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A capsular polysaccharide, namely CPS-2, was isolated from Lactobacillus fermentum GBJ, purified using DEAE-52 anion exchange chromatography, and structurally characterized. We found that CPS-2 is homogenous, has an average molecular weight of 377 KDa, and is mainly composed of galactose and glucose at a molar ratio of 1.54:1.00. Its backbone comprises α-D-Galp-(1 → 3), α-D-Galp-(1 → 3,6), β-D-Glcp-(1 → 2), β-D-Galp-(1 → 6), and α-D-Galp-(1 → 4) residues with a side chain of β-D-Glcp-(1→). CPS-2 exerts an immunomodulatory effect by improving the proliferation and phagocytosis of macrophage RAW264.7 and promoting the secretion of NO and cytokines. The maximum secretion levels of IL-1β, IL-6, IL-10, and TNF-α were 1.96-, 0.11-, 0.22-, and 0.46-fold higher than those of the control, respectively. Furthermore, CPS-2 could significantly enhance the antioxidant system, extend lifespan, and improve stress tolerance of Caenorhabditis elegans at both exposure doses of 31.25 and 62.5 μg/mL. The average lifespan of nematodes reached a maximum in the 62.5 μg/mL-treated group after 10.39 days, 6.56 h, and 23.56 h in normal, oxidative stress, and heat shock environment, with extension percentages of 16.61 %, 43.23 %, and 15.77 %, respectively; therefore, CPS-2 displays an anti-aging effect. The significant bioactivity of CPS-2 promotes its application as a promising immunomodulatory and anti-aging ingredient in the food or pharmaceutical field., Competing Interests: Declaration of competing interest None., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Pectin, a kind of dietary fiber, has attracted much attention owing to its beneficial effect on human health in recent years. In this study, the effects of both 'Ganzhou' navel orange peel pectin (GOP) and modified GOP (MGOP) on type 2 diabetes (T2DM) were investigated. The results indicated that GOP and MGOP intervention had positive effects on T2DM in C57BL/6 mice. After modification, pectin can be changed into low methoxy pectin (LMP) and the content of GalA can increase, which endow MGOP with significant effects on improving lipid metabolism (TC, TG, and LDL-C decreased by 30.46%, 50%, and 37.56%, respectively, and HDL-C increased by 56%) and OGTT, further reducing insulin resistance (insulin decreased by 74.35%). In addition, MGOP was superior to GOP in improving oxidative stress (GSH and GSH-Px increased by 52.05% and 29.08% respectively, and MDA decreased by 84.02%), inhibiting inflammation and promoting SCFA synthesis. 16S rRNA analysis showed that MGOP changed the composition of intestinal microbiota in diabetic mice, decreased the abundance of Alistipes , Helicobacter and Oscillibacter , and increased the relative abundance of Dubosiella , Akkermansiaceae , and Atopobiaceae . The phenotypes of the gut microbiome also changed accordingly, which showed that MGOP significantly inhibited the growth of Gram-negative bacteria and potential pathogenic bacteria and reversed the related complications. Taken together, our findings revealed that MGOP intake regulated lipid metabolism and oxidative stress and improved the gut health of mice, with promising effects against T2DM and related complications.

Objective: To develop an automated landmark location system applicable to the case of landmark missing. Methods: Four and eighty-one lateral cephalograms, which contained 240 males and 241 females, with an average age of (24.5±5.6) years, taken from January 2015 to January 2021 in the Department of Orthodontics, Capital Medical University School of Stomatology, and met the inclusion criteria were collected. Five postgraduate orthodontic students were the annotators to manually locate 61 possible landmarks in 481 lateral cephalograms. Two assistant professors in the department as reviewers performed calibration. Two professors as arbitrators, made final decision. Data sets were established (341 were used as training set, 40 as validation set, and 100 as test set). In this paper, an automatic landmarks identification and location model based on convolutional neural networks (CNN), CephaNET, was developed. The model was trained by feeding the original image into the feature extraction module and convolutional pose machine (CPM) module to locate landmarks with high accuracy using deep supervision. Training set was enhanced to 1 684 images by histogram equalization, cropping, and adjustment of brightness. The model was trained to compare the Gaussian heat maps output from the network with the set threshold to identify landmark missing cases. Test set of 100 lateral cephalograms was used to test the accuracy of the model. The evaluation criteria used were success detection rate of missing landmark, mean radial error (MRE) and success detection rate (SDR) in the range of 2.0, 2.5, 3.0, 3.5 and 4.0 mm. Results: The model identified and located 61 commonly used landmarks in 0.13 seconds on average. It had an average accuracy of 93.5% in identifying missing landmarks. The MRE of our testing set was (1.19±0.91) mm. SDR of 2.0, 2.5, 3.0, 3.5 and 4.0 mm were 85.4%, 90.2%, 93.5%, 95.4%, 97.0% respectively. Conclusions: The model proposed in this paper could adapt to the absence of landmark in lateral cephalograms and locate 61 commonly used landmarks with high accuracy to meet the requirements of different cephalometric analysis methods.

α-Carbonyl cations are the umpolung forms of the synthetically fundamental α-carbonyl carbanions. They are highly reactive yet rarely studied and utilized species and their precursors were rather limited. Herein, we report the catalyst-controlled divergent generations of α-carbonyl cations from single alkyne functionalities and the interception of them via Wagner-Meerwein rearrangement. Two chemodivergent catalytic systems have been established, leading to two different types of α-carbonyl cations and, eventually, two different types of products, i.e. the α,β- and β,γ-unsaturated carbonyl compounds. Broad spectrum of alkynes including aryl alkyne, ynamide, alkynyl ether, and alkynyl sulfide could be utilized and the migration priorities of different groups in the Wagner-Meerwein rearrangement step was elucidated. Density functional theory calculations further supported the intermediacy of α-carbonyl cations via the N-O bond cleavage in both the two catalytic systems. Another key feature of this methodology was the fragmentation of synthetically inert tert-butyl groups into readily transformable olefin functionalities. The synthetic potential was highlighted by the scale-up reactions and the downstream diversifications including the formal synthesis of nicotlactone B and galbacin., (© 2023 Wiley-VCH GmbH.)

Interfacial catalysis occurs ubiquitously in electrochemical systems, such as batteries, fuel cells, and photocatalytic devices. Frequently, in such a system, the electrode material evolves dynamically at different operating voltages, and this electrochemically driven transformation usually dictates the catalytic reactivity of the material and ultimately the electrochemical performance of the device. Despite the importance of the process, comprehension of the underlying structural and compositional evolutions of the electrode material with direct visualization and quantification is still a significant challenge. In this work, we demonstrate a protocol for studying the dynamic evolution of the electrode material under electrochemical processes by integrating microscopic and spectroscopic analyses, operando magnetometry techniques, and density functional theory calculations. The presented methodology provides a real-time picture of the chemical, physical, and electronic structures of the material and its link to the electrochemical performance. Using Co(OH) 2 as a prototype battery electrode and by monitoring the Co metal center under different applied voltages, we show that before a well-known catalytic reaction proceeds, an interfacial storage process occurs at the metallic Co nanoparticles/LiOH interface due to injection of spin-polarized electrons. Subsequently, the metallic Co nanoparticles act as catalytic activation centers and promote LiOH decomposition by transferring these interfacially residing electrons. Most intriguingly, at the LiOH decomposition potential, electronic structure of the metallic Co nanoparticles involving spin-polarized electrons transfer has been shown to exhibit a dynamic variation. This work illustrates a viable approach to access key information inside interfacial catalytic processes and provides useful insights in controlling complex interfaces for wide-ranging electrochemical systems., Competing Interests: Competing interests statement:The authors declare no competing interest.

Glass microlens arrays (MLAs) have tremendous prospects in the fields of optical communication, sensing and high-sensitivity imaging for their excellent optical properties, high mechanical robustness and physicochemical stability. So far, glass MLAs are primarily fabricated using femtosecond laser modification assisted etching, in which the preparation procedure is time-consuming, with each concave-shaped microlens being processed using a femtosecond laser point by point. In this paper, a new method is proposed for implementing large-scale glass MLAs using glass particle sintering with the assistance of ultraviolet (UV) lithography. The glass particles are dispersed into the photoresist at first, and then immobilized as large-scaled micropillar arrays on quartz glass substrate using UV lithographing. Subsequently, the solidified photoresist is debinded and the glass particles are melted by means of sintering. By controlling the sintering conditions, the convex microlens will be self-assembled, attributed to the surface tension of the molten glass particles. Finally, MLAs with different focal lengths (0.12 to 0.2 mm) are successfully fabricated by utilizing different lithography masks. Meanwhile, we also present the optimization of the sintering parameter for eliminating the bubbles in the microlenses. The main factors that affect the focal length of the microlens and the image performance of the MLAs have been studied in detail.

