RESUMO
Studies have shown silver nanoparticles (AgNPs) exposure can result in a series of toxic effects in fish gills. However, it is still unclear how AgNPs affect metabolite expression and their related molecular metabolic pathways in fish gills. In this study, we employed untargeted metabolomics to study the effects of AgNPs and silver supernatant ions on fish gill metabolites. The results showed that AgNPs can induce significant changes in 96 differentially expressed metabolites, which mainly affect amino acid metabolism and energy metabolism in fish gills. Among these metabolites, AgNPs specifically induce significant changes in 72 differentially expressed metabolites, including L-histidine, L-isoleucine, L-phenylalanine, and citric acid. These metabolites were significantly enriched in the pathways of aminoacyl-tRNA biosynthesis, ABC transporters, and the citrate cycle. In contrast, Ag+ supernatant exposure can specifically induce significant changes in 14 differentially expressed metabolites that mainly interfere with sphingolipid metabolism in fish gills. These specifically regulated fish gill metabolites include sphinganine, sphingosine, and phytosphingosine, which were significantly enriched in the sphingolipid metabolism pathway. Our results clearly reveal the effects and potential toxicity mechanisms of AgNPs on fish gill metabolites. Furthermore, our study further determined the unique functions of released silver ions in AgNPs toxicity in fish gills.
Assuntos
Carpas , Nanopartículas Metálicas , Animais , Brânquias , Metabolômica , Nanopartículas Metálicas/toxicidade , Prata/toxicidadeRESUMO
BACKGROUND: Brain-derived estrogen is implicated in pain-related aversion; however, which estrogen receptors mediate this effect remains unclear. This study hypothesized that the different estrogen receptors in the rostral anterior cingulate cortex play distinct roles in pain-related aversion. METHODS: Formalin-induced conditioned place avoidance and place escape/avoidance paradigms were used to evaluate pain-related aversion in rodents. Immunohistochemistry and Western blotting were used to detect estrogen receptor expression. Patch-clamp recordings were used to examine N-methyl-D-aspartate-mediated excitatory postsynaptic currents in rostral anterior cingulate cortex slices. RESULTS: The administration of the estrogen receptor-ß antagonist 4-(2-phenyl-5,7-bis [trifluoromethyl] pyrazolo [1,5-a] pyrimidin-3-yl) phenol (PHTPP) or the G protein-coupled estrogen receptor-1 antagonist (3aS*,4R*,9bR*)-4-(6-bromo-1,3-benzodioxol-5-yl)-3a,4,5,9b-3H-cyclopenta [c] quinolone (G15) but not the estrogen receptor-α antagonist 1,3-bis (4-hydroxyphenyl)-4-methyl-5-[4-(2-piperidinylethoxy) phenol]-1H-pyrazole dihydrochloride (MPP) into the rostral anterior cingulate cortex blocked pain-related aversion in rats (avoidance score, mean ± SD: 1,3-bis [4-hydroxyphenyl]-4-methyl-5-(4-[2-piperidinylethoxy] phenol)-1H-pyrazole dihydrochloride (MPP): 47.0 ± 18.9%, 4-(2-phenyl-5,7-bis [trifluoromethyl] pyrazolo [1,5-a] pyrimidin-3-yl) phenol (PHTPP): -7.4 ± 20.6%, and [3aS*,4R*,9bR*]-4-[6-bromo-1,3-benzodioxol-5-yl]-3a,4,5,9b-3H-cyclopenta [c] quinolone (G15): -4.6 ± 17.0% vs. vehicle: 46.5 ± 12.2%; n = 7 to 9; P < 0.0001). Consistently, estrogen receptor-ß knockdown but not estrogen receptor-α knockdown by short-hairpin RNA also inhibited pain-related aversion in mice (avoidance score, mean ± SD: estrogen receptor-α-short-hairpin RNA: 26.0 ± 7.1% and estrogen receptor-ß-short-hairpin RNA: 6.3 ± 13.4% vs. control short-hairpin RNA: 29.1 ± 9.1%; n = 7 to 10; P < 0.0001). Furthermore, the direct administration of the estrogen receptor-ß agonist 2,3-bis (4-hydroxyphenyl)-propionitrile (DPN) or the G protein-coupled estrogen receptor-1 agonist (±)-1-([3aR*,4S*,9bS*]-4-(6-bromo-1,3-benzodioxol-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopenta [c]quinolin-8-yl)-ethanone (G1) into the rostral anterior cingulate cortex resulted in conditioned place avoidance (avoidance score, mean ± SD: 2,3-bis (4-hydroxyphenyl)-propionitrile (DPN): 35.3 ± 9.5% and (±)-1-([3aR*,4S*,9bS*]-4-(6-bromo-1,3-benzodioxol-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopenta [c]quinolin-8-yl)-ethanone (G1): 43.5 ± 22.8% vs. vehicle: 0.3 ± 14.9%; n = 8; P < 0.0001) but did not affect mechanical or thermal sensitivity. The activation of the estrogen receptor-ß/protein kinase A or G protein-coupled estrogen receptor-1/protein kinase B pathway elicited the long-term potentiation of N-methyl-D-aspartate-mediated excitatory postsynaptic currents. CONCLUSIONS: These findings indicate that estrogen receptor-ß and G protein-coupled estrogen receptor-1 but not estrogen receptor-α in the rostral anterior cingulate cortex contribute to pain-related aversion by modulating N-methyl-D-aspartate receptor-mediated excitatory synaptic transmission.
