Lacticaseibacillus rhamnosus HF01 fermented yogurt alleviated high-fat diet-induced obesity and hepatic steatosis via the gut microbiota-butyric acid-hepatic lipid metabolism axis.

The objective of this study was to investigate the anti-obesity effects and underlying mechanism of Lacticaseibacillus rhamnosus HF01 fermented yogurt (HF01-Y). Herein, obesity was induced in mice through a high-fat diet and the changes in the gut microbiota were evaluated using 16S rRNA gene sequencing, combined with the expression levels of the liver AMPK signaling pathway to analyze the potential relationship between HF01-Y-mediated gut microbiota and obesity. The results showed that supplementation with HF01-Y improved obesity-related phenotypes in mice, including reduced body weight, improved serum lipid profiles, and decreased hepatic lipid droplet formation. In addition, HF01-Y altered the composition of the gut microbiota in obese mice, significantly upregulated norank_f__Muribaculaceae, unclassified_c__Clostridia, Blautia, unclassified_o__Bacteroidales, and Rikenellaceae_RC9_gut_group, while downregulating unclassified_f__Desulfovibrionaceae, Colidextribacter, and unclassified_f__Oscillospiraceae. These alterations led to an increase of the cecum butyric acid content, which in turn indirectly promoted the activation of the AMPK signaling pathway, subsequently, inhibited fat synthesis, and promoted fatty acid oxidation related gene expression. Therefore, HF01-Y was likely to alleviate hepatic fat and relieve obesity by modulating the gut microbiota-butyric acid-hepatic lipid metabolism axis, ultimately promoting host health.

Dynamics of antibiotic resistance genes in the sediments of a water-diversion lake and its human exposure risk behaviour.

The dynamics and exposure risk behaviours of antibiotic resistance genes (ARGs) in the sediments of water-diversion lakes remain poorly understood. In this study, spatiotemporal investigations of ARG profiles in sediments targeting non-water (NWDP) and water diversion periods (WDP) were conducted in Luoma Lake, a typical water-diversion lake, and an innovative dynamics-based risk assessment framework was constructed to evaluate ARG exposure risks to local residents. ARGs in sediments were significantly more abundant in the WDP than in the NWDP, but there was no significant variation in their spatial distribution in either period. Moreover, the pattern of ARG dissemination in sediments was unchanged between the WDP and NWDP, with horizontal gene transfer (HGT) and vertical gene transfer (VGT) contributing to ARG dissemination in both periods. However, water diversion altered the pattern in lake water, with HGT and VGT in the NWDP but only HGT in the WDP, which were critical pathways for the dissemination of ARGs. The significantly lower ARG sediment-water partition coefficient in the WDP indicated that water diversion could shift the fate of ARGs and facilitate their aqueous partitioning. Risk assessment showed that all age groups faced a higher human exposure risk of ARGs (HERA) in the WDP than in the NWDP, with the 45-59 age group having the highest risk. Furthermore, HERA increased overall with the bacterial carrying capacity in the local environment and peaked when the carrying capacity reached three (NWDP) or four (WDP) orders of magnitude higher than the observed bacterial population. HGT and VGT promoted, whereas ODF covering gene mutation and loss mainly reduced HERA in the lake. As the carrying capacity increased, the relative contribution of ODF to HERA remained relatively stable, whereas the dominant mechanism of HERA development shifted from HGT to VGT.

Predicting ICU Interventions: A Transparent Decision Support Model Based on Multivariate Time Series Graph Convolutional Neural Network.