To address the issue of floating and loss of floc sludge caused by gas production in anaerobic ammonia oxidation (anammox) reactors, this study proposes a limited nitrite supply strategy to regulate gas production during the settling and enhance sludge retention. Results indicate that the effluent suspended solids in the anammox reactor can be reduced to as low as 0.11 g/L under specific feast-starvation conditions. Even under long-term intermittent nitrite-starvation stress, the maximum growth rate of Candidatus_Kuenenia can still reach 0.085d -1 , with its abundance increasing from 0.47% to 8.83% within 69 days. Although the combined effects of starvation and sedimentation would lead to a temporary decrease in anammox activity, this reversible inhibition can be fully restored through substrate intervention. The limited nitrite supply strategy promotes the sedimentation of anammox sludge without significantly affecting its growth rate, and effective sludge retention is crucial for enriching anammox sludge during initial cultivation., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

We report the application of ancestral sequence reconstruction on coronavirus spike protein, resulting in stable and highly soluble ancestral scaffold antigens (AnSAs). The AnSAs interact with plasma of patients recovered from COVID-19 but do not bind to the human angiotensin-converting enzyme 2 (ACE2) receptor. Cryo-EM analysis of the AnSAs yield high resolution structures (2.6-2.8 Å) indicating a closed pre-fusion conformation in which all three receptor-binding domains (RBDs) are facing downwards. The structures reveal an intricate hydrogen-bonding network mediated by well-resolved loops, both within and across monomers, tethering the N-terminal domain and RBD together. We show that AnSA-5 can induce and boost a broad-spectrum immune response against the wild-type RBD as well as circulating variants of concern in an immune organoid model derived from tonsils. Finally, we highlight how AnSAs are potent scaffolds by replacing the ancestral RBD with the wild-type sequence, which restores ACE2 binding and increases the interaction with convalescent plasma., (© 2023. Springer Nature Limited.)

Background: Sepsis is a common severe complication in major burn victims and is characterized by a dysregulated systemic response to inflammation. YTH domain family 2 (YTHDF2), a well-studied N6-methyladenosine (m6A) reader that specifically recognizes and binds to m6A-modified transcripts to mediate their degradation, is connected to pathogenic and physiological processes in eukaryotes, but its effect on sepsis is still unknown. We aimed to discover the effects and mechanisms of YTHDF2 in sepsis., Methods: Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and western blot analyses were used to measure the expression of YTHDF2, the interleukin 6 receptor (IL-6R), high-mobility group box-1 (HMGB1), Janus kinase 2 (JAK2) and signal transducer and activator of transcription 1 (STAT1) under different in vitro conditions. Enzyme-linked immunosorbent assays were utilized to evaluate the expression of HMGB1, IL-6, IL-1β and tumor necrosis factor-α. To confirm that YTHDF2 specifically targets IL-6R mRNA, RNA immunoprecipitation and dual-luciferase reporter assays were performed. Finally, we utilized a mouse model of lipopolysaccharide (LPS)-induced sepsis to verify the effects of YTHDF2 in vivo ., Results: According to our findings, YTHDF2 was expressed at a low level in peripheral blood mononuclear cells from septic mice and patients as well as in LPS-induced RAW264.7 cells. Overexpression of YTHDF2 alleviated the inflammatory response by inhibiting HMGB1 release and JAK2/STAT1 signalling in LPS-stimulated cells. Mechanistically, YTHDF2 suppressed JAK2/STAT1 signalling by directly recognizing the m6A-modified site in IL-6R and decreasing the stability of IL-6R mRNA, thereby inhibiting HMGB1 release. In vivo experiments showed that YTHDF2 played a protective role in septic mice by suppressing the IL-6R/JAK2/STAT1/HMGB1 axis., Conclusions: In summary, these findings demonstrate that YTHDF2 plays an essential role as an inhibitor of inflammation to reduce the release of HMGB1 by inhibiting the IL-6R/JAK2/STAT1 pathway, indicating that YTHDF2 is a novel target for therapeutic interventions in sepsis., Competing Interests: The authors state no conflict of interest., (© The Author(s) 2023. Published by Oxford University Press.)

Fluorescent RNAs, aptamers that bind and activate small fluorogenic dyes, have provided a particularly attractive approach to visualizing RNAs in live cells. However, the simultaneous imaging of multiple RNAs remains challenging due to a lack of bright and stable fluorescent RNAs with bio-orthogonality and suitable spectral properties. Here, we develop the Clivias, a series of small, monomeric and stable orange-to-red fluorescent RNAs with large Stokes shifts of up to 108 nm, enabling the simple and robust imaging of RNA with minimal perturbation of the target RNA's localization and functionality. In combination with Pepper fluorescent RNAs, the Clivias enable the single-excitation two-emission dual-color imaging of cellular RNAs and genomic loci. Clivias can also be used to detect RNA-protein interactions by bioluminescent imaging both in live cells and in vivo. We believe that these large Stokes shift fluorescent RNAs will be useful tools for the tracking and quantification of multiple RNAs in diverse biological processes., (© 2023. The Author(s), under exclusive licence to Springer Nature America, Inc.)

Nitrogen from ammonia is one of the most common pollutants toxics to aquatic species in aquatic environment. The intestinal mucosa is one of the key mucosal defenses of aquatic species, and the accumulation of ammonia nitrogen in water environment will cause irreversible damage to intestinal function. In this study, histology, immunohistochemistry, ultrastructural pathology, enzyme activity analysis and qRT-PCR were performed to reveal the toxic effect of ammonia nitrogen stress on the intestine of Pelteobagrus fulvidraco . According to histological findings, ammonia nitrogen stress caused structural damage to the intestine and reduced the number of mucous cells. Enzyme activity analysis revealed that the activity of bactericidal substances (Lysozyme, alkaline phosphatase, and ACP) had decreased. The ultrastructure revealed sparse and shortened microvilli as well as badly degraded tight junctions. Immunohistochemistry for ZO-1 demonstrated an impaired intestinal mucosal barrier. Furthermore, qRT-PCR revealed that tight junction related genes ( ZO-1, Occludin, Claudin-1 ) were downregulated, while the pore-forming protein Claudin-2 was upregulated. Furthermore, as ammonia nitrogen concentration grew, so did the positive signal of Zap-70 (T/NK cell) and the expression of inflammation-related genes ( TNF , IL-1β , IL-8, IL-10) . In light of the above findings, we conclude that ammonia nitrogen stress damages intestinal mucosal barrier of Pelteobagrus fulvidraco and induces intestinal inflammation., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Liu, Luo, Zuo, Huang, Zhong, Liu, Geng, Ou, Chen, Cai and Deng.)

Calcium, magnesium and phosphate are predominant constituents in the human bone. In this study, magnesium-calcium phosphate composite bioceramic scaffolds were fabricated utilizing Mg 3 (PO 4 ) 2 and β-Ca 3 (PO 4 ) 2 as starting materials, and their pore structure was constructed by 3D printing. The porosity and compressive strength of the composite bioceramic scaffolds could be adjusted by altering the sintering temperature and the formula of starting materials. The composite bioceramic scaffolds prepared from 60 wt% Mg 3 (PO 4 ) 2 and 40 wt% β-Ca 3 (PO 4 ) 2 were dominated by the Ca 3 Mg 3 (PO 4 ) 4 phase, and this Ca 3 Mg 3 (PO 4 ) 4 -based bioceramic scaffolds possessed the highest compressive strength (12.7 - 92.4 MPa). Moreover, the Ca 3 Mg 3 (PO 4 ) 4 -based bioceramic scaffolds stimulated cellular growth and osteoblastic differentiation of bone marrow stromal cells. The Ca 3 Mg 3 (PO 4 ) 4 -based bioceramic scaffolds as bone regenerative biomaterials are flexible to the requirement of bone defects at various sites., Competing Interests: Declaration of Competing Interest The authors report no declarations of interest., (Copyright © 2023 Elsevier B.V. All rights reserved.)

