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1.
J Colloid Interface Sci ; 559: 51-64, 2020 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-31610305

RESUMO

Aiming at the inefficiency and toxicity in traditional antitumor therapy, a novel multifunctional nanoplatform was constructed based on hollow mesoporous carbon (HMC) to achieve triple stimuli response and dual model antitumor therapy via chemo-photothermal synergistic effect. HMC was used as an ideal nanovehicle with a high drug loading efficiency as well as a near-infrared (NIR) photothermal conversion agent for photothermal therapy. Acid-dissoluble, luminescent ZnO quantum dots (QDs) were used as the proper sealing agents for the mesopores of HMC, conjugated to HMC via disulfide linkage to prevent drug (doxorubicin, abbreviated as Dox) premature release from Dox/HMC-SS-ZnO. After cellular endocytosis, the Dox was released in a pH, GSH and NIR laser triple stimuli-responsive manner to realize accurate drug delivery. Moreover, the local hyperthermia effect induced by NIR irradiation could promote the drug release, enhance cell sensitivity to chemotherapeutic agents, and also directly kill cancer cells. As expected, Dox/HMC-SS-ZnO exhibited a high drug loading capacity of 43%, well response to triple stimuli and excellent photothermal conversion efficiency η of 29.7%. The therapeutic efficacy in 4T1 cells and multicellular tumor spheroids (MCTSs) demonstrated that Dox/HMC-SS-ZnO + NIR had satisfactory chemo-photothermal synergistic effect with a combination index (CI) of 0.532. The cell apoptosis rate of the combined treatment group was more than 95%. The biodistribution and pharmacodynamics studies showed its biosecurity to normal tissues and synergistic inhibition effect to tumor cells. These distinguished results indicated that the Dox/HMC-SS-ZnO nanoplatform is potential to realize efficient triple stimuli-responsive drug delivery and dual model chemo-photothermal synergistic antitumor therapy.


Assuntos
Antineoplásicos/química , Carbono/química , Terapia Combinada/métodos , Portadores de Fármacos/química , Nanopartículas/química , Pontos Quânticos/química , Óxido de Zinco/química , Animais , Antineoplásicos/farmacocinética , Apoptose/efeitos dos fármacos , Materiais Biocompatíveis/química , Linhagem Celular Tumoral , Permeabilidade da Membrana Celular , Sobrevivência Celular/efeitos dos fármacos , Doxorrubicina/química , Liberação Controlada de Fármacos , Corantes Fluorescentes/química , Humanos , Raios Infravermelhos , Camundongos Endogâmicos BALB C , Fototerapia/métodos , Porosidade , Propriedades de Superfície , Distribuição Tecidual , Óxido de Zinco/farmacocinética
2.
Colloids Surf B Biointerfaces ; 184: 110532, 2019 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-31590051

RESUMO

In this work, a multi-stimuli responsive drug delivery system (MCHP) was designed for combinational chemotherapy and photothermal therapy (PTT). Mesoporous carbon nanoparticles (MCN) with a high loading efficiency were used as near-infrared (NIR)-responsive drug carriers. Human serum albumin (HSA) was attached to the pore openings of MCN via disulfide bonds to serve as a gatekeeper due to its biocompatibility and appropriate molecular size. To improve the dispersity and biocompatibility, the surface of the MCN was modified with polyethylene glycol (PEG). In vitro photothermal effect results showed that MCHP exhibited a power and concentration-dependent photothermal conversion capacity and a good photothermal stability. The doxorubicin (DOX) release from the MCHP/DOX system exhibited NIR/pH/reduction-responsive release properties. A cytotoxicity assay demonstrated that, under NIR irradiation, the MCHP/DOX exhibited chemo-photothermal synergistic effects with a combination index (CI) of 0.643. The biodistribution of DOX in vivo indicated that an NIR laser can prolong the retardation time of DOX in tumor sites. In vivo antitumor experiments showed that MCHP/DOX with NIR irradiation had the highest tumor inhibition rate against 4T1 tumors in mice. This work suggested that MCHP could be explored as a multi-responsive drug release platform for combinational photothermo-chemotherapy.

