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1.
Front Immunol ; 11: 531785, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33013923

RESUMO

Interleukin-2 (IL-2) is an important immunomodulatory cytokine that primarily promotes the activation, proliferation, and differentiation of CD4+ T helper subsets and CD4+ T regulatory cells. In our previous studies, IL-2 and IL-2 receptor beta (IL-2Rß) genes of flounder (Paralichthys olivaceus) were cloned, and IL-2Rß molecules expressed on both B and T lymphocytes were identified. In the present study, the interaction of flounder IL-2 (fIL-2) with the IL-2 receptor beta (fIL-2Rß) was investigated. The proportion of CD4+ T lymphocytes and IL-2Rß+ cells were detected both in vivo and in vitro. Firstly, the binding of recombinant flounder IL-2 protein (rfIL-2) and rfIL-2Rß was verified by pull-down assay and enzyme-linked immunosorbent assay. Indirect immunofluorescence assay showed that rfIL-2 enhanced the proliferation of CD4+ and IL-2Rß+ cells in the gill and spleen. Furthermore, CD4-1+, CD4-2+ T lymphocytes and IL-2Rß+ cells were significantly upregulated in cultured peripheral blood lymphocytes (PBLs) with addition of rfIL-2, as shown by Flow cytometry. The related genes were examined by Q-PCR in cultured PBLs with added rfIL-2. The results showed that the IL-2-IL-2R interaction induced upregulated expression of T lymphocyte surface makers, Th1-related cytokines or transcription factors, and critical genes of the IL-2 signaling pathway. In addition, these IL-2-elicited biological functions and immune responses were downregulated by blocked with anti-rfIL-2Rß and anti-rfIL-2 Abs, showing that IL-2Rß plays an indispensable role in IL-2 elicited biological function. Our results demonstrated that the interaction between IL-2 and IL-2Rß showed its potential to enhance the proliferation of CD4+ T lymphocytes in flounder. As found in mammals, a Th1-mediated mechanism regulated by this interaction exists in teleost.

2.
Nature ; 2020 Sep 23.
Artigo em Inglês | MEDLINE | ID: mdl-32968279

RESUMO

Cell death in human diseases is often a consequence of disrupted cellular homeostasis. If cell death is prevented without restoring cellular homeostasis, it may lead to a persistent dysfunctional and pathological state. Although mechanisms of cell death have been thoroughly investigated1-3, it remains unclear how homeostasis can be restored after inhibition of cell death. Here we identify TRADD4-6, an adaptor protein, as a direct regulator of both cellular homeostasis and apoptosis. TRADD modulates cellular homeostasis by inhibiting K63-linked ubiquitination of beclin 1 mediated by TRAF2, cIAP1 and cIAP2, thereby reducing autophagy. TRADD deficiency inhibits RIPK1-dependent extrinsic apoptosis and proteasomal stress-induced intrinsic apoptosis. We also show that the small molecules ICCB-19 and Apt-1 bind to a pocket on the N-terminal TRAF2-binding domain of TRADD (TRADD-N), which interacts with the C-terminal domain (TRADD-C) and TRAF2 to modulate the ubiquitination of RIPK1 and beclin 1. Inhibition of TRADD by ICCB-19 or Apt-1 blocks apoptosis and restores cellular homeostasis by activating autophagy in cells with accumulated mutant tau, α-synuclein, or huntingtin. Treatment with Apt-1 restored proteostasis and inhibited cell death in a mouse model of proteinopathy induced by mutant tau(P301S). We conclude that pharmacological targeting of TRADD may represent a promising strategy for inhibiting cell death and restoring homeostasis to treat human diseases.

3.
Sci Rep ; 10(1): 13827, 2020 Aug 14.
Artigo em Inglês | MEDLINE | ID: mdl-32796864

RESUMO

CD4+ T lymphocytes play crucial roles in the adaptive immune system. CD4, as the most effective marker to delineate the T-helper subsets, was identified in many fish species. Two CD4 homologs, CD4-1 and CD4-2, have been reported in flounder (Paralichthys olivaceus). In this study, monoclonal antibodies (mAbs) against CD4-1 and CD4-2 of flounder were produced, CD4+ T lymphocytes were isolated and identified, and the variations in CD4+ and CD8+ T lymphocytes and IgM+ B lymphocytes after Poly I:C, PMA or ß-glucan stimulation were investigated. Then, the expression of transcription factors and cytokines in sorted CD4+ T lymphocytes was analyzed. The results showed that the mAbs were specific to flounder CD4-1+ and CD4-2+ T cells. CD4-1+ and CD4-2+ cells responded to all three stimulants, while CD8+ T lymphocytes only give a strong response to Poly I:C, and the percentages of IgM+ B lymphocytes showed a tendency to increase. After stimulation, the expression of transcription factors and cytokines of Th1, Th2 and Th17 cells varied in CD4+ T cells. These results will provide crucial foundations for the differentiation and function of teleost CD4+ T lymphocytes.

