RESUMEN
Pancreatic ductal adenocarcinoma (PDAC) is a common cause of cancer-related death, and most patients are with advanced disease when diagnosed. At present, despite a variety of treatments have been developed for PDAC, few effective treatment options are available; on the other hand, PDAC shows significant resistance to chemoradiotherapy, targeted therapy, and immunotherapy due to its heterogeneous genetic profile, molecular signaling pathways, and complex tumor immune microenvironment. Nevertheless, over the past decades, there have been many new advances in the key theory and understanding of the intrinsic mechanisms and complexity of molecular biology and molecular immunology in pancreatic cancer, based on which more and more diverse new means and reasonable combination strategies for PDAC treatment have been developed and preliminary breakthroughs have been made. With the continuous exploration, from surgical local treatment to comprehensive medical management, the research-diagnosis-management system of pancreatic cancer is improving. This review focused on the variety of treatments for advanced PDAC, including traditional chemotherapy, targeted therapy, immunotherapy, microenvironment matrix regulation as well as the treatment targeting epigenetics, metabolism and cancer stem cells. We pointed out the current research bottlenecks and future exploration directions.
Asunto(s)
Carcinoma Ductal Pancreático/terapia , Terapia Molecular Dirigida , Neoplasias Pancreáticas/terapia , Medicina de Precisión/métodos , Carcinoma Ductal Pancreático/genética , Humanos , Neoplasias Pancreáticas/genética , Microambiente TumoralRESUMEN
Diabetic mellitus is a chronic metabolic disorder that is associated with several complications including testicular dysfunction. This research investigated the protective action of the ethyl acetate extract from Shorea roxburghii (SRE) on diabetes induced testicular damage in rats. Diabetic rats were orally administered with SRE at doses of 100 and 400 mg/kg for 4 weeks. SRE improved the body weight gain, testes weight, testes index and increased serum concentration of testosterone. Furthermore, SRE increased the testicular antioxidant enzymes including superoxide dismutase, catalase and glutathione peroxidase. In addition, SRE ameliorated testicular inflammatory mediators such as myeloperoxidase, tumor necrosis factor alpha, interleukin 6, p38 MAPK and nuclear factor kappa B activation and decreased testicular cell apoptosis in the treated diabetic rats. SRE also raised sperm parameters after treatment of diabetic rats. Conclusively, our results suggested that SRE ameliorated diabetes induced testicular damage by inhibiting oxidative stress and inflammation.
Asunto(s)
Dipterocarpaceae , Extractos Vegetales/farmacología , Testículo/efectos de los fármacos , Acetatos , Animales , Antioxidantes/metabolismo , Apoptosis/efectos de los fármacos , Catalasa/metabolismo , Diabetes Mellitus Experimental/complicaciones , Diabetes Mellitus Experimental/tratamiento farmacológico , Glutatión Peroxidasa/metabolismo , Inflamación/metabolismo , Mediadores de Inflamación/metabolismo , Masculino , Estrés Oxidativo/efectos de los fármacos , Extractos Vegetales/uso terapéutico , Ratas , Espermatozoides/efectos de los fármacos , Superóxido Dismutasa/metabolismo , Testosterona/metabolismoRESUMEN