In this study, we present a novel approach for predicting interventions for patients in the intensive care unit using a multivariate time series graph convolutional neural network. Our method addresses two critical challenges: the need for timely and accurate decisions based on changing physiological signals, drug administration information, and static characteristics; and the need for interpretability in the decision-making process. Drawing on real-world ICU records from the MIMIC-III dataset, we demonstrate that our approach significantly improves upon existing machine learning and deep learning methods for predicting two targeted interventions, mechanical ventilation and vasopressors. Our model achieved an accuracy improvement from 81.6% to 91.9% and a F1 score improvement from 0.524 to 0.606 for predicting mechanical ventilation interventions. For predicting vasopressor interventions, our model achieved an accuracy improvement from 76.3% to 82.7% and a F1 score improvement from 0.509 to 0.619. We also assessed the interpretability by performing an adjacency matrix importance analysis, which revealed that our model uses clinically meaningful and appropriate features for prediction. This critical aspect can help clinicians gain insights into the underlying mechanisms of interventions, allowing them to make more informed and precise clinical decisions. Overall, our study represents a significant step forward in the development of decision support systems for ICU patient care, providing a powerful tool for improving clinical outcomes and enhancing patient safety.

A snail species identification method based on deep learning in food safety.

In daily life, snail classification is an important mean to ensure food safety and prevent the occurrence of situations that toxic snails are mistakenly consumed. However, the current methods for snail classification are mostly based on manual labor, which is inefficient. Therefore, a snail detection and classification method based on improved YOLOv7 was proposed in this paper. First, in order to reduce the FLOPs of the model, the backbone of the original model was improved. Specifically, the original 3×3 regular convolution was replaced with 3×3 partial convolution, and the Conv2D_BN_SiLU module in the partial convolution was replaced with the Conv2D_BN_FReLU module. FReLU could enhance the model's representational capacity without increasing the number of parameters. Then, based on the specific features of snail images, in order to solve the problems of small and dense targets of diverse shapes, a receptive field enhancement module was added to the head to learn the different receptive fields of the feature maps and enhance the feature pyramid representation. In addition, the CIoU was replaced with the WIoU to make the model pay more attention to targets at the edge or difficult-to-regress accurate bounding boxes. Finally, the images of nine common types of snails were collected, including the Pomacea canaliculata, the Viviparidae, the Nassariidae, and so on. These images were then labeled using LabelImg software to create a snail image dataset. Experiments were conducted based on the dataset, and the results showed that the proposed method demonstrated the best performance compared to other state-of-the-art methods.

Link between triglyceride-glucose-body mass index and future stroke risk in middle-aged and elderly chinese: a nationwide prospective cohort study.

OBJECTIVE: Current literature is deficient in robust evidence delineating the correlation between the triglyceride glucose-body mass index (TyG-BMI) and the incidence of stroke. Consequently, this investigation seeks to elucidate the potential link between TyG-BMI and stroke risk in a cohort of middle-aged and senior Chinese individuals. METHODS: This study employs longitudinal data from four waves of the China Health and Retirement Longitudinal Study (CHARLS) conducted in 2011, 2013, 2015, and 2018, encompassing 8,698 participants. The CHARLS cohort was assembled using a multistage probability sampling technique. Participants underwent comprehensive evaluations through standardized questionnaires administered via face-to-face interviews. Our analytic strategy involved the application of Cox proportional hazards regression models to investigate the association between TyG-BMI and the risk of stroke. To discern potential non-linear relationships, we incorporated Cox proportional hazards regression with smooth curve fitting. Additionally, we executed a battery of sensitivity and subgroup analyses to validate the robustness of our findings. RESULTS: Our study utilized a multivariate Cox proportional hazards regression model and found a significant correlation between the TyG-BMI and the risk of stroke. Specifically, a 10-unit increase in TyG-BMI corresponded to a 4.9% heightened risk of stroke (HR = 1.049, 95% CI 1.029-1.069). The analysis also uncovered a non-linear pattern in this relationship, pinpointed by an inflection point at a TyG-BMI value of 174.63. To the left of this inflection point-meaning at lower TyG-BMI values-a 10-unit hike in TyG-BMI was linked to a more substantial 14.4% rise in stroke risk (HR 1.144; 95% CI 1.044-1.253). Conversely, to the right of the inflection point-at higher TyG-BMI values-each 10-unit increment was associated with a smaller, 3.8% increase in the risk of stroke (HR 1.038; 95% CI 1.016-1.061). CONCLUSIONS: In the middle-aged and elderly Chinese population, elevated TyG-BMI was significantly and positively associated with stroke risk. In addition, there was also a specific non-linear association between TyG-BMI and stroke (inflection point 174.63). Further reduction of TyG-BMI below 174.63 through lifestyle changes and dietary control can significantly reduce the risk of stroke.