Assuntos
Giro do Cíngulo/fisiopatologia , Dor/fisiopatologia , Dor/psicologia , Receptores de Estrogênio , Animais , Aprendizagem da Esquiva , Proteínas Quinases Dependentes de AMP Cíclico/genética , Proteínas Quinases Dependentes de AMP Cíclico/metabolismo , Proteínas Quinases Dependentes de GMP Cíclico/genética , Proteínas Quinases Dependentes de GMP Cíclico/metabolismo , Antagonistas de Estrogênios/farmacologia , Receptor beta de Estrogênio/efeitos dos fármacos , Receptor beta de Estrogênio/genética , Potenciais Pós-Sinápticos Excitadores/efeitos dos fármacos , Feminino , Técnicas de Silenciamento de Genes , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Técnicas de Patch-Clamp , RNA Interferente Pequeno , Ratos , Ratos Sprague-Dawley , Receptores de Estrogênio/efeitos dos fármacos , Receptores de Estrogênio/genéticaRESUMO
Silver nanoparticles (AgNPs) are increasingly being used as antimicrobial agents and drug carriers in biomedical fields. However, toxicological information on their effects on red blood cells (RBCs) and the mechanisms involved remain sparse. In this article, we examined the size dependent nanotoxicity of AgNPs using three different characteristic sizes of 15 nm (AgNPs15), 50 nm (AgNPs50), and 100 nm (AgNPs100) against fish RBCs. Optical microscopy and transmission electron microscopy observations showed that AgNPs exhibited a size effect on their adsorption and uptake by RBCs. The middle sized AgNPs50, compared with the smaller or bigger ones, showed the highest level of adsorption and uptake by the RBCs, suggesting an optimal size of â¼50 nm for passive uptake by RBCs. The toxic effects determined based on the hemolysis, membrane injury, lipid peroxidation, and antioxidant enzyme production were fairly size and dose dependent. In particular, the smallest sized AgNPs15 displayed a greater ability to induce hemolysis and membrane damage than AgNPs50 and AgNPs100. Such cytotoxicity induced by AgNPs should be attributed to the direct interaction of the nanoparticle with the RBCs, resulting in the production of oxidative stress, membrane injury, and subsequently hemolysis. Overall, the results suggest that particle size is a critical factor influencing the interaction between AgNPs and the RBCs.
Assuntos
Eritrócitos/efeitos dos fármacos , Nanopartículas Metálicas/toxicidade , Prata/toxicidade , Adsorção , Animais , Transporte Biológico , Células Cultivadas , Eritrócitos/química , Eritrócitos/metabolismo , Eritrócitos/ultraestrutura , Carpa Dourada , Hemólise/efeitos dos fármacos , Nanopartículas Metálicas/química , Nanopartículas Metálicas/ultraestrutura , Microscopia Eletrônica de Transmissão , Tamanho da Partícula , Prata/químicaRESUMO
Anxiety and depression are frequently observed in patients suffering from trigeminal neuralgia (TN), but neural circuits and mechanisms underlying this association are poorly understood. Here, we identified a dedicated neural circuit from the ventral hippocampus (vHPC) to the medial prefrontal cortex (mPFC) that mediates TN-related anxiodepression. We found that TN caused an increase in excitatory synaptic transmission from vHPCCaMK2A neurons to mPFC inhibitory neurons marked by the expression of corticotropin-releasing hormone (CRH). Activation of CRH+ neurons subsequently led to feed-forward inhibition of layer V pyramidal neurons in the mPFC via activation of the CRH receptor 1 (CRHR1). Inhibition of the vHPCCaMK2A-mPFCCRH circuit ameliorated TN-induced anxiodepression, whereas activating this pathway sufficiently produced anxiodepressive-like behaviors. Thus, our studies identified a neural pathway driving pain-related anxiodepression and a molecular target for treating pain-related psychiatric disorders.
