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1.
Exp Cell Res ; 439(1): 114076, 2024 Jun 01.
Article in English | MEDLINE | ID: mdl-38719174

ABSTRACT

Glioblastoma (GBM) is a common primary central nervous system tumor. The molecular mechanisms of glioma are unknown, and the prognosis is poor. Therefore, exploring the underlying mechanisms and screening for new prognostic markers and therapeutic targets is crucial. We utilized the weighted gene co-expression network analysis (WGCNA), Differentially Expressed Genes (DEGs), and LASSO-COX analysis to identify three target genes. Next, we constructed and evaluated a prognostic model, screening out COL8A1 as a risk gene. Through a sequence of cellular functional experiments, in vivo studies, and RNA sequencing, we delved into exploring the functional effects and molecular mechanisms of COL8A1 on GBM cells. Finally, the correlation between COL8A1 and tumor immune cells and different inflammatory responses was analyzed. Immunohistochemistry experiments revealed the influence of COL8A1 on macrophage polarization. The COL8A1 expression level was associated with the grade, prognosis, and tumor microenvironment (TME) of glioma. Functional experiments showed that COL8A1 inhibited GBM cell apoptosis and promoted migration, invasion, and proliferation in vitro and in vivo. We also found that COL8A1 promotes the epithelial-mesenchymal transition process and may mediate the activation of the ERK pathway through SHC1. In addition, immune infiltration analysis showed that COL8A1 was closely associated with macrophages in glioma tissues, significantly suppressing the signaling of M1-like -type macrophages and enhancing the signaling of M2-like -type macrophages. COL8A1 was first found to be associated with prognosis, progression, and immune microenvironment of glioma and may serve as a new marker of prognosis and a therapeutic target.


Subject(s)
Biomarkers, Tumor , Brain Neoplasms , Cell Movement , Cell Proliferation , Gene Expression Regulation, Neoplastic , Glioma , Tumor Microenvironment , Animals , Female , Humans , Male , Mice , Apoptosis/genetics , Biomarkers, Tumor/genetics , Biomarkers, Tumor/metabolism , Brain Neoplasms/pathology , Brain Neoplasms/genetics , Brain Neoplasms/metabolism , Cell Line, Tumor , Cell Movement/genetics , Cell Proliferation/genetics , Epithelial-Mesenchymal Transition/genetics , Glioblastoma/pathology , Glioblastoma/genetics , Glioblastoma/metabolism , Glioma/genetics , Glioma/pathology , Glioma/metabolism , Mice, Nude , Prognosis , Tumor Microenvironment/genetics
2.
Article in English | MEDLINE | ID: mdl-38557632

ABSTRACT

Few-shot learning (FSL) is a challenging yet promising technique that aims to discriminate objects based on a few labeled examples. Learning a high-quality feature representation is key with few-shot data, and many existing models attempt to extract general information from the sample or task levels. However, the common sample-level means of feature representation limits the models generalizability to different tasks, while task-level representation may lose class characteristics due to excessive information aggregation. In this article, we synchronize the class-specific and task-shared information from the class and task levels to obtain a better representation. Structure-based contrastive learning is introduced to obtain class-specific representations by increasing the interclass distance. A hierarchical class structure is constructed by clustering semantically similar classes using the idea of granular computing. When guided by a class structure, it is more difficult to distinguish samples in different classes that have similar characteristics than those with large interclass differences. To this end, structure-guided contrastive learning is introduced to study class-specific information. A hierarchical graph neural network is established to transfer task-shared information from coarse to fine. It hierarchically infers the target sample based on all samples in the task and yields a more general representation for FSL classification. Experiments on four benchmark datasets demonstrate the advantages of our model over several state-of-the-art models.

3.
Life Sci ; 341: 122504, 2024 Mar 15.
Article in English | MEDLINE | ID: mdl-38354973

ABSTRACT

Cingulin and its paralog paracingulin are vital components of the apical junctional complex in vertebrate epithelial and endothelial cells. They are both found in tight junctions (TJ), and paracingulin is also detectable in adherens junctions (AJ) as TJ cytoplasmic plaque proteins. Cingulin and paracingulin interact with other proteins to perform functions. They interact with cytoskeletal proteins, modulate the activity of small GTPases, such as RhoA and Rac1, and regulate gene expression. In addition, cingulin and paracingulin regulate barrier function and many pathological processes, including inflammation and tumorigenesis. In this review, we summarize the discovery and structure, expression and subcellular distribution, and molecular interactions of cingulin family proteins and discuss their role in development, physiology, and pathological processes.