Discovery of Unique Bis-Substituted Aromatic Amide Derivatives as Novel Highly Potent Antibiotics for Combating Methicillin-Resistant Staphylococcus aureus (MRSA).

Due to the increasing antibiotic resistance, developing novel antimicrobials to fight infections caused by resistant bacteria is imperative. Herein, a series of novel bis-substituted aromatic amides were designed and synthesized through modifying the hit compound 1, and their antimicrobial activities were evaluated. Among them, compound 4t, as the most potent lead, exhibited excellent antimicrobial activities against Gram-positive bacteria, including clinical methicillin-resistant Staphylococcus aureus (MRSA) isolates, while keeping weak hemolytic and mammalian cytotoxic activities. Furthermore, compound 4t displayed rapid bactericidal capabilities, low tendency to produce resistance, and favorable capacities to destroy bacterial biofilms. Further explorations indicated that compound 4t induces bacterial death by binding to cardiolipin (CL) on the bacterial membrane, disrupting the cell membrane, and facilitating the accumulation of reactive oxygen species (ROS). Additionally, compound 4t showed remarkable anti-MRSA activity in vivo, demonstrating compound 4t could be developed as a potential candidate to combat MRSA infections.

Limosilactobacillus fermentum HF06-derived paraprobiotic and postbiotic alleviate intestinal barrier damage and gut microbiota disruption in mice with ulcerative colitis.

BACKGROUND: Paraprobiotics and postbiotics have shown potential in the treatment of ulcerative colitis (UC). However, their in vivo application is still in its infancy and their mechanisms of action are not well understood. RESULTS: Here, we investigated the mitigation effects of Limosilactobacillus fermentum HF06-derived paraprobiotic (6-PA) and postbiotic (6-PS) on dextran sulfate sodium induced UC and the potential mechanisms. Results indicated that the administration of 6-PA and 6-PS resulted in the inhibition of weight loss and colon shortening in mice with UC. Furthermore, they led to a significant reduction in both fecal moisture content and the levels of proinflammatory cytokines and oxidative stress in the intestine of the mice. 6-PA and 6-PS treatment strengthened the intestinal mucosal barrier by dramatically upregulating the levels of zonula occludens-1 and occludin proteins. In addition, 6-PA and 6-PS restored intestinal dysbiosis by regulating abundances of certain bacteria, such as Bifidobacterium, Faecalibaculum, Muribaculaceae, Corynebacterium, Escherichia-Shigella and Clostridium_sensu_stricto_1, and regulated the level of short-chain fatty acids. CONCLUSION: These findings illustrated for the first time that L. fermentum HF06-derived paraprobiotic and postbiotic enhanced the intestinal barrier function, and restored gut microbiota alterations. © 2023 Society of Chemical Industry.

Genetic Engineering and Biosynthesis Technology: Keys to Unlocking the Chains of Phage Therapy.

Phages possess the ability to selectively eliminate pathogenic bacteria by recognizing bacterial surface receptors. Since their discovery, phages have been recognized for their potent bactericidal properties, making them a promising alternative to antibiotics in the context of rising antibiotic resistance. However, the rapid emergence of phage-resistant strains (generally involving temperature phage) and the limited host range of most phage strains have hindered their antibacterial efficacy, impeding their full potential. In recent years, advancements in genetic engineering and biosynthesis technology have facilitated the precise engineering of phages, thereby unleashing their potential as a novel source of antibacterial agents. In this review, we present a comprehensive overview of the diverse strategies employed for phage genetic engineering, as well as discuss their benefits and drawbacks in terms of bactericidal effect.

Phenotypic Characterization and Comparative Genomic Analyses of Mycobacteriophage WIVsmall as A New Member Assigned to F1 Subcluster.

