Chinese horseshoe crab,
, is an ancient marine arthropod with a long evolutionary history. As a kind of living fossil species, the pathogen defenses of horseshoe crabs entirely depend on the innate ...immune system. Although, there are abundant immune molecules found in the horseshoe crab hemolymph, the biological mechanisms underlying their abilities of distinguishing and defending against invading microbes are still unclear. In this study, we used high-throughput sequencing at mRNA and protein levels and bioinformatics analysis methods to systematically analyze the innate immune response to Gram-negative bacteria in hemolymph of Chinese horseshoe crab. These results showed that many genes in the complement and coagulation cascades, Toll, NF-κB, C-type lectin receptor, JAK-STAT, and MAPK signaling pathways, and antimicrobial substances were activated at 12 and 24 h post-infection, suggesting that Gram-negative bacteria could activate the hemolymph coagulation cascade and antibacterial substances release
the above pathways. In addition, we conjectured that Toll and NF-κB signaling pathway were most likely to participate in the immune response to Gram-negative bacteria in hemolymph of horseshoe crab through an integral signal cascade. These findings will provide a useful reference for exploring the ancient original innate immune mechanism.
A large number of studies have indicated that Astragalus Membranaceus (AM) possesses growth-promoting effects in animal husbandry. However, the function of AM in common carp growth performance has ...not meant fully evaluated. In this work, to investigate the potential effects of AM and Lactobacillus plantarum-fermented AM (FAM) on growth performance, intestinal function, and intestinal microenvironment of common carp, fish with an average weight of 10.10 ± 0.25 g were divided into seven groups, and fed daily with an additive-free basal diet (BD); 0.1%, 0.2%, and 0.4% (w/w) of AM (L-AM, M-AM, and H-AM); 0.1%, 0.2%, and 0.4% (w/w) of FAM (L-FAM, M-FAM and H-FAM) for an eight-week period. The results showed that incorporating L-AM or L-FAM in the diet significantly increased (P < 0.05) the final weight, weight gain ratio, and specific growth rate compared with the BD group, but significantly decreased (P < 0.05) the feed conversion ratio. In L-AM and L-FAM groups, the villus height was considerably increased (P < 0.05), and crypt depth remained unaffected. L-AM and L-FAM both changed the abundance of intestinal microbiota with strikingly different dominant microbiota. Dietary L-FAM supplementation increased the abundance of Firmicutes, Verrucomicrobia, Cyanobacteria, TM7, WS3, Chlamydiae, TM6, Fusobacteria, Spirochaetes, Acidobacteria, Gemmatimonadetes, Actinobacteria, Chloroflexi, Thermi, and Nitrospirae, whereas decreased Bacteroidetes and Proteobacteria abundances. Intestinal metabolome analyses demonstrated that L-AM altered metabolic composition and function, which were linked to immunity and host metabolism. Furthermore, L-FAM showed superior beneficial metabolic effects than L-AM in fish. On growth performance improvement and intestinal microenvironment modulation, dietary L-FAM supplementation was superior to AM in common carp. We speculate that L-FAM may improve the intestinal morphology and microenvironment, which strengthens the growth of fish. In addition, FAM might be promising antibiotics’ alternatives for promoting fish growth and controlling pathogens that has immense economic value in aquaculture.
•0.1% (w/w) of dietary Astragalus Membranaceus was beneficial to the growth of common carp.•Dietary Astragalus Membranaceus could promote the villus height of intestine in common carp.•Dietary Astragalus Membranaceus could change the abundance of intestinal microbiota and the metabolic composition in common carp.•Fermentation could heighten growth-promoting effects of Astragalus Membranaceus in common carp.
Purpose
To evaluate the diagnostic performance of LR‐5 for diagnosing poorly differentiated hepatocellular carcinoma (p‐HCC). To build a contrast‐enhanced ultrasound (CEUS) signature for improving ...the differential diagnostic performance between p‐HCC and intrahepatic cholangiocarcinoma (ICC).
Methods
The B‐mode ultrasound (BUS) and CEUS features of 60 p‐HCCs and 56 ICCs were retrospectively analyzed. The CEUS LI‐RADS category was assigned according to CEUS LI‐RADS v2017. A diagnostic CEUS signature was built based on the independent significant features. An ultrasound (US) signature combining both BUS and CEUS features was also built. The diagnostic performances of the CEUS signature, US signature, and LR‐5 were evaluated by receiver operating characteristic (ROC) analysis.