Subject(s)
Endothelial Cells , Membrane Proteins , Membrane Proteins/metabolism , Endothelial Cells/metabolism , Clinical Relevance , Cytoskeletal Proteins/genetics , Tight Junctions/metabolism , Epithelial Cells/metabolism
4.
IUBMB Life ; 76(3): 140-160, 2024 Mar.
Article in English | MEDLINE | ID: mdl-37728571

ABSTRACT

The molecular mechanisms of glioblastoma (GBM) are unclear, and the prognosis is poor. Spinster homolog 2 (SPNS2) is reportedly involved in pathological processes such as immune response, vascular development, and cancer. However, the biological function and molecular role of SPNS2 in GBM are unclear. SPNS2 is aberrantly low expressed in glioma. Survival curves, risk scores, prognostic nomograms, and univariate and multifactorial Cox regression analyses showed that SPNS2 is an independent prognostic indicator significantly associated with glioma progression and prognosis. Cell function assays and in vivo xenograft transplantation were performed that downregulation of SPNS2 promoted GBM cell growth, migration, invasion, epithelial-mesenchymal transition (EMT), anti-apoptosis, drug resistance, and stemness, while overexpression of SPNS2 had the opposite effect. Meanwhile, the functional enrichment and signaling pathways of SPNS2 in the Cancer Genome Atlas (TCGA), Chinese Glioma Genome Atlas (CGGA), and RNA sequencing were analyzed by Gene ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and Gene set enrichment analysis (GSEA). The above results were related to the inhibition of the PTEN/PI3K/AKT pathway by SPNS2. In addition, we predicted that SPNS2 is closely associated with immune infiltration in the tumor microenvironment by four immune algorithms, ESTIMATE, TIMER, CIBERSORT, and QUANTISEQ. In particular, SPNS2 was negatively correlated with the infiltration of most immune cells, immunomodulators, and chemokines. Finally, single-cell sequencing analysis also revealed that SPNS2 was remarkably correlated with macrophages, and downregulation of SPNS2 promotes the expression of M2-like macrophages. This study provides new evidence that SPNS2 inhibits malignant progression, stemness, and immune infiltration of GBM cells through PTEN/PI3K/AKT pathway. SPNS2 may become a new diagnostic indicator and potential immunotherapeutic target for glioma.


Subject(s)
Glioblastoma , Glioma , Humans , Glioblastoma/pathology , Proto-Oncogene Proteins c-akt/genetics , Proto-Oncogene Proteins c-akt/metabolism , Phosphatidylinositol 3-Kinases/genetics , Phosphatidylinositol 3-Kinases/metabolism , Gene Expression Regulation, Neoplastic , Glioma/pathology , Tumor Microenvironment/genetics , PTEN Phosphohydrolase/genetics , PTEN Phosphohydrolase/metabolism
5.
BMC Med Imaging ; 23(1): 31, 2023 02 10.
Article in English | MEDLINE | ID: mdl-36765284

ABSTRACT

OBJECTIVES: To investigate the value of preoperative positron emission tomography/computed tomography (PET/CT) in differentiating the invasive degree of hypometabolic lung adenocarcinoma. METHODS: We retrospectively analyzed the data of patients who underwent PET/CT examination, high-resolution computed tomography, and surgical resection for low-metabolism lung adenocarcinoma in our hospital between June 2016 and December 2021. We also investigated the relationship between the preoperative PET/CT findings and the pathological subtype of hypometabolic lung adenocarcinoma. RESULTS: A total of 128 lesions were found in 113 patients who underwent resection for lung adenocarcinoma, including 20 minimally invasive adenocarcinomas (MIA) and 108 invasive adenocarcinomas (IAC), whose preoperative PET/CT showed low metabolism. There were significant differences in the largest diameter (Dmax), lesion type, maximum standard uptake value (SUVmax), SUVindex (the ratio of SUVmax of lesion to SUVmax of contralateral normal lung paranchyma), fasting blood glucose, lobulation, spiculation, and pleura indentation between the MIA and IAC groups (p < 0.05). Multivariate logistic regression analysis showed that the Dmax (odds ratio (OR) = 1.413, 95% confidence interval (CI: 1.155-1.729, p = 0.001)) and SUVmax (OR = 12.137, 95% CI: 1.068-137.900, p = 0.044) were independent risk factors for predicting the hypometabolic IAC (p < 0.05). Receiver operating characteristic (ROC) curve analysis showed that the Dmax ≥ 10.5 mm and SUVmax ≥ 0.85 were the cut-off values for differentiating MIA from IAC, with high sensitivity (84.3% and 75.9%, respectively) and specificity (84.5% and 85.0%, respectively), the Combined Diagnosis showed higher sensitivity (91.7%) and specificity (85.0%). CONCLUSIONS: The PET/CT findings correlated with the subtype of hypometabolic lung adenocarcinoma. The parameters Dmax and SUVmax were independent risk factors for predicting IAC, and the sensitivity of Combined Diagnosis prediction is better.