Assuntos
Hormônio Liberador da Corticotropina , Neuralgia do Trigêmeo , Humanos , Hormônio Liberador da Corticotropina/metabolismo , Neuralgia do Trigêmeo/etiologia , Neuralgia do Trigêmeo/metabolismo , Neurônios/metabolismo , Hipocampo/fisiologia , Dor/metabolismoRESUMO
Although the toxicity of silver nanoparticles (AgNPs) has been widely reported, the persistence and reversibility of AgNPs toxicity are poorly understood. In the present work, AgNPs with particle sizes of 5 nm, 20 nm, and 70 nm (AgNPs5, AgNPs20, and AgNPs70) were selected to investigate the nanotoxicity and recovery effects of Chlorella vulgaris in the exposure (72 h) and recovery (72 h) stages using non-targeted metabolomics techniques. The exposure of AgNPs exerted size-dependent effects on several aspects of C. vulgaris physiology, including growth inhibition, chlorophyll content, intracellular silver accumulation, and differential expression of metabolites, and most of these adverse effects were reversible. Metabolomics revealed that AgNPs with small sizes (AgNPs5 and AgNPs20) mainly inhibited glycerophospholipid and purine metabolism, and the effects were reversible. In contrast, AgNPs with large sizes (AgNPs70) reduced amino acid metabolism and protein synthesis by inhibiting aminoacyl-tRNA biosynthesis, and the effects were irreversible, demonstrating the persistence of nanotoxicity of AgNPs. The size-dependent persistence and reversibility of AgNPs toxicity provides new insights to further understand the mechanisms of toxicity of nanomaterials.
Assuntos
Chlorella vulgaris , Nanopartículas Metálicas , Poluentes Químicos da Água , Nanopartículas Metálicas/toxicidade , Prata/toxicidade , Poluentes Químicos da Água/toxicidade , Água DoceRESUMO
BACKGROUND AND PURPOSE: Patients suffering from trigeminal neuralgia are often accompanied by anxiety and depression. Microglia-mediated neuroinflammation is involved in the development of neuropathic pain and anxiodepression pathogenesis. Whether and how microglia are involved in trigeminal neuralgia-induced anxiodepression remains unclear. EXPERIMENTAL APPROACH: Unilateral constriction of the infraorbital nerve (CION) was performed to establish trigeminal neuralgia in rat and mouse models. Mechanical allodynia and anxiodepressive-like behaviours were measured. Optogenetic and pharmacological manipulations were employed to investigate the role of hippocampal microglia in anxiety and depression caused by trigeminal neuralgia. KEY RESULTS: Trigeminal neuralgia activated ipsilateral but not contralateral hippocampal microglia, up-regulated ipsilateral hippocampal ATP and interleukin-1ß (IL-1ß) levels, impaired ipsilateral hippocampal long-term potentiation (LTP) and induced anxiodepressive-like behaviours in a time-dependent manner in rodents. Pharmacological or optogenetic inhibition of ipsilateral hippocampal microglia completely blocked trigeminal neuralgia-induced anxiodepressive-like behaviours. Activation of unilateral hippocampal microglia directly elicited an anxiodepressive state and impaired hippocampal LTP. Knockdown of ipsilateral hippocampal P2X7 receptors prevented trigeminal neuralgia-induced microglial activation and anxiodepressive-like behaviours. Furthermore, we demonstrated that microglia-derived IL-1ß mediated microglial activation-induced anxiodepressive-like behaviours and LTP impairment. CONCLUSION AND IMPLICATIONS: These findings suggest that priming of microglia with ATP/P2X7 receptors in the ipsilateral hippocampus drives pain-related anxiodepressive-like behaviours via IL-1ß. An asymmetric role of the bilateral hippocampus in trigeminal neuralgia-induced anxiety and depression was uncovered. The approaches targeting microglia and P2X7 signalling might offer novel therapies for trigeminal neuralgia-related anxiety and depressive disorder.