In the last decade, oil-based titanium dioxide nanofluids (TiO 2 NFs) have gained immense interest due to their unique insulating properties and excellent thermal performance, which endow them with the potential for application in the field of modified insulating oils. A timely comparison, analysis and summary of recent advances in the preparation, characterization, and properties of different oil-based TiO 2 NFs for oil-immersed power transformers will contribute to provide a useful reference for the subsequent development of such materials. Preparation methods are reviewed along with their merits and demerits. Characterization techniques including scanning electron microscopy (SEM), transmission electron microscopy (TEM), optical microscopy (OM), X-ray diffraction (XRD), energy-dispersive X-ray spectroscopy (EDX), dynamic light scattering (DLS), Fourier transform infrared (FTIR) spectroscopy, thermally stimulated current (TSC), pulse electroacoustic technique (PEA), finite element analysis (FEA), fluorescence spectroscopy, Raman spectroscopy and zeta potential analysis are all applied to determine the crystal structure, particle size, surface function, surface charge and stability. Stabilization mechanisms are also discussed in detail. Some critical properties of oil-based TiO 2 NFs under the application of different influencing factors such as base oils, crystal structure, size of nanoparticles, surface modifiers, mixing percentage, and aging environment are highlighted. Finally, the existing challenges and perspectives are presented for future research.

This study aimed to investigate the mechanisms of nature and modified 'Guanximiyou' pummelo peel pectin (GGP and MGGP) in alleviating T2DM through in vitro and in vivo. After modification, pectin was transformed from high methoxy pectin (HMP) to low methoxy pectin (LMP), and the content of galacturonic acid was increased. These made MGGP have stronger antioxidant capacity and better inhibition effect on corn starch digestion in vitro. In vivo experiments have shown that both GGP and MGGP inhibited the development of diabetes after 4 weeks of ingestion. However, MGGP can more effectively reduce blood glucose and regulate lipid metabolism, and has significant antioxidant capacity and the ability to promote SCFAs secretion. In addition, 16S rRNA analysis showed that MGGP changed the composition of intestinal microbiota in diabetic mice, decreased the abundance of Proteobacteria, and increased the relative abundance of Akkermansia, Lactobacillus, Oscillospirales and Ruminococcaceae. The phenotypes of the gut microbiome also changed accordingly, indicating that MGGP can inhibit the growth of pathogenic bacteria, alleviate intestinal functional metabolic disorders and reverse the potential risk of related complications. Altogether, our findings demonstrate that MGGP, as a dietary polysaccharide, may inhibit the development of diabetes by reversing the imbalance of gut microbiota., Competing Interests: Declaration of competing interest The authors declare that there are no conflicts of interest., (Copyright © 2023. Published by Elsevier B.V.)

The structure properties, stability and β-carotene slow-release mechanism of soybean protein isolate-citrus pectin-gallic acid complex (SPI-CP-GA) stabilized high-internal phase Pickering emulsion (HIPPE) were investigated. The results showed that compared with the SPI-CP binary complex, the turbidity of the SPI-CP-GA ternary complex increased from 2.174 ± 0.001 to 3.027 ± 0.001, the surface wettability was increased, the infrared peaks was blue-shifted, changed from hydrophilic to hydrophobic, and the equilibrium interfacial tension of particles increased from 10.77 ± 0.02 mN/m to 13.46 ± 0.03 mN/m, the complex was more stable. When the GA was 2.0 mg/mL, the encapsulation efficiency of β-carotene was higher. With increased GA concentration and oil phase volume fraction (φ), the apparent viscosity and viscoelastic behavior of HIPPE performed well, forming a stable gel network structure. After 30 days of storage, there was no oil separation in the sample group with GA concentration of 2.0 mg/mL and φ = 0.7, and the stability was strong. After gastrointestinal digestion, the particle size of the HIPPE decreased from 13.51 ± 0.86 μm to 7.70 ± 0.68 μm, the free fatty acid (FFA) release rate was 22.03%, and the bioaccessibility of β-carotene was 6.67 ± 0.19%, and the sustained-release effect was obvious. These results indicated that the SPI-CP-GA ternary complex is a potential stabilizer for HIPPE, and providing theoretical guidance for the design of protein-polysaccharide-polyphenol stabilized HIPPE., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Glucaric acid is a valuable chemical with applications in the detergent, polymer, pharmaceutical and food industries. In this study, two key enzymes for glucaric acid biosynthesis, MIOX4 ( myo -inositol oxygenase) and Udh (uronate dehydrogenase), were fused and expressed with different peptide linkers. It was found that a strain harboring the fusion protein MIOX4-Udh linked by the peptide (EA 3 K) 3 produced the highest glucaric acid titer and thereby resulted in glucaric acid production that was 5.7-fold higher than that of the free enzymes. Next, the fusion protein MIOX4-Udh linked by (EA 3 K) 3 was integrated into delta sequence sites of the Saccharomyces cerevisiae opi1 mutant, and a strain, GA16, that produced a glucaric acid titer of 4.9 g/L in a shake flask fermentation was identified by a high-throughput screening method using an Escherichia coli glucaric acid biosensor. Strain improvement by further engineering was performed to regulate the metabolic flux of myo -inositol to increase the supply of glucaric acid precursors. The downregulation of ZWF1 and the overexpression of INM1 and ITR1 increased glucaric acid production significantly, and glucaric acid production was increased to 8.49 g/L in the final strain GA-ZII in a shake flask fermentation. Finally, in a 5-L bioreactor, GA-ZII produced a glucaric acid titer of 15.6 g/L through fed-batch fermentation. IMPORTANCE Glucaric acid is a value-added dicarboxylic acid that was synthesized mainly through the oxidation of glucose chemically. Due to the problems of the low selectivity, by-products, and highly polluting waste of this process, producing glucaric acid biologically has attracted great attention. The activity of key enzymes and the intracellular myo -inositol level were both rate-limiting factors for glucaric acid biosynthesis. To increase glucaric acid production, this work improved the activity of the key enzymes in the glucaric acid biosynthetic pathway through the expression of a fusion of Arabidopsis thaliana MIOX4 and Pseudomonas syringae Udh as well as a delta sequence-based integration. Furthermore, intracellular myo -inositol flux was optimized by a series of metabolic strategies to increase the myo -inositol supply, which improved glucaric acid production to a higher level. This study provided a way for constructing a glucaric acid-producing strain with good synthetic performance, making glucaric acid production biologically in yeast cells much more competitive., Competing Interests: The authors declare no conflict of interest.

Naturally occurring fluorescent proteins (FPs) are the most widely used tools for tracking cellular proteins and sensing cellular events. Here, we chemically evolved the self-labeling SNAP-tag into a palette of SNAP-tag mimics of fluorescent proteins (SmFPs) that possess bright, rapidly inducible fluorescence ranging from cyan to infrared. SmFPs are integral chemical-genetic entities based on the same fluorogenic principle as FPs, i.e., induction of fluorescence of non-emitting molecular rotors by conformational locking. We demonstrate the usefulness of these SmFPs in real-time tracking of protein expression, degradation, binding interactions, trafficking, and assembly, and show that these optimally designed SmFPs outperform FPs like GFP in many important ways. We further show that the fluorescence of circularly permuted SmFPs is sensitive to the conformational changes of their fusion partners, and that these fusion partners can be used for the development of single SmFP-based genetically encoded calcium sensors for live cell imaging., (© 2023. The Author(s).)

Objective: To establish a comprehensive diagnostic classification model of lateral cephalograms based on artificial intelligence (AI) to provide reference for orthodontic diagnosis. Methods: A total of 2 894 lateral cephalograms were collected in Department of Orthodontics, Capital Medical University School of Stomatology from January 2015 to December 2021 to construct a data set, including 1 351 males and 1 543 females with a mean age of (26.4± 7.4) years. Firstly, 2 orthodontists (with 5 and 8 years of orthodontic experience, respectively) performed manual annotation and calculated measurement for primary classification, and then 2 senior orthodontists (with more than 20 years of orthodontic experience) verified the 8 diagnostic classifications including skeletal and dental indices. The data were randomly divided into training, validation, and test sets in the ratio of 7∶2∶1. The open source DenseNet121 was used to construct the model. The performance of the model was evaluated by classification accuracy, precision rate, sensitivity, specificity and area under the curve (AUC). Visualization of model regions of interest through class activation heatmaps. Results: The automatic classification model of lateral cephalograms was successfully established. It took 0.012 s on average to make 8 diagnoses on a lateral cephalogram. The accuracy of 5 classifications was 80%-90%, including sagittal and vertical skeletal facial pattern, mandibular growth, inclination of upper incisors, and protrusion of lower incisors. The acuracy rate of 3 classifications was 70%-80%, including maxillary growth, inclination of lower incisors and protrusion of upper incisors. The average AUC of each classification was ≥0.90. The class activation heat map of successfully classified lateral cephalograms showed that the AI model activation regions were distributed in the relevant structural regions. Conclusions: In this study, an automatic classification model for lateral cephalograms was established based on the DenseNet121 to achieve rapid classification of eight commonly used clinical diagnostic items.