Subject(s)
Adenocarcinoma of Lung , Adenocarcinoma , Lung Neoplasms , Humans , Positron Emission Tomography Computed Tomography/methods , Lung Neoplasms/diagnostic imaging , Lung Neoplasms/surgery , Lung Neoplasms/pathology , Retrospective Studies , Adenocarcinoma of Lung/diagnostic imaging , Adenocarcinoma of Lung/surgery , Adenocarcinoma of Lung/pathology , Adenocarcinoma/diagnostic imaging , Adenocarcinoma/surgery , Adenocarcinoma/pathology , Fluorodeoxyglucose F18
6.
Clin Cancer Res ; 29(13): 2525-2539, 2023 07 05.
Article in English | MEDLINE | ID: mdl-36729148

ABSTRACT

PURPOSE: Pancreatic ductal adenocarcinoma (PDA) resists immunotherapy of adoptive cell transfer (ACT) and immune checkpoint inhibitors. Understanding the mechanisms underlying this resistance will improve PDA immunotherapy. This study investigated therapeutic effects and underlying mechanisms of anticoagulants on immunotherapy in PDA. EXPERIMENTAL DESIGN: The antitumor activity of immunotherapy was evaluated in mouse models of desert, excluded, and inflamed tumors. The underlying mechanisms were investigated by analyzing immune cell infiltration by immunofluorescence imaging and tumor microcirculation by interstitial fluid pressure and coagulation status measurement. RESULTS: Combined use of heparin and ACT inhibited tumor growth and metastasis, whereas neither heparin nor ACT had any therapeutic effect. The combination of heparin and ACT significantly increased the intratumor infiltration of CD8+ T cells and M1 macrophages and reduced the infiltration of immunosuppressive M2 macrophages and FOXP3+/CD4+ regulatory T cells (Treg). Assessments of tumor microenvironment revealed that heparin promoted tumor vascular regression and normalized the remaining blood vessels, facilitating the extravasation and perivascular accumulation of activated CD8+ T cells in tumors. Mechanistically, tumor microvessel hemodynamic properties were significantly improved by heparin, which is consistent with its inhibitory effects on tumor angiogenesis. Similarly, the combination of heparin and anti-PD1 also produced a pronounced antitumor activity, whereas neither heparin nor anti-PD1 treatment had appreciable antitumor activity. CONCLUSIONS: Combined treatment of heparin and ACT or anti-PD1 produced synergistic antitumor effects, which were at least in part through tumor vascular normalization, hence increased antitumor T-cell responses due to reduced Treg infiltration and increased M1 macrophage polarization. This synergistic combination therapy warrants clinical evaluation. See related commentary by Korc, p. 2348.


Subject(s)
Carcinoma, Pancreatic Ductal , Pancreatic Neoplasms , Mice , Animals , Heparin/pharmacology , Anticoagulants/pharmacology , Microcirculation/drug effects , Pancreatic Neoplasms/pathology , Immunotherapy , Carcinoma, Pancreatic Ductal/pathology , Tumor Microenvironment/immunology , Pancreatic Neoplasms
7.
FASEB J ; 37(1): e22688, 2023 01.
Article in English | MEDLINE | ID: mdl-36468780

ABSTRACT

Colorectal cancer (CRC) is one of the most common gastrointestinal malignancies. Vasorin (VASN) has been reported to be critical in tumor development and angiogenesis. However, VASN has not been reported in CRC, and its role is unclear. In this study, VASN expression is upregulated in CRC compared with the normal tissues, and VASN expression positively correlates with N stage and poor overall survival by analysis of different datasets and 32 CRC clinicopathologic samples. Overexpression of VASN significantly promotes CRC cell progression, including proliferation, migration, invasion, and epithelial-mesenchymal transition (EMT), while knockdown of VASN inhibits CRC progression. We found that VASN was associated with the YAP/TAZ and PI3K/AKT pathways by gene set enrichment analysis (GSEA) and gene ontology (GO) analysis. Notably, western blotting, immunofluorescence staining and co-immunofluorescence (co-IP) confirmed that VASN could interact with YAP and activate the YAP/TAZ and PTEN/PI3K/AKT pathways, and knockdown of YAP reversed this effect. Importantly, our findings indicate that VASN interacts with YAP to inhibit YAP phosphorylation and stimulates CRC proliferation, migration, and invasion through activation of the YAP/TAZ-TEAD target gene CTGF and PTEN/PI3K/AKT pathways. Our results also show that knockdown of YAP reverses the cellular phenotype induced by increased VASN. In conclusion, our study reveals that VASN acts as an oncogene to stimulate tumor progression in CRC, providing new insights into the molecular mechanisms of CRC development and representing a possible novel biomarker for CRC.