Results
One (1.7%) p‐HCC and 26 (46.4%) ICC patients presented cholangiectasis or cholangiolithiasis (P < .001). Fifty‐four (90.0%) p‐HCCs and 8 (14.3%) ICCs showed clear boundaries in the artery phase (P < .001). The washout times of p‐HCCs and ICCs were 81.0 ± 42.5 s and 34.7 ± 8.6 s, respectively (P < .001). The AUC, sensitivity, and specificity of the CEUS signature, US signature, and LR‐5 were 0.955, 91.67%, and 90.57% versus 0.976, 96.67%, and 92.45% versus 0.758, 51.67%, and 100%, respectively. The AUC and sensitivity of CEUS LI‐RADS were much lower than those of the CEUS and US signatures (P < .001).
Conclusion
LR‐5 had high specificity but low sensitivity in diagnosing p‐HCC. When the washout time and tumor boundary were included in the CEUS signature, the sensitivity and AUC were remarkably increased in the differentiation between p‐HCC and ICC.
Multi-target stance detection is the detection of the stance of multiple targets in text. Currently, most multi-target stance detection methods only detect the stance of two targets individually and ...do not make the two targets complement each other to take full advantage of the relevant semantic information between the two targets. In this paper, we propose a comparative learning based stance agreement detection framework. We applied contrastive learning to stance agreement detection, it enabled the model to learn more information about the features of the target and to strengthen the links between the semantic information of the targets so that they assist each other in stance detection. In addition, we fine-tuned a new model as our encoder to more fully exploit the semantic information between hidden contexts. We also apply joint training as a multi-task learning approach, allowing models to share domain-specific information based on the dataset. By comparing different methods, experimental results show that our method achieves state-of-the-art results on multi-target benchmark datasets. In the concluding sections of our paper, we conducted error analysis experiments on the proposed methodology, elucidating its inherent limitations and furnishing invaluable insights conducive to future enhancements.
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•Nitrite exposure decreased hemocyanin contents and caused severe oxidative stress.•Nitrite exposure influenced immune defense, cytoskeleton remodeling and apoptosis.•Amino acid ...metabolism, lipid metabolism and nucleotide metabolism were disrupted.•Higher antioxidative ability and enhanced fatty acids were observed in LV-NT.•Increased detoxification gene and HIF-1α expression were related to tolerance.
Nitrite accumulation in aquatic environments is a potential risk factor that disrupts multiple physiological functions in aquatic animals. In this study, the physiology, transcriptome and metabolome of the control group (LV-C), nitrite-tolerance group (LV-NT) and nitrite-sensitive group (LV-NS) were investigated to identify the stress responses and mechanisms underlying the nitrite tolerance of Litopenaeus vannamei. After LV-NT and LV-NS were subjected to nitrite stress, the hemocyanin contents were significantly decreased, and hepatopancreas showed severe histological damage compared with LV-C. Likewise, the antioxidant enzymes were also significantly changed after nitrite exposure. The transcriptome data revealed differentially expressed genes associated with immune system, cytoskeleton remodeling and apoptosis in LV-NT and LV-NS. The combination of transcriptomic and metabolomic analysis revealed nitrite exposure disturbed metabolism processes in L. vannamei, including amino acid metabolism, nucleotide metabolism and lipid metabolism. The multiple comparative analysis implicated that higher nitrite tolerance of LV-NT than LV-NS may be attributed to enhanced hypoxia inducible factor-1α expression to regulate energy supply and gaseous exchange. Moreover, LV-NT showed higher antioxidative ability, detoxification gene expression and enhanced fatty acids contents after nitrite exposure in relative to LV-NS. Collectively, all these results will greatly provide new insights into the molecular mechanisms underlying the stress responses and tolerance of nitrite exposure in L. vannamei.