Objective: To develop a multi-classification orthodontic image recognition system using the SqueezeNet deep learning model for automatic classification of orthodontic image data. Methods: A total of 35 000 clinical orthodontic images were collected in the Department of Orthodontics, Capital Medical University School of Stomatology, from October to November 2020 and June to July 2021. The images were from 490 orthodontic patients with a male-to-female ratio of 49∶51 and the age range of 4 to 45 years. After data cleaning based on inclusion and exclusion criteria, the final image dataset included 17 453 face images (frontal, smiling, 90° right, 90° left, 45° right, and 45° left), 8 026 intraoral images [frontal occlusion, right occlusion, left occlusion, upper occlusal view (original and flipped), lower occlusal view (original and flipped) and coverage of occlusal relationship], 4 115 X-ray images [lateral skull X-ray from the left side, lateral skull X-ray from the right side, frontal skull X-ray, cone-beam CT (CBCT), and wrist bone X-ray] and 684 other non-orthodontic images. A labeling team composed of orthodontic doctoral students, associate professors, and professors used image labeling tools to classify the orthodontic images into 20 categories, including 6 face image categories, 8 intraoral image categories, 5 X-ray image categories, and other images. The data for each label were randomly divided into training, validation, and testing sets in an 8∶1∶1 ratio using the random function in the Python programming language. The improved SqueezeNet deep learning model was used for training, and 13 000 natural images from the ImageNet open-source dataset were used as additional non-orthodontic images for algorithm optimization of anomaly data processing. A multi-classification orthodontic image recognition system based on deep learning models was constructed. The accuracy of the orthodontic image classification was evaluated using precision, recall, F1 score, and confusion matrix based on the prediction results of the test set. The reliability of the model's image classification judgment logic was verified using the gradient-weighted class activation mapping (Grad-CAM) method to generate heat maps. Results: After data cleaning and labeling, a total of 30 278 orthodontic images were included in the dataset. The test set classification results showed that the precision, recall, and F1 scores of most classification labels were 100%, with only 5 misclassified images out of 3 047, resulting in a system accuracy of 99.84%(3 042/3 047). The precision of anomaly data processing was 100% (10 500/10 500). The heat map showed that the judgment basis of the SqueezeNet deep learning model in the image classification process was basically consistent with that of humans. Conclusions: This study developed a multi-classification orthodontic image recognition system for automatic classification of 20 types of orthodontic images based on the improved SqueezeNet deep learning model. The system exhibitted good accuracy in orthodontic image classification.

Misregulation of spermatogenesis transcription factors (TF) in hybrids can lead to misexpression, which is a mechanism for hybrid male sterility (HMS). We used dzo (male offspring of Bos taurus ♂ × Bos grunniens ♀) in bovines to investigate the relationship of the key TF with HMS via RNA sequencing and assay for transposase-accessible chromatin with high-throughput sequencing analyses. RNA sequencing showed that the widespread misexpression in dzo was associated with spermatogenesis-related genes and somatic or progenitor genes. The transition from leptotene or zygotene spermatocytes to pachytene spermatocytes may be the key stage for meiosis arrest in dzo. The analysis of TF-binding motif enrichment revealed that the male meiosis-specific master TF MYB proto-oncogene like 1 (MYBL1, known as A-MYB) motif was enriched on the promoters of downregulated pachytene spermatocyte genes in dzo. Assay for transposase-accessible chromatin with high-throughput sequencing revealed that TF-binding sites for MYBL1, nuclear transcription factor Y, and regulatory factor X were enriched in the low-chromatin accessibility region of dzo. The target genes of the MYBL1-binding motif were associated with meiosis-specific genes and significantly downregulated in dzo testis. The transcription factor MYBL1 may be the candidate master regulator for pachytene spermatocyte genes dysregulated in interspecific HMS dzo. This study reported that a few upstream TF regulation changes might exert a cascading effect downstream in a regulatory network as a mechanism for HMS., (The Authors. Published by Elsevier Inc. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Due to the similarity in the grain and difference in the market value among many rice varieties, deliberate mislabeling and adulteration has become a serious problem. To check the authenticity, we aimed to discriminate rice varieties based on their volatile organic compounds (VOCs) composition by headspace solid phase microextraction (HS-SPME) coupled with gas chromatography mass spectrometry (GC-MS). The VOC profiles of Wuyoudao 4 from nine sites in Wuchang were compared to 11 rice cultivar from other regions. Multivariate analysis and unsupervised clustering showed an unambiguous distinction between Wuchang rice and non-Wuchang rice. Partial least squares discriminant analysis (PLS-DA) demonstrated a goodness of fit of 0.90 and a goodness of prediction of 0.85. The discriminating ability of volatile compounds is also supported by Random forest analysis. Our data revealed eight biomarkers including 2-acetyl-1-pyrroline (2-AP) that can be used for variation identification. Taken together, the current method can readily distinguish Wuchang rice from other varieties which it holds great potential in checking the authenticity of rice., (© 2023. The Author(s).)

The COVID-19 pandemic has greatly impacted lifestyles and travel patterns, revealing existing societal and transportation gaps and introducing new challenges. In the context of an aging population, this study investigated how the travel behaviors of older adults (aged 60+) in New York City were affected by COVID-19, using an online survey and analyzing younger adult (aged 18-59) data for comparative analysis. The purpose of the study is to understand the pandemic's effects on older adults' travel purpose and frequency, challenges faced during essential trips, and to identify potential policies to enhance their mobility during future crises. Descriptive analysis and Wilcoxon signed-rank tests were used to summarize the changes in employment status, trip purposes, transportation mode usage, and attitude regarding transportation systems before and during the outbreak and after the travel restrictions were lifted. A Natural Language Processing model, Gibbs Sampling Dirichlet Multinomial Mixture, was adopted to open-ended questions due to its advantage in extracting information from short text. The findings show differences between older and younger adults in telework and increased essential-purpose trips (e.g., medical visits) for older adults. The pandemic increased older adults' concern about health, safety, comfort, prices when choosing travel mode, leading to reduced transit use and walking, increased driving, and limited bike use. To reduce travel burdens and maintain older adults' employment, targeted programs improving digital skills (telework, telehealth, telemedicine) are recommended. Additionally, safe, affordable, and accessible transportation alternatives are necessary to ensure mobility and essential trips for older adults, along with facilitation of walkable communities., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2023 The Authors.)

This study reports the isolation and characterization of a human monoclonal antibody (mAb) called 19n01. This mAb was isolated by using single-cell RNAseq of B cells from donors infected with the ancestral strain. This mAb possesses a potent and broad capacity to bind and neutralize all previously circulating variants of concern (VOCs), including Omicron sublineages BA.1, BA.2, and BA.4/5. The pseudovirus neutralization assay revealed robust neutralization capacity against the G614 strain, BA.1, BA.2, and BA.4/5, with inhibitory concentration (IC50) values ranging from 0.0035 to 0.0164 μg/mL. The microneutralization assay using the G614 strain and VOCs demonstrated IC50 values of 0.013-0.267 μg/mL. Biophysical and structural analysis showed that 19n01 cross-competes with ACE2 binding to the receptor-binding domain (RBD) and the kinetic parameters confirmed the high affinity against the Omicron sublineages (K D of 61 and 30 nM for BA.2 and BA.4/5, respectively). These results suggest that the 19n01 is a remarkably potent and broadly reactive mAb., Competing Interests: M.G.V., M.R.S., E.A.M.G., V.M.H., and J.H. are coinventors of patent applications describing the neutralizing mAbs. The other authors declare no competing interests., (© 2023 The Author(s).)

Introduction: Amplitude spectrum area (AMSA) is a well-established measure than can predict defibrillation outcome and guiding individualized resuscitation of ventricular fibrillation (VF) patients. However, accurate AMSA can only be calculated during cardiopulmonary resuscitation (CPR) pause due to artifacts produced by chest compression (CC). In this study, we developed a real-time AMSA estimation algorithm using a convolutional neural network (CNN). Methods: Data were collected from 698 patients, and the AMSA calculated from the uncorrupted signals served as the true value for both uncorrupted and the adjacent corrupted signals. An architecture consisting of a 6-layer 1D CNN and 3 fully connected layers was developed for AMSA estimation. A 5-fold cross-validation procedure was used to train, validate and optimize the algorithm. An independent testing set comprised of simulated data, real-life CC corrupted data, and preshock data was used to evaluate the performance. Results: The mean absolute error, root mean square error, percentage root mean square difference and correlation coefficient were 2.182/1.951 mVHz, 2.957/2.574 mVHz, 22.887/28.649% and 0.804/0.888 for simulated and real-life testing data, respectively. The area under the receiver operating characteristic curve regarding predicting defibrillation success was 0.835, which was comparable to that of 0.849 using the true value of the AMSA. Conclusions: AMSA can be accurately estimated during uninterrupted CPR using the proposed method., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Zuo, Dai, Wei, Gong, Yin and Li.)