3.
J Colloid Interface Sci ; 552: 639-650, 2019 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-31173992

RESUMO

In this work, a tumor-targeted and multi-stimuli responsive drug delivery system combining infrared thermal imaging of cells with thermo-chemotherapy was developed. Oxidized mesoporous carbon nanoparticles (MCNs-COOH) with high photothermal conversion ability (photothermal transduction efficiency η = 27.4%) in near-infrared (NIR) region were utilized to encapsulate doxorubicin (DOX). The outer surfaces of MCNs-COOH were capped with multifunctional carbon dots (CDHA) as simultaneous smart gatekeepers, a tumor targeting moiety and a fluorescent probe. NIR laser irradiation killed cancer cells through NIR-light induced hyperthermia, facilitated chemotherapeutic drug release and enhanced the sensitivity of tumor cells to drugs. The therapeutic efficacy in two-dimensional (2D) and three-dimensional (3D) cells demonstrated that MC-CDHA loading DOX (MC-CDHA/DOX) had good chemo-photothermal synergistic antitumor effects (combination index of CI = 0.448). The biodistribution and pharmacodynamics experiments of MC-CDHA/DOX in the 4T1 tumor model indicated that MCNs-COOH prolonged the residence time of DOX in tumor tissues and therefore actualized effective synergistic photothermal chemotherapy. By combining these excellent capabilities, the tumor-targeted and multi-stimuli responsive drug delivery system can be utilized as a visible nanoplatform for chemophotothermal synergistic therapy.

4.
Fish Shellfish Immunol ; 88: 318-327, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30853654

RESUMO

A variety of combinations of leucine-rich repeat (LRR) and immunoglobulin-like (Ig) domains have been found and discovered in invertebrates and vertebrates, but the functions remain largely unexplored. In the present study, a novel LRR and Ig domain-containing protein (LRRIG), CgLRRIG-3, was identified and characterized from oyster Crassostrea gigas. It contained two typical LRR motifs, a LRRNT motif and an Ig domain and PSI-BALST and phylogeny analysis revealed that the sequence of CgLRRIG-3 was most related with leucine-rich repeat neuronal 1 proteins from vertebrate. Its mRNA transcripts were constitutively expressed in muscle, gill, hepatopancreas, mantle, gonad and hemocytes with the highest level in hepatopancreas. The mRNA expression level of CgLRRIG-3 in hemocytes could respond to the stimulations of variety PAMPs including lipopolysaccharide (LPS), peptidoglycan (PGN), glucan (GLU) and polyinosinic-polycytidylic acid (poly I:C). The recombinant proteins exhibited a wide PAMP binding repertoire to four typical PAMPs and could significantly induce the expression of CgTNF-1 and CgIL17-5 as well as increase phagocytosis in primary cultured oyster hemocytes. In hepatopancreas, CgLRRIG-3 was mainly distributed in the basolateral membrane of digestive tubule and the hemocoel sinusoid between the digestive tubules. And in hemocytes, the positive signal was mainly distributed in a special group of granulocytes. These results collectively indicated that CgLRRIG-3 could not only function as an immune effector.


Assuntos
Crassostrea/genética , Crassostrea/imunologia , Imunidade Inata , Receptores de Reconhecimento de Padrão/genética , Receptores de Reconhecimento de Padrão/imunologia , Sequência de Aminoácidos , Animais , Células Cultivadas , Hemócitos/metabolismo , Domínios de Imunoglobulina , Padrões Moleculares Associados a Patógenos/farmacologia , Filogenia , Domínios Proteicos , Receptores de Reconhecimento de Padrão/química , Alinhamento de Sequência
5.
Heart Surg Forum ; 22(1): E019-E023, 2019 01 28.
Artigo em Inglês | MEDLINE | ID: mdl-30802192

RESUMO

BACKGROUND: To study the effect of miR-29b on myocardial infarction via Notch signaling pathway in rats. METHODS: The rat acute myocardial infarction (AMI) models were established and were divided into AMI group, sham group and normal group (N = 10 in each group). HE (Hemotoxylin and eosin) staining was used to detect whether the model was constructed successfully. MiR-29b mimics, inhibitors, mimics negative control (NC) were transfected into H9c2 (2-1) cells. Then, cells were divided into a mimics group, inhibitor group, NC group, and blank group. The relative expression levels of miR-29b, Notch1, DII4 and Hesl were detected by qRT-PCR. The expression of NICD1 was detected by Western blotting. RESULTS: The rat AMI model was successfully constructed. Compared with normal and sham groups, the miR-29b expression was down-regulated, while the expression of Notch1, DII4 and Hesl was increased, and the NICD1 protein expression was increased in the myocardial infarction area of the AMI group (P < .05). Compared with the NC and blank groups, the relative expression of Notch1, DII4, Hesl and NICD1 were upregulated in the mimics group (P < .05), whereas the expression of Notch1, DII4, Hesl and NICD1 in the inhibitor group was decreased (P < .05). CONCLUSION: MiR-29b inhibited myocardial fibrosis and cardiac hypertrophy by activating the Notch signaling pathway and protected myocardium against myocardial infarction.


