NIK (NF-κB inducing kinase) belongs to the mitogen-activated protein kinase family, which activates NF-κB and plays a vital role in immunology, inflammation, apoptosis, and a series of pathological responses. In NF-κB noncanonical pathway, NIK and IKKα have been often studied in mammals and zebrafish. However, few have explored the relationship between NIK and other subunits of the IKK complex. As a classic kinase in the NF-κB canonical pathway, IKKß has never been researched with NIK in fish. In this paper, the full-length cDNA sequence of grass carp (Ctenopharyngodon idella) NIK (CiNIK) was first cloned and identified. The expression level of CiNIK in grass carp cells was increased under GCRV stimuli. Under the stimulation of GCRV, poly (I:C), and LPS, the expression of NIK in various tissues of grass carp was also increased. This suggests that CiNIK responds to viral stimuli. To study the relationship between CiNIK and CiIKKß, we co-transfected CiNIK-FLAG and CiIKKB-GFP into grass carp cells in coimmunoprecipitation and immunofluorescence experiments. The results revealed that CiNIK interacts with CiIKKß. Besides, the degree of autophosphorylation of CiNIK was enhanced under poly (I:C) stimulation. CiIKKß was phosphorylated by CiNIK and then activated the activity of p65. The activity change of p65 indicates that NF-κB downstream inflammatory genes will be functioning. CiNIK or CiIKKß up-regulated the expression of IL-8. It got higher when CiNIK and CiIKKß coexisted. This paper revealed that NF-κB canonical pathway and noncanonical pathway are not completely separated in generating benefits.
Amino Acid Sequence , Carps , Fish Proteins , Interleukin-8 , NF-kappa B , Protein Serine-Threonine Kinases , Up-Regulation , Animals , Carps/genetics , Carps/immunology , Fish Proteins/genetics , Fish Proteins/immunology , Fish Proteins/chemistry , NF-kappa B/genetics , NF-kappa B/metabolism , Interleukin-8/genetics , Interleukin-8/metabolism , Interleukin-8/immunology , Protein Serine-Threonine Kinases/genetics , Protein Serine-Threonine Kinases/immunology , Protein Serine-Threonine Kinases/metabolism , Fish Diseases/immunology , Signal Transduction , Reoviridae/physiology , Phylogeny , NF-kappaB-Inducing Kinase , Gene Expression Regulation/immunology , Poly I-C/pharmacology , Lipopolysaccharides/pharmacology , Reoviridae Infections/immunology , Reoviridae Infections/veterinary , Sequence Alignment/veterinary , Immunity, Innate/genetics , Base Sequence , Gene Expression Profiling/veterinary
As a member of Mex3 (muscle excess protein-3) family, Mex3B (Mex-3 RNA binding family member B) is crucial in cell proliferation and migration in mammals. In this study, an ortholog of mammalian Mex3B (denominated CiMex3B, MT276802.1) was cloned and identified in grass carp (Ctenopharyngodon idella). CiMex3B is 1578 bp in length and encodes a polypeptide of 525 amino acids. Consistent with its mammalian counterpart, CiMex3B also contains one C-terminal RING domain and two N-terminal conserved tandem KH domains. CiMex3B up-regulates the expressions of IFN1, ISG15, MX2, as well as the expressions of inflammatory cytokines such as IL6, IL8 and TNFα in response to poly(I:C). A screening test for identifying potential targets indicated that CiMex3B is associated with TLR3 and TRIF. CiMex3B co-localizes with TLR3 in the late endosome, mitochondria and endoplasmic reticulum after poly(I:C) stimulation, whereas they are rarely discovered in the lysosomes. CiMex3B serves as a positive regulator in the phosphorylation of IRF3 and induces IFN1 expression. In addition, two truncation mutants of CiMex3B (1-220 and 221-525) were constructed to better understand the molecular mechanism of CiMex3B-mediated ubiquitination of TLR3. In line with wild-type protein, CiMex3B mutant (1-220) was found mainly in the cytoplasm; however, CiMex3B mutant (221-525) resided in the cytoplasm and the nucleus as well, and it was further confirmed that CiMex3B mutant (221-525) still interacts with TLR3. We also observed that CiMex3B promotes the K63-linked ubiquitination of TLR3, while neither of the truncation mutants (1-220 or 221-525) retains this activity. To sum up, this study revealed that CiMex3B potentiates the K63-linked ubiquitination of TLR3, and then elicits the IRF3-mediated antiviral innate immune responses.
Carps , Toll-Like Receptor 3 , Animals , Toll-Like Receptor 3/genetics , Carps/genetics , Carps/metabolism , Immunity, Innate , Cytokines/genetics , Poly I-C/pharmacology , Ubiquitination , Fish Proteins , Mammals/metabolism
With the advent of emerging markets, the need for low-cost innovation to meet the rising demands of people at the base of the pyramid has increased significantly. Although the critical influence of customer participation on new product development has been recognized, there have been few studies on the effects of customer participation on low-cost innovation. This study builds a moderated mediation model and explores the roles of customer participation on low-cost innovation. Based on the exaptation and strategic flexibility theories, the mediating role of exaptation and the moderating role of strategic flexibility are emphasized. A survey of 348 firms revealed that customer participation positively impacted both exaptation and low-cost innovation. In addition, exaptation mediated the correlation between customer participation and low-cost innovation. Resource flexibility negatively moderated the correlation between customer participation and exaptation and negatively moderated the mediating effect of exaptation. Furthermore, coordinate flexibility positively moderated the correlation between customer participation and exaptation and positively moderated the mediating effect of exaptation.
PURPOSE: To explore the effect of external approach microsurgery in retinal dialysis. METHODS: Consecutive 30 eyes of 28 patients with retinal dialysis were enrolled for this study. The progresses of the external approach microsurgery were following. Under the surgical microscopy, the preplacement of mattress sutures for buckling and/or encircling following retrobulbar anesthesia and scleral exposure, draining subretinal fluid, the cryotherapy of retinal breaks, checking the position of breaks on scleral buckle and gases injection were performed in turn. RESULTS: After drainage of subretinal fluid, with scleral depression cryotherapy reaction around breaks could be observed clearly under the microscopy. All breaks were located on anterior slope of the buckle. Intraoperative complications were mild subretinal hemorrhage at drainage site and corneal epithelium exfoliation in 3 eyes, respectively. Postoperative complications were mainly secondary glaucoma and retinal redetachment. The one-operation reattachmentl rate was 96.7% (29 eyes), and the final reattachment rate was 100% after one eye had a second external approach microsurgery. The postoperative vision acuity (VA) was significantly better than the preoperative VA (chi(2)= 9.529, P< 0.01). CONCLUSION: External approach microsurgery has favourable effect on the surgery of retinal dialysis.
Microsurgery/methods , Retinal Detachment/surgery , Adolescent , Adult , Child , Female , Humans , Male , Middle Aged , Treatment Outcome , Visual Acuity , Young Adult
