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Abstract Breast cancer is now the most deadly cancer worldwide. Mammography screening is the most effective method for early detection and diagnosis of breast cancer. Due to the lack of labeled mammograms, building an AI system for mammography screening often relies heavily on human-designed data augmentation, which doesn’t always perform robustly when applied to clinical scenarios. This paper presents a novel framework of Supervised Contrastive Pre-training followed by Supervised Fine-tuning (SCP+SF) for mammography screening. Unlike the previous approaches, the proposed supervised contrastive pre-training does not need a data augmentation module. We apply the SCP+SF framework to two challenging and important mammography screening tasks for breast cancer: mammographic abnormality screening and mammographic malignancy screening. Our extensive experiments on a large-scale dataset show that the supervised contrastive pre-training (SCP) can substantially improve the final model performance compared with the traditional direct supervised training approach. Superior results of AUC and specificity/sensitivity have been achieved on two clinically significant mammographic screening tasks in comparison with previously reported State-Of-The-Art approaches. We believe this work is the first to show that supervised contrastive pre-training (SCP) followed by supervised fine-tuning (SF) can outperform the supervised counterpart on these two critical medical imaging tasks.

IntroductionBacillus species are probiotics commonly utilized in aquaculture to enhance aquatic animal growth, inhibit pathogens, and strengthen immunity. However, research comparing the effects of probiotic bacterial cells and cultures is limited. This study aimed to evaluate the probiotic potential of Bacillus licheniformis strain HN318 and compare its impact on growth, immunity, and disease resistance in hybrid grouper when administered as bacterial cells or cultures.MethodsThe study involved assessing the auto-aggregation capability, gastrointestinal stress resistance, and safety of HN318. Enzymatic activities and antibacterial properties of HN318 cultures and bacterial cells were also compared. Hybrid grouper were fed with HN318 cultures and bacterial cells for 8 weeks, and their growth, feed utilization, immune responses, and survival rates upon challenge with Vibrio harveyi were evaluated. Additionally, the expression of immune-related genes was analyzed.ResultsHN318 exhibited high auto-aggregation and gastrointestinal stress resistance, and was found to be safe for use in aquaculture. Cultures of HN318 displayed higher protease, amylase, and antibacterial activities compared to bacterial cells. Both forms significantly improved growth and feed utilization in hybrid grouper. Notably, HN318 cultures induced higher levels of immune enzymes and activities, and provided better protection against V. harveyi challenge, with a higher relative percent survival compared to bacterial cells. Furthermore, HN318 cultures upregulated the expression of immune-related genes more than bacterial cells.DiscussionThis study highlights the potential of both HN318 cultures and bacterial cells as immune enhancers for hybrid grouper. However, HN318 cultures demonstrated superior probiotic properties, including higher enzymatic activities, antibacterial properties, and immunomodulatory effects. These findings provide new insights and references for the diverse application forms of probiotics in aquaculture, suggesting that cultures may be more effective than bacterial cells in enhancing the health and performance of aquatic animals.

Plectropomus leopardus is an economically valuable marine farmed fish. However, diseases have seriously restricted the healthy development of its breeding industry. Cell line is an important in vitro research system in the fields of disease control, environmental toxicology, nanotechnology, and so on. However, the application of marine fish cell lines is far from the expected level, and many species have not yet been reported to establish cell lines. In our study, we developed a brain cell line derived from P. leopardus, designated as PLB, and investigated its susceptibility to bacterial agents and heavy metal exposure. The findings indicated that PLB cell lines grew fastest in L-15 medium supplemented with 20 % fetal bovine serum (FBS) at 26 °C. The established cell line was identified by 18 S rRNA gene sequencing as being derived from P. leopardus. Chromosome analysis revealed that the PLB cell line had a chromosome count of 48. The transfection efficiency of PLB cells was about 27 % by liposome transfection method, indicating that PLB cell lines offer utility for conducting functional investigations of exogenous genes. In addition, the bacterial sensitivity analysis results indicated that PLB cells exhibited susceptibility to both Vibrio harveyi and Edwardsiella tarda, furthermore, PLB cells displayed heightened sensitivity towards V. harveyi in comparison to E. tarda. The outcomes from the heavy metal toxicity experiments demonstrated significant toxicity of three heavy metals (Hg2+, Cu2+ and Cd2+) towards PLB cells, with Hg2+ exhibiting the highest degree of toxicity among them. Moreover, there was a dose-dependent relationship between cell survival rate of PLB and heavy metal concentration. In summary, PLB cell lines represent a promising tool for conducting in vitro analyses of foreign gene functionality, bacterial susceptibility, and heavy metal toxicity. It helps ensure the safety of aquatic breeding environments and also supports genetic enhancements in fish fry, promoting the development of traits such as increased growth and disease resistance, which are vital for aquaculture progress.