Assuntos
Regulação da Expressão Gênica , MicroRNAs/genética , Infarto do Miocárdio/genética , RNA/genética , Receptor Notch1/genética , Remodelação Ventricular , Animais , Western Blotting , Modelos Animais de Doenças , Masculino , MicroRNAs/biossíntese , Infarto do Miocárdio/metabolismo , Infarto do Miocárdio/fisiopatologia , Ratos , Reação em Cadeia da Polimerase em Tempo Real , Receptor Notch1/metabolismo , Transdução de Sinais
6.
Dev Comp Immunol ; 91: 132-142, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30389518

RESUMO

As an important post-translational protein modification, ubiquitination has been demonstrated to play a vital role in immune response of vertebrates. Ubiquitin (Ub)-conjugating enzyme E2 is the "heart" of ubiquitination, which is responsible for Ub cellular signaling and substrate modification. In the present study, an Ub-conjugating enzyme E2 (designed as CgUbe2g1) was identified from oyster Crassostrea gigas, and its regulation in the immune response against lipopolysaccharide (LPS) stimulation was investigated. CgUbe2g1 encoded a polypeptide of 168 amino acids with the predicted molecular mass of 19.20 kDa and contained conserved catalytic 'Ubc' domains. It shared a higher similarity with the known UBC2G1 type E2s and was closely clustered with the type E2s identified from invertebrates in the phylogenetic assay. The mRNA transcripts of CgUbe2g1 were mainly distributed in hemocyte, mantle, hepatopancreas and male gonad of C. gigas. CgUbe2g1 protein was found to be colocalized with Ub around the nucleus of oyster hemocyte. The recombinant CgUbe2g1 protein (rCgUbe2g1) could activate the ubiquitination in vitro by binding both activated and un-activated Ub. The expressions of inflammation-related factors TNF-α and NF-κB in CgUbe2g1 transfected cells were both significantly up-regulated after LPS stimulation, which were 12.9-fold at 3 h (p < 0.01) and 2.3-fold at 6 h (p < 0.01) of that in negative control group, respectively. The phagocytic rate of hemocyte and the ROS level in hemocyte were both significantly decreased (p < 0.01), while the apoptosis rate was significantly increased (p < 0.01) after CgUbe2g1 mRNA was interfered. These results demonstrated that Ub-conjugating enzyme CgUbe2g1 was involved in the innate immune response of oyster against invading pathogen, which might play important roles in the activation of inflammatory response and regulation of cellular immune response.


Assuntos
Crassostrea/imunologia , Hemócitos/fisiologia , Enzimas de Conjugação de Ubiquitina/genética , Animais , Apoptose , Células Cultivadas , Clonagem Molecular , Humanos , Imunidade Inata , NF-kappa B/metabolismo , Filogenia , Transcriptoma , Fator de Necrose Tumoral alfa/metabolismo , Enzimas de Conjugação de Ubiquitina/metabolismo , Ubiquitinação
7.
J Colloid Interface Sci ; 535: 380-391, 2019 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-30316125

RESUMO

"Gate" engineered mesoporous silica nanoparticles (MSN) have been extensively applied in cancer theranostics. Due to the complexity of tumor development and progression, with chemotherapy alone, it has often been difficult to achieve a good therapeutic effect. Currently, it has been shown that the combination with photothermal therapy overcomes the shortcoming of chemotherapy. In most studies, the photothermal effect has proven to accelerate drug release from nanocarriers and ablate malignant cells directly, but the influence on the intracellular fate of nanocarriers remains unknown. Herein, a lipophilic cyanine dye Cypate acting as a photothermal converting agent was conjugated on the external surface of MSN through a disulfide bond (MSN-Cy) and d-α-tocopherol polyethylene glycol 1000 succinate (TPGS) was coated on the outside of the MSN-Cy via a hydrophobic interaction (TCMSN) to cover the pores, preventing drug preleakage in the circulation. The TCMSN underwent exocytosis through the lysosome-mediated pathway. Moderate heat induced by near-infrared light promoted lysosome disruption, which thus partly inhibited lysosome-mediated particle exocytosis. In the meantime, TPGS, as a P-glycoprotein inhibitor, blocked the drug efflux. This research elaborated the photothermal effect from a new perspective-inhibiting particle exocytosis. The as-designed "gate" engineered MSN realized a double inhibition of drug efflux and particle exocytosis from cancer cells, thus sustaining the drug action time and enhancing the antitumor activity.