Anaerobic ammonium oxidation (anammox) is a potential process to achieve the neutralization of energy and carbon. Due to the low temperature and variation of municipal sewage, the application of mainstream anammox is hard to be implemented. For spreading mainstream anammox in practice, several key issues and bottlenecks including the start-up, stable NO 2 - -N supply, maintenance and dominance of AnAOB with high activity, prevention of NO 3 - -N buildup, reduction of sludge loss, adaption to the seasonal temperature and alleviation of COD impacts on AnAOB are discussed and summarized in this review in order to improve its startup, stable operation and resilience of mainstream anammox. Hence a combined biological nitrogen removal (CBNR) system based on conventional denitrification, shortcut nitrification-denitrification, Partial Nitritation and partial Denitrification combined Anammox (PANDA) process through the management of organic matter and nitrate is proposed correspondingly aiming at adaptation to the variations of seasonal temperature and pollutants in influent., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

During the outbreak of COVID-19, people's reliance on social media for pandemic-related information exchange, daily communications, and online professional interactions increased because of self-isolation and lockdown implementation. Most of the published research addresses the performance of nonpharmaceutical interventions (NPIs) and measures on the issues impacted by COVID-19, such as health, education, and public safety; however, not much is known about the interplay between social media use and travel behaviors. This study aims to determine the effect of social media on human mobility before and after the COVID-19 outbreak, and its impact on personal vehicle and public transit use in New York City (NYC). Apple mobility trends and Twitter data are used as two data sources. The results indicate that Twitter volume and mobility trend correlations are negative for both driving and transit categories in general, especially at the beginning of the COVID-19 outbreak in NYC. A significant time lag (13 days) between the online communication rise and mobility drop can be observed, thereby providing evidence of social networks taking quicker reactions to the pandemic than the transportation system. In addition, social media and government policies had different impacts on vehicular traffic and public transit ridership during the pandemic with varied performance. This study provides insights on the complex influence of both anti-pandemic measures and user-generated content, namely social media, on people's travel decisions during pandemics. The empirical evidence can help decision-makers formulate timely emergency responses, prepare targeted traffic intervention policies, and conduct risk management in similar outbreaks in the future., Competing Interests: The author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article., (© National Academy of Sciences: Transportation Research Board 2021.)

Ammonia nitrogen is extremely toxic to aquatic animals, and is also the most common pollutant in the aquatic environment. In order to investigate the effect of high concentration of ambient ammonia nitrogen on fish gills, two groups, including a high ammonia group (T group: TAN = 2.5 mg/L, 10 % 96 h LC50) and a control group (Z group: total ammonia nitrogen (TAN) = 0 mg/L) were set up in this study. The effects of chronic ammonia stress on the gills of Pelteobagrus fulvidraco were investigated by histopathological, enzymatic, transcriptomic and proteomic analyses after 28 d of stress at different ammonia nitrogen concentrations. Histopathological observations revealed significant inflammatory cell infiltration, necrotic and abscission at the base of the gill filaments, and massive proliferation of cells at the base of the gill lamellae. Ammonia nitrogen stress led to increased reactive oxygen species (ROS) content and decreased catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GSH-PX) activities in gills, indicating significant oxidative stress in gills. And further transcriptomic analysis revealed that 807 differential expression genes (DEGs) were generated in the gills, of which 587 DEGs were up-regulated and 220 DEGs were down-regulated. In addition, proteomics analysis identified 1073 differential expression proteins (DEPs) in gills, including 983 up- and 90 down-regulated DEPs. Pathway enrichment analysis of the DEGs and DEPs revealed that multiple inflammation-related signaling pathways were activated in the gill, including the significantly enriched IL17 signaling pathway. This suggests that IL17 signaling pathway might have a significant impact during signaling transduction. Further analysis of network regulation by mapping DEGs and DEPs to KEGG pathway revealed that IL17 signaling pathway mediated inflammation and cell proliferation in gills under ammonia stress. The results of this study provided new insights into the response of fish gills to ammonia nitrogen stress, and the IL17 signaling pathway may be a potential therapeutic target for reducing ammonia nitrogen gill toxicity., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier B.V.)

Competing Interests: X.S.X. and Y.C. are the inventors of the provisional patent applications for BD series neutralizing antibodies and are founders of Singlomics Biopharmaceuticals. All other authors declare no competing interests.

Catechol-based hydrogels have good adhesion properties; however, since the concentration of catechol is low and it can be easily oxidized to quinone, the adhesion performance of the hydrogels is reduced, which limits their application as self-adhesive flexible wearable sensors. In this work, a dopamine: poly(sodium 4-styrenesulfonate) (DA:PSS)-initiated strategy was proposed to construct adhesive hydrogels, where the semiquinone radicals present in DA:PSS were used to initiate radical polymerization to obtain the DA:PSS/poly(acrylic acid) (DA:PSS/PAA) hydrogel. This hydrogel exhibited good stretchability and adhesion with various substrates. We observed that, even after exposure to air for 21 days under certain relative humidity (76%), the catechol groups hardly oxidized and the DA:PSS/PAA hydrogel presented good adhesion. The DA:PSS/PAA hydrogel also showed good electrical conductivity and fast response ability. Thus, the general strategy of triggering monomer polymerization to form hydrogels based on the semiquinone radical present in DA:PSS offers great potential for their application in flexible electronic devices and wearable sensors., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (This journal is © The Royal Society of Chemistry.)

Sepsis comprises a lethal immunologic response due to infection. Increasingly, evidence has demonstrated the important role of long non-coding RNA growth arrest-specific transcript 5 (GAS5) in the regulation of sepsis. Nevertheless, the mechanisms by which GAS5 participates in the progression of sepsis remain unclear. Our study demonstrated the role and underlying mechanism of GAS5 in regulating lipopolysaccharide (LPS)-induced inflammation. In this study, GAS5 expression was found to be markedly decreased in serum samples of sepsis patients and a sepsis mouse model, and was negatively related with HMGB1 expression. GAS5 overexpression inhibited cell inflammatory responses by decreasing HMGB1 release. Furthermore, GAS5 inhibited LPS-mediated hyperacetylation and the release of HMGB1 by increasing the expression of sirtuin1 (SIRT1). Additionally, upregulated GAS5 attenuated inflammatory responses in vitro and vivo, and the knockdown of a miR-155-5p mimic and SIRT1 rescued the effects of GAS5 upregulation. Mechanistically, GAS5 sponged miR-155-5p to upregulate SIRT1, thereby inhibiting HMGB1 acetylation and release. In conclusion, our findings indicate that GAS5 suppresses inflammatory responses by modulating the miR-155-5p/SIRT1/HMGB1 axis in sepsis, providing a novel therapeutic target for inflammation in sepsis., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Circulating tumor cells (CTCs), shed by primary malignancies, function as "seeds" for distant metastasis. However, it is still largely unknown how CTCs escape immune surveillance. Here, we characterize the transcriptomes of human pancreatic ductal adenocarcinoma CTCs, primary, and metastatic lesions at single-cell scale. Cell-interaction analysis and functional studies in vitro and in vivo reveal that CTCs and natural killer (NK) cells interact via the immune checkpoint molecule pair HLA-E:CD94-NKG2A. Disruption of this interaction by blockade of NKG2A or knockdown of HLA-E expression enhances NK-mediated tumor cell killing in vitro and prevents tumor metastasis in vivo. Mechanistic studies indicate that platelet-derived RGS18 promotes the expression of HLA-E through AKT-GSK3β-CREB signaling, and overexpression of RGS18 facilitates pancreatic tumor hepatic metastasis. In conclusion, platelet-derived RGS18 protects CTCs from NK-mediated immune surveillance by engaging the immune checkpoint HLA-E:CD94-NKG2A. Interruption of the suppressive signaling prevents tumor metastasis in vivo by immune elimination of CTCs., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Introduction: This study aims to compare the differences in circulating adiponectin levels and their relationships to regional adiposity, insulin resistance, serum lipid, and inflammatory factors in young, healthy Japanese women with different physical activity statuses., Methods: Adipokines (adiponectin and leptin), full serum lipid, and inflammatory factors [white blood cell counts, C-reactive protein, tumor necrosis factor-α, tissue plasminogen activator inhibitor-1 (PAI-1)] were measured in 101 sedentary and 100 endurance-trained healthy Japanese women (aged 18-23 years). Insulin sensitivity was obtained through a quantitative insulin-sensitivity check index (QUICKI). Regional adiposity [trunk fat mass (TFM), lower-body fat mass (LFM), and arm fat mass (AFM)] was evaluated using the dual-energy X-ray absorptiometry method., Results: No significant difference was observed between the sedentary and trained women in terms of adiponectin levels. The LFM-to-TFM ratio and the high-density lipoprotein cholesterol (HDL-C) were the strong positive determinants for adiponectin in both groups. Triglyceride in the sedentary women was closely and negatively associated with adiponectin, as well as PAI-1 in the trained women. The QUICKI level was higher in the trained than sedentary women. However, no significant correlation between adiponectin and insulin sensitivity was detected in both groups. Furthermore, LFM was associated with a favorable lipid profile against cardiovascular diseases (CVDs) in the whole study cohort, but this association became insignificant when adiponectin was taken into account., Conclusions: These findings suggest that adiponectin is primarily associated with regional adiposity and HDL-C regardless of insulin sensitivity and physical activity status in young, healthy women. The associations among adiponectin, lipid, and inflammatory factors are likely different in women with different physical activity statuses. The correlation of LFM and a favorable lipid profile against CVD and adiponectin is likely involved in this association., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Guan, Zuo, Zhao, Nian, Shi, Xu, Huang, Kazumi and Wu.)