Humpback grouper (Cromileptes altivelis) is an economically important fish, but the increasing density of its farming has led to more severe disease outbreaks. To address this challenge, we established brain (CAB) and kidney (CAK) cell lines in our laboratory previously, providing a valuable tool for in vitro studies on immune responses. In this study, we used quantitative real-time polymerase chain reaction (qRT-PCR) to identify the optimal reference gene from six reference genes for CAB and CAK cells, under both normal conditions and after stimulation with LPS or Poly I: C. The qRT-PCR data were analyzed using geNorm, NormFinder, and BestKeeper software (Version 3.5) to ensure comprehensve evaluation. The results showed that RPL13 was the most stable reference gene for both CAB and CAK cells under normal conditions. Following LPS stimulation, TTLL1 was the best reference gene for CAB cells, while RPL13 remained the most suitable for CAK cells. For Poly I: C stimulation, EF1A and Actin were identified as the most stable reference genes for CAB and CAK cells, respectively. To confirm the reliability of the selected reference genes, we analyzed the expression of the cytokine genes IL-6 and IFN-h, demonstrating the dependability of these reference genes. This study lays a solid foundation for exploring gene expression patterns in humpback grouper cell lines under various experimental conditions, providing essential insights for future research into immune processes and disease control strategies in aquaculture.

Abstract To explore the clinical application value of optical coherence microscopy (OCM) in Hirschsprung’s disease. 109 HSCR patients were recuited in a Chinese hospital from January 2018 to July 2021. All the recruited patients underwent barium enema angiography preoperatively and the resected diseased intestinal tubes were evaluated intraoperatively. The OCM and the histopathological examination were performed successively on the surgical specimens, and the OCM images were compared with the relevant tissue sections to characterize different lesions. 10 non-HSCR fetal colorectal tissues at the same period were retained for OCM, the characteristics of which with and without HSCR under OCM imaging were analyzed. In the OCM images of in vitro tissue, it can be clearly observed that the scattering degree of HSCR narrow segment mucosal is high, glands and crypt structures are reduced or even atrophy, and the scattering degree of submucosal and intermuscular is low; In the dilated segment, the low scattering and high scattering are complex, and the muscle layer is obviously hypertrophy and structural disorder. Compared with the pathological findings, the OCM sensitivity, Kappa value, and AUC area reached 92.66%, 0.63, and 0.91, respectively. OCM can quickly and clearly display the structure of all layers of colorectal tissue, which is highly consistent with the corresponding histopathological examination results and has high sensitivity. which will provide a more reliable basis for OCM diagnosis of early HSCR, targeted biopsy and location of operative treatment, and has a certain potential for clinical application.