Assuntos
Antineoplásicos/farmacologia , Nanopartículas/química , Dióxido de Silício/química , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/antagonistas & inibidores , Animais , Antineoplásicos/química , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Portadores de Fármacos/química , Ensaios de Seleção de Medicamentos Antitumorais , Exocitose/efeitos dos fármacos , Feminino , Lisossomos/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos BALB C , Estrutura Molecular , Tamanho da Partícula , Porosidade , Propriedades de Superfície
8.
Zookeys ; (794): 85-94, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30416341

RESUMO

One new leafhopper genus Paracodilia gen. n. with one new species P.geniculata sp. n., and two other new species in different genera, Bolivielaexpanda and Armaturolidiasymmetrica spp. n., are described in the tribe Coelidiini (Cicadellidae: Coelidiinae) from the Neotropical region. Photographs and illustrations are provided.

9.
Zootaxa ; 4415(3): 591-600, 2018 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-30313619

RESUMO

Three new species from Republic of the Congo, Brasura sinistra, B. piscinura and Tialidia hama spp. nov., and one new species Limentinus declinatus sp. nov. from Madagascar are described and illustrated in this paper.


Assuntos
Hemípteros , Animais , Congo , Madagáscar
10.
ACS Appl Mater Interfaces ; 10(23): 19386-19397, 2018 Jun 13.
Artigo em Inglês | MEDLINE | ID: mdl-29793337

RESUMO

Thermochemotherapy exhibits a synergistic therapeutic efficiency for cancer, and the sensitivity of cancer cells to chemical drugs could be increased to a large extent at elevated temperature. In this work, a biocompatible nanocomposite thermosensitive mesoporous carbon nanoparticles (TSMCN) was prepared by covering a liposome on mesoporous carbon nanoparticles (MCN). The TSMCN had good photothermal efficiency and photostability. The doxorubicin (DOX)-loaded TSMCN (DOX/TSMCN) showed a slower release than the DOX-loaded MCN-COOH (DOX/MCN-COOH) both in simulated tumor environment and physiological environment. And release curves of DOX/TSMCN exposed to NIR laser exhibited the fast release property. The confocal laser scanning microscopy results illustrated that cellular uptake of DOX for DOX/TSMCN can be enhanced by NIR laser. The temperature of the tumor site reached up to 51.9 °C within 3 min after exposure to laser at 1.25 W/cm2 power density, which is above the phase transition temperature ( Tm) of liposome (40.7 °C). The biodistribution of DOX in vivo indicated that NIR laser can prolong the retardation time of DOX in the tumor site. The results of both 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and antitumor efficiency elucidated that the DOX/TSMCN under NIR irradiation had a synergistic therapeutic effect for cancer. Thus, the TSMCN could be explored as a powerful nanoplatform that shows great prospect in thermochemotherapy of tumor therapy.


Assuntos
Nanopartículas , Carbono , Doxorrubicina , Sistemas de Liberação de Medicamentos , Humanos , Bicamadas Lipídicas , Neoplasias , Distribuição Tecidual
11.
Fish Shellfish Immunol ; 77: 419-428, 2018 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-29609030