MicroRNAs (miRNAs) play significant roles in both embryonic hematopoiesis and hematological malignancy. Zebrafish miR-462-731 cluster is orthologous of miR-191-425 in human which regulates ...proliferation and tumorigenesis. In our previous work, miR-462-731 was found highly and ubiquitously expressed during early embryogenesis. In this study, by loss-of-function analysis (morpholino knockdown combined with CRISRP/Cas9 knockout) and mRNA profiling, we suggest that miR-462-731 is required for normal embryonic development by regulating cell survival. We found that loss of miR-462/miR-731 caused a remarkable decrease in the number of erythroid cells as well as an ectopic myeloid cell expansion at 48 hpf, suggesting a skewing of myeloid-erythroid lineage differentiation. Mechanistically, miR-462-731 provides an instructive input for pu.1-dependent primitive myelopoiesis through regulating etsrp/scl signaling combined with a novel pu.1/miR-462-731 feedback loop. On the other hand, morpholino (MO) knockdown of miR-462/miR-731 resulted in an expansion of posterior blood islands at 24 hpf, which is a mild ventralization phenotype resulted from elevation of BMP signaling. Rescue experiments with both BMP type I receptor inhibitor dorsomorphin and alk8 MO indicate that miR-462-731 acts upstream of alk8 within the BMP/Smad signaling pathway and functions as a novel endogenous BMP antagonist. Besides, an impairment of angiogenesis was observed in miR-462/miR-731 morphants. The specification of arteries and veins was also perturbed, as characterized by the irregular patterning of efnb2a and flt4 expression. Our study unveils a previously unrecognized role of miR-462-731 in BMP/Smad signaling mediated hematopoietic specification of mesodermal progenitors and demonstrates a miR-462-731 mediated regulatory mechanism driving primitive myelopoiesis in the ALPM. We also show a requirement for miR-462-731 in regulating arterial-venous specification and definitive hematopoietic stem cell (HSC) production. The current findings might provide further insights into the molecular mechanistic basis of miRNA regulation of embryonic hematopoiesis and hematological malignancy.
The seepage property of low-permeability rock is of significant importance for the design and safety analysis of underground cavities. By using a self-developed test system, both permeability and ...porosity of granite from an underground oil storage depot were measured. In order to study the influence of rock types on permeability, a tight sandstone was selected as a contrast. The experimental results suggested that the porosity of this granite is less than 5% and permeability is low to 10
–20
m
2
within the range of effective stress. During the loading process, both exponential relationship and power law can be utilized to describe the relationship between effective stress and permeability. However, power law matches the experimental data better during the unloading condition. The stress dependent porosity of granite during loading process can be described via an exponential relationship while the match between the model and experimental data can be improved by a power law in unloading paths. The correlation of permeability and porosity can be described in a power law form. Besides, granite shows great different evolution rules in permeability and porosity from sandstone. It is inferred that this difference can be attributed to the preparing of samples and different movements of microstructures subjected to effective stress.
DTHP, a novel derivative of DTG, inhibits cell proliferation and induces apoptosis of NSCLC cells via the Ca2+ signaling pathway, leading to the activation of CaMKK / AMPK proteins, excessive ROS ...production and reduction of mitochondria membrane potential.
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Non-small cell lung cancer (NSCLC) is the most prevalent type of lung cancer. However, there has been little improvement in its cure rate in the last 30 years, due to its intricate heterogeneity and drug resistance. Accumulating evidences have demonstrated that dysregulation of calcium (Ca2+) homeostasis contributes to oncogenesis and promotes tumor development. Inhibitors of Ca2+ channels/transporters to restore intracellular Ca2+ level were found to arrest tumor cell division, induce apoptosis, and suppress tumor growth both in vitro and in vivo. Dolutegravir (DTG), which is a first-line drug for Acquired Immune Deficiency Syndrome (AIDs) treatment, has been shown to increase intracellular Ca2+ levels and Reactive oxygen species (ROS) levels in human erythrocytes, leading to suicidal erythrocyte death or eryptosis. To explore the potential of DTG as an antitumor agent, we have designed and synthesized a panel of compounds based on the principle of biologically active substructure splicing of DTG. Our data demonstrated that 7-methoxy-4-methyl-6,8-dioxo-N-(3-(1-(2-(trifluoromethyl)phenyl)-1H-1,2,3-triazol-4-yl)phenyl)-3,4,6,8,12,12a-hexahydro-2H-pyrido1′,2′:4,5pyrazino2,1-b1,3oxazine-9-carboxamide (DTHP), a novel derivative of DTG, strongly inhibited the colony-forming ability and proliferation of NSCLC cells, but displayed no cytotoxicity to normal lung cells. DTHP treatment also induced apoptosis and upregulate intracellular Ca2+ level in NSCLC cells significantly. Inhibiting Ca2+ signaling alleviated DTHP-induced apoptosis, suggesting the perturbation of intracellular Ca2+ is responsible for DTHP-induced apoptosis. We further discovered that DTHP activates AMPK signaling pathway through binding to SERCA, a Ca2+-ATPase. On the other hand, DTHP treatment promoted mitochondrial ROS production, causing mitochondrial dysfunction and cell death. Finally, DTHP effectively inhibited tumor growth in the mouse xenograft model of lung cancer with low toxicity to normal organs. Taken together, our work identified DTHP as a superior antitumor agent, which will provide a novel strategy for the treatment of NSCLC with potential clinical application.