Groupers are valuable economic fish in the southern sea area of China, but the threat of disease is becoming more and more serious. Vibrio harveyi, V. parahaemolyticus, and Singapore grouper iridovirus (SGIV) are three important pathogens that cause disease in groupers, and infection with either a single one or a mix of these pathogens poses a serious threat to the healthy development of grouper culture. To enhance the rapid diagnosis and screening in the early stages, it is necessary to develop rapid detection methods of these pathogens. To simultaneously and rapidly detect the three pathogens, in this study, we utilized the TolC of V. harveyi, DNAJ of V. parahaemolyticus, and RAD2 of SGIV as the target genes and established a triple visual loop-mediated isothermal amplification (LAMP) method. This LAMP method showed a detection time as fast as 30 min and a high sensitivity of 100 fg/μL. Moreover, this method exhibited strong specificity and no cross-reaction with seven types of Vibrio and Staphylococcus aureus, as well as five common viruses in aquatic animals. Then, the LAMP products were enzymically cut, and three characteristic strips were used to identify the pathogen species. The results of the clinical trials demonstrated that the method could accurately and specifically detect V. harveyi, V. parahaemolyticus, and SGIV in grouper tissues. In summary, this study successfully established a triple visual LAMP rapid detection method for V. harveyi, V. parahaemolyticus, and SGIV. The method offers several advantages including simple equipment, easy operation, rapid reaction, high specificity, high sensitivity, and visual results. It is suitable for the early and rapid diagnosis of groupers infected with V. harveyi, V. parahaemolyticus, and SGIV, thereby providing useful technical support for further application in the large-scale disease surveillance of aquaculture animals.

IntroductionB-cell lymphoma-2 (Bcl-2) is the first identified member of the Bcl-2 family that performs an anti-apoptotic function in mammals. However, its role in teleosts is not fully understood. In this study, Bcl-2 of Trachinotus ovatus (TroBcl2) was cloned, and its role in apoptosis was investigated.MethodsIn this study, Bcl-2 of Trachinotus ovatus (TroBcl2) was cloned by PCR. Quantitative real-time PCR (qRT-PCR) was used to detect its mRNA expression level in healthy condition and after LPS stimulation. Subcellular localization was performed by transfecting the pTroBcl2-N3 plasmid into golden pompano snout (GPS) cells and observed under an inverted fluorescence microscope DMi8 and further verified by immunoblotting. In vivo overexpression and RNAi knockdown method were performed to evaluate the role of TroBcl2 in apoptosis. The anti-apoptotic activity of TroBcl2 was detected by flow cytometry. The effect of TroBcl2 on the mitochondrial membrane potential (MMP) was measured by an enhanced mitochondrial membrane potential assay kit with JC-1. The terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) method was performed to evaluate the role of TroBcl2 in the DNA fragmentation. Immunoblotting was used to verify whether TroBcl2 inhibits the release of cytochrome c from mitochondria into the cytoplasm. The Caspase 3 and Caspase 9 Activity Assay Kits were used to investigate the effect of TroBcl2 on caspase 3 and caspase 9 activities. The effects of TroBcl2 on the expression of apoptosis-related and nuclear factor- κB (NF-κB) signaling pathway-related genes in vitro were evaluated by qRT-PCR and Enzyme linked immunosorbent assay (ELISA). Luciferase reporter assay was used to evaluate the activity in NF-κB signaling pathway.Results and discussionThe full-length coding sequence of TroBcl2 contains 687 bp and encodes a protein containing 228 amino acids. Four conserved Bcl-2 homology (BH) domains and one invariant “NWGR” motif located in BH1 were identified in TroBcl2. In healthy T. ovatus, TroBcl2 was widely distributed in the eleven tested tissues, and higher expression levels were found in immune-related tissues, such as spleen and head kidney tissues. After stimulation with lipopolysaccharide (LPS), the expression of TroBcl2 in the head kidney, spleen, and liver was significantly upregulated. In addition, subcellular localization analysis revealed that TroBcl2 was localized in both the cytoplasm and nucleus. Functional experiments showed that TroBcl2 inhibited apoptosis, possibly by reducing mitochondrial membrane potential loss, decreasing DNA fragmentation, preventing cytochrome c release into cytoplasm, and reducing the caspase 3 and caspase 9 activations. Moreover, upon LPS stimulation, overexpression of TroBcl2 suppressed the activation of several apoptosis-related genes, such as BOK, caspase-9, caspase-7, caspase-3, cytochrome c, and p53. Furthermore, knockdown of TroBcl2 significantly increased the expression of those apoptosis-related genes. In addition, TroBcl2 overexpression or knockdown induced or inhibited, respectively, the transcription of NF-κB and regulated the expression of genes (such as NF-κB1 and c-Rel) in the NF-κB signaling pathway as well as the expression of the downstream inflammatory cytokine IL-1β. Overall, our study suggested that TroBcl2 performs its conserved anti-apoptotic function via the mitochondrial pathway and may serve as an anti-apoptotic regulator in T. ovatus.