Subject(s)
Colorectal Neoplasms , Proto-Oncogene Proteins c-akt , Humans , Phosphatidylinositol 3-Kinases , Oncogenes , Signal Transduction , Colorectal Neoplasms/genetics , Carrier Proteins , Membrane Proteins
8.
Mar Biotechnol (NY) ; 23(2): 294-307, 2021 Apr.
Article in English | MEDLINE | ID: mdl-33570690

ABSTRACT

In mammals, mature miR-122 is 22 nucleotides long and can be involved in regulating a variety of physiological and biological pathways. In this study, the expression profile and effects of grouper Epinephelus coioides miR-122 response to Singapore grouper iridovirus (SGIV) infection were investigated. The sequences of mature microRNAs (miRNAs) from different organisms are highly conserved, and miR-122 from E. coioides exhibits high similarity to that from mammals and other fish. The expression of miR-122 was up-regulated during SGIV infection. Up-regulation of miR-122 could significantly enhance the cytopathic effects (CPE) induced by SGIV, the transcription levels of viral genes (MCP, VP19, LITAF and ICP18), and viral replication; reduce the expression of inflammatory factors (TNF-a, IL-6, and IL-8), and the activity of AP-1 and NF-κB, and miR-122 can bind the target gene p38α MAPK to regulate the SGIV-induced cell apoptosis and the protease activity of caspase-3. The results indicated that SGIV infection can up-regulate the expression of E. coioides miR-122, and up-regulation of miR-122 can affect the activation of inflammatory factors, the activity of AP-1 and NF-κB, and cell apoptosis to regulate viral replication and proliferation.


Subject(s)
Bass/metabolism , Fish Diseases/virology , Iridovirus/metabolism , MicroRNAs/metabolism , Animals , Apoptosis , Bass/genetics , DNA Virus Infections/virology , Genes, Viral , Iridovirus/genetics , MicroRNAs/genetics , NF-kappa B , Transcription Factor AP-1 , Virus Replication
9.
Dev Comp Immunol ; 119: 104013, 2021 06.
Article in English | MEDLINE | ID: mdl-33465381

ABSTRACT

Programmed cell death 4 (PDCD4) in mammals, a gene closely associated with apoptosis, is involved in many biological processes, such as cell aging, differentiation, regulation of cell cycle, and inflammatory response. In this study, grouper Epinephelus coioides PDCD4, EcPDCD4-1 and EcPDCD4-2, were obtained. The open reading frame (ORF) of EcPDCD4-1 is 1413 bp encoding 470 amino acids with a molecular mass of 52.39 kDa and a theoretical pI of 5.33. The ORF of EcPDCD4-2 is 1410 bp encoding 469 amino acids with a molecular mass of 52.29 kDa and a theoretical pI of 5.29. Both EcPDCD4-1 and EcPDCD4-2 proteins contain two conserved MA3 domains, and their mRNA were detected in all eight tissues of E. coioides by quantitative real-time PCR (qRT-PCR) with the highest expression in liver. The expressions of two EcPDCD4s were significantly up-regulated after Singapore grouper iridovirus (SGIV) or Vibrio alginolyticus infection. In addition, over-expression of EcPDCD4-1 or EcPDCD4-2 can inhibit the activity of the nuclear factor-κB (NF-κB) and activator protein-1 (AP-1), and regulate SGIV-induced apoptosis. The results demonstrated that EcPDCD4s might play important roles in E. coioides tissues during pathogen-caused inflammation.


Subject(s)
Apoptosis Regulatory Proteins/immunology , Fish Proteins/immunology , Gene Expression Regulation/immunology , Iridovirus/immunology , Perciformes/immunology , Vibrio alginolyticus/immunology , Amino Acid Sequence , Animals , Apoptosis/genetics , Apoptosis/immunology , Apoptosis Regulatory Proteins/chemistry , Apoptosis Regulatory Proteins/genetics , Cloning, Molecular , Fish Diseases/immunology , Fish Diseases/microbiology , Fish Diseases/virology , Fish Proteins/classification , Fish Proteins/genetics , Gene Expression Profiling , Host-Pathogen Interactions/immunology , Iridovirus/physiology , NF-kappa B/genetics , NF-kappa B/immunology , NF-kappa B/metabolism , Perciformes/microbiology , Perciformes/virology , Phylogeny , Protein Conformation , Reverse Transcriptase Polymerase Chain Reaction , Sequence Homology, Amino Acid , Transcription Factor AP-1/genetics , Transcription Factor AP-1/immunology , Transcription Factor AP-1/metabolism , Vibrio alginolyticus/physiology
10.
Dev Comp Immunol ; 119: 104020, 2021 06.
Article in English | MEDLINE | ID: mdl-33476669