In this study, we conducted the morphological observation, biological and genomic characterization, evolutionary analysis, comparative genomics description, and proteome identification of a recently isolated mycobacteriophage, WIVsmall. Morphologically, WIVsmall is classified as a member of the Siphoviridae family, characterized by a flexible tail, measuring approximately 212 nm in length. The double-stranded phage genome DNA of WIVsmall spans 53,359 base pairs, and exhibits a G + C content of 61.01%. The genome of WIVsmall comprises 103 protein-coding genes, while no tRNA genes were detected. The genome annotation unveiled the presence of functional gene clusters responsible for mycobacteriophage assembly and maturation, replication, cell lysis, and functional protein synthesis. Based on the analysis of the phylogenetic tree, the genome of WIVsmall was classified as belonging to subgroup F1. A comparative genomics analysis indicated that the WIVsmall genome exhibited the highest similarity to the phage SG4, with a percentage of 64%. The single-step growth curve analysis of WIVsmall revealed a latent period of 120 min, and an outbreak period of 200 min.

Design of High-Capacity MoS3 Decorated Nitrogen Doped Carbon Coated Cu2 S Electrode Structures with Dual Heterogenous Interfaces for Outstanding Sodium-Ion Storage.

The hierarchical Cu2 S@NC@MoS3 heterostructures have been firstly constructed by the high-capacity MoS3 and high-conductive N-doped carbon to co-decorate the Cu2 S hollow nanospheres. During the heterostructure, the middle N-doped carbon layer as the linker facilitates the uniform deposition of MoS3 and enhances the structural stability and electronic conductivity. The popular hollow/porous structures largely restrain the big volume changes of active materials. Due to the cooperative effect of three components, the new Cu2 S@NC@MoS3 heterostructures with dual heterogenous interfaces and small voltage hysteresis for sodium ion storage display a high charge capacity (545 mAh g-1 for 200 cycles at 0.5 A g-1 ), excellent rate capability (424 mAh g-1 at 15 A g-1 ) and ultra-long cyclic life (491 mAh g-1 for 2000 cycles at 3 A g-1 ). Except for the performance test, the reaction mechanism, kinetics analysis, and theoretical calculation have been performed to explain the reason of excellent electrochemical performance of Cu2 S@NC@MoS3 . The rich active sites and rapid Na+ diffusion kinetics of this ternary heterostructure is beneficial to the high efficient sodium storage. The assembled full cell matched with Na3 V2 (PO4 )3 @rGO cathode likewise displays remarkable electrochemical properties. The outstanding sodium storage performances of Cu2 S@NC@MoS3 heterostructures indicate the potential applications in energy storage fields.

Lactobacillus plantarum HF02 alleviates lipid accumulation and intestinal microbiota dysbiosis in high-fat diet-induced obese mice.

BACKGROUND: Obesity is closely associated with lipid accumulation and intestinal microbiota dysbiosis. It has been proved that probiotics supplement contributes to alleviate obesity. The objective of this study was to investigate the mechanism by which Lactobacillus plantarum HF02 (LP-HF02) alleviated lipid accumulation and intestinal microbiota dysbiosis in high-fat diet-induced obese mice. RESULTS: Our results showed that LP-HF02 ameliorated body weight, dyslipidemia, liver lipid accumulation, and liver injury in obese mice. As expected, LP-HF02 inhibited pancreatic lipase activity in small intestinal contents and increased fecal triglyceride levels, thereby reducing dietary fat hydrolysis and absorption. Moreover, LP-HF02 ameliorated the intestinal microbiota composition, as evidenced by the enhanced ratio of Bacteroides to Firmicutes, the decreased abundance of pathogenic bacteria (including Bacteroides, Alistipes, Blautia, and Colidextribacter) and the increased abundance of beneficial bacteria (including Muribaculaceae, Akkermansia, Faecalibaculum, and Rikenellaceae_RC9_gut_group). LP-HF02 also increased fecal short-chain fatty acids (SCFAs) levels and colonic mucosal thickness, and subsequently decreased serum lipopolysaccharide (LPS), interleukin-1ß (IL-1ß), and tumor necrosis factor-α (TNF-α) levels in obese mice. Additionally, reverse transcription quantitative polymerase chain reaction (RT-qPCR) and Western blot results demonstrated that LP-HF02 ameliorated hepatic lipid accumulation via activating the adenosine monophosphate (AMP)-activated protein kinase (AMPK) pathway. CONCLUSION: Therefore, our results indicated that LP-HF02 could be considered as a probiotic preparation for preventing obesity. © 2023 Society of Chemical Industry.