IntroductionThe transcription factor interferon regulatory factor 3 (IRF3) plays an important role in host defence against viral infections. However, its role during bacterial infection in teleosts remains unclear. In the present study, we evaluated the antibacterial effects of Trachinotus ovatus IRF3 (TroIRF3) and how it regulates type I interferon (IFN).MethodsSubcellular localisation experiments, overexpression, and quantitative real-time PCR (qRT-PCR) were performed to examine the nuclear localisation signal (NLS) of TroIRF3 and its role in the antibacterial regulatory function of TroIRF3. We assessed the binding activity of TroIRF3 to the IFNa3 promoter by luciferase reporter assay.Results and DiscussionThe results showed that TroIRF3 was constitutively expressed at high levels in the gill and liver. TroIRF3 was significantly upregulated and transferred from the cytoplasm to the nucleus after Vibrio harveyi infection. By overexpressing TroIRF3, the fish were able to inhibit the replication of V. harveyi, whereas knocking it down increased bacterial replication. Moreover, the overexpression of TroIRF3 increased type I interferon (IFNa3) production and the IFN signalling molecules. The NLS, which is from the 64–127 amino acids of TroIRF3, contains the basic amino acids KR74/75 and RK82/84. The results proved that NLS is required for the efficient nuclear import of TroIRF3 and that the NLS domain of TroIRF3 consists of the key amino acids KR74/75 and RK82/84. The findings also showed that NLS plays a key role in the antibacterial immunity and upregulation of TroIFNa3 induced by TroIRF3. Moreover, TroIRF3 induces TroIFNa3 promoter activity, whereas these effects are inhibited when the NLS domain is deficient. Overall, our results suggested that TroIRF3 is involved in the antibacterial immunity and regulation of type I IFN in T. ovatus and that the NLS of TroIRF3 is vital for IRF3-mediated antibacterial responses, which will aid in understanding the immune role of fish IRF3.

IntroductionInsulin-like growth factor binding protein 5 (IGFBP5) exerts an essential biological role in many processes, including apoptosis, cellular differentiation, growth, and immune responses. However, compared to mammalians, our knowledge of IGFBP5 in teleosts remains limited.MethodsIn this study, TroIGFBP5b, an IGFBP5 homologue from golden pompano (Trachinotus ovatus) was identified. Quantitative real-time PCR (qRT-PCR) was used to check its mRNA expression level in healthy condition and after stimulation. In vivo overexpression and RNAi knockdown method were performed to evaluate the antibacterial profile. We constructed a mutant in which HBM was deleted to better understand the mechanism of its role in antibacterial immunity. Subcellular localization and nuclear translocation were verified by immunoblotting. Further, proliferation of head kidney lymphocytes (HKLs) and phagocytic activity of head kidney macrophages (HKMs) were detected through CCK-8 assay and flow cytometry. Immunofluorescence microscopy assay (IFA) and dual luciferase reporter (DLR) assay were used to evaluate the activity in nuclear factor-κB (NF-κβ) pathway.ResultsThe TroIGFBP5b mRNA expression level was upregulated after bacterial stimulation. In vivo, TroIGFBP5b overexpression significantly improved the antibacterial immunity of fish. In contrast, TroIGFBP5b knockdown significantly decreased this ability. Subcellular localization results showed that TroIGFBP5b and TroIGFBP5b-δHBM were both present in the cytoplasm of GPS cells. After stimulation, TroIGFBP5b-δHBM lost the ability to transfer from the cytoplasm to the nucleus. In addition, rTroIGFBP5b promoted the proliferation of HKLs and phagocytosis of HKMs, whereas rTroIGFBP5b-δHBM, suppressed these facilitation effects. Moreover, the in vivo antibacterial ability of TroIGFBP5b was suppressed and the effects of promoting expression of proinflammatory cytokines in immune tissues were nearly lost after HBM deletion. Furthermore, TroIGFBP5b induced NF-κβ promoter activity and promoted nuclear translocation of p65, while these effects were inhibited when the HBM was deleted.DiscussionTaken together, our results suggest that TroIGFBP5b plays an important role in golden pompano antibacterial immunity and activation of the NF-κβ signalling pathway, providing the first evidence that the HBM of TroIGFBP5b plays a critical role in these processes in teleosts.