4.
Int J Mol Sci ; 21(13)2020 Jul 02.
Artigo em Inglês | MEDLINE | ID: mdl-32630682

RESUMO

In previous research, voltage-dependent anion channel protein 2 (VDAC2) and the receptor of activated protein C kinase 1 (RACK1) in flounder (Paralichthys olivaceus) were confirmed as functional receptors for lymphocystis disease virus (LCDV) entry; however, the underlying mechanism of VDAC2- and RACK1-mediated LCDV entry remains unclear. In this study, we elucidated the endocytosis pathway of LCDV entry into flounder gill (FG) cells by treatment with specific inhibitory agents, siRNAs, and co-localization analysis. LCDV entry was significantly inhibited by the disruption of caveolae-mediated endocytosis, dynamin, and microtubules, and the knockdown of caveoline-1 and dynamin expression, but was not inhibited by the disruption of clathrin-mediated endocytosis, micropinocytosis, or low-pH conditions. The disruption of caveolae-mediated and clathrin-mediated endocytosis was verified by the internalization of cholera toxin subunit B (CTB) and transferrin, respectively. Confocal immunofluorescence assay demonstrated that LCDV was co-localized with VDAC2 and RACK1, CTB was co-localized with VDAC2 and RACK1 and partially with LCDV, but transferrin was not co-localized with LCDV, VDAC2, or RACK1, indicating that LCDV utilized the same pathway as CTB, i.e., caveolae-mediated endocytosis. This was different from the pathway of transferrin, which used clathrin-mediated endocytosis. Furthermore, caveolin-1 was co-localized with LCDV, VDAC2, and RACK1, suggesting that caveolin-1 was involved in LCDV entry. These results revealed for the first time that LCDV entered into FG cells via caveolae-mediated endocytosis facilitated by VDAC2 and RACK1 receptors, relying on dynamin and microtubules in a pH-independent manner, which provided new insight into the molecular mechanisms of LCDV entry and potential for the development of antiviral agents, expanding our understanding of iridovirus infection.

5.
Fish Shellfish Immunol ; 106: 296-306, 2020 Jul 24.
Artigo em Inglês | MEDLINE | ID: mdl-32717325

RESUMO

In our previous work, prohibitin1 (PHB1) was identified to be only expressed in granulocytes of Fenneropenaeus chinensis. In order to elucidate the potential immunological properties of prohibitins in hemocyte subpopulations, in this paper, the full-length cDNAs of PHB1 and PHB2 were firstly cloned from F. chinensis using rapid amplification of cDNA ends approach, and they were designated FcPHB1 and FcPHB2, respectively. Based on the sequence analysis and multiple sequence alignment, FcPHB1 and FcPHB2 were members of SPFH protein family. By quantitative real-time RT-PCR, the higher mRNA transcription levels of FcPHB1 and FcPHB2 were detected in intestine and hemocytes of F. chinensis, and these two genes in hemocytes were significantly up-regulated upon WSSV infection. The FcPHB1 and FcPHB2 were recombinantly expressed in Escherichia coli BL21 (DE3), and employed as immunogens to produce the polyclonal antibodies (PAbs) in rabbits. Indirect immunofluorescence assay (IFA) revealed that the FcPHB1 and FcPHB2 were located both in the cytoplasm and nuclei of hemocytes, which could also be specifically recognized by the PAbs against FcPHB1 or FcPHB2 in Western blot. Interestingly, it was found that FcPHB1 and FcPHB2 were only expressed in the granulocytes of heathy shrimp and highly expressed in the WSSV-infected granulocytes, however only weak expressions of FcPHB1 and FcPHB2 were observed in the hyalinocytes of WSSV-infected shrimp. Meanwhile, silencing of FcPHB1 and FcPHB2 genes were performed by small interfering RNA, and the results showed that the WSSV copies in hemocytes were increased by knockdown of either FcPHB1 or FcPHB2, and the cumulative mortalities of shrimp in the silenced groups were also markedly increased. These results demonstrated that FcPHB1 and FcPHB2 played important roles in anti-WSSV infection, and their differential expression characteristics in hemocyte subpopulations provided a further understanding of the immune functions of granulocytes and hyalinocytes.