Assuntos
Neuralgia , Neuralgia do Trigêmeo , Camundongos , Ratos , Animais , Neuralgia do Trigêmeo/patologia , Microglia/patologia , Roedores , Hipocampo , Hiperalgesia , Trifosfato de AdenosinaRESUMO
Although nanoparticles have been widely used as optical contrasts for cell imaging, the complicated prefunctionalized steps and low labeling efficiency of nanoprobes greatly inhibit their applications in cellular protein imaging. In this study, we developed a novel and general strategy that employs an aptamer not only as a recognizer for protein recognition but also as a linker for nanoreporter targeting to specifically label membrane proteins of interest and track their endocytic pathway. With this strategy, three kinds of nanoparticles, including gold nanoparticles, silver nanoparticles, and quantum dots (QDs), have been successfully targeted to the membrane proteins of interest, such as nucleolin or prion protein (PrP(C)). The following investigations on the subcellular distribution with fluorescent immunocolocalization assay indicated that PrP(C)-aptamer-QD complexes most likely internalized into cytoplasm through a classical clathrin-dependent/receptor-mediated pathway. Further single-particle tracking and trajectory analysis demonstrated that PrP(C)-aptamer-QD complexes exhibited a complex dynamic process, which involved three types of movements, including membrane diffusion, vesicle transportation, and confined diffusion, and all types of these movements were associated with distinct phases of PrP(C) endocytosis. Compared with traditional multilayer methods, our proposed aptamer-mediated strategy is simple in procedure, avoiding any complicated probe premodification and purification. In particular, the new double-color labeling strategy is unique and significant due to its superior advantages of targeting two signal reporters simultaneously in a single protein using only one aptamer. What is more important, we have constructed a general and versatile aptamer-mediated protein labeling nanoplatform that has shown great promise for future biomedical labeling and intracellular protein dynamic analysis.
Assuntos
Aptâmeros de Nucleotídeos/metabolismo , Endocitose , Espaço Intracelular/metabolismo , Imagem Molecular/métodos , Nanopartículas/química , Proteínas PrPC/metabolismo , Coloração e Rotulagem/métodos , Biotina/metabolismo , Sobrevivência Celular , Cor , Células HeLa , Humanos , Modelos Moleculares , Proteínas PrPC/química , Conformação Proteica , Pontos Quânticos , Estreptavidina/metabolismo , Especificidade por SubstratoRESUMO
Microplastic exposure leads to various toxic effects in Daphnia magna; however, the effects of microplastics on the metabolic processes in D. magna and the corresponding molecular toxicity mechanisms remain unclear. In the present study, the effects of acute exposure to polyethylene microplastics with different particle sizes (20 µm [MPs-20] and 30 µm [MPs-30]) on metabolites in D. magna and the mechanisms of toxicity were investigated by combining metabolomics and traditional toxicology techniques. Exposure to both MPs-20 and MPs-30 resulted in significant accumulation of microplastics in the gut of D. magna and significantly reduced D. magna survival and heart rate. Metabolomics analysis revealed that MPs-20 and MPs-30 induced significant changes in up to 88 and 91 differential metabolites, respectively, and collectively induced significant changes in 75 metabolites in D. magna. Among lipid metabolites, MPs-20 specifically downregulated phosphatidylcholine and upregulated phosphatidylethanolamine, which mainly affected phospholipid metabolism, whereas MPs-30 specifically downregulated amino acid metabolites l-glutamine, l-glutamate and malic acid, which mainly interfered with energy metabolism. The results of this study provide novel insights into the mechanism of effects of microplastics on metabolic processes in D. magna.