Pathological conditions in diabetic feet cause surface temperature variations, which can be captured quantitatively using infrared thermography. Thermal images captured during recovery of diabetic feet after active cooling may reveal richer information than those from passive thermography, but diseased foot regions may exhibit very small temperature differences compared with the surrounding area, complicating plantar foot segmentation in such cold-stressed active thermography. In this study, we investigate new plantar foot segmentation methods for thermal images obtained via cold-stressed active thermography without the complementary information from color or depth channels. To better deal with the temporal variations in thermal image contrast when planar feet are recovering from cold immersion, we propose an image pre-processing method using a two-stage adaptive gamma transform to alleviate the impact of such contrast variations. To improve upon existing deep neural networks for segmenting planar feet from cold-stressed infrared thermograms, a new deep neural network, the Plantar Foot Segmentation Network (PFSNet), is proposed to better extract foot contours. It combines the fundamental U-shaped network structure, a multi-scale feature extraction module, and a convolutional block attention module with a feature fusion network. The PFSNet, in combination with the two-stage adaptive gamma transform, outperforms multiple existing deep neural networks in plantar foot segmentation for single-channel infrared images from cold-stressed infrared thermography, achieving an accuracy of 97.3% and 95.4% as measured by Intersection over Union (IOU) and Dice Similarity Coefficient (DSC) respectively.

The corrosion of offshore reinforced concrete structures under combined chloride and sulfate ions attack is a complex corrosion phenomenon. The purpose of this study is to develop numerical models in conjunction with experimental studies for long-term durability assessment of offshore concrete structures. A numerical model was developed to describe the simultaneously transport of chloride and sulfate ions considering the competitive binding by hardened cement pastes effect. The model was validated by a series of experimental studies. The validated model was implemented to predict the service life of the offshore RC structures under various chloride and sulfate solutions. The results show that chloride ions induced corrosion of steel bars in offshore RC structures is highly influenced by the concentration of sulfate ions. The sulfate ions induced concrete expansion and cracking from ettringite formation could potentially accelerate chloride ions induced corrosion of steel bars in concrete, ultimately the premature failure of the offshore RC structures.

The spread of invasive species (IS) has the potential to upset ecosystem balances. In extreme cases, this can hinder economical utilization of both aquatic (fisheries) and terrestrial (agricultural) systems. As a result, many countries regard risk assessment of IS as an important process for solving the problem of biological invasion. Yet, some IS are purposefully introduced for what is seen as their potential economic benefits. Thus, conducting IS risk assessments and then formulating policies based on scientific information will allow protocols to be developed that can reduce problems associated with IS incursions, whether occurring purposefully or not. However, the risk assessment methods currently adopted by most countries use qualitative or semiquantitative methodologies. Currently, there is a mismatch between qualitative and quantitative assessments. Moreover, most assessment systems are for terrestrial animals. What is needed is an assessment system for aquatic animals; however, those currently available are relatively rudimentary. To fill this gap, we used the analytic hierarchy process (AHP) to build a risk assessment model system for aquatic IS. Our AHP has four primary indexes, twelve secondary indexes, and sixty tertiary indexes. We used this AHP to conduct quantitative risk assessments on five aquatic animals that are typically introduced in China, which have distinct biological characteristics, specific introduction purposes, and can represent different types of aquatic animals. The assessment results show that the risk grade for Pterygoplichthys pardalis is high; the risk grade for Macrobrachium rosenbergii, Crassostrea gigas, and Trachemys scripta elegans is medium; and the grade risk for Ambystoma mexicanum is low. Risk assessment of the introduction of aquatic animals using our AHP is effective, and it provides support for the introduction and healthy breeding of aquatic animals. Thus, the AHP model can provide a basis for decision-making risk management concerning the introduction of species.