6.
Fish Shellfish Immunol ; 104: 279-288, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32505718

RESUMO

Glycoprotein is an important immunogenic protein of Hirame novirhabdovirus (HIRRV). In this study, the full-length and N-/C-terminal portions of glycoprotein were recombinantly expressed (rG, rGn and rGc protein), and the induced immune responses were investigated in flounder (Paralichthys olivaceus) model. The results showed that compared to PBS control, rG, rGn and rGc proteins and inactivated HIRRV suspension (iVS) could all stimulate significant increases of flounder CD4-1+, CD4-2+ T lymphocytes and surface IgM positive (sIgM+) B lymphocytes in peripheral blood, spleen and head kidney (p < 0.05). However, no significant differences of the percentages of CD4-1+ or CD4-2+ T lymphocytes were observed among three protein vaccination groups (p > 0.05). iVS could induce the highest mean levels of CD4+ T lymphocytes in peripheral blood and spleen. For sIgM+ B lymphocytes, the average peak percentages in rG and rGc groups were higher than rGn group. Moreover, significant increases of specific serum IgM against HIRRV or rG protein were observed in iVS, rG, rGn and rGc groups, but rG group exhibited the highest mean level. Furthermore, rG protein induced the highest titer of neutralizing antibodies against HIRRV, followed by iVS. Meanwhile, the challenge test showed that the relative percent survival (RPS) of rG, rGn, rGc and iVS groups were 75.0%, 35.7%, 53.6% and 60.7%, respectively. These results revealed that the full-length G protein would be a more effective subunit vaccine candidate against HIRRV infection.

7.
Int J Nanomedicine ; 15: 2647-2658, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32368046

RESUMO

Purpose: Myocardial ischemia-reperfusion injury primarily causes myocardial infarction (MI), which is manifested by cell death. Angiogenesis is essential for repair and regeneration in cardiac tissue after MI. In this study, we aimed to investigate the effect of exosomes derived from the serum of MI patients in angiogenesis and its related mechanism. Patients and Methods: Exosomes, isolated from serum, were collected from MI (MI-exosome) and control (Con-exosome) patients. After coculturing with human umbilical vein endothelial cells, MI-exosome promoted cell proliferation, migration, and tube formation. Results: The results revealed that the production and release of MI-exosome were associated with cardiomyocytes. Moreover, microarray assays demonstrated that miRNA-143 was significantly decreased in MI-exosome. Meanwhile, the overexpression and knockdown of miRNA-143 could inhibit and enhance angiogenesis, respectively. Furthermore, the effect of exosomal miRNA-143 on angiogenesis was mediated by its targeting gene, insulin-like growth factor 1 receptor (IGF-IR), and was associated with the production of nitric oxide (NO). Conclusion: Taken together, exosomes derived from the serum of patients with MI promoted angiogenesis through the IGF-IR/NO signaling pathway. The results provide novel understanding of the function of exosomes in MI.


Assuntos
Vasos Coronários/metabolismo , Exossomos/metabolismo , MicroRNAs/metabolismo , Infarto do Miocárdio/sangue , Neovascularização Fisiológica , Receptor IGF Tipo 1/metabolismo , Animais , Linhagem Celular , Movimento Celular , Proliferação de Células , Células Endoteliais da Veia Umbilical Humana/metabolismo , Humanos , Masculino , Camundongos Endogâmicos C57BL , Camundongos Nus , Óxido Nítrico/biossíntese , Transdução de Sinais
8.
Colloids Surf B Biointerfaces ; 190: 110896, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32114270

RESUMO

Magnetic fluid hyperthermia has been one clinical treatment method in malignancy on account of the sufficient heat generation, which originates from hysteresis loss of magnetic nanomedicine in alternating magnetic field. Magnetic nanomedicine can also be employed as drug carrier for chemotherapy. Nevertheless few magnetic nanocarries has been approved in clinic, owing to the high pharmaceutical demand. For broadening the clinical application of current magnetic nanomedicine, novel magnetic hydrogel complex (DOX@FMT-MC) constituted by Doxorubicin, Ferumoxytol and Medical Chitosan was produced for hyperthermia and chemo synergistic therapy. The three materials were approved in clinic. Heat induction in vitro and rheology mesurements suggested this complex succeed in transforming into physical hydrogel when reaching hyperthermia temperature in alternating magnetic field. Drug release experiment implied the complex has the temperature-dependent slow drug release behaviour. Cell apoptosis assay presented that DOX@FMT-MC complex gave enhanced synergistic efficacy with 32.4 % on colon carcinoma cell treatment in vitro, compared to other therapeutic groups. Heat induction in mice subcutaneous xenografted tumour demonstrated the better heating performance of the complex than that of DOX@FMT. The novel hydrogel complex incorporated with three clinical available drugs promises the great potential in tumour synergistic treatment, motivating the clinical indication development of magnetic nanomedicine.