This study aims to predict the material basis and mechanism of Dachengqi Decoction in the treatment of sepsis based on network pharmacology. The chemical constituents and targets of Dachengqi Decoction were retrieved from TCMSP, UniPot and DrugBank and the targets for the treatment of sepsis from OMIM and GeneCards. The potential targets of Dachengqi Decoction for the treatment of sepsis were screened by OmicShare. STRING database and Cytoscape 3.7.2 were used to construct the Chinese medicinal-active component-target-disease, active component-key target-key pathway, and protein-protein interaction(PPT) networks. The gene ontology(GO) term enrichment analysis and Kyoto encyclopedia of genes and genomes(KEGG) pathway enrichment analysis were performed by DAVID(P<0.05). Finally, the animal experiment was conducted to verify some targets and pathways. A total of 40 active components and 157 targets of the Dachengqi Decoction, 2 407 targets for the treatment of sepsis, and 91 common targets of the prescription and the disease were also obtained. The key targets were prostaglandin G/H synthase 2(PTGS2), prostaglandin G/H synthase 1(PTGS1), protein kinase cAMP-dependent catalytic-α(PRKACA), coagulation factor 2 receptor(F2 R), phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic gamma subunit(PIK3 CG), dipeptidyl peptidase 4(DPP4), etc. A total of 533 terms and 125 pathways were obtained for the 91 targets. The main terms were the response to drug, negative regulation of apoptotic process, positive regulation of nitric oxide biosynthetic process and lipopolysaccharide-mediated signaling pathway, and the pathways included pathways in cancer, hepatitis B, and phosphatidylinositol 3-kinase and protein kinase B(PI3 K/Akt) signaling pathway. The animal experiment confirmed that Dachengqi Decoction can down-regulate inflammatory cytokines interleukin-1ß(IL-1ß), IL-6 and tumor necrosis factor α(TNF-α)(P<0.01). It could also reduce the wet/dry weight ratio of lung tissue, the level of myeloperoxidase(MPO) and the phosphorylation of PI3 K and Akt(P<0.01). These results indicated that Dchengqi Decoction could act on inflammation-related targets and improve sepsis by inhibiting PI3 K/Akt signaling pathway. The animal experiment supported the predictions of network pharmacology. Dachengqi Decoction intervenes sepsis via multiple components, multiple targets, and multiple pathways. The result lays a foundation for further research on the mechanism of Dachengqi Decoction in the treatment of sepsis.
Asunto(s)
Medicamentos Herbarios Chinos , Sepsis , Animales , Ontología de Genes , Extractos Vegetales , Sepsis/tratamiento farmacológico , Sepsis/genéticaRESUMEN
Evidence to date suggests that ß-arrestins act beyond their role as adapter proteins. Arginine vasopressin (AVP) may be a factor in inflammation and fibrosis in the pathogenesis of heart failure. In the present study we investigated the effect of AVP on inflammatory cytokine IL-6 production in murine hearts and the impact of ß-arrestin 2-dependent signaling on AVP-induced IL-6 production. We found that administration of AVP (0.5 U/kg, iv) markedly increased the levels of IL-6 mRNA in rat hearts with the maximum level occurred at 6 h. In ß-arrestin 2 KO mouse hearts, deletion of ß-arrestin 2 decreased AVP-induced IL-6 mRNA expression. We then performed in vitro experiments in adult rat cardiac fibroblasts (ARCFs). We found that AVP (10-9-10-6 M) dose-dependently increased the expression of IL-6 mRNA and protein, activation of NF-κB signaling and ERK1/2 phosphorylation, whereas knockdown of ß-arrestin 2 blocked AVP-induced IL-6 increase, NF-κB activation and ERK1/2 phosphorylation. Pharmacological blockade of ERK1/2 using PD98059 diminished AVP-induced NF-κB activation and IL-6 production. The selective V1A receptor antagonist SR49059 effectively blocked AVP-induced NF-κB phosphorylation and activation as well as IL-6 expression in ARCFs. In AVP-treated mice, pre-injection of SR49059 (2 mg/kg, iv) abolished AVP-induced NF-κB activation and IL-6 production in hearts. The above results suggest that AVP induces IL-6 induction in murine hearts via the V1A receptor-mediated ß-arrestin2/ERK1/2/NF-κB pathway, thus reveal a novel mechanism of myocardial inflammation in heart failure involving the V1A/ß-arrestin 2/ERK1/2/NF-κB signaling pathway.