Assuntos
Microplásticos , Poluentes Químicos da Água , Animais , Daphnia , Ácido Glutâmico , Glutamina/farmacologia , Fosfatidilcolinas/farmacologia , Fosfatidiletanolaminas/farmacologia , Plásticos/toxicidade , Polietileno/toxicidade , Poluentes Químicos da Água/análiseRESUMO
Mitochondria are recognized as an important target organelle for the toxicity of nanomaterials. Although the toxic effects of silver nanoparticles (AgNPs) on mitochondria have been widely reported, the mechanism behind the toxicity remains unclear. In this study, the effects of two forms of silver (AgNPs and AgNO3) on carp gill mitochondria were investigated by analyzing the mitochondrial ultrastructure, physicochemical properties of mitochondrial membrane, and mitochondrial proteomics. After exposure of common carp to AgNPs (0.75 mg/L) and AgNO3 (0.05 mg/L) for 96 h, both forms of silver were shown to cause gill mitochondrial lesions, including irregular shape, loss of mitochondrial cristae, and increased mitochondrial membrane permeability. Proteomics results revealed that AgNPs and AgNO3 induced 362 and 297 differentially expressed proteins (DEPs) in gill mitochondria, respectively. Among the DEPs, 244 were shared between AgNPs and AgNO3 treatments. These shared proteins were mainly distributed in the mitochondrial membrane and matrix, and were significantly enriched in the tricarboxylic acid (TCA) cycle and oxidative phosphorylation pathway. The functional annotation of DEPs induced by both silver forms was mainly involved in energy production and conversion. These results indicated that the toxic mechanism of AgNPs and AgNO3 on gill mitochondria were comparable and the two forms of silver caused mitochondrial dysfunction in fish gills by inhibiting the TCA cycle and disrupting the electron transport chain.
Assuntos
Carpas , Nanopartículas Metálicas , Poluentes Químicos da Água , Animais , Nitrato de Prata/toxicidade , Brânquias , Nanopartículas Metálicas/química , Proteômica , Poluentes Químicos da Água/toxicidade , Prata/toxicidade , Prata/metabolismo , Corantes/metabolismo , Corantes/farmacologia , Ácidos Tricarboxílicos/metabolismo , Ácidos Tricarboxílicos/farmacologiaRESUMO
The trophodynamics of toxic trace metals is significant for assessing the threat of toxic trace metals to the aquatic ecosystem and human safety. However, due to the difficulty of accurately calculating the trophic positions of freshwater aquatic organisms in the food web, the comprehensive process of trophodynamics of toxic trace metals in freshwater ecosystems was still rarely known. By integrating the compound-specific nitrogen stable isotopic analysis of amino acids (CSIA-AAs) and the Bayesian stable isotope mixing model (SIMM) as a novel approach, the present study investigated the trophodynamics of five toxic trace metals (Zn, As, Cr, Cu, and Hg) in the food web of the YangZong Lake, a plateau freshwater lake that was once heavily polluted by arsenic in Yunnan Province, China. The results revealed that Hg tended to be efficiently biomagnified in the food web with a trophic magnification factor of 1.75; As, Cr, and Cu were biodiluted significantly, while Zn showed no biomagnification or biodilution trends. The dietary health risk assessment indicated the potential health risk of toxic trace metals for the local residents of long-term fish consumption. The present work highlights the accuracy and reliability of the novel CSIA-AAS+SIMM approach in the calculation of the trophic positions of freshwater organisms.
Assuntos
Mercúrio , Oligoelementos , Poluentes Químicos da Água , Animais , Organismos Aquáticos/metabolismo , Teorema de Bayes , China , Ecossistema , Monitoramento Ambiental/métodos , Cadeia Alimentar , Humanos , Lagos/química , Mercúrio/metabolismo , Reprodutibilidade dos Testes , Medição de Risco , Oligoelementos/metabolismo , Poluentes Químicos da Água/análiseRESUMO
Microplastics, a new class of environmental pollutants, accumulates in the environment at an uncontrollable rate, which threatens aquatic organisms. Plankton are the basis of food webs and play a significant role in the material circulation and energy flow of aquatic ecosystems. Plankton are sensitive to various environmental pollutants. It is necessary to investigate the impacts of microplastics on plankton. Here, we analzyed the sources and characteristics of microplastics, and the current state of microplastic pollution in aquatic ecosystem. The direct and indirect harmful effects of microplastics on aquatic organisms were elaborated. Then, we focused on the potential consequences of microplastics on phytoplankton and zooplankton species from different scales, ranging from individual, population, to community level. With respect to plankton organisms, few studies were carried out on genomics and proteomics from the microcosmic perspective, and on popu-lation and community responses from the macroscopic aspect. This review would provide references for further studies.