ABSTRACT

Mitogen-activated protein kinase 4 (MKK4), a member of the MAP kinase family, play important roles in response to many environmental and cellular stresses in mammals. In this study, three MKK4 subtypes, EcMKK4-1, EcMKK4-2 and EcMKK4-3, were obtained from grouper Epinephelus coioides. The open reading frame (ORF) of EcMKK4s are obtained and the EcMKK4s proteins contain highly conserved domains: a S_TKc domain, a canonical diphosphorylation group and two conserved MKKK ATP binding motifs, Asp-Phe-Gly (DFG) and Ala-Pro-Glu (APE). EcMKK4s could be found both in the cytoplasmic and nuclear. The EcMKK4s mRNA were detected in all E. coioides tissues examined with the different expression levels, and the expression were up-regulated during SGIV (Singapore grouper iridescent virus) or Vibrio alginolyticus infection. EcMKK4 could significantly reduce the activation of AP-1 reporter gene. The results suggested that EcMKK4s might play important roles in pathogen-caused inflammation.


Subject(s)
Fish Diseases/immunology , Fish Proteins/immunology , Gene Expression Regulation/immunology , Iridovirus/immunology , MAP Kinase Kinase 4/immunology , Perciformes/immunology , Vibrio alginolyticus/immunology , Amino Acid Sequence , Animals , Apoptosis/genetics , Apoptosis/immunology , Cell Line , Cloning, Molecular , Fish Diseases/microbiology , Fish Diseases/virology , Fish Proteins/classification , Fish Proteins/genetics , Gene Expression Profiling , Host-Pathogen Interactions/immunology , Iridovirus/physiology , MAP Kinase Kinase 4/classification , MAP Kinase Kinase 4/genetics , Perciformes/microbiology , Perciformes/virology , Phylogeny , RNA, Messenger/genetics , RNA, Messenger/immunology , RNA, Messenger/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Sequence Homology, Amino Acid , Up-Regulation/immunology , Vibrio alginolyticus/physiology
11.
Inorg Chem ; 59(8): 5368-5376, 2020 Apr 20.
Article in English | MEDLINE | ID: mdl-32233424

ABSTRACT

Exploration of rare-earth (RE)-based Kagomé lattice magnets with spin-orbital entangled jeff = 1/2 moments will provide a new platform for investigating the exotic magnetic phases. Here, we report a new family of RE3BWO9 (RE = Pr,Nd,Gd-Ho) boratotungstates with magnetic RE3+ ions arranged on Kagomé lattice and perform its structure and magnetic characterizations. These serial compounds crystallize in a hexagonal coordinated structure with space group P63 (no. 173), where magnetic RE3+ ions have distorted Kagomé lattice connections within the ab plane and stacked in an AB-type fashion along the c axis. The interlayer RE-RE separation is comparable with that of the intralayer distance, forming 3-dimensional (3D) exchange coupled magnetic framework of RE3+ ions. The magnetic susceptibility data of RE3BWO9 (RE = Pr, Nd, Gd-Ho) reveal dominant antiferromagnetic interactions between magnetic RE3+ ions, but without visible magnetic ordering down to 2 K. The magnetization analyses for different RE3+ ions show diverse anisotropic behaviors, making RE3BWO9 as an appealing Kagomé-lattice antiferromagnet to explore exotic magnetic phases.

12.
RSC Adv ; 10(27): 15784-15793, 2020 Apr 21.
Article in English | MEDLINE | ID: mdl-35493661

ABSTRACT

A single device with extensive new functionality is highly attractive for the increasing demands for complex and multifunctional optoelectronics. Multi-field coupling has been drawing considerable attention because it leads to materials that can be simultaneously operated under several external stimuli (e.g. magnetic field, electric field, electric current, light, strain, etc.), which allows each unit to store multiple bits of information and thus enhance the memory density. In this work, we report an electro-opto-mechano-driven reversible multi-state memory device based on photocurrent in Bi0.9Eu0.1FeO3 (BEFO)/La0.67Sr0.33MnO3 (LSMO)/0.7Pb(Mg1/3Nb2/3)O3-0.3PbTiO3 (PMN-PT) heterostructures. It is found that the short-circuit current density (J sc) can be switched by the variation of the potential barrier height and depletion region width at the Pt/BEFO interface modulated by light illumination, external strain, and ferroelectric polarization reversal. This work opens up pathways toward the emergence of novel device design features with dynamic control for developing high-performance electric-optical-mechanism integrated devices based on the BiFeO3-based heterostructures.