The variability in characteristics of concrete material, such as pore size and chemical composition, has significant impact on the long-term performance of concrete structures under sulfate attack. This paper presents a reliability-based numerical model for assessing the long-term corrosion behaviour of concrete subject to sulfate attack. Using a reliability approach, the model can describe the reactive transport behaviour of sulfate ions and concrete degradation process. Specially, the variables in pore characteristics in interfacial transition zone (ITZ) and concrete chemical composition were quantified using a large amount of experimental data to determine the time-dependent reliability index (β) under long-term sulfate attack. The results show that the filling of ettringite and monosulfate after sulfate attack could modify the physical pore structure of ITZ at early stage. The time for concrete to reach the reliability index limit state shorten remarkably with the increase of sulfate concentration. For example, a four-fold increase of sulfate concentration (e.g. from 57 mol/m3 to 14.25 mol/m3) could shorten the time to reach the reliability index limit (i.e. β = 4.7) by 70 %. In addition, aggregate surface treatment especially with silica fume could improve the reliability of deteriorating concrete significantly by prolonging the time to reach the reliability index limit by 40 % in this study.

The golden pompano snout (GPS) and head kidney (GPHK) cell lines have been proven to be meaningful tools for the study on pathogenic infections in vitro. In this study, we aimed to select the most stable reference genes from seven housekeeping genes (Actin, B2M, GAPDH, RPL13, EF1A, 18S and UBCE) applied to two cell lines of golden pompano (GPS and GPHK) under both normal physiological conditions and stimulated conditions of the lipopolysaccharide (LPS) or polyinosinic:polycytidylic acid (Poly I:C) relying on quantitative real-time PCR (qRT-PCR). Additionally, the raw Ct value resulting from the qRT-PCR was analyzed by the geNorm, NormFinder and BestKeeper algorithm, and the results indicated that expression for all candidate genes exhibited some discrepancy under different experimental conditions or cell types. As for the non-stimulated group, 18S and RPL13 were the most appropriate reference genes in GPS and GPHK cells, respectively. Nevertheless, the most suitable reference genes in GPS and GPHK cells, under the stimulation of LPS, were RPL13 and 18S, respectively, whereas after being stimulated with Poly I:C, UBCE and EF1A were recommended as the optimal candidates for GPS and GPHK cells, respectively. To be sure of the reliability of the selected reference genes, immune-related genes (ISG15, BCL2, IRF1 and IRF7) were chosen as target genes to normalize. The study will provide a direction for various golden pompano cell lines to screen appropriate reference genes, and will set the stage for the application of these cell lines in relevant research areas.

MicroRNAs (miRNAs) could regulate various biological processes. Nervous necrosis virus (NNV) is one of the primary germs of the Humpback grouper (Cromileptes altivelis), a commercial fish of great importance for Asian aquaculture. However, there is limited available information on the host-virus interactions of C. altivelis. miRNAs have been shown to play key roles in the host response to infection by a variety of pathogens. To better understand the regulatory mechanism of miRNAs, we constructed miRNA transcriptomes and identified immune-related miRNAs of C. altivelis spleen in response to NNV infection. Reads from the three libraries were mapped onto the Danio rerio reference genome. As a result, a total of 942 mature miRNAs were determined, with 266 known miRNAs and 676 novel miRNAs. Among them, thirty-two differentially expressed miRNAs (DEmiRs) were identified compared to the PBS control. These DEmiRs were targeted on 895 genes, respectively, by using miRanda v3.3a. Then, 14 DEmiRs were validated by qRT-PCR and showed consistency with those obtained from high-throughput sequencing. In order to study the relationship between viral infection and host miRNA, a cell line from C. altivelis brain (CAB) was used to examine the expressions of five known DEmiRs (miR-132-3p, miR-194a, miR-155, miR-203b-5p, and miR-146) during NNV infection. The results showed that one miRNA, cal-miRNA-155, displayed significantly increased expression in response to the virus infection. Subsequently, it was proved that overexpression of cal-miR-155 enhanced cell apoptosis with or without NNV infection and inhibited virus replication in CAB cells. Oppositely, the cal-miRNA-155 inhibitor markedly suppressed apoptosis in CAB cells. The results of the apoptosis-related genes mRNA expression also showed the regulation of cal-miR-155 on the apoptosis process in CAB cells. These findings verify that miR-155 might exert a function as a pro-apoptotic factor in reply to NNV stimulation in CAB cells and help us further study the molecular mechanisms of the pathogenesis of NNV in C. altivelis.

