved in YycFG-associated cell wall biosynthesis, biofilm development and polysaccharide intercellular adhesin (PIA) accumulation. Two YycFG-associated regulatory mechanisms, accessory gene regulator (agr) and staphylococcal accessory regulator (SarA), were also discussed. We highlighted the importance of biofilm formation in the development of antimicrobial drug resistance in S. aureus infections. Data revealed that inhibition of the YycFG pathway reduced PIA production, biofilm formation and bacterial pathogenicity, which provides a potential target for the management of MRSA-induced infections.

Hepatitis B virus (HBV) establishes chronic infection in only some patients, but the mechanisms underlying clearance failure in these patients are not fully understood. Here, the authors identify and characterize an HBV strain that can persist in mice and show that IL-21 and IL-33 responses contribute to clearance.

Primary duck hepatocytes (PDH) displays differential susceptibility to duck hepatitis B virus when maintained in the media supplemented with fetal bovine serum or dimethyl sulfoxide (DMSO) which has been widely used for the maintenance of hepatocytes, and prolonging susceptibility to hepadnavirus. However the mechanism underlying maintenance of susceptibility to hepadnavirus by DMSO treatment remains unclear. In this study, a global transcriptome analysis of PDHs under different culture conditions was conducted for investigating the effects of DMSO on maintenance of susceptibility of PDH to DHBV in vitro. The 384 differential expressed genes (DEGs) were identified by comparisons between each library pair (PDHs cultured with or without DMSO or fresh isolated PDH). We analyzed canonical pathways in which the DEGs were enriched in Hepatic Fibrosis / Hepatic Stellate Cell Activation, Bile Acid Biosynthesis and Tight Junction signaling. After re-annotation against human genome data, the 384 DEGs were pooled together with proteins belonging to hepatitis B pathway to construct a protein-protein interaction network. The combination of decreased expression of liver-specific genes (CYP3A4, CYP1E1, CFI, RELN and GSTA1 et al) with increased expression of hepatocyte-dedifferentiation-associated genes (PLA2G4A and PLCG1) suggested that in vitro culture conditions results in the fading of hepatocyte phenotype in PDHs. The expression of seven DEGs associated with tight junction formation (JAM3, PPP2R2B, PRKAR1B, PPP2R2C, MAGI2, ACTA2 and ACTG2) was up-regulated after short-term culture in vitro, which was attenuated in the presence of DMSO. Those results could shed light on DHBV infection associated molecular events affected by DMSO.

Prince Rupprecht's larch (Larix principis-rupprechtii Mayr) is a native high-value forest tree species in North China whose clonal propagation through somatic embryogenesis (SE) has the potential to rapidly capture the benefits of breeding or genetic engineering programs and to improve raw material uniformity and quality. To date, research has focused on clarifying the molecular mechanism of SE, but proteomic studies are still in the early stages. In this study, isobaric tags for relative and absolute quantitation (iTRAQ) analysis was performed on three developmental stages of SE in L. principis-rupprechtii in an attempt to identify a wide range of proteins that are regulated differentially during this process. Proteins were extracted and analyzed from the pro-embryogenic mass (PEM), globular embryo (GE), and cotyledon embryo (CE) stages of embryo development. We detected 503 proteins in total and identified 96 proteins expressed differentially during different developmental stages. The identified proteins were analyzed further to provide information about their expression patterns and functions during SE. Four clusters of proteins based on shared expression profiles were generated. Functional analysis showed that proteins involved in primary metabolism, phosphorylation, and oxidation reduction were upregulated during somatic embryo development. This work provides novel insights into the process of larch embryo development in vitro and a basis for further study of the biological process and opportunities for practical application of this knowledge.

Hepatitis B virus (HBV) takes humans as its sole natural host, and productive infection in vivo is restricted exclusively to hepatocytes in the liver. Consequently, HBV-derived viral vectors are attractive candidates for liver-targeting gene therapies. Previously, we developed a novel recombinant HBV vector, designated 5c3c, from a highly replicative clinical isolate. 5c3c was demonstrated to be capable of efficiently delivering protein or RNA expression into infected primary tupaia hepatocytes (PTH), but the design of 5c3c imposes stringent restrictions on inserted sequences, which have limited its wider adoption. In this work, we addressed issues with 5c3c by re-designing the insertion strategy. The resultant vector, designated 5dCG, was more replicative than parental 5c3c, imposed no specific restrictions on inserted sequences, and allowed insertion of a variety of cargo genes without significant loss of replication efficiency. 5dCG-based recombinant HBV effectively delivered protein and RNA expression into infected PTH. Furthermore, due to the loss of functional core ORF, 5dCG vectors depend on co-infecting wild type HBV for replication and efficient expression of cargo genes. Development of the improved 5dCG vector makes wider applications of recombinant HBV possible, while dependence on co-infecting wild type HBV results in improved safety for certain in vivo applications.