13.
Psychophysiology ; 57(10): e13640, 2020 10.
Article in English | MEDLINE | ID: mdl-33460208

ABSTRACT

Top-down spatial attention can modulate contingent attentional capture, but the underlying mechanism is still not clear. Using variants of spatial cueing paradigms, our previous event-related potential study showed that peripheral color singleton cues with task-relevant features captured attention (indexed by cue-elicited N2pc) even when the targets appeared at central locations, but the magnitude of attentional capture was smaller than when the targets appeared at same peripheral locations. One reasonable explanation is that the modulation effect is due to spatial relevance of cues. However, several other confounding factors might also explain the modulation effect, such as task difficulty, spread of attentional window, and inside/outside relation between cue and attentional window. In the present study, we rearranged the relative locations between cues and targets to control these factors and to further examine whether inequivalence of attentional capture across attended and unattended locations was a common phenomenon. In two experiments, color singleton cues elicited apparent N2pc components when participants were searching for targets possessing the same color, which replicated typical findings of contingent attentional capture. More importantly, the modulation effect of top-down spatial attention on the cue-elicited N2pc still existed even when the factors of task difficulty (Experiment 1), spread of attentional window (Experiment 1), or inside/outside relation between cue and attentional window (Experiment 2) were controlled. These results consistently demonstrate that attentional capture by a color singleton is stronger at spatially relevant locations than at spatially irrelevant locations, suggesting an important role of spatial relevance on contingent attentional capture.


Subject(s)
Attention/physiology , Color Perception/physiology , Evoked Potentials/physiology , Pattern Recognition, Visual/physiology , Space Perception/physiology , Adult , Cues , Electroencephalography , Female , Humans , Male , Psychomotor Performance/physiology , Young Adult
14.
Fish Shellfish Immunol ; 97: 125-134, 2020 Feb.
Article in English | MEDLINE | ID: mdl-31809835

ABSTRACT

Heat shock protein 22 (Hsp22) is an important regulatory factor response to various stresses in mammals. In this study, the full length cDNA of Epinephelus coioides Hsp22, which was 1680bp in length, with a 289 bp 5' UTR, a 725 bp 3'UTR, and a 666 bp open reading frame encoding 221 amino acids, was obtained. E. coioides Hsp22 contains a highly conserved α-crystallin domain. E. coioides Hsp22 mRNA was detected in all tissues examined by quantitative real-time PCR, with the highest expression in blood, followed by the spleen, skin, gill, head kidney, muscle, heart, liver, trunk kidney, stomach, pyloric caeca, intestine, brain and thymus. The expression patterns of E. coioides Hsp22 response to infection with Singapore grouper iridovirus (SGIV) and Vribro alginolyticus, the important pathogens of E. coioides, were studied. The expression levels of the gene were up-regulated in the tissues examined. Subcellular localization analysis demonstrated that E. coioides Hsp22 was distributed in both the cytoplasm and nucleus. In addition, E. coioides Hsp22 significantly inhibited the SGIV-induced cell apoptosis. In summary, the E. coioides Hsp22 might play a critical role in pathogenic stimulation.


Subject(s)
Bass/immunology , Fish Proteins/genetics , Heat-Shock Proteins/genetics , Vibrio Infections/veterinary , Virus Diseases/veterinary , Animals , Bass/microbiology , Bass/virology , Cloning, Molecular , Fish Diseases/immunology , Fish Diseases/microbiology , Fish Diseases/virology , Gene Expression , Heat-Shock Proteins/immunology , Iridovirus , Vibrio Infections/immunology , Vibrio alginolyticus , Virus Diseases/immunology
15.
Fish Shellfish Immunol ; 92: 500-507, 2019 Sep.
Article in English | MEDLINE | ID: mdl-31247318

ABSTRACT

Mitogen-activated protein kinase 6 (MKK6) is one of the major important central regulatory proteins response to environmental and physiological stimuli. In this study, a novel MKK6, EcMKK6, was isolated from Epinephelus coioides, an economically important cultured fish in China and Southeast Asian counties. The open reading frame (ORF) of EcMKK6 is 1077 bp encoding 358 amino acids. EcMKK6 contains a serine/threonine protein kinase (S_TKc) domain, a tyrosine kinase catalytic domain, a conserved dual phosphorylation site in the SVAKT motif and a conserved DVD domain. By in situ hybridization (ISH) with Digoxigenin-labeled probe, EcMKK6 mainly located at the cytoplasm of cells, and a little appears in the nucleus. EcMKK6 mRNA can be detected in all eleven tissues examined, but the expression level is different in these tissues. After challenge with Vibrio alginolyticus and Singapore grouper iridovirus (SGIV), the transcription level of EcMKK6 was apparently up-regulated in the tissues examined. The data demonstrated that the sequence and the characters of EcMKK6 were conserved, EcMKK6 showed tissue-specific expression profiles in healthy grouper, and the expression was significantly varied after pathogen infection, indicating that EcMKK6 may play important roles in E. coioides during pathogen-caused inflammation.