Influence of storage time on protein composition and simulated digestion of UHT milk and centrifugation presterilized UHT milk in vitro.

The centrifugation presterilizing UHT (C-UHT) sterilization method removes 90% of the microorganism and somatic cells from raw milk using high-speed centrifugation following UHT treatment. This study aimed to study the changes in protein composition and plasmin in the UHT and C-UHT milk. The digestive characteristics, composition, and peptide spectrum of milk protein sterilized with the 2 technologies were studied using a dynamic digestive system of a simulated human stomach. The Pierce bicinchoninic acid assay, laser scanning confocal microscope, liquid chromatography-tandem mass spectrometry, and AA analysis were used to study the digestive fluid at different time points of gastric digestion in vitro. The results demonstrated that C-UHT milk had considerably higher protein degradation than UHT milk. Different processes resulted during the cleavage of milk proteins at different sites during digestion, resulting in different derived peptides. The results showed there was no significant effect of UHT and C-UHT on the peptide spectrum of milk proteins, but C-UHT could release relatively more bioactive peptides and free AA.

Systematic Characterization and Regulatory Role of lncRNAs in Asian Honey Bees Responding to Microsporidian Infestation.

Long noncoding RNAs (lncRNAs) are pivotal regulators in gene expression and diverse biological processes, such as immune defense and host-pathogen interactions. However, little is known about the roles of lncRNAs in the response of the Asian honey bee (Apis cerana) to microsporidian infestation. Based on our previously obtained high-quality transcriptome datasets from the midgut tissues of Apis cerana cerana workers at 7 days post inoculation (dpi) and 10 dpi with Nosema ceranae (AcT7 and AcT10 groups) and the corresponding un-inoculated midgut tissues (AcCK7 and AcCK10 groups), the transcriptome-wide identification and structural characterization of lncRNAs were conducted, and the differential expression pattern of lncRNAs was then analyzed, followed by investigation of the regulatory roles of differentially expressed lncRNAs (DElncRNAs) in host response. Here, 2365, 2322, 2487, and 1986 lncRNAs were, respectively, identified in the AcCK7, AcT7, AcCK7, and AcT10 groups. After removing redundant ones, a total of 3496 A. c. cerana lncRNAs were identified, which shared similar structural characteristics with those discovered in other animals and plants, such as shorter exons and introns than mRNAs. Additionally, 79 and 73 DElncRNAs were screened from the workers' midguts at 7 dpi and 10 dpi, respectively, indicating the alteration of the overall expression pattern of lncRNAs in host midguts after N. ceranae infestation. These DElncRNAs could, respectively, regulate 87 and 73 upstream and downstream genes, involving a suite of functional terms and pathways, such as metabolic process and Hippo signaling pathway. Additionally, 235 and 209 genes co-expressed with DElncRNAs were found to enrich in 29 and 27 terms, as well as 112 and 123 pathways, such as ABC transporters and the cAMP signaling pathway. Further, it was detected that 79 (73) DElncRNAs in the host midguts at 7 (10) dpi could target 321 (313) DEmiRNAs and further target 3631 (3130) DEmRNAs. TCONS_00024312 and XR_001765805.1 were potential precursors for ame-miR-315 and ame-miR-927, while TCONS_00006120 was the putative precursor for both ame-miR-87-1 and ame-miR-87-2. These results together suggested that DElncRNAs are likely to play regulatory roles in the host response to N. ceranae infestation through the regulation of neighboring genes via a cis-acting effect, modulation of co-expressed mRNAs via trans-acting effect, and control of downstream target genes' expression via competing endogenous RNA networks. Our findings provide a basis for disclosing the mechanism underlying DElncRNA-mediated host N. ceranae response and a new perspective into the interaction between A. c. cerana and N. ceranae.