9.
Gene ; 741: 144552, 2020 May 30.
Artigo em Inglês | MEDLINE | ID: mdl-32165297

RESUMO

Hypoxia, as a form of stress, plays a critical role in oncogenesis, including metabolic reprogramming. Mitochondrial, the centers of energy production, re-balance mitochondria dynamic to maintain cell survival during high levels of environmental stresses. NDRG1 is a hypoxia-inducible protein that is involved in various human cancers, including HCC. However, little is known about whether NDRG1 participants in the quality control of mitochondrial in times of stress. Here, we firstly showed that how NDRG1 exerted its role through mediating mitochondrial dynamic in HCC cells under hypoxia. Initially, we identified that NDRG1 expression varies with oxygen content. NDRG1 silencing notably induced cell apoptosis under hypoxia, while no obviously change of wildtype cells in hypoxia compared with that in normoxia. Further analysis revealed that NDRG1 silencing in HCC cells led to increase of pro apoptotic protein BAX and decrease in anti-apoptotic proteins Bcl-2 and Bclx, which meant mitochondrial damage were induced. In the analysis of mitochondria, we found that more released cytochrome c located in cytosolic with NDRG1 knockdown in hypoxia, which may be due to mitochondria division. And the following experiment proved that more fragmented mitochondria were presented in NDRG1 silencing cells, as well as destroyed mitochondrial membrane potential with evidence by JC-1 was verified. Moreover, these trends could be reversed by Mdivi1. Further research showed that NDRG1 silencing disrupt hypoxia-enhanced aerobic glycolysis through effectively decreased glucose uptake, lactate output and ECAR value. In sum, we provide the first direct evidence that NDRG1-driven change in mitochondrial dynamics and aerobic glycolysis maintain cells survival in HCC during hypoxia.


Assuntos
Carcinoma Hepatocelular/genética , Proteínas de Ciclo Celular/genética , Reprogramação Celular/genética , Peptídeos e Proteínas de Sinalização Intracelular/genética , Neoplasias Hepáticas/genética , Apoptose/genética , Carcinoma Hepatocelular/patologia , Linhagem Celular Tumoral , Proliferação de Células/genética , Sobrevivência Celular/genética , Citocromos c/genética , Regulação Neoplásica da Expressão Gênica/genética , Glicólise/genética , Humanos , Neoplasias Hepáticas/patologia , Potencial da Membrana Mitocondrial/genética , Dinâmica Mitocondrial/genética , Proteínas Proto-Oncogênicas c-bcl-2/genética , Hipóxia Tumoral/genética , Proteína bcl-X/genética
10.
Sensors (Basel) ; 20(6)2020 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-32183461

RESUMO

This paper presents a line matching method based on multiple intensity ordering with uniformly spaced sampling. Line segments are extracted from the image pyramid, with the aim of adapting scale changes and addressing fragmentation problem. The neighborhood of line segments was divided into sub-regions adaptively according to intensity order to overcome the difficulty brought by various line lengths. An intensity-based local feature descriptor was introduced by constructing multiple concentric ring-shaped structures. The dimension of the descriptor was reduced significantly by uniformly spaced sampling and dividing sample points into several point sets while improving the discriminability. The performance of the proposed method was tested on public datasets which cover various scenarios and compared with another two well-known line matching algorithms. The experimental results show that our method achieves superior performance dealing with various image deformations, especially scale changes and large illumination changes, and provides much more reliable correspondences.