Ethnopharmacological Relevance: With high mortality and morbidity rates, lung cancer (LC) has become one of the major threats to human health. The treatment strategies for LC currently face issues, such as drug resistance and body tolerance. Traditional Chinese medicine (TCM) is characterized by novel pharmacological mechanisms, low toxicity, and limited side effects. TCM includes a substantial number of biologically active ingredients, several of which are effective monomeric agents against LC. An increasing number of researchers are focusing their efforts on the discovery of active anti-cancer ingredients in TCM., Aim of the Review: In this review, we summarized the anti-LC mechanisms of five types of TCM monomeric compounds. Our goal is to provide research ideas for the identification of new prospective medication candidates for the treatment of LC., Materials and Methods: We collected reports on the anti-LC effects of TCM monomers from web databases, including PubMed, Science Direct, Web of Science, and Europe PubMed Central. Among the keywords used were "lung cancer," "traditional Chinese medicine," "pharmacology," and their combinations thereof. Then, we systematically summarized the anti-LC efficacy and related mechanisms of TCM monomers., Results: Based on the available literature, this paper reviewed the therapeutic effects and mechanisms of five types of TCM monomers on LC. The characteristics of TCM monomers include the capabilities to suppress the tumor cell cycle, inhibit proliferation, induce apoptosis, promote autophagy, inhibit tumor cell invasion and metastasis, and enhance efficacy or reduce drug resistance when combined with cytotoxic agents and other methods to arrest the progression of LC and prolong the survival of patients., Conclusions: TCM contains numerous flavonoids, alkaloids, terpenoids, polyphenols, and other active compounds that are effective against LC. Given their chemical structure and pharmacological properties, these monomers are suitable as candidate drugs for the treatment of LC., Competing Interests: Declaration of competing interest The authors declare no potential conflict of interest. Figures were created by Figdraw (www.figdraw.com)., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Background: Severe burn injury can be a life-threatening experience and can also lead to financial issues for suffers. The purpose of the current study was to analyze the direct hospitalization costs of severe burn inpatients in Southwest China., Methods: Data related to all inpatients admitted with severe burns [total body surface area (TBSA) ≥30%] pooled from 2015 to 2021 were reviewed retrospectively at the Institute of Burn Research of Army Medical University. Demographic parameters, medical economics, and clinical data were obtained from medical records., Results: A total of 668 cases were identified. The average age was 37.49 ± 21.00 years, and 72.3% were men. The average TBSA was 51.35 ± 19.49%. The median length of stay of inpatients in the burn intensive care unit was 14 [interquartile range (IQR): 5.0-34.8] days, and the median length of stay (LOS) was 41 (IQR: 22.0-73.8) days. The mortality rate was 1.6%. The median total cost was 212,755.45 CNY (IQR: 83,908.80-551,621.57 CNY) per patient varying from 3,521.30 to 4,822,357.19 CNY. The direct cost of scald burns was dramatically lower compared with that of other types of burns, with 11,213.43 to 2,819,019.14 CNY. Medical consumables presented the largest portion of total costs, with a median cost of 65,942.64 CNY (IQR: 18,771.86-171,197.97 CNY). The crucial risk factors for medical cost in our study were TBSA, surgical frequency, LOS, depth of burn, and outcome., Conclusion: We conclude that an effective burn prevention program, shorter hospital stays, and facilitating the healing of wounds should be focused on with tailored precautionary protocols to reduce the medical costs of inpatients with severe burns., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Zeng, Li, Yang, Feng, Zuo, Luo, Peng and Yuan.)

Proper calibration process is of considerable importance for traffic safety evaluations using simulation models. Allowing for a pure with and without comparison under identical circumstances that is not directly testable in the field, microsimulation-based approach has drawn considerable attention for the performance evaluation of emerging technologies, such as connected vehicle (CV) safety applications. Different from the traditional approaches to evaluate mobility impacts, safety evaluations of such applications demand the simulation models to be well calibrated to match real-world safety conditions. This paper proposes a novel calibration framework which combines traffic conflict techniques and multi-objective stochastic optimization so that the operational and safety measures can be calibrated simultaneously. The conflict distribution of different severity levels categorized by time-to-collision (TTC) is applied as the safety performance measure. Simultaneous perturbation stochastic approximation (SPSA) algorithm, which can efficiently approximate the gradient of the multi-objective stochastic loss function, is used for model parameters optimization that minimizes the total simulation error of both operational and safety performance measures. The proposed calibration methodology is implemented using an open-source software SUMO on a simulation network of the Flatbush Avenue corridor in Brooklyn, NY. 17 key parameters are calibrated using the SPSA algorithm and are compared with the real-world traffic conflicts extracted using vehicle trajectories from 14 h' high-resolution aerial and traffic surveillance videos. Representative days are identified to create variation envelopes for performance measures. Four acceptability criteria, including control for time-variant outliers and inliers, bounded dynamic absolute and system errors are adopted for results analysis. The results show that the calibrated parameters can significantly improve the performance of the simulation model to represent real-world safety conditions (i.e., traffic conflicts) as well as operational conditions. The case study also demonstrates the usefulness of aerial imagery and the applicability of the proposed model calibration framework, so the calibrated model can be used to evaluate the safety benefits of CV applications more accurately., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

This study explored the role of the long non-coding RNA (lncRNA) XIST (X-inactive specific transcript) as a driver of RA pathogenesis, with a particular focus on the ability of this lncRNA to interact with GATA1 and CCN6. The GSE83147and GSE181614 datasets were downloaded for analysis. XIST and CCN6 expression were assessed in synovial fibroblasts (SFs) and in both normal cartilage samples and those from RA patients, with the relationship between XIST and CCN6 additionally being examined. XIST and CCN6 were respectively knocked down or overexpressed in SFs to establish their regulatory roles in these cells in the context of RA. Further studies of the regulatory interplay between XIST, GATA1, and CCN6 were then performed through RNA immunoprecipitation, RNA pull-down, gain-of-function, loss-of-function, and luciferase reporter assays. In addition, RA model rats were established and used to measure the production of TNF-α, IL-6, and IL-8 and to subject tissues from these animals to histopathological examination. RA patient synovial tissues and SFs exhibited XIST and CCN6 upregulation. The knockdown of XIST suppressed SF migratory, proliferative, invasive, and angiogenic activity, while CCN6 knockdown partially reversed the ability of XIST to influence these phenotypic outcomes in vitro and in vivo. XIST bound to GATA1 within SFs, thus promoting enhanced CCN6 transcription. Knocking down XIST alleviated RA-related pathological damage, synovial injury, and inflammatory response induction in rats. The binding of XIST to GATA1 leads to CCN6 upregulation, driving RA pathogenesis by altering SF proliferation and angiogenic activity, suggesting that this pathway may represent a viable target for therapeutic intervention., Competing Interests: Conflict of Interest The authors declare that they have no conflict of interest., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Cancer is a leading cause of death and a severe threat to global public health. Organoid, as a novel 3D in vitro model, has been applied in various tumor related studies due to its apparent advantages. The organoid is mainly constructed by Matrigel-depended 3D culture system, Air-Liquid Interface (ALI) culture, and Microfluidic culture or Organ-on-chips platform. For the application in carcinogenesis studies, the organoid model may favor depicting initiative hallmarks and identifying potential intervening targets, investigating driver genes of carcinogenesis, and identifying known or unknown risk or protective factors. In this review, we discussed different organoid construction methods and their properties. We also noted that tumor organoids can portray initiative hallmarks and identify possible intervening targets, as well as explore carcinogenesis driver genes and uncover known or unknown risks or protective factors. Organoid systems have been used to identify tumor-preventive drugs such as oligomeric proanthocyanidins, Vitamin D, n-3 PUFAs, and pomegranate. The current evidence underscores the organoid model's potential importance in developing innovative tumorprevention techniques., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