Tandem mass tag-based quantitative proteomics analysis reveals the effects of the α-lactalbumin peptides GINY and DQW on lipid deposition and oxidative stress in HepG2 cells.

The objective of this study was to investigate the mechanism by which the α-lactalbumin peptides Gly-Ile-Asn-Tyr (GINY) and Asp-Gln-Trp (DQW) ameliorate free fatty acid-induced lipid deposition in HepG2 cells. The results show that GINY and DQW reduced triglyceride, total cholesterol, and free fatty acid levels significantly in free fatty acid-treated HepG2 cells. Based on proteomic analysis, GINY and DQW alleviated lipid deposition and oxidative stress mainly through the peroxisome proliferator-activated receptor (PPAR) pathway, fatty acid metabolism, oxidative phosphorylation, and response to oxidative stress. In vitro experiments confirmed that GINY and DQW upregulated the mRNA and protein expression of fatty acid ß-oxidation-related and oxidative stress-related genes, and downregulated the mRNA and protein expression of lipogenesis-related genes by activating peroxisome proliferator-activated receptor α (PPARα). Meanwhile, GINY and DQW reduced free fatty acid-induced lipid droplet accumulation and reactive oxygen species generation, and enhanced the mitochondrial membrane potential and ATP levels. Furthermore, GINY and DQW enhanced carnitine palmitoyl-transferase 1a (CPT-1a) and superoxide dismutase activities, and diminished acetyl-coenzyme A carboxylase 1 (ACC1) and fatty acid synthase (FASN) activities in a PPARα-dependent manner. Interestingly, GW6471 (a PPARα inhibitor) weakened the effects of GINY and DQW on the PPARα pathway. Hence, our findings suggest that GINY and DQW have the potential to alleviate nonalcoholic fatty liver disease by activating the PPARα pathway.

Triglyceride glucose-body mass index and the risk of progression to diabetes from prediabetes: A 5-year cohort study in Chinese adults.

Objective: Evidence regarding the relationship between the triglyceride glucose-body mass index (TyG-BMI) and the risk of progression from prediabetes to diabetes remains limited. Our study aimed to investigate the relationship between them in patients with prediabetes. Methods: In this retrospective cohort study, data were collected from 25,279 patients with prediabetes who received health checks between 2010 and 2016. We used a Cox proportional-hazards regression model to examine the relationship between TyG-BMI and diabetes risk. We used Cox proportional hazards regression with cubic spline functions and smooth curve fitting to identify the nonlinear relationship between them. In addition, A series of sensitivity and subgroup analyses were also conducted. Results: The mean age of the included participants was 49.29 ± 13.82 years old, and 1,6734 (66.2%) were male. The mean TyG-BMI was 219.47. The median follow-up time was 2.89 years, and 2,687 (10.63%) individuals had a final diagnosis of diabetes. After adjusting for covariates, TyG-BMI was positively linked with incident diabetes in patients with prediabetes (HR = 1.011, 95%CI 1.010-1.012). TyG-BMI had a non-linear connection with diabetes risk, and its inflection point was 231.66. Right and left effects sizes (HR) at the inflection point were 1.017 (95%CI:1.014-1.019) and 1.007 (95%CI:1.005-1.009), respectively. The sensitivity analysis demonstrated the robustness of these results. Conclusion: This study demonstrated a positive, non-linear relationship between the TyG-BMI and diabetes risk in Chinese patients with prediabetes. When the TyG-BMI was <231.66, there was a significant positive association between TyG-BMI and the risk of progression from prediabetes to diabetes. This study serves as a reference to promote clinical consultation and optimize diabetes prevention decisions for patients with prediabetes.

Large-pose facial makeup transfer based on generative adversarial network combined face alignment and face parsing.