11.
Int J Neurosci ; : 1-9, 2020 Apr 05.
Artigo em Inglês | MEDLINE | ID: mdl-32186218

RESUMO

Aim/Purpose: Dental pulp stem cells (DPSCs) were widely used as seed cells in the field of tissue engineering and regenerative medicine, including spinal cord injury (SCI) repair and other neuronal degenerative diseases, due to their easy isolation, multiple differentiation potential, low immunogenicity and low rates of rejection during transplantation. Various studies have shown that bFGF can enhance peripheral nerve regeneration after injury, and phospho-ERK (p-ERK) activation as a major mediator may be involved in this process. Previous studies also have proved that a suitable biomaterial scaffold can carry and transport the therapeutic cells effectively to the recipient area. It has showed in our earlier experiments that 3D porous chitosan scaffolds exhibited a suitable circumstance for survival and neural differentiation of DPSCs in vitro. The purpose of the study was to evaluate the influence of chitosan scaffolds and bFGF on differentiation of DPSCs.Materials and methods: In current study, DPSCs were cultured in chitosan scaffolds and treated with neural differentiation medium for 7 days. The neural genes and protein markers were analyzed by western blot and immunofluorescence. Meanwhile, the relevant signaling pathway involved in this process was also tested.Results: Our study revealed that the viability of DPSCs was not influenced by co-culture with the chitosan scaffolds as well as bFGF. Compared with the control and DPSC/chitosan-scaffold groups, the levels of GFAP, S100ß and ß-tubulin III significantly increased in the DPSC/chitosan-scaffold+bFGF group.Conclusion: Chitosan scaffolds were non-cytotoxic to the survival of DPSCs, and chitosan scaffolds combined with bFGF facilitated the neural differentiation of DPSCs. The transplantation of DPSCs/chitosan-scaffold+bFGF might be a secure and effective method of treating SCI and other neuronal diseases.

12.
Int Immunopharmacol ; 81: 106195, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32028242

RESUMO

Baicalin (BA), a flavone glycoside, is the constituent of Scutellaria baicalensis, a Chinese herbal medicine used to treat non-alcoholic steatohepatitis (NASH). However, the mechanism of BA on NASH is still not clear. Here, the improving effect of BA on hepatocyte through inhibition of pyroprosis was investigated in vitro. With a cell model of NASH exposing HepG2 cells in free fatty acids (FFA), we revealed that BA could improve hepatocyte from FFA-induced morphological damage and death. And then through transcriptomes screening, a significant down-regulation of NLR pyrin domain containing 3 (Nlrp3), gasdermin D (Gsdmd), andinterleukin-1 beta (IL-1ß) expression were found after BA treatment. Further analysis confirmed that BA could decrease the levels of NLRP3 and GSDMD, as well as the release of IL-1ß and IL-18, resulting in the reduction of pyroptosis. Moreover, the improving effect of BA could be attenuated by Gsdmd knockdown. In conclusion, BA can reduce pyroptosis of hepatocyte by blocking NLRP3-GSDMD signaling in vitro.

13.
J Med Chem ; 63(3): 1337-1360, 2020 02 13.
Artigo em Inglês | MEDLINE | ID: mdl-31910017

RESUMO

p300 and CREB-binding protein (CBP) are ubiquitously expressed pleiotropic lysine acetyltransferases and play a key role as transcriptional co-activators that are essential for a multitude of cellular processes. Despite great importance, there is a lack of highly selective, potent, druglike p300/CBP inhibitors. Through the artificial-intelligence-assisted drug discovery pipeline and further optimization, we reported the discovery of novel, highly selective, potent small-molecule inhibitors of p300/CBP histone acetyltransferases (HAT) with desired druglike properties, exemplified by B026. Our data demonstrated that B026, with half maximal inhibitory concentration (IC50) values of 1.8 nM to p300 and 9.5 nM to CBP enzyme inhibitory activity, is the most potent, selective p300/CBP HAT inhibitor. Moreover, B026 achieves significant and dose-dependent tumor growth inhibition in an animal model of human cancer, suggesting that B026 is a highly promising p300/CBP HAT inhibitor and warrants extensive preclinical investigation as a potential clinical development candidate.


Assuntos
Antineoplásicos/uso terapêutico , Proteína de Ligação a CREB/antagonistas & inibidores , Proteína p300 Associada a E1A/antagonistas & inibidores , Inibidores Enzimáticos/uso terapêutico , Neoplasias/tratamento farmacológico , Compostos de Espiro/uso terapêutico , Animais , Antineoplásicos/síntese química , Antineoplásicos/farmacocinética , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Conjuntos de Dados como Assunto , Descoberta de Drogas , Inibidores Enzimáticos/síntese química , Inibidores Enzimáticos/farmacocinética , Feminino , Humanos , Masculino , Camundongos Endogâmicos BALB C , Microssomos Hepáticos/metabolismo , Simulação de Acoplamento Molecular , Estrutura Molecular , Redes Neurais de Computação , Ratos Sprague-Dawley , Compostos de Espiro/síntese química , Compostos de Espiro/farmacocinética , Relação Estrutura-Atividade
14.
Dev Comp Immunol ; 106: 103615, 2020 05.
Artigo em Inglês | MEDLINE | ID: mdl-31956084