Assuntos
Microplásticos , Poluentes Químicos da Água , Organismos Aquáticos , Ecossistema , Monitoramento Ambiental , Plâncton , Plásticos/toxicidade , Poluentes Químicos da Água/análise , Poluentes Químicos da Água/toxicidadeRESUMO
Toxic effects of silver nanoparticles (AgNPs) on fish gills have been widely reported but the recoverability of AgNPs-induced fish gill injuries is still unknown. In this study, combined multiomics and conventional toxicological analytical methods were used to investigate the changes in the gills of common carp responses to AgNPs (0.1 mg/L) toxicity after 24 h exposure and 7-day recovery. Conventional toxicological results showed that AgNPs exposure significantly increased silver content in gills and caused epithelial hyperplasia and lamellar fusion. After the recovery period, the silver content in fish gills significantly decreased; accompanied by the disappearance of histopathological characteristics in fish gills. Multiomics results revealed that AgNPs exposure resulted in the differential expression of 687 genes and 96 metabolites in fish gills. These differentially expressed genes (DEGs) and metabolites mainly participate in amino acid, carbohydrate, and lipid metabolisms, and are significantly enriched in the tricarboxylic acid (TCA) cycle. After the recovery period, the number of DEGs and metabolites in gills decreased to 33 and 90, respectively. Moreover, DEGs and metabolites in the TCA cycle recovered to control levels. In summary, the present study found that AgNPs-induced fish gill toxicity showed potential recoverability at molecular and phenotype levels.
Assuntos
Carpas , Nanopartículas Metálicas , Poluentes Químicos da Água , Animais , Brânquias/química , Metabolômica , Nanopartículas Metálicas/toxicidade , Prata/análise , Prata/toxicidade , Transcriptoma , Poluentes Químicos da Água/toxicidadeRESUMO
PURPOSE: To use three-dimensional reconstruction measurement, preoperative diagnosis, surgical design, surgical simulation, guide plate production, navigation verification and effect evaluation of orthognathic surgery assisted by digital technology, in order to explore more scientific and reasonable programs and procedures of orthognathic surgery. METHODS: Twenty-five patients with congenital dental and maxillofacial deformity were selected as the experimental subjects, craniofacial spiral CT was conducted before surgery and CT data were imported into Mimics 20.0 software to establish a 3D head digital model. The bone landmarks in three-dimensional reconstruction digital model were selected, measured, analyzed and diagnosed, and the design of the surgical plan and the production of the guide plates were performed. Surgical navigation system was used to confirm the maxillary position, verify the bone retention and guide precise bone grinding during operation. Craniofacial spiral CT was conducted 1 week after surgery for postoperative validation of the surgical design protocol. Statistical analysis was performed using SPSS 24.0 software package. RESULTS: All 25 patients were operated according to the digital orthognathic surgery design and procedure.There were no significant differences in X, Y and Z three-dimensional directions in 10 actual landmarks between the postoperative actual head model and the preoperative predictive head modelï¼P>0.05ï¼. CONCLUSIONS: Orthognathic surgery assisted by digital technology has the advantages of precision and minimal invasiveness.
Assuntos
Cirurgia Ortognática , Procedimentos Cirúrgicos Ortognáticos , Cirurgia Assistida por Computador , Tecnologia Digital , Humanos , Imageamento Tridimensional , Maxila/diagnóstico por imagem , Maxila/cirurgia , Tomografia Computadorizada EspiralRESUMO
BACKGROUND: Oral lichen planus (OLP) is a chronic inflammatory disorder, and it can affect normal oral function. The conventional treatments for OLP are not always effective, and relapse easily occurs. Therefore, treatment of OLP is difficult and challenging. In this study, we evaluated over a long period the clinical efficacy of surgical excision and acellular dermal matrix (ADM) grafting in patients with refractory OLP. CASE SUMMARY: Eleven patients with refractory OLP underwent a standardized protocol of surgical excision and ADM grafting. The condition of the area of the grafted wound, the intraoperative maximum mouth opening, pain, and clinical healing were assessed at postoperative follow-up visits. All patients had a flat surgical area with similar mucosal tissue coverage and local scar formation. Patients had no irritation and pain in their mucous membranes when eating acidic and spicy food. All patients' mouth openings returned to normal within 2-6 mo after surgery. During follow-up, none of the patients had recurrence of OLP after surgery. The longest follow-up was 11 yr and the shortest was 6 mo, and none of the patients relapsed during follow-up. CONCLUSION: Surgical excision and ADM grafting could be an effective method to treat refractory OLP.