Facial makeup transfer is a special form of image style transfer. For the reference makeup image with large-pose, improving the quality of the image generated after makeup transfer is still a challenging problem worthy of discussion. In this paper, a large-pose makeup transfer algorithm based on generative adversarial network (GAN) is proposed. First, a face alignment module (FAM) is introduced to locate the key points, such as the eyes, mouth and skin. Secondly, a face parsing module (FPM) and face parsing losses are designed to analyze the source image and extract the face features. Then, the makeup style code is extracted from the reference image and the makeup transfer is completed through integrating facial features and makeup style code. Finally, a large-pose makeup transfer (LPMT) dataset is collected and constructed. Experiments are carried out on the traditional makeup transfer (MT) dataset and the new LPMT dataset. The results show that the image quality generated by the proposed method is better than that of the latest method for large-pose makeup transfer.

A robust and high-precision edge segmentation and refinement method for high-resolution images.

Limited by GPU memory, high-resolution image segmentation is a highly challenging task. To improve the accuracy of high-resolution segmentation, High-Resolution Refine Net (HRRNet) is proposed. The network is divided into a rough segmentation module and a refinement module. The former improves DeepLabV3+ by adding the shallow features of 1/2 original image size and the corresponding skip connection to obtain better rough segmentation results, the output of which is used as the input of the latter. In the refinement module, first, the global context information of the input image is obtained by a global process. Second, the high-resolution image is divided into patches, and each patch is processed separately to obtain local details in a local process. In both processes, multiple refine units (RU) are cascaded for refinement processing, and two cascaded residual convolutional units (RCU) are added to the different output paths of RU to improve the mIoU and the convergence speed of the network. Finally, according to the context information of the global process, the refined patches are pieced to obtain the refined segmentation result of the whole high-resolution image. In addition, the regional non-maximum suppression is introduced to improve the Sobel edge detection, and the Pascal VOC 2012 dataset is enhanced, which improves the segmentation accuracy and robust performance of the network. Compared with the state-of-the-art semantic segmentation models, the experimental results show that our model achieves the best performance in high-resolution image segmentation.

Effect of Ascosphaera apis Infestation on the Activities of Four Antioxidant Enzymes in Asian Honey Bee Larval Guts.

Ascosphaera apis infects exclusively bee larvae and causes chalkbrood, a lethal fungal disease that results in a sharp reduction in adult bees and colony productivity. However, little is known about the effect of A. apis infestation on the activities of antioxidant enzymes in bee larvae. Here, A. apis spores were purified and used to inoculate Asian honey bee (Apis cerana) larvae, followed by the detection of the host survival rate and an evaluation of the activities of four major antioxidant enzymes. At 6 days after inoculation (dpi) with A. apis spores, obvious symptoms of chalkbrood disease similar to what occurs in Apis mellifera larvae were observed. PCR identification verified the A. apis infection of A. cerana larvae. Additionally, the survival rate of larvae inoculated with A. apis was high at 1−2 dpi, which sharply decreased to 4.16% at 4 dpi and which reached 0% at 5 dpi, whereas that of uninoculated larvae was always high at 1~8 dpi, with an average survival rate of 95.37%, indicating the negative impact of A. apis infection on larval survival. As compared with those in the corresponding uninoculated groups, the superoxide dismutase (SOD) and catalase (CAT) activities in the 5- and 6-day-old larval guts in the A. apis−inoculated groups were significantly decreased (p < 0.05) and the glutathione S-transferase (GST) activity in the 4- and 5-day-old larval guts was significantly increased (p < 0.05), which suggests that the inhibition of SOD and CAT activities and the activation of GST activity in the larval guts was caused by A. apis infestation. In comparison with that in the corresponding uninoculated groups, the polyphenol oxidase (PPO) activity was significantly increased (p < 0.05) in the 5-day-old larval gut but significantly reduced (p < 0.01) in the 6-day-old larval gut, indicating that the PPO activity in the larval guts was first enhanced and then suppressed. Our findings not only unravel the response of A. cerana larvae to A. apis infestation from a biochemical perspective but also offer a valuable insight into the interaction between Asian honey bee larvae and A. apis.

ame-miR-34 Modulates the Larval Body Weight and Immune Response of Apis mellifera Workers to Ascosphara apis Invasion.