Asunto(s)
Arginina Vasopresina/farmacología , Corazón/fisiopatología , Interleucina-6/metabolismo , Arrestina beta 2/genética , Animales , Arginina Vasopresina/administración & dosificación , Relación Dosis-Respuesta a Droga , Fibroblastos/metabolismo , Técnicas de Silenciamiento del Gen , Insuficiencia Cardíaca/fisiopatología , Masculino , Ratones , Ratones Noqueados , Proteína Quinasa 1 Activada por Mitógenos/metabolismo , Proteína Quinasa 3 Activada por Mitógenos/metabolismo , FN-kappa B/metabolismo , Ratas , Ratas Sprague-Dawley , Receptores de Vasopresinas/metabolismoRESUMEN
The main goal of this work is to develop an economical, portable, disposable, and reliable point of care paper biosensor based on visualization, which can be used to detect viruses, bacteria, and proteins. However, the sensitivity of immunochromatography test (ICT) strips based on nitrocellulose to target detection has always been a problem. Here, we use an electrospun nitrocellulose (ENC) fiber membrane instead of traditional nitrocellulose fiber membrane to construct ICT strips for early pregnancy detection. By proper selection of the diameter of the ENC fiber to adjust the pore size, porosity, and morphology of the membrane, ICT strips with low flow rate and high protein loading were obtained. Based on these properties, a convenient and sensitive method for the colorimetric determination of human chorionic gonadotropin was developed. Under the optimal conditions, the detection limit of ICT based on ENC membrane is 10 mIU mL-1 (S/N = 3), the linear detection range is 5-1000 mIU mL-1, and the linear relationship is Y = 0.0434 X - 0.0136 (R2 = 0.9802). In addition, the test strip has good specificity and stability, and will not produce false-positive results. Graphical abstract.
Asunto(s)
Cromatografía de Afinidad/métodos , Colodión/química , Gonadotropina Coriónica/análisis , Humanos , Límite de Detección , Sistemas de Atención de Punto , Tiras ReactivasRESUMEN
Ethylene regulates climacteric fruit ripening, and EIN3-LIKE1 (EIL1) plays an important role in this process. In apple (Malus domestica), fruit coloration is accompanied by ethylene release during fruit ripening, but the molecular mechanism that underlies these two physiological processes is unknown. In this study, we found that ethylene treatment markedly induced fruit coloration as well as the expression of MdMYB1, a positive regulator of anthocyanin biosynthesis and fruit coloration. In addition, we found that MdEIL1 directly bound to the promoter of MdMYB1 and transcriptionally activated its expression, which resulted in anthocyanin biosynthesis and fruit coloration. Furthermore, MdMYB1 interacted with the promoter of ETHYLENE RESPONSE FACTOR3, a key regulator of ethylene biosynthesis, thereby providing a positive feedback for ethylene biosynthesis regulation. Overall, our findings provide insight into a mechanism involving the synergistic interaction of the ethylene signal with the MdMYB1 transcription factor to regulate ethylene biosynthesis and fruit coloration in apple.
Asunto(s)
Antocianinas/metabolismo , Etilenos/metabolismo , Malus/genética , Reguladores del Crecimiento de las Plantas/metabolismo , Proteínas de Plantas/metabolismo , Frutas/genética , Frutas/metabolismo , Regulación de la Expresión Génica de las Plantas , Malus/metabolismo , Proteínas de Plantas/genética , Factores de Transcripción/genética , Factores de Transcripción/metabolismoRESUMEN
BACKGROUND: The insecticide exposure has been linked to Parkinson's disease (PD). In the present study, we used a most widely used cell line in study of PD, the SH-SY5Y cells, to investigate mechanisms of chlorpyrifos (CPF) induced cell toxicity and the possible roles of cell pyroptosis and oxidative stress in SH-SY5Y cells, as well as role of miR-181/SIRT1/PGC-1α/Nrf2 signaling pathway in this process. METHODS: SH-SY5Y cells were treated with different concentrations of CPF. Cell viability was measured using CCK-8 assay. Cell pyroptosis was determined by immunofluorescence of caspase-1 and TUNEL assay. The miR-181 (has-miR-181-5p) level was determined by qRT-PCR. Expression of SIRT1, PGC-1α, Nrf2, and pyroptosis related proteins NLRP3, caspase-1, IL-1ß, and IL-18 was determined by both qRT-PCR and Western blotting. RESULTS: Cell viability was found to be decreased with the increased CPF concentrations. The pyroptosis related proteins, ROS levels, as well as level of caspase-1 and the TUNEL positive cells were all significantly up-regulated by CPF. Meanwhile, expression of miR-181 and pyroptosis proteins was also enhanced, while the SIRT1/PGC-1α/Nrf2 signaling was inhibited by CPF. Knockdown of Nrf2 significantly up-regulated the expression of pyroptosis related proteins, ROS level, caspase-1, and the TUNEL positive cells, while over-expression of Nrf2 resulted in opposite results. The expression of PGC-1α and Nrf2 was significantly down-regulated when SIRT1 was inhibited, while over-expressed SIRT1 led to increased PGC-1α and Nrf2 levels. Besides, miR-181 promoted the CPF induced activation of pyroptosis and oxidative stress, as well as down-regulated SIRT1/PGC-1α/Nrf2 signaling, while inhibition of miR-181 led to opposite results. CONCLUSIONS: Chlorpyrifos could inhibit cell proliferation, activate cell pyroptosis and increase susceptibility on oxidative stress-induced toxicity by elevating miR-181 through down-regulation of the SIRT1/PGC-1α/Nrf2 pathway in human neuroblastoma SH-SY5Y cells. This study might give deeper insights for mechanisms of CPF induced toxicity and might give some novel research targets for PD treatment.