RESUMO
Although holding the advantages of both an aptamer and a molecular beacon (MB), a molecular aptamer beacon (MAB) needs complicated and expensive modifications at both of its ends and usually has a high background signal because of the low energy transfer efficiency between the donor and the acceptor. To overcome these shortcomings, in this study, we develop a long-range resonance energy transfer (LrRET) system by separating the donor from the acceptor, wherein only one end of the MAB is fluorescently labeled and acts as the energy donor and multiwalled carbon nanotubes (MWCNTs) are introduced as the energy acceptor. To test the feasibility of the newly designed MAB system, adenosine triphosphate (ATP) has been employed as a proof-of-concept target. It is found that the fluorescence of the designed MAB is completely quenched by MWCNTs, supplying a very low background signal. Then the quenched fluorescence is recovered significantly with the addition of ATP, so that ATP can be detected in the range of 0.8-80 µM with a limit of detection of 0.5 µM (3σ). Compared with the conventional fluorescence resonance energy transfer, the efficiency of LrRET between the dye and MWCNTs is much higher. Since only one end of the MAB needs the modification, the present strategy is simple and cost-effective. Furthermore, the use of MWCNTs can greatly reduce the fluorescence background of the MAB and supply a high sensitivity, showing its generality for detection of a variety of targets.
Assuntos
Aptâmeros de Nucleotídeos/análise , Transferência Ressonante de Energia de Fluorescência/métodos , Nanotubos de Carbono/químicaRESUMO
The major challenge of prion disease diagnosis at the presymptomatic stage is how to sensitively or selectively discriminate and detect the minute quantity of disease-associated prion protein isoform (PrP(Res)) in complex biological systems such as serum and brain homogenate. In this contribution, we developed a dual-aptamer strategy by taking the advantages of aptamers, the excellent separation ability of magnetic microparticles (MMPs), and the high fluorescence emission features of quantum dots (QDs). Two aptamers (Apt1 and Apt2), which can recognize their two corresponding distinct epitopes of prion proteins (PrP), were coupled to the surfaces of MMPs and QDs, respectively, to make MMPs-Apt1 and QDs-Apt2 ready at first, which then could be coassociated together through the specific recognitions of the two aptamers with their two corresponding distinct epitopes of PrP, forming a sandwich structure of MMPs-Apt1-PrP-Apt2-QDs and displaying the strong fluorescence of QDs. Owing to the different binding affinities of the two aptamers with PrP(Res) and cellular prion protein (PrP(C)), both of which have distinct denaturing detergent resistance, our dual-aptamer strategy could be applied to discriminate PrP(Res) and PrP(C) successfully in serum. Further identifications showed that the present dual-aptamer assay could be successfully applied to the detection of PrP in 0.01% brain homogenate, about 1000-fold lower than that of commonly applied antibody-mediated assays, which can detect PrP just in 10% brain homogenate, indicating that the present designed dual-aptamer assay is highly sensitive and adequate for clinical diagnosis without isolation of target protein prior to assay.
Assuntos
Aptâmeros de Nucleotídeos/química , Magnetismo , Príons/análise , Pontos Quânticos , Espectrometria de Fluorescência/métodos , Animais , Encéfalo/metabolismo , Humanos , Camundongos , Doenças Priônicas/diagnóstico , Príons/sangue , Isoformas de Proteínas/análise , Isoformas de Proteínas/sangueRESUMO
In this contribution, we report a molecular recognition between adenosine and its aptamer, which leads to the formation of a four-stranded tetraplex structures (G-quartet) of the aptamer. It is found that the formed G-quartet could induce the side-by-side self-assembly of gold nanorods (AuNRs) owing to the electrostatic interaction between the positive charge of cetyltrimethylammonium bromide (CTAB) on the AuNR surface and the negative charge of the formed G-quartet. Furthermore, the side-by-side self-assembly of AuNRs is characterized by the enhancement of plasmon resonance light scattering (PRLS) signals and the blue-shift of the longitudinal plasmon resonance absorption (LPRA) band owing to the plasmon resonance coupling. Then, based on the enhanced PRLS signals, a simple, highly selective and sensitive detection method for adenosine was developed in the range of 4.0-80.0 nM with the limit of determination of 2.0 nM, which is up to now the best sensitive optical detection method to our knowledge. This method has been successfully applied to the detection of adenosine phosphates in the brains of SD rats, which was in good agreement with a high-performance liquid chromatographic (HPLC) method.