Subject(s)
Bass/genetics , Bass/immunology , Fish Diseases/immunology , Gene Expression Regulation/immunology , Immunity, Innate/genetics , MAP Kinase Kinase 6/genetics , MAP Kinase Kinase 6/immunology , Amino Acid Sequence , Animals , DNA Virus Infections/immunology , DNA Virus Infections/veterinary , Fish Proteins/chemistry , Fish Proteins/genetics , Fish Proteins/immunology , Gene Expression Profiling/veterinary , MAP Kinase Kinase 6/chemistry , Phylogeny , Ranavirus/physiology , Sequence Alignment/veterinary , Vibrio Infections/immunology , Vibrio Infections/veterinary , Vibrio alginolyticus/physiology
16.
ACS Appl Bio Mater ; 2(10): 4583-4593, 2019 Oct 21.
Article in English | MEDLINE | ID: mdl-35021417

ABSTRACT

Layer-by-layer assembled multilayer films with antifouling and pH induced self-cleaning activities were constructed by polyurethane micelles with dense PEG brush coronas (PEG-g-PUM) and polyethylenimine (bPEI). The dense PEG brush coronas and acidic induced surface charge transform activities of PEG-g-PUM rendered the multilayer films (PEG-g-PUM/bPEI)n with antifouling and self-cleaning activities, respectively. Multilayer films constructed from polyurethane micelles without surface charge switchable properties (PEG-c-PUM, PEG-b-PUM), showed negligible pH induced surface release of PU micelles. Furthermore, Triclosan encapsulation in the multilayer films ((PEG-g-PUM-TLS/bPEI)n) further improved the antibacterial performance, as a result of pH and lipase triggered release of payloads from the surfaces. The antifouling properties of PEG-g-PUM and the on-demand release of payloads and PU micelles, may account for their excellent multiple antibacterial performance and pH-induced self-cleaning activities.

17.
Fish Shellfish Immunol ; 84: 1090-1097, 2019 Jan.
Article in English | MEDLINE | ID: mdl-30419398

ABSTRACT

MEK dual-specificity protein kinases are a group of mitogen-activated protein kinase kinases, which act as an integration point by transferring extracellular signals to the nucleus. To investigate the function of MEK in teleost fish, we cloned MEK1 and MEK2 cDNA sequences from the orange-spotted grouper (Epinephelus coioides). EcMEK1 and EcMEK2 shared 80% amino acid identity with each other. EcMEK1 had 89-99% amino acid identity with teleosts or mammals, whereas EcMEK2 shared 85-97% amino acid identity. The exon structures of the grouper MEK1/2 genes were conserved with zebrafish and human MEK1/2. Tissue distribution analysis showed that EcMEK1 and EcMEK2 had a similar expression pattern in grouper tissues and was mainly transcribe in systemic immune organs. Both EcMEK1 and EcMEK2 were distributed throughout the cytoplasm of transfected GS or HEK293T cells. Overexpression of EcMEK1 or EcMEK2 activated Activator protein 1 dependent luciferase. The phosphorylation levels of EcMEK1/2 and EcERK1/2 were significantly increased in head kidney leukocytes by stimulation with PMA treatment. The grouper MEK1/2-ERK1/2 axis was activated in Cryptocaryon irritans infection and showed an enhanced phosphorylation after immunization.


Subject(s)
Bass/genetics , Bass/immunology , Fish Diseases/immunology , Fish Proteins/genetics , Fish Proteins/immunology , Gene Expression Regulation/immunology , Immunity, Innate/genetics , Amino Acid Sequence , Animals , Fish Proteins/chemistry , Gene Expression Profiling/veterinary , MAP Kinase Kinase 1/genetics , MAP Kinase Kinase 1/immunology , MAP Kinase Kinase 2/genetics , MAP Kinase Kinase 2/immunology , Phylogeny , Sequence Alignment/veterinary
18.
Fish Shellfish Immunol ; 86: 1081-1087, 2019 Mar.
Article in English | MEDLINE | ID: mdl-30593900

ABSTRACT

Antimicrobial peptides (AMPs) are small proteins showing broad-spectrum antimicrobial activity that have been known to be powerful agents against a variety of pathogens (bacteria, fungi and viruses). In this study, the effects of AMPs from Bacillus subtilis on Epinephelus coioides were examined. E. coioides were fed with diets containing AMPs (0, 100, 200, 400 or 800 mg/kg) for four weeks. Results showed that the levels of total protein (TP), albumin (ALB), alanine aminotransferase (ALT), aspartate aminotransferase (AST), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C) and blood glucose (GLU) and lipopolysaccharide (LPS) in the serum of E. coioides changed than those of the control group; compared to the control group, the levels of total antioxidant capacity (T-AOC), superoxide dismutase (SOD), catalase (CAT), malondialdehyde (MDA) and lysozyme (LZM) levels in E. coioides fed with different dosages AMP diets were also different; in addition, the mRNA expression of tumor necrosis factor alpha (TNF-α), interleukin-1-beta (IL-1ß), and heat shock protein 90 (Hsp90) in the tissues of E. coioides were measured, the three genes in the tissues examined were significantly upregulated. The results demonstrated that diets containing AMPs can enhance the antioxidant capacity and innate immune ability of E. coioides, indicating that AMPs might be a potential alternative to antibiotics in E. coioides.