With biotechnological advancements, innovative omics technologies are constantly emerging that have enabled researchers to access multi-layer information from the genome, epigenome, transcriptome, proteome, metabolome, and more. A wealth of omics technologies, including bulk and single-cell omics approaches, have empowered to characterize different molecular layers at unprecedented scale and resolution, providing a holistic view of tumor behavior. Multi-omics analysis allows systematic interrogation of various molecular information at each biological layer while posing tricky challenges regarding how to extract valuable insights from the exponentially increasing amount of multi-omics data. Therefore, efficient algorithms are needed to reduce the dimensionality of the data while simultaneously dissecting the mysteries behind the complex biological processes of cancer. Artificial intelligence has demonstrated the ability to analyze complementary multi-modal data streams within the oncology realm. The coincident development of multi-omics technologies and artificial intelligence algorithms has fuelled the development of cancer precision medicine. Here, we present state-of-the-art omics technologies and outline a roadmap of multi-omics integration analysis using an artificial intelligence strategy. The advances made using artificial intelligence-based multi-omics approaches are described, especially concerning early cancer screening, diagnosis, response assessment, and prognosis prediction. Finally, we discuss the challenges faced in multi-omics analysis, along with tentative future trends in this field. With the increasing application of artificial intelligence in multi-omics analysis, we anticipate a shifting paradigm in precision medicine becoming driven by artificial intelligence-based multi-omics technologies., Competing Interests: Conflict of interest statement The authors declare that there are no conflicts of interest., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Passive administration of neutralizing antibodies (Abs) is an attractive strategy for the control of gastrointestinal infections. However, an unanswered practical concern is the need to assure the stability of sufficient amounts of orally administered neutralizing Abs against intestinal pathogens (e.g., norovirus) in the harsh environment of the gastrointestinal tract. To this end, we expressed a single-domain Ab (VHH, nanobody) against norovirus on the cell surface of Lactobacillus , a natural and beneficial commensal component of the gut microbiome. First, we used intestinal epithelial cells generated from human induced pluripotent stem cells to confirm that VHH 1E4 showed neutralizing activity against GII.17 norovirus. We then expressed VHH 1E4 as a cell-wall-anchored form in Lactobacillus paracasei BL23. Flow cytometry confirmed the expression of VHH 1E4 on the surface of lactobacilli, and L. paracasei that expressed VHH 1E4 inhibited the replication of GII.17 norovirus in vitro. We then orally administered VHH 1E4-expressing L. paracasei BL23 to germ-free BALB/c mice and confirmed the presence of lactobacilli with neutralizing activity in the intestine for at least 10 days after administration. Thus, cell-wall-anchored VHH-displaying lactobacilli are attractive oral nanobody deliver vectors for passive immunization against norovirus infection.

Vascular smooth muscle cells (VSMCs), which provides structural integrity and regulates the diameter of vasculature, are of great potential for modeling vascular-associated diseases and tissue engineering. Here, we presented a detailed comparison of differentiating human pluripotent stem cells (hPSCs) into VSMCs (hPSCs-VSMCs) in four different culture methods, including 2-dimensional (2D) culture, 3-dimensional (3D) PNIPAAm-PEG hydrogel culture, 3-dimensional (3D) alginate hydrogel culture, and transferring 3-dimensional alginate hydrogel culture to 2-dimensional (2D) culture. Both hydrogel-based culture methods could mimic in vivo microenvironment to protect cells from shear force, and avoid cells agglomeration, resulting in the extremely high culture efficiency (e.g., high viability, high purity and high yield) compared with 2D culture. We demonstrated hPSC-VSMCs produced from hydrogel-based culture methods had better contractile phenotypes and the potential of vasculature formation. The transcriptome analysis showed the hPSC-VSMCs derived from hydrogel-based culture methods displayed more upregulated genes in vasculature development, angiogenesis and blood vessel development, extracellular matrix compared with 2D culture. Taken together, hPSC-VSMCs produced from hydrogel-based culture system could be applied in various biomedical fields, and further indicated the suitable development of alginate hydrogel for industrial production by taking all aspects into consideration., Competing Interests: The authors declare no competing financial interest., (© 2022 The Japanese Society for Regenerative Medicine. Production and hosting by Elsevier B.V.)

Esophageal cancer (EC) is an aggressive malignancy raising a healthcare concern worldwide. Standard treatment options include surgical resection, chemotherapy, radiation therapy, and targeted molecular therapy. The five-year survival rate for all stages of EC is approximately 20%, ranging from 5% to 47%, with a high recurrence rate and poor prognosis after treatment. Immunotherapy has shown better efficacy and tolerance than conventional therapies for several malignancies. Immunotherapy of EC, including immune checkpoint inhibitors, cancer vaccines, and adoptive cell therapy, has shown clinical advantages. In particular, monoclonal antibodies against PD-1 have a satisfactory role in combination therapy and are recommended for first- or second-line treatments. Here, we present a systematic summary and analysis of immunotherapy-based combination therapies for EC., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Wang, Xu, Zuo, Liu and Yang.)

Helicobacter pylori infection triggers inflammation that may lead to gastritis, stomach ulcers and cancer. Probiotic bacteria, such as Lactobacillus , have been of interest as treatment options, however, little is known about the molecular mechanisms of Lactobacillus -mediated inhibition of H. pylori pathogenesis. In this work, we investigated the effect of Lactobacillus culture supernatants, so-called conditioned medium (CM), from two gastric isolates, L. gasseri and L. oris , on the expression of transcriptional regulators in H. pylori . Among the four known two-component systems (TCSs), i.e., ArsRS, FlgRS, CheAY and CrdRS, the flagellar regulator gene flgR and the acid resistance associated arsS gene were down-regulated by L. gasseri CM, whereas expression of the other TCS-genes remained unaffected. L. gasseri CM also reduced the motility of H. pylori , which is in line with reduced flgR expression. Furthermore, among six transcription factors of H. pylori only the ferric uptake regulator gene fur was regulated by L. gasseri CM. Deletion of fur further led to dramatically increased sensitivity to the antimicrobial peptide LL-37. Taken together, the results highlight that released/secreted factors of some lactobacilli, but not all, downregulate transcriptional regulators involved in motility, acid tolerance and LL-37 sensitivity of H. pylori .

Although recent studies have indicated that mutations in the gene encoding diacylglycerol kinase epsilon (DGKE) result in some proteinuria related hereditary kidney diseases, the DGKE expression pattern in the kidney and its contribution to acute kidney injury (AKI) remain unknown. Therefore, the present study was designed to detect the role of DGKE in mice with AKI. DGKE expression was time-dependently altered in the kidneys of mice with renal ischemia/reperfusion injury (IRI). Compared with wild-type (WT) mice, DGKE- overexpressing mice ( Rosa26-Dgke +/+ ) exhibited protective effects against renal IRI, including reduced serum creatinine, blood urea concentration, tubular cell death and inflammatory responses as well as improved morphological injuries. Consistently, in vitro , DGKE overexpression in human renal proximal tubule (HK-2) cells also protected against oxygen-glucose deprivation (OGD)/reoxygenation-induced cell death. Mechanistically, DGKE regulated Klotho expression, at least partly via the transcription factor Krüppel-like factor (KLF) 15. Moreover, a significant reduction in DGKE was also found in kidneys from patients with ischemia-associated acute tubular necrosis (ATN). Collectively, our studies demonstrate that DGKE protects against AKI in mice at least partly through KLF15/Klotho signaling pathway, indicating that DGKE may present an innovative therapeutic strategy for treating patients with AKI.