Assuntos
Adenosina/análise , Aptâmeros de Nucleotídeos/química , Encéfalo , Ouro/química , Modelos Animais , Nanotubos/química , Animais , Cromatografia Líquida de Alta Pressão , Quadruplex G , Ratos , Ratos Sprague-Dawley , Sensibilidade e Especificidade , Ressonância de Plasmônio de SuperfícieRESUMO
A simple, one-pot and controllable strategy is reported in this contribution for biomimetic synthesis and self-assembly of gold nanoparticles (Au-NPs). It involves our synthesized polyaldehyde dextran (PAD), which has been proved to be a biomacromolecule with excellent biocompatibility and biodegradability, acting as both a reducing agent and a stabilizer. The morphology of the as-prepared Au-NP assemblies can be controlled by adjusting the reaction conditions, such as the concentration of aldehyde in PAD, the reaction time and the temperature. Investigations of the mechanism suggest that stabilizers may distribute on different crystal facets of NPs non-uniformly owing to the different binding forces, and dipole-dipole interaction of NPs could be the main driving force for the assembly of Au-NPs. In addition, intermolecular hydrogen bonding interaction of stabilizers could also act as a possible driving force. The excellent biocompatibility of the Au-NP assemblies makes them promising candidates for fabricating future optical nanodevices and application in biological systems.
Assuntos
Biomimética/métodos , Ouro/química , Nanopartículas Metálicas/química , Nanotecnologia/métodos , Dextranos/química , Nanopartículas Metálicas/ultraestrutura , Tamanho da Partícula , Soluções , Espectrofotometria UltravioletaRESUMO
Although the toxic effects of silver nanoparticles (AgNPs) on fish gills have been reported, the underlying mechanism of toxicity remains unclear. The present study aimed to elucidate the mechanism of toxicity in the gills of common carp following exposure to AgNPs and silver nitrate (AgNO3) using histopathology and proteomics. Histopathological findings revealed that both AgNPs and AgNO3 caused telangiectasia and epithelial cell hyperplasia in fish gills; however, the pathological features and location of lesions caused by the two forms of silver were markedly different. Proteomics revealed that AgNPs and AgNO3 induced 139 and 185 differential expression proteins (DEPs) in gills, respectively, and the two forms of silver induced only 42 shared proteins. AgNPs specifically induced 87 DEPs which mainly involved signaling mechanisms, cytoskeleton, and the arachidonic acid metabolism processes. AgNO3 specifically induced 125 DEPs that were mainly clustered in the glutathione metabolism and protease processes. These results suggested that the toxic effects of AgNPs and AgNO3 were dramatically different in terms of protein expression in fish gills, which may provide novel perspectives for understanding the toxicity mechanism of silver nanoparticles in fish gills.
Assuntos
Proteínas de Peixes/metabolismo , Brânquias/efeitos dos fármacos , Nanopartículas Metálicas/toxicidade , Proteoma/metabolismo , Nitrato de Prata/toxicidade , Animais , Carpas , Regulação para Baixo , Brânquias/patologia , Hiperplasia , Mapas de Interação de Proteínas , Proteômica , Transdução de Sinais/efeitos dos fármacos , Prata/química , Telangiectasia/induzido quimicamente , Regulação para CimaRESUMO
This study concentrated on the 15 perfluoroalkyl substances (PFASs) in the coastal areas of China and South Korea, an urbanized area with intensive human activities. In total, 126 water samples and 125 sediment samples were collected and determined by HPLC-MS/MS. Fifteen PFASs were detected at least once in all the sites. The total concentrations of PFASs in water and sediment samples ranged from 6.75 ng·L-1 to 20982 ng·L-1 and from 0.229 ng·g-1 to 53.8 ng·g-1 (dw), respectively. The concentrations of PFASs in waters and sediments were relatively high in China, and PFOA was the predominant PFAS. In contrast, short chain compounds such as PFBA and PFPeA played a major role in water, and PFOS, PFBA, and PFOA dominated the sediment samples in South Korea. The partition coefficients of PFASs were closely related to the fraction of organic carbon in the sediment and the carbon chain length of PFASs. Among the 15 compounds, the partition coefficients of 9 were significantly correlated with the fraction of organic carbon (r>0.21, P<0.05), and the partition coefficient steadily increased with the increase of the carbon chain length. The results of ecological risk assessment suggested that the coastal urbanized areas of China and South Korea are still at a low risk. However, some specific areas also showed PFAS pollution. More attention should be paid to the potential health risks.