Humpback grouper Chromileptes altivelis (HG), red-spotted grouper Epinephelus akaara (RG) and black seabream Acanthopagrus schlegelii (BS) are three popular perciform fishes with an increasingly important farming industry. The prices of BS are much lower than other grouper species; however, the differences in the nutritive values of these three perciform fishes with commercial specifications have not been reported. In this study, the biochemical composition and non-volatile taste active compounds of adult HG, RG and BS were investigated. Moisture contents in BS were significantly higher than in HG and RG (p < 0.05), and relatively lower crude protein contents in BS were observed. Lipid contents of back muscle were lower than that of abdomen muscle in the three fish species. C22:6n-3 (DHA) was the major poly-unsaturated fatty acid (PUFA) in HG and BS, while the main PUFA in RG was C18:2n-6. The total healthy omega-3 fatty acid (Σn-3) profiles in HG were the highest (24.08–24.59%), followed by RG (18.24–19.06%) and BS (13.63–15.91%) (p < 0.05). Glycine was the most abundant free amino acid (FAA) in HG and RG, while lysine was the major FAA in BS. Equivalent umami concentration (EUC) values in BS were the highest, followed by HG and RG (p < 0.05). Lactic acid and PO43− were the major organic acids and inorganic ions, respectively. In conclusion, HG and RG provided more protein and healthy omega-3 fatty acids than BS, while BS had a stronger umami taste according to the EUC values.

R848 is an imidazoquinoline compound that is a specific activator of toll-like receptor (TLR) 7/8 and is often used in immunological research in mammals and teleosts. However, the immune responses initiated by R848 through the TLR7/8 pathway in response to bacterial infection remain largely unexplored in teleosts. In the current study, we investigated the antibacterial response and the participating signaling pathway initiated by R848 in golden pompano (Trachinotus ovatus). We found that R848 could stimulate the proliferation of head kidney lymphocytes (HKLs) in a dose-dependent manner, enhance the survival rate of HKLs, and inhibit the replication of bacteria in vivo. However, these effects induced by R848 were significantly reduced when chloroquine (CQ) was used to blocked endosomal acidification. Additionally, an in vivo study showed that R848 strengthened the antibacterial immunity of fish through a TLR7/8 and Myd88-dependent signaling pathway. A cellular experiment showed that Pepinh-MYD (a Myd88 inhibitor) significantly reduced the R848-mediated proliferation and survival of HKLs. Luciferase activity analysis showed that R848 enhanced the nuclear factor kappa B (NF-κB) activity, whereas this activity was reduced when CQ and Pepinh-MYD were present. Additionally, when an NF-κB inhibitor was present, the R848-mediated pro-proliferative and pro-survival effects on HKLs were significantly diminished. An in vivo study showed that knockdown of TLR7, TLR8, and Myd88 expression in golden pompano via siRNA following injection of R848 resulted in increased bacterial dissemination and colonization in fish tissues compared to that of fish injection of R848 alone, suggesting that R848-induced antibacterial immunity was significantly reduced. In conclusion, these results indicate that R848 plays an essential role in the antibacterial immunity of golden pompano via the TLR7/8-Myd88-NF-κB- signaling pathway.