RESUMO

Astakine is a cytokine-like factor containing a prokineticin domain, which directly participates in hematopoiesis and blood cell differentiation. In the present study, a novel Astakine gene was identified from Chinese mitten crab Eriocheir sinensis (designated as EsAst). The full-length cDNA of EsAst was of 1163 bp, consisting of a 5' untranslated region (UTR) of 120 bp, a 3' UTR of 656 bp, and an open reading frame (ORF) of 387 bp encoding a polypeptide of 128 amino acids. There were a signal peptide and a prokineticin domain with nine conserved cysteine residues in the deduced amino acid sequence of EsAst. EsAst shared higher similarity with Astakines from Penaeus monodon and Pacifastacus leniusculus, and it was closely clustered with the Astakine from shrimp P. monodon in the phylogenetic tree. The EsAst mRNA transcript was higher expressed in hemocytes and hepatopancreas. The relative expression level of EsAst in hemocytes was continuously increased from 1.5 to 48 h after Vibro anguillarum challenge compared that in the untreated control group. After Pichia pastoris GS115 challenge, the relative expression level of EsAst in hemocytes was also up-regulated. After rEsAst injection, ROS levels in HPT cells were also increased at 12 and 24 h, and the total hemocyte counts were also significantly increased at 6, 9, 12, and 24 h post rEsAst injection. The interference of EsAst expression with dsRNA injection could delay the recovery of hemocytes production post A. hydrophila stimulation. When mitochondrial complexes I was knock down by dsRNA, ROS levels were decreased and THCs were also decreased. Recovery of hemocyte production inducing by A. hydrophila stimulation and rEsAst injection were delayed with dsEsbc1 injection. When ROS levels were increased after RNAi of Lon protease, THCs were also increased. The expression levels of five genes (EsJNK, EsSTAT, EsPI3K, EsAKT1, EsP70S6K) involved in SAPK-JNK and mTOR signaling pathways were up-regulated at 12 and 24 h in rEsAst group and EsLon dsRNA group compared with that in EGFP dsRNA group, and were similar to the trend of ROS levels. These results collectively suggested that EsAst should be a novel Astakine to promote the production of hemocytes in a ROS-dependent way in E. sinensis.


Assuntos
Braquiúros/genética , Braquiúros/imunologia , Regulação da Expressão Gênica/imunologia , Imunidade Inata/genética , Fator de Crescimento do Endotélio Vascular Derivado de Glândula Endócrina/genética , Fator de Crescimento do Endotélio Vascular Derivado de Glândula Endócrina/imunologia , Aeromonas hydrophila/fisiologia , Sequência de Aminoácidos , Animais , Proteínas de Artrópodes/química , Proteínas de Artrópodes/genética , Proteínas de Artrópodes/imunologia , Sequência de Bases , Perfilação da Expressão Gênica , Filogenia , Pichia/fisiologia , Distribuição Aleatória , Alinhamento de Sequência , Fator de Crescimento do Endotélio Vascular Derivado de Glândula Endócrina/química , Vibrio/fisiologia
12.
Dev Comp Immunol ; 82: 94-103, 2018 05.
Artigo em Inglês | MEDLINE | ID: mdl-29307815

RESUMO

Reactive oxygen species (ROS) produced in vivo during various electron transfer reactions are generally kept at a certain level since they are harmful to cells. However, it can sensitize hematopoietic progenitors to differentiation, and plays a signaling role in the regulation of hematopoietic cell fate. In the present study, the transcriptomes of crab HPT and hemocytes were sequenced using the Ion Torrent Proton sequencing platform. A total of 51,229,690 single end reads were obtained from six single-end libraries, which were assembled into 31346 unireads as reference. After mapping and transcript assembling, 362 differently expressed genes were identified and 301 of them were deemed to be more abundant in HPT. GO annotation revealed that they were mostly implicated in DNA, RNA and protein synthesis, cell division, mitochondria activities and energy metabolism. The expression level of mitochondrial complexes I (mitochondrial NADH-ubiquinone oxidoreductase) which was the main natural producers of mitochondrial ROS was found to be 8.6-fold (p < 0.01) higher in HPT than that in hemocytes. In hemocytes, the proteinase genes associated with proPO activation from the 61 up-regulated genes in hemocytes were the main up-regulated genes which might be the potential markers for mature hemocytes. ROS level in HPT cells was relatively higher which was confirmed with the high expression level of mitochondria related genes identified by transcriptome sequencing. After the ROS level was depressed by N-acetyl-l-cysteine (NAC), the production of hemocytes from HPT was inhibited, and the recovery of the total hemocytes counts was delayed. These results collectively indicated that the genes in redox system were more active in HPT, and ROS could function as an important modulator in the hematopoiesis of crab and promote the production of hemocytes from HPT.