Asunto(s)
Cloropirifos/toxicidad , Insecticidas/toxicidad , Factor 2 Relacionado con NF-E2/metabolismo , Estrés Oxidativo/efectos de los fármacos , Enfermedad de Parkinson Secundaria/inducido químicamente , Piroptosis/efectos de los fármacos , Transducción de Señal/efectos de los fármacos , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Humanos , MicroARNs/genética , MicroARNs/metabolismo , Enfermedad de Parkinson Secundaria/metabolismo , Enfermedad de Parkinson Secundaria/patología , Coactivador 1-alfa del Receptor Activado por Proliferadores de Peroxisomas gamma/genética , Coactivador 1-alfa del Receptor Activado por Proliferadores de Peroxisomas gamma/metabolismo , Sirtuina 1/genética , Sirtuina 1/metabolismoRESUMEN
OBJECTIVES: The arsenic trioxide (ATO) plus all-trans retinoic acid (ATRA) therapy has demonstrated a tremendous success in the first-line treatment of acute promyelocytic leukemia (APL). Actually, early death (ED) is currently thought as a major challenge in APL. ATO has been reported to inhibit platelet function in vitro, and whether it increases the ED rate by exacerbating the hemorrhagic symptoms remains to be investigated. METHODS: Effects of ATO on platelet aggregation and adhesion were evaluated in vitro and in thirty-two complete remission (CR) and four newly diagnosed APL patients. Furthermore, concentrations of plasma total arsenic were monitored in APL patients via ICP-MS. RESULTS: The inhibition of platelet function, either aggregation or adhesion, did occur in vitro when the concentration of ATO reached 2 µmol/L. However, in CR APL patients receiving ATO with normal platelet count, the platelets responded normally when being activated and so did those in the newly diagnosed patients with thrombocytopenia. Our data further showed that the conventional dosage of ATO reached a plasma concentration substantially below the required concentration to inhibit platelets. CONCLUSIONS: In the first-line treatment of APL, the use of ATO is safe and effective and does not compromise the hemostatic potential that may eventually increase ED rate.
Asunto(s)
Antineoplásicos/administración & dosificación , Arsenicales/administración & dosificación , Hemorragia/etiología , Leucemia Promielocítica Aguda/complicaciones , Leucemia Promielocítica Aguda/tratamiento farmacológico , Óxidos/administración & dosificación , Adolescente , Adulto , Antineoplásicos/efectos adversos , Antineoplásicos/farmacocinética , Protocolos de Quimioterapia Combinada Antineoplásica/efectos adversos , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Trióxido de Arsénico , Arsenicales/efectos adversos , Arsenicales/farmacocinética , Coagulación Sanguínea/efectos de los fármacos , Femenino , Hemorragia/mortalidad , Humanos , Leucemia Promielocítica Aguda/sangre , Quimioterapia de Mantención , Masculino , Persona de Mediana Edad , Óxidos/efectos adversos , Óxidos/farmacocinética , Adhesividad Plaquetaria/efectos de los fármacos , Agregación Plaquetaria/efectos de los fármacos , Pruebas de Función Plaquetaria , Inducción de Remisión , Resultado del Tratamiento , Adulto JovenRESUMEN
Galactoside-binding lectins, also known as galectins, play crucial roles in innate immune response in invertebrates. In this study, three cDNA sequences from Hyriopsis cumingii were identified and collectively called HcGalec genes. Each of the three deduced HcGalec proteins contained a galactose-binding lectin domain or a GLECT domain. All the three HcGalec genes are mainly present in the hepatopancreas and gills, and their expression is induced at 24 h after bacterial challenge. Three recombinant HcGalec proteins can bind and agglutinate (Ca2+-dependent) various microorganisms, including Gram-positive and Gram-negative bacteria. These proteins can attach to mannan and peptidoglycan. Meanwhile, the expression of the three HcGalec genes in the gills were significantly down-regulated after dsRNA interference (HcGalec1-RNAi, HcGalec2-RNAi, and HcGalec3-RNAi) and Vibrio parahaemolyticus injection. The expression levels of some antimicrobial peptides, including lysozyme 1 and lysozyme 2, were also markedly decreased after dsRNA interference. Overall, these results suggested that these three HcGalec proteins may function as potential receptors participating in the innate immune responses of H. cumingii against bacterial infection.