Subject(s)
Antimicrobial Cationic Peptides/pharmacology , Antioxidants/metabolism , Bass/immunology , Immunity, Innate , Animal Feed/analysis , Animals , Antimicrobial Cationic Peptides/administration & dosage , Bacillus subtilis/chemistry , Bass/metabolism , Blood Chemical Analysis/veterinary , Diet/veterinary
19.
Mater Sci Eng C Mater Biol Appl ; 93: 921-930, 2018 Dec 01.
Article in English | MEDLINE | ID: mdl-30274129

ABSTRACT

Three different kinds of polyurethane (PU) micelles, i.e. PEG-c-PU, PEG-g-PU and PEG-b-PU, with hydrophobic PCL core and hydrophilic PEG corona were prepared by self-assembly method. DLS studies illustrated that PEG-g-PU micelles showed pH dependent surface charge switching properties while no obvious surface charge switching activities were found for PEG-b-PU and PEG-c-PU micelles. Triclosan was loaded into PCL core by dialysis method with pretty high encapsulate content and efficiency and the payloads were released at an accelerate rate in the presence of lipase. MIC and MBC studies demonstrated an enhanced antibacterial activity of encapsulated Triclosan against planktonic bacteria than free Triclosan. CLSM images of S. aureus biofilms treated with Nile red loaded PU micelles illustrated the penetration and accumulation of PEG-g-PU micelles inside the bacterial biofilms at an acidic environment. In addition, Triclosan loaded into PEG-g-PU micelles showed more potent antibiofilm activities than that loaded into PEG-c-PEG and PEG-b-PU micelles. Therefore, the PEG-g-PU micelles can be potentially used as hydrophobic antibiotic carriers to treat bacterial infections and biofilms.


Subject(s)
Anti-Bacterial Agents , Biofilms/drug effects , Lipase , Micelles , Polyurethanes/chemistry , Staphylococcus aureus/physiology , Triclosan , Anti-Bacterial Agents/chemistry , Anti-Bacterial Agents/pharmacokinetics , Anti-Bacterial Agents/pharmacology , Biofilms/growth & development , Hydrogen-Ion Concentration , Lipase/chemistry , Lipase/pharmacokinetics , Lipase/pharmacology , Triclosan/chemistry , Triclosan/pharmacokinetics , Triclosan/pharmacology
20.
Colloids Surf B Biointerfaces ; 156: 87-94, 2017 Aug 01.
Article in English | MEDLINE | ID: mdl-28527361

ABSTRACT

Choline phosphate (CP) containing polymers modified surfaces have been shown good resist to the adhesion of proteins while prompt the attaching of mammalian cells due to the dipole pairing between the CP groups of the polymer and the phosphorylcholine (PC) groups on the cell membrane. However, the antifouling activities of CP modified surface against microbes have not been investigated at present. In addition, CP containing polymers modified surface with different molecular architectures has not been prepared and studied. To this end, glass slides surface modified with two different 2-(meth-acryloyloxy)ethyl cholinephosphate (MCP) containing polymer (PMCP) structures, i.e. brush-like (Glass-PMCP) and bottle brush-like (Glass-PHEMA-g-PMCP) architectures, were prepared in this work by surface-initiated atom transfer radical polymerization (SI-ATRP). The surface physichemical and antifouling properties of the prepared surfaces were characterized and studied. The Glass-PMCP shows improved antifouling properties against proteins and bacteria as compared to pristine glass slides (Glass-OH) and glass slides grafted with poly(2-hydroxyethyl methacrylate) (Glass-PHEMA). Notably, a synergetic fouling resistant properties of PHEMA and PMCP is presented for Glass-PHEMA-g-PMCP, which shows superior antifouling activities over Glass-PHEMA and Glass-PMCP. Furthermore, glass slides containing PMCP, i.e. Glass-PMCP and Glas-PHEMA-g-PMCP, decrease platelet adhesion and prevent their activation significantly. Therefore, the combination of antifouling PHEMA and PMCP into one system holds potential for prevention of bacterial fouling and biomaterial-centered infections.


Subject(s)
Phosphorylcholine/chemistry , Polymers/chemistry , Animals , Bacteria/drug effects , Blood Platelets/cytology , Phosphorylcholine/pharmacology , Photoelectron Spectroscopy , Platelet Activation , Rabbits , Surface Properties
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