Assuntos
Braquiúros/fisiologia , Hematopoese , Células-Tronco Hematopoéticas/fisiologia , Hemócitos/fisiologia , Mitocôndrias/metabolismo , NADH Desidrogenase/genética , Acetilcisteína/farmacologia , Animais , Diferenciação Celular , Células Cultivadas , Metabolismo Energético , Perfilação da Expressão Gênica , Ontologia Genética , NADH Desidrogenase/metabolismo , Oxirredução , Espécies Reativas de Oxigênio/metabolismo
13.
Fish Shellfish Immunol ; 74: 332-340, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29305333

RESUMO

Clip-domain serine proteinase is an important serine proteinase family involved in many biological processes, which is only found in invertebrates. In the present study, the full-length cDNA of a clip domain serine proteinase (designed as EsCDSP) gene was cloned from Chinese mitten crab Eriocheir sinensis using rapid amplification of cDNA ends (RACE) technique. It was of 1488 bp with an open reading frame (ORF) of 1134 bp encoding a polypeptide of 377 amino acids. There were a signal peptide, a clip domain, and a Tryp_SPc domain in the deduced amino acid sequence of EsCDSP. Highly conserved cysteine residues were identified in the clip domain and Tryp_SPc domain. EsCDSP shared similarities of 40%-61% with CDSPs from Penaeus monodon (ACP19562.1), Scylla paramamosain (CCW43200.1), Drosophila melanogaster (NP_649734.2) and Delia antiqua (AAW57295.1). It was clustered with other CDSPs from crabs in the phylogenetic tree. EsCDSP transcript was highly expressed in hemocytes and it could response to the stimulations of Vibro anguillarum and Pichia pastoris. rEsCDSP could activate proPO system and significantly increase the PO activity of HLS. In addition, rEsCDSP could bond to Aeromonas hydrophila, Vibro anguillarum and Vibro alginolyticus, and reduced the mortality rate causing by pathogen infection. All the results suggested that EsCDSP was an important immune response participator involved in activation of the proPO system of crab.


Assuntos
Braquiúros/genética , Braquiúros/imunologia , Regulação da Expressão Gênica/imunologia , Imunidade Inata/genética , Serina Proteases/genética , Serina Proteases/imunologia , Aeromonas hydrophila/fisiologia , Sequência de Aminoácidos , Animais , Proteínas de Artrópodes/química , Proteínas de Artrópodes/genética , Proteínas de Artrópodes/imunologia , Sequência de Bases , Perfilação da Expressão Gênica , Filogenia , Pichia/fisiologia , Alinhamento de Sequência , Serina Proteases/química , Vibrio/fisiologia
14.
Anal Biochem ; 545: 38-42, 2018 03 15.
Artigo em Inglês | MEDLINE | ID: mdl-29355484

RESUMO

A strand exchange amplification (SEA) method to detect foodborne pathogen Listeria monocytogenes was developed. SEA is a novel nucleic acid amplification method that only requires one pair of primers. The specie-specific primers were designed by targeting the 16S rRNA gene and the amplification reaction was performed as short as 60 min at 61 °C. Notably, SEA method could not only detect genomic DNA but also detect RNA by one step without requiring extra reverse transcription. The result could be visualized by naked eyes so that water bath pot would be the only equipment needed. Moreover, culture fluids and bacteria colony could be successfully detected without any pretreatment and the method displayed good specificity and strong anti-jamming capacity. These features greatly simplified the operating procedure and made SEA method be potential for developing point-of-care testing (POCT) devices to detect viable L. monocytogenes.


Assuntos
Listeria monocytogenes/isolamento & purificação , Técnicas de Amplificação de Ácido Nucleico/métodos , Microbiologia de Alimentos , Humanos , Listeria monocytogenes/genética , Testes Imediatos , RNA Ribossômico 16S/genética
15.
Drug Deliv ; 24(sup1): 94-107, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-29124979

RESUMO

Recent development of nano-technology provides highly efficient and versatile treatment methods to achieve better therapeutic efficacy and lower side effects of malignant cancer. The exploration of drug delivery systems (DDSs) based on nano-material shows great promise in translating nano-technology to clinical use to benefit patients. As an emerging inorganic nanomaterial, mesoporous carbon nanomaterials (MCNs) possess both the mesoporous structure and the carbonaceous composition, endowing them with superior nature compared with mesoporous silica nanomaterials and other carbon-based materials, such as carbon nanotube, graphene and fullerene. In this review, we highlighted the cutting-edge progress of carbon nanomaterials as drug delivery systems (DDSs), including immediate/sustained drug delivery systems and controlled/targeted drug delivery systems. In addition, several representative biomedical applications of mesoporous carbon such as (1) photo-chemo synergistic therapy; (2) delivery of therapeutic biomolecule and (3) in vivo bioimaging are discussed and integrated. Finally, potential challenges and outlook for future development of mesoporous carbon in biomedical fields have been discussed in detail.