MiRNAs are critical regulators of numerous physiological and pathological processes. Ascosphaera apis exclusively infects bee larvae and causes chalkbrood disease. However, the function and mechanism of miRNAs in the bee larval response to A. apis infection is poorly understood. Here, ame-miR-34, a previously predicted miRNA involved in the response of Apis mellifera larvae to A. apis invasion, was subjected to molecular validation, and overexpression and knockdown were then conducted to explore the regulatory functions of ame-miR-34 in larval body weight and immune response. Stem-loop RT-PCR and Sanger sequencing confirmed the authenticity of ame-miR-34 in the larval gut of A. mellifera. RT-qPCR results demonstrated that compared with that in the uninfected larval guts, the expression level of ame-miR-34 was significantly downregulated (p < 0.001) in the guts of A. apis-infected 4-, 5-, and 6-day-old larvae, indicative of the remarkable suppression of host ame-miR-34 due to A. apis infection. In comparison with the corresponding negative control (NC) groups, the expression level of ame-miR-34 in the larval guts in the mimic-miR-34 group was significantly upregulated (p < 0.001), while that in the inhibitor-miR-34 group was significantly downregulated (p < 0.01). Similarly, effective overexpression and knockdown of ame-miR-34 were achieved. In addition, the body weights of 5- and 6-day-old larvae were significantly increased compared with those in the mimic-NC group; the weights of 5-day-old larvae in the inhibitor-miR-34 group were significantly decreased in comparison with those in the inhibitor-NC group, while the weights of 4- and 6-day-old larvae in the inhibitor-miR-34 group were significantly increased, indicating the involvement of ame-miR-34 in modulating larval body weight. Furthermore, the expression levels of both hsp and abct in the guts of A. apis-infected 4-, 5-, and 6-day-old larvae were significantly upregulated after ame-miR-34 overexpression. In contrast, after ame-miR-34 knockdown, the expression levels of the aforementioned two key genes in the A. apis-infected 4-, 5-, and 6-day-old larval guts were significantly downregulated. Together, the results demonstrated that effective overexpression and knockdown of ame-miR-34 in both noninfected and A. apis-infected A. mellifera larval guts could be achieved by the feeding method, and ame-miR-34 exerted a regulatory function in the host immune response to A. apis invasion through positive regulation of the expression of hsp and abct. Our findings not only provide a valuable reference for the functional investigation of bee larval miRNAs but also reveal the regulatory role of ame-miR-34 in A. mellifera larval weight and immune response. Additionally, the results of this study may provide a promising molecular target for the treatment of chalkbrood disease.

Periodontal Ligament Stem Cell-Derived Extracellular Vesicles Enhance Tension-Induced Osteogenesis.

Tension-induced osteogenesis has great significance in maintaining bone homeostasis and ensuring the efficiency and stability of orthodontic treatment. Recently, extracellular vesicles (EVs) have shown great potential in regulating bone remodeling. Here, we aimed to explore the effects of periodontal ligament stem cell (PDLSC)-derived EVs on tension-induced osteogenesis and the potential mechanism. PDLSC-derived EVs were extracted by ultracentrifugation. In vitro, PDLSC-derived EVs of 10 µg/mL significantly improved the proliferation of MC3T3-E1 cells and enhanced the osteogenic differentiation of osteoblasts under a tensile strain of 2000 uε. Next, a mouse model of orthodontic tooth movement (OTM) was established and treated with subperiosteal injection of PDLSC-derived EVs (1 mg/kg) on the tension side. The results showed that treatment with PDLSC-derived EVs effectively enhanced OTM and promoted osteogenesis on the tension side, including increasing trabecular bone parameters and promoting the expression of osteogenic-related biomarkers (OCN and OPN). More interestingly, we identified several mechano-sensitive miRNAs enriched in PDLSC-derived EVs by high-throughput miRNA sequencing. Bioinformatics analysis indicated that they were related to various osteogenesis-related signaling pathways. Therefore, PDLSC-derived EVs could improve the efficiency of OTM by enhancing tension-induced osteogenesis of osteoblasts. Our study may provide potential evidence for the promoting effects of PDLSC-derived EVs on osteogenesis and offer new insights into the development of treatment strategies for enhancing osteogenesis in orthodontic treatment and other metabolic bone diseases.