RESUMO

In our previous study, we cloned and explored the biological functions of flounder (Paralichthys olivaceus) interleukin-2 (poIL-2), and showed that poIL-2 might have adjuvant potential for fish vaccines. In this study, the adjuvant effects of recombinant and molecular forms of poIL-2 (rIL-2 and pcIL-2) were comparatively analyzed and evaluated in flounder from several aspects by co-vaccination with the recombinant E. tarda OmpV (rOmpV). The results showed that co-vaccination with rOmpV plus rIL-2 or pcIL-2 resulted in a relative percent survival of 71% and 57% respectively, which was significantly higher than the control groups, rOmpV plus rHis (40%) or pcN3 (36%). Immunological analysis showed that: (1) the levels of specific serum antibodies and sIg + lymphocytes in head kidney, spleen and peripheral blood induced by rOmpV plus rIL-2 or pcIL-2 were significantly higher than that in the two control groups; (2) Compared to the two control groups, CD4-1, CD4-2, CD8α, CD8ß, MHCIα, MHCIIα, IgM and IFN-γ mRNA levels were also significantly induced by rOmpV plus rIL-2 or pcIL-2; (3) the rOmpV plus rIL-2 could induce higher levels of sIg + lymphocytes, specific serum antibodies and the expressions of all investigated genes than rOmpV plus pcIL-2. These results demonstrated that co-vaccination with rOmpV with rIL-2 or pcIL-2 could induce stronger humoral and cellular immune responses, and evoked higher immune protective efficacy against E. tarda infection, suggesting that poIL-2 could be served as a promising candidate adjuvant and have a potential application in the control of flounder diseases.

15.
Dev Comp Immunol ; 103: 103492, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-31494219

RESUMO

The magnitude of the immune response induced by DNA vaccines depends on the amount and type of antigen-presenting cells attracted to the injection site. In our previous study, a DNA plasmid encoding the VAA gene of Vibrio anguillarum was constructed and shown to confer moderate protection against V. anguillarum challenge. To augment the protective efficacy of the VAA DNA vaccine and compare the adjuvant effects of CCL3, CCL4, CCL19 and CCL21, four bicistronic DNA plasmids containing the VAA gene of V. anguillarum together with the gene encoding the CCL3/CCL4/CCL19/CCL21 chemokines of flounder were successfully constructed and administered to fish, and the immune response of the animals and the enhancement of immunoprotection by the four chemokines were investigated. Vaccinated with pCCL3-VAA, pCCL4-VAA, pCCL19-VAA and pCCL21-VAA, flounder showed relative percent survivals of 62.16%, 83.78%, 78.38% and 72.97%, respectively, higher than the relative survival of flounder immunized with pVAA (40.54%). Compared with the pVAA group, the percentages of sIgM+, CD4-1+, and CD4-2+ lymphocytes and the levels of specific antibodies increased in pCCL3-VAA, pCCL4-VAA, pCCL19-VAA and pCCL21-VAA injection groups; CCL4 and CCL19 induced significantly higher levels of these parameters than CCL3 and CCL21 did. The amount of V. anguillarum in liver, spleen and kidney of pCCL3-VAA-, pCCL4-VAA-, pCCL19-VAA- and pCCL21-VAA-immunized flounder after V. anguillarum challenge was reduced compared to that in the pVAA group. Moreover, the co-expression of CCL3/CCL4/CCL19/CCL21 up-regulated immune-related gene expression associated with the local immune response. Our results indicate that CCL4 and CCL19 are promising adjuvants for use in VAA DNA vaccine against V. anguillarum.

16.
J Nanosci Nanotechnol ; 20(2): 668-672, 2020 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-31383061

RESUMO

Currently, optical probes with near-infrared (NIR) fluorescence are of great interest in chemical biology. In the present study, we designed and synthesized a novel NIR fluorescent probe, IR789. IR789 has high selectivity and sensitivity for living cells imaging. The stronger excitation and emission characteristics suggested its dominant optical properties over ICG. IR789 also showed a high affinity and inconspicuous cytotoxicity at the cellular level. The results of fluorescent image in living A549 cells (human lung adenocarcinoma epithelial cell line) further demonstrated its potential applications for biomedical diagnosis in biological systems utilization of nanotechnology.