Assuntos
Carbono/química , Nanoestruturas/química , Animais , Sistemas de Liberação de Medicamentos/métodos , Grafite/química , Humanos , Nanopartículas/química , Porosidade , Dióxido de Silício/química
16.
Fish Shellfish Immunol ; 70: 308-318, 2017 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-28889011

RESUMO

Leucine-rich repeat (LRR) domain and immunoglobulin (Ig) domain are both competent immune recognition modules, and the immunological roles of LRR and Ig domain containing- proteins (LRRIGs) are speculated to be multifunctional and worth investigating. In the present study, two novel LRRIGs, CgLRRIG-1 and CgLRRIG-2, were identified and characterized from oyster Crassostrea gigas. Both of them contained an N-terminal LRR region, an Ig domain, a transmembrane region, and a C-terminal cytoplasmic tail. The mRNA transcripts of CgLRRIG-1 and CgLRRIG-2 were constitutively expressed in muscle, gill, hepatopancreas, mantle, gonad and hemocytes with the highest expression level in hepatopancreas. Their mRNA expression levels in hemocytes were significantly up-regulated after the stimulations with four PAMPs including peptidoglycan (PGN), lipopolysaccharide (LPS), glucan (GLU) and polyinosinic-polycytidylic acid (poly I:C) and one bacteria Vibrio anguillarum. The recombinant proteins, rCgLRRIG-1 and rCgLRRIG-2, could bind to PGN, LPS, GLU and poly I:C, and rCgLRRIG-2 exhibited higher binding affinity. Additionally, rCgLRRIG-1 and rCgLRRIG-2 could significantly induce the expression of CgTNF-1 and CgIL17-5 in cultured oyster hemocytes, and the activity of rCgLRRIG-2 was higher than that of rCgLRRIG-1. All these results indicated that CgLRRIG-1 and CgLRRIG-2 could function as immune effectors or pro-inflammatory factors as well as PRRs in oyster.


Assuntos
Crassostrea/genética , Crassostrea/imunologia , Citocinas/genética , Imunidade Inata , Receptores de Reconhecimento de Padrão/genética , Receptores de Reconhecimento de Padrão/imunologia , Sequência de Aminoácidos , Animais , Sequência de Bases , Citocinas/imunologia , Expressão Gênica , Padrões Moleculares Associados a Patógenos/farmacologia , Filogenia , Receptores de Reconhecimento de Padrão/química , Alinhamento de Sequência , Vibrio/fisiologia
17.
Chem Commun (Camb) ; 53(77): 10696-10699, 2017 Sep 26.
Artigo em Inglês | MEDLINE | ID: mdl-28913529

RESUMO

We developed a novel method to control carryover contamination in loop-mediated isothermal amplification (LAMP) by primer engineering to carry recognition sites for a restriction endonuclease, providing a robust ability to eliminate carryover contaminants.


Assuntos
Primers do DNA , Enzimas de Restrição do DNA/metabolismo , DNA/química , Técnicas de Amplificação de Ácido Nucleico/métodos , Sequência de Bases , DNA/genética , Enzimas de Restrição do DNA/química
18.
Dev Comp Immunol ; 77: 188-199, 2017 12.
Artigo em Inglês | MEDLINE | ID: mdl-28807724

RESUMO

Leucine-rich repeat (LRR)-only proteins are involved in the innate immune responses as they mediate protein-ligand interactions. In the present study, three novel LRR-only proteins, CfLRRop-4, CfLRRop-5 and CfLRRop-6, were identified and characterized from Zhikong scallop Chlamys farreri. They all contained LRR motifs with consensus signature sequences of LxxLxLxxNxL or LxxLxLxxCxxL. All the mRNA transcripts of three CfLRRops were high abundant in hepatopancreas, gills and gonads, and their mRNA transcripts in hemocytes could respond to the stimulations of different microbes, including Vibrio anguillarum, Micrococcus luteus and Pichia pastoris. These three CfLRRops exhibited similar ligand binding and recognition characteristics as Toll-like receptors (TLRs) and NOD-like receptors (NLRs). The immune effectors, including tumor necrosis factor α, superoxide dismutase, catalase and lysozyme, varied significantly after the scallops were stimulated by recombinant LRR-only proteins. All these results indicated that LRR-only proteins are functionally differentiated and exhibit different immunomodulation activities on various downstream immune effectors.