This study examined the impact of varying doses of mannan oligosaccharide (MOS) supplementation on antioxidant defense, non-specific immunity, resistance to typhoon stress, and resistance to disease in Trachinotus ovatus. Fish groups C, M1, M2, and M3 were fed 1%, 0.3%, 0.6%, and 1% MOS-supplemented feed, respectively. On the 28th and 56th day of the feeding trial, samples from all groups were taken to measure the antioxidant and non-specific immunity parameters and challenged with Vibrio harvey. On the 32nd day, a typhoon hit the experimental site, and the survival rates of all groups were calculated. Results showed that on the 28th day, groups M2 and M3 showed considerably higher levels of antioxidant abilities (T-AOC), and groups M1 and M2 showed significantly enhanced non-specific immunity (AKP, LZM, and TP) levels than those of group C (p < 0.05). All MOS groups had significantly lower cumulative mortality following the challenge test and typhoon stress than the control group (p < 0.05). On the 56th day, groups M2 and M3 showed significantly enhanced antioxidant ability (CAT) compared to the control group. Only one non-specific immunity parameter increased significantly in groups M2 and M3 (p < 0.05). After the challenge test, only group M3 exhibited a significantly declined cumulative mortality rate compared to the control groups (p < 0.05). In summary, supplementation of MOS at dose 0.6% (M2) for 28 days showed the best overall improving effects on the antioxidant capacity, non-specific immunity, disease resistance, and typhoon stress resistance in T. ovatus juveniles. [ABSTRACT FROM AUTHOR]

Introduction: Insulin-like growth factor binding protein 5 (IGFBP5) exerts an essential biological role in many processes, including apoptosis, cellular differentiation, growth, and immune responses. However, compared to mammalians, our knowledge of IGFBP5 in teleosts remains limited. Methods: In this study, TroIGFBP5b, an IGFBP5 homologue from golden pompano (Trachinotus ovatus) was identified. Quantitative real-time PCR (qRT-PCR) was used to check its mRNA expression level in healthy condition and after stimulation. In vivo overexpression and RNAi knockdown method were performed to evaluate the antibacterial profile. We constructed a mutant in which HBM was deleted to better understand the mechanism of its role in antibacterial immunity. Subcellular localization and nuclear translocation were verified by immunoblotting. Further, proliferation of head kidney lymphocytes (HKLs) and phagocytic activity of head kidney macrophages (HKMs) were detected through CCK-8 assay and flow cytometry. Immunofluorescence microscopy assay (IFA) and dual luciferase reporter (DLR) assay were used to evaluate the activity in nuclear factor-kB (NFkb) pathway. Results: The TroIGFBP5b mRNA expression level was upregulated after bacterial stimulation. In vivo, TroIGFBP5b overexpression significantly improved the antibacterial immunity of fish. In contrast, TroIGFBP5b knockdown significantly decreased this ability. Subcellular localization results showed that TroIGFBP5b and TroIGFBP5b-dHBM were both present in the cytoplasm of GPS cells. After stimulation, TroIGFBP5b-dHBM lost the ability to transfer from the cytoplasm to the nucleus. In addition, rTroIGFBP5b promoted the proliferation of HKLs and phagocytosis of HKMs, whereas rTroIGFBP5b-dHBM, suppressed these facilitation effects. Moreover, the in vivo antibacterial ability of TroIGFBP5b was suppressed and the effects of promoting expression of proinflammatory cytokines in immune tissues were nearly lost after HBM deletion. Furthermore, TroIGFBP5b induced NFkb promoter activity and promoted nuclear translocation of p65, while these effects were inhibited when the HBM was deleted. Discussion: Taken together, our results suggest that TroIGFBP5b plays an important role in golden pompano antibacterial immunity and activation of the NF-kb signalling pathway, providing the first evidence that the HBM of TroIGFBP5b plays a critical role in these processes in teleosts. [ABSTRACT FROM AUTHOR]

The selection of intelligent logistics service model has become an important factor in the competition of the entire social logistics industry. Using the technologies such as the Internet of Things (IoT) and cloud computing, the service platform of cloud intelligent logistics (CIL) virtualizes and accesses to the distributed physical logistics resources and logistics capabilities, and relies on its powerful processing and control capabilities to obtain the best service portfolio of CIL. The paper proposes the service combinatorial optimization algorithm (COA). Based on this, it studies the cloud intelligent logistics service and service combination method. The results show that compared with clustering algorithm and differential evolution algorithm, COA algorithm has greater superiority and stability in selection and combination of CIL service; the CIL service has the characteristics of dynamicity and diversity, heterogeneity and distribution, abstraction and similarity; the service portfolio of CIL is divided into three stages: service classification, service negotiation and optimal combination. The application of COA in the CIL selection can greatly reduce the time consumption of combined logistics service and improve the overall service quality of combinatorial service. [ABSTRACT FROM AUTHOR]