The ATP-binding cassette (ABC) transporters have crucial roles in various biological processes such as growth, development and immune defense in eukaryotes. However, the roles of ABC transporters in ...the immune system of crustaceans remain elusive. In this study, 38 ABC genes were systematically identified and characterized in Penaeus vannamei. Bioinformation analysis revealed that PvABC genes were categorized into ABC A-H eight subfamilies with 17 full-transporters, 11 half transporters and 10 soluble proteins, and multiple immunity-related cis-elements were found in gene promoter regions. Expression analysis showed that most PvABC genes were widely and highly expressed in immune-related tissues and responded to the stimulation of Vibrio parahaemolyticus. To investigate whether PvABC genes mediated innate immunity, PvABCC5, PvABCF1 and PvABCB4 were selected for dsRNA interference experiment. Knockdown of PvABCF1 and PvABCC5 not PvABCB4 increased the cumulative mortality of P. vannamei and bacterial loads in hepatopancreas after infection with V. parahaemolyticus. Further analysis showed that the PvABCF1 and PvABCC5 knockdown decreased expression levels of NF-κB pathway genes and antimicrobial peptides (AMPs). Collectively, these findings indicated that PvABCF1 and PvABCC5 might restrict V. parahaemolyticus challenge by positively regulating NF-κB pathway and then promoting the expression of AMPs, which would contribute to overall understand the function of ABC genes in innate immunity of invertebrates.
•A total of 38 ABC genes were systematically identified and characterized in P. vannamei genome.•Most PvABC genes were ubiquitously expressed in all examined tissues with high expression in hemocytes, gill, hepatopancreas and intestine.•The mRNA expression of selected PvABC genes was obviously altered after V. parahemolyticus challenge.•PvABCC5 and PvABCF1 knockdown increased shrimp mortality and the bacteria loads in the hepatopancreas upon V. parahemolyticus challenge.•PvABCC5 and PvABCF1 might restrict V. parahaemolyticus challenge by positively regulating NF-κB pathway, and then promoting the expression of AMPs in P. vannamei.
The economic and biological significance of blunt snout bream (Megalobrama amblycephala) makes this species important to explore the underlying molecular mechanism of hypoxia response. In the present ...study, we compared the transcriptional responses to serious hypoxia in skeletal muscle among hypoxia tolerant (MT), sensitive (MS) and control (without hypoxia treatment, MC) M. amblycephala obtained according to the time difference of losing balance after hypoxia treatment. A total of 88,200,889 clean reads were generated and assembled into 44,493 unigenes. Transcriptomic comparison revealed 463 genes differentially expressed among different groups. A similar hypoxia-induced transcription patterns suggested a common hypoxia response involved in cell cycle, p53 signaling pathway, apoptosis, heart contraction and blood circulation. Interesting, four genes, heat shock protein beta-8 (hspb8), cysteine/serine-rich nuclear protein 1 (csrnp1), salt-inducible kinase 1 (sik1), and visinin-like 1a (vsnl1a) were up-regulated in MT Vs MC but down-regulated in MS Vs MC. Additionally, FoxO signaling pathway was significantly enriched only in MT Vs MC. These results not only provided the first insights into the mechanism that muscle tissue coped with the hypoxia stress in cyprinid species, but offered a theory base for breeding of M. amblycephala with hypoxia-resistant traits.
•463 DEGs were found among hypoxia tolerant, sensitive and control M. amblycephala.•Similar hypoxia-induced patterns were in fish with different hypoxia tolerance.•FoxO signal pathway was significantly enriched only in hypoxia tolerant fish.
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