Assuntos
Infecções por Bactérias Gram-Positivas/imunologia , Hepatopâncreas/fisiologia , Micrococcus luteus/imunologia , Micoses/imunologia , Pectinidae/imunologia , Pichia/imunologia , Proteínas Repressoras/genética , Vibrioses/imunologia , Vibrio/imunologia , Animais , Imunidade Inata , Imunomodulação , Leucina/genética , Ligação Proteica , Receptores de Reconhecimento de Padrão/metabolismo , Proteínas Repressoras/metabolismo , Fator de Necrose Tumoral alfa/metabolismo
19.
J Colloid Interface Sci ; 507: 410-420, 2017 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-28806660

RESUMO

An efficient and intelligent nano-carrier that combines cell imaging with near infrared (NIR) light and redox dual-responsive drug delivery was successfully prepared. The hollow mesoporous carbon (HMC) nanoparticles with high photothermal conversion ability were developed to increase the drug loading efficiency and realize chemotherapy and photothermal synergetic therapy. The photo-stable and luminescent carbon dots (CDs) were prepared from branched polyethyleneimine (PEI) by hydrothermal reaction. The PEI CDs (CDPEI) were grafted on the openings of HMC as the "gatekeepers" via disulfide units (HMC-SS-CDPEI) to prevent the premature release of doxorubicin (DOX). In the presence of GSH, the CDPEI separated from HMC due to the breakage of disulfide bonds, thus triggering the rapid release of the encapsulated drug. In addition, the release rate of DOX could be further accelerated by NIR light irradiation due to the increased temperature which would decrease the interaction between HMC and DOX. The fluorescence of the CDPEI is quenched when being attached to the HMC, while it is recovered when the CDPEI breaking away from HMC. Hence, the fluorescent CDPEI not only act as a gatekeeper to control drug release but also play a vital role in monitoring the process of the drug delivery. The developed HMC-SS-CDPEI showed dual-responsive drug release property and could be used as visible nano-platforms for chemo-photothermal synergistic therapy.

20.
Aquat Toxicol ; 189: 216-228, 2017 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-28666131

RESUMO

Ocean acidification (OA) could decrease the shells and skeletons formation of mollusk by reducing the availability of carbonate ions at calcification sites. Carbonic anhydrases (CAs) convert CO2 to HCO3- and play important roles in biomineralization process from invertebrate to vertebrate. In the present study, a CA (designated as CgCA) was identified and characterized in Pacific oyster C. gigas. The cDNA of CgCA was of 927bp encoding a predicted polypeptide of 308 amino acids with a signal peptide and a CA catalytic function domain. The mRNA transcripts of CgCA were constitutively expressed in all tested tissues with the highest levels in mantle and hemocytes. During the early development period, the mRNA transcripts of CgCA could be detected in all the stages with the highest level in D-veliger larvae. Elevated CO2 increased the mRNA transcripts of CgCA in muscle, mantle, hepatopancreas, gill and hemocytes significantly (p<0.05) and induced the translocation of CgCA in hemocytes and mantle. Moreover, elevated CO2 also caused the decrease of intracellular Ca2+ in hemocytes (p<0.05). The inhibition of CA by acetazolamide and suppression of CgCA gene via RNA interference could increase the intracellular Ca2+ in hemocytes (p<0.05). Besides, the decrease of intracellular Ca2+ content caused by Ca2+ reagent ionomycin could affect localization of CgCA in mantle tissue. The results indicated CgCA played essential roles in calcification and elevated CO2 accelerated the mutual modulation between calcium and CgCA, implying reduced calcification rate and dissolved shells under OA.


Assuntos
Exoesqueleto/efeitos dos fármacos , Cálcio/metabolismo , Dióxido de Carbono/toxicidade , Anidrases Carbônicas/metabolismo , Crassostrea/efeitos dos fármacos , Poluentes Químicos da Água/toxicidade , Exoesqueleto/metabolismo , Animais , Anidrases Carbônicas/genética , Crassostrea/genética , Crassostrea/metabolismo , Monitoramento Ambiental , Brânquias/efeitos dos fármacos , Brânquias/metabolismo , Hemócitos/efeitos dos fármacos , Hemócitos/metabolismo , Hepatopâncreas/efeitos dos fármacos , Hepatopâncreas/metabolismo , Larva , Oceanos e Mares , RNA Mensageiro/genética
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