17.
Mol Immunol ; 118: 40-51, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-31841966

RESUMO

To further elucidate the roles of T and B lymphocytes in fish subunit and DNA candidate vaccines for immunisation, the immune responses of T and B lymphocytes to recombinant protein (rOmpK) and plasmid OmpK (pOmpK) from Vibrio anguillarum plus cyclosporine A (CsA) were investigated in flounder (Paralichthys olivaceus). The results showed that in the rOmpK-immunised groups, the percentages of CD4-1+ and CD4-2+ T (PCD4-1+ and PCD4-2+ T) lymphocytes significantly increased to a peak on days 5 or 7. The percentages of IgM+ B (PIgM+ B) lymphocytes and specific antibodies markedly increased to a peak at weeks 4 or 5. The nine immune-related genes were significantly up-regulated and the expression levels of CD4-1, CD4-2 and MHC II genes were higher than that of CD8α, CD8ß and MHC I genes. The CD4+ T lymphocytes, IgM+ B lymphocytes, and specific antibodies were significantly inhibited by CsA. Therefore, the responses of CD4+ T lymphocytes influenced the responses of the B lymphocytes and antibodies. In the pOmpK-immunised groups, the PCD4-1+, PCD4-2+, and PCD8ß+ T lymphocytes significantly increased to a peak on days 11 or 14, days 9 or 11, and days 7 or 9, respectively. The PIgM+ B lymphocytes and specific antibodies significantly increased to a peak at weeks 5 or 6. Immune related genes upregulated, and CD4+ and CD8+ T lymphocytes, IgM+ B lymphocytes and specific antibodies all suppressed by CsA, suggesting that the responses of T lymphocytes subpopulations influenced B lymphocytes and antibodies responses. Therefore, the subpopulations of T lymphocytes played an important role in the immune responses induced by subunit and DNA candidate vaccines of OmpK and regulated the immune responses of B lymphocytes in flounder.


Assuntos
Linfócitos B/imunologia , Proteínas da Membrana Bacteriana Externa/imunologia , Linguado/imunologia , Subpopulações de Linfócitos T/imunologia , Vacinas de DNA/imunologia , Vacinas de Subunidades/imunologia , Vibrio/imunologia , Animais , Anticorpos Antibacterianos/imunologia , Antígenos de Bactérias/imunologia , Proteínas de Bactérias/imunologia , Vacinas Bacterianas/imunologia , Doenças dos Peixes/imunologia , Doenças dos Peixes/microbiologia , Linguado/microbiologia , Imunização/métodos , Vacinação/métodos , Vibrioses/imunologia , Vibrioses/microbiologia
18.
Zhong Nan Da Xue Xue Bao Yi Xue Ban ; 44(9): 996-1002, 2019 Sep 28.
Artigo em Chinês | MEDLINE | ID: mdl-31645488

RESUMO

OBJECTIVE: To explore the relationship between paediatric early warning score (PEWS) and the occurrence of mechanical ventilation complications in children with acute respiratory distress syndrome (ARDS).
 Methods: A total of 110 children with ARDS diagnosed in First Affiliated Hospital of Hebei North University, who underwent mechanical ventilation, were selected. The baseline data, blood gas analysis index, laboratory test index, ventilator parameters, pediatric critical illness score (PCIS) and PEWS in the children were recorded. With reference to ventilatory treatment results, the children with ventilator-associated complications were included in the trial group (n=20), while the patients with good cohort status were included in the control group (n=40) according to the nested case-control study. Independent sample t-test and multivariate logistic regression analysis were used to analyze the factors affecting the occurrence of complications after ventilatory treatment.
 Results: There were statistically significant differences in multiple organ dysfunction syndrome (MODS), partial pressure of oxygen/fraction of inspired oxygen (PaO2/FiO2), partial pressure of carbon dioxide (PaCO2), serum creatinine (SCr), albumin (ALB), blood urea nitrogen (BUN), mechanical ventilation time, mean article pressure (MAP), tidal volume (VT), positive end-expiratory pressure (PEEP), PCIS, PEWS between the control group and the experimental group (all P<0.05). Multivariate logistic regression analysis showed that MODS, PaO2/FiO2, PaCO2, VT, PEEP and PEWS had influence on complications after mechanical ventilation in children with ARDS (all P<0.05).
 Conclusion: The MODS, PaO2/FiO2, PaCO2, VT, PEEP, and PEWS exert effects on complications after mechanical ventilation in children with ARDS. PEWS combined with other indicators can assess the risk of complications in children with ARDS after mechanical ventilation.