Chronic inflammation induced by Helicobacter pylori is strongly associated with gastric cancer development, which is influenced by both bacterial virulence and host genetics. The sialic acid-binding adhesin SabA and the MUC5AC-binding adhesin LabA are important H. pylori virulence factors that facilitate adhesion of the bacterium, which is a crucial step in colonization. Lactate utilization has been reported to play a key role in the pathogenicity of different bacterial species. However, this is poorly understood in H. pylori. In this study, we investigated the effect of lactate on H. pylori adhesin gene expression and the regulation of host inflammatory cytokines. We show that the bacterial adhesins SabA and LabA were downregulated at the transcriptional level during incubation of H. pylori with lactate. Downregulation of sabA required the involvement of the two-component system ArsRS, while labA was regulated via the CheA/CheY system, indicating differences in the regulation of these genes in response to lactate. The levels of the proinflammatory cytokines TNF and IL-6 in H. pylori-stimulated macrophages were reduced when lactate was present. Interestingly, glucose did not prevent the secretion of these cytokines. Taken together, our data suggest that lactate affects H. pylori adhesin gene expression and the host response upon infection., (© 2022. The Author(s).)

Rechargeable aluminum-ion batteries have attracted increasing attention owing to the advantageous multivalent ion storage mechanism thus high theoretical capacity as well as inherent safety and low cost of using aluminum. However, their development has been largely impeded by the lack of suitable positive electrodes to provide both sufficient energy density and satisfactory rate capability. Here we report a candidate positive electrode based on ternary metal oxides, Fe 2 (MoO 4 ) 3 , which was assembled by cross-stacking of porous nanosheets, featuring superior rate performance and cycle stability, and most importantly a well-defined discharge voltage plateau near 1.9 V. Specifically, the positive electrode is able to deliver reversible capacities of 239.3 mA h g -1 at 0.2 A g -1 and 73.4 mA h g -1 at 8.0 A g -1 , and retains 126.5 mA h g -1 at 1.0 A g -1 impressively, after 2000 cycles. Furthermore, the aluminum-storage mechanism operating on Al 3+ intercalation in this positive electrode is demonstrated for the first time via combined in situ and ex situ characterization studies and density functional theory calculations. This work not only explores potential positive electrodes for aluminum-based batteries but also sheds light on the fundamental charge storage mechanism within the electrode., Competing Interests: The authors declare no conflict of interest., (This journal is © The Royal Society of Chemistry.)

The protein hydrolysate of purple speckled kidney bean (PSKB) was used as the raw material in this study, and the antioxidant peptide of the PSKB protein hydrolysate was purified using macroporous resin. The XAD-7HP macroporous resin was selected as the best purification material, and the static adsorption-desorption of the purified PSKB antioxidant peptide was optimized. The optimum static adsorption and desorption conditions were as follows: the adsorption capacity reached 11.93 ± 0.11 mg/ml at pH 7 for 24 h, and the desorption capacity was 5.24 ± 0.04 mg/ml with 60% ethanol for 30 min. Under this condition, the amount of antioxidant peptide obtained by adsorption-desorption was the highest. The optimum process conditions were as follows: the appropriate flow rate was 1 ml/min, and the optimal injection volume was 40 ml. The adsorption amount at this time can reach 12.19 ± 0.15 mg/ml. The components with an elution time of 10-30 min were separated using the reversed-phase high-performance liquid chromatography (RP-HPLC) technique to obtain three main components, namely, RP 1 , RP 2 , and RP 3 . The DPPH free radical scavenging ability reached 56.26 ± 0.56, 66.42 ± 0.56, and 78.57 ± 0.56%, respectively, which were 36.65, 46.34 ± 0.56, and 54.39 ± 0.56% higher than those before purification. The amino acid sequences of the three components were identified as Phe-Leu-Val-Asp-Arg-Ile, Phe-Leu-Val-Ala-Pro-Asp-Asp, and Lys-Asp-Arg-Val-Ile-Ser-Glu-Leu., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Li, Xu, Yang, Wu, Ma, Zuo and Zhang.)

The performance of electrode materials depends intensively on the lithium (Li)-ion storage mechanisms correlating ultimately with the Coulombic efficiency, reversible capacity, and morphology variation of electrode material upon cycling. Transition metal nitrides anode materials have exhibited high-energy density and superior rate capability; however, the intrinsic mechanism is largely unexplored and still unclear. Here, a typical 3D porous Fe 2 N micro-coral anode is prepared and, an intercalation-conversion-heterogeneity hybrid Li-ion storage mechanism that is beyond the conventional intercalation or conversion reaction is revealed through various characterization techniques and thermodynamic analysis. Interestingly, using advanced in situ magnetometry, the ratio (ca. 24.4%) of the part where conversion reaction occurs to the entire Fe 2 N can further be quantified. By rationally constructing a Li-ion capacitor comprising 3D porous Fe 2 N micro-corals anode and commercial AC cathode, the hybrid full device delivers a high energy-density (157 Wh kg -1 ) and high power-density (20 000 W kg -1 ), as well as outstanding cycling stability (93.5% capacitance retention after 5000 cycles). This research provides an original and insightful method to confirm the reaction mechanism of material related to transition metals and a fundamental basis for emerging fast charging electrode materials to be efficiently explored for a next-generation battery., (© 2022 The Authors. Advanced Science published by Wiley-VCH GmbH.)

Background: Cerebral protection remains the cornerstone of successful aortic surgery; however, there is no consensus as to the optimal strategy., Objective: To compare the safety and efficacy of innominate to axillary artery cannulation for delivering antegrade cerebral protection during proximal aortic arch surgery., Methods: This randomized controlled trial (The Aortic Surgery Cerebral Protection Evaluation CardioLink-3 Trial, ClinicalTrials.gov Identifier: NCT02554032), conducted across 6 Canadian centers between January 2015 and June 2018, allocated 111 individuals to innominate or axillary artery cannulation. The primary safety outcome was neuroprotection per the appearance of new severe ischemic lesions on the postoperative diffusion-weighted-magnetic resonance imaging. The primary efficacy outcome was the difference in total operative time. Secondary outcomes included 30-day all-cause mortality and postoperative stroke., Results: One hundred two individuals (mean age, 63 ± 11 years) were in the primary safety per-protocol analysis. Baseline characteristics between the groups were similar. New severe ischemic lesions occurred in 19 participants (38.8%) in the axillary versus 18 (34%) in the innominate group (P for noninferiority = .0009). Total operative times were comparable (median, 293 minutes; interquartile range, 222-411 minutes) for axillary versus (298 minutes; interquartile range, 231-368 minutes) for innominate (P for superiority = .47). Stroke/transient ischemic attack occurred in 4 (7.1%) participants in the axillary versus 2 (3.6%) in the innominate group (P = .43). Thirty-day mortality, seizures, delirium, and duration of mechanical ventilation were similar in both groups., Conclusions: diffusion-weighted magnetic resonance imaging assessments indicate that antegrade cerebral protection with innominate cannulation is safe and affords similar neuroprotection to axillary cannulation during aortic surgery, although the burden of new neurological lesions is high in both groups., (Copyright © 2020 The American Association for Thoracic Surgery. Published by Elsevier Inc. All rights reserved.)

Background: Most severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-infected individuals are asymptomatic or only exhibit mild disease. In about 10% of cases, the infection leads to hypoxemic pneumonia, although it is much more rare in children., Objective: We evaluated 31 young patients aged 0.5 to 19 years who had preexisting inborn errors of immunity (IEI) but lacked a molecular diagnosis and were later diagnosed with coronavirus disease 2019 (COVID-19) complications., Methods: Genetic evaluation by whole-exome sequencing was performed in all patients. SARS-CoV-2-specific antibodies, autoantibodies against type I IFN (IFN-I), and inflammatory factors in plasma were measured. We also reviewed COVID-19 disease severity/outcome in reported IEI patients., Results: A potential genetic cause of the IEI was identified in 28 patients (90.3%), including mutations that may affect IFN signaling, T- and B-cell function, the inflammasome, and the complement system. From tested patients 65.5% had detectable virus-specific antibodies, and 6.8% had autoantibodies neutralizing IFN-I. Five patients (16.1%) fulfilled the diagnostic criteria of multisystem inflammatory syndrome in children. Eleven patients (35.4%) died of COVID-19 complications. All together, at least 381 IEI children with COVID-19 have been reported in the literature to date. Although many patients with asymptomatic or mild disease may not have been reported, severe presentation of COVID-19 was observed in 23.6% of the published cases, and the mortality rate was 8.7%., Conclusions: Young patients with preexisting IEI may have higher mortality than children without IEI when infected with SARS-CoV-2. Elucidating the genetic basis of IEI patients with severe/critical COVID-19 may help to develop better strategies for prevention and treatment of severe COVID-19 disease and complications in pediatric patients., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)