Assuntos
Síndrome do Desconforto Respiratório do Adulto , Estudos de Casos e Controles , Criança , Humanos , Respiração com Pressão Positiva , Respiração Artificial , Volume de Ventilação Pulmonar
19.
Fish Shellfish Immunol ; 94: 907-915, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31604147

RESUMO

Previous studies have demonstrated that white spot syndrome virus (WSSV) could induce hemocytes apoptosis in shrimps, however the inter-relationship between apoptotic process and the WSSV infection status is still currently underexplored. In the present work, the apoptosis and the viral proliferation in hemocytes of Litopenaeus vannamei were simultaneously investigated post WSSV infection by two-color immunofluorescence flow cytometry and real-time quantitative PCR. The apoptotic hemocytes of WSSV-infected shrimp was significantly increased at 12 h post infection (hpi), whereas underwent a slight decline at 24 hpi subsequently. Since 24 hpi, the apoptotic rate of hemocytes in the WSSV-infected shrimp exhibited a rapid and significant increase, and reached the peak level at 48 hpi with the ratio of 18.1 ±â€¯2.0%. Meanwhile, the percentage of WSSV-infected hemocytes and WSSV copies in hemocytes significantly increased at 24 hpi and maintained at a high level afterwards. With the rapid increase of hemocytes apoptosis, hemocyte density in hemolymph decreased dramatically to less than 20% of the mean value of control. Co-localization assay showed that the apoptotic WSSV-infected hemocytes occupied the dominant proportion of total apoptotic hemocytes, which reached the peak at 48 hpi with 12.6 ±â€¯1.5%. The expression profiles of seven pro-apoptotic genes and two apoptosis-inhibiting genes showed significant differential responses at different stages of WSSV infection, reflecting the interplay between the virus and the host immune response. Our results demonstrated that the apoptotic response of shrimp hemocytes could be significantly influenced by the WSSV infection process, which might provide an insight into deeper relationships between viral infection and apoptosis.


Assuntos
Apoptose , Proteínas de Artrópodes/genética , Hemócitos/imunologia , Penaeidae/imunologia , Vírus da Síndrome da Mancha Branca 1/fisiologia , Animais , Proteínas de Artrópodes/imunologia
20.
Antioxidants (Basel) ; 8(9)2019 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-31480577

RESUMO

BACKGROUND: Although kidney injury caused by cisplatin has attracted much attention, cisplatin-induced cardiotoxicity is elusive. Our previous studies have confirmed that saponins (ginsenosides) from Panax quinquefolius can effectively reduce acute renal injuries. Our current study aimed to identify the potential effects of saponins from leaves of P. quinquefolius (PQS) on cisplatin-evoked cardiotoxicity. METHODS: Mice were intragastrically with PQS at the doses of 125 and 250 mg/kg daily for 15 days. The mice in cisplatin group and PQS + cisplatin groups received four times intraperitoneal injections of cisplatin (3 mg/kg) two days at a time from the 7th day, respectively. All mice were killed at 48 h following final cisplatin injection. Body weights, blood and organic samples were collected immediately. RESULTS: Our results showed that cisplatin-challenged mice experienced a remarkable cardiac damage with obvious histopathological changes and elevation of lactate dehydrogenase (LDH), creatine kinase (CK), creatine kinase isoenzyme MB (CK-MB) and cardiac troponin T (cTnT) concentrations and viabilities in serum. Cisplatin also impaired antioxidative defense system in heart tissues manifested by a remarkable reduction in reduced glutathione (GSH) content and superoxide dismutase (SOD) activity, demonstrating the overproduction of reactive oxygen species (ROS) and oxidative stress. Interestingly, PQS (125 and 250 mg/kg) can attenuate cisplatin-evoked changes in the above-mentioned parameters. Additionally, PQS administration significantly alleviated the oxidation resulted from inflammatory responses and apoptosis in cardiac tissues via inhibition of overexpressions of TNF-α, IL-1ß, Bax, and Bad as well as the caspase family members like caspase-3, and 8, respectively. CONCLUSION: Findings from our present research clearly indicated that PQS exerted significant effects on cisplatin-induced cardiotoxicity in part by inhibition of the NF-κB activity and regulation of PI3K/Akt/apoptosis mediated signaling pathways.

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