Asunto(s)
Galectinas/genética , Galectinas/inmunología , Inmunidad Innata/genética , Receptores de Reconocimiento de Patrones/genética , Receptores de Reconocimiento de Patrones/inmunología , Unionidae/genética , Unionidae/inmunología , Animales , Perfilación de la Expresión Génica , Bacterias Gramnegativas/fisiología , Bacterias Grampositivas/fisiologíaRESUMEN
The immune deficiency (IMD) signal pathway mediates innate immunity against Gram-negative bacteria in crustaceans. In the present study, an IMD homolog (MnIMD) from the oriental river prawn, Macrobrachium nipponense was identified. The full-length cDNA of MnIMD was 782bp with an open reading frame of 549 bp that encodes a putative protein of 182 amino acids including a death domain at the C-terminus. Multiple alignment analysis showed that IMDs in prawn M. nipponense and other crustaceans shared high similarity. The recombinant protein of MnIMD was expressed and purified for further functional analyses. Western blot analysis indicated that MnIMD was present in many tissues, but with the highest level in the gills, which was consistent with the qRT-PCR results. After Vibrio parahaemolyticus challenge, MnIMD was significantly induced in gills. RNA interference analysis showed that the IMD pathway was involved in regulating the expression of different antimicrobial peptide (AMP) genes, including Cru4 and Cru6. These results are helpful in promoting research on the innate immunity in M. nipponense.
Asunto(s)
Proteínas de Artrópodos/inmunología , Inmunidad Innata , Palaemonidae/genética , Palaemonidae/inmunología , Transducción de Señal , Animales , Péptidos Catiónicos Antimicrobianos/genética , Péptidos Catiónicos Antimicrobianos/inmunología , Proteínas de Artrópodos/genética , Clonación Molecular , ADN Complementario , Regulación de la Expresión Génica , Branquias/metabolismo , Palaemonidae/microbiología , Filogenia , Alineación de Secuencia , Vibriosis/inmunología , Vibrio parahaemolyticus/fisiologíaRESUMEN
To establish the ultra performance liquid chromatography (UPLC) fingerprint of Dandeng Tongnao Ruanjiaonang and conduct a systemic, comprehensive quality evaluation of the drug by combining with a chemical pattern recognition method. In this study, Waters UPLC ultra-high performance liquid chromatography instrument and ACQUITY UPLCîHSS T3 chromatographic colum n were employed to perform the separation with acetonitrile-0.1% formic acid aqueous solution as the mobile phase for gradient elution; and the detection wavelength was set at 256 nm to establish the UPLC fingerprint of 10 batches of Dandeng Tongnao Ruanjiaonang. Then, the further quality assessment of the drug was carried out by similarity evaluation, Cluster Analysis(CA), Principal Component Analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA). Finally, 77 peaks were recognised as common peaks in the fingerprint, and 15 peaks of them were identified using standard references. The similarity value of these 10 batches of drugs was all above 0.960, indicating a relatively stable quality. But minor differences were still discovered between the batches of the drug by CA and PCA. Finally, 6 common peaks were recognised as the quality makers using OPLS-DA method. The analysis method established in this study was scientific, accurate, reliable and simple; fingerprint combined with chemical pattern recognition technique can be used to systematically and comprehensively evaluate the drug quality of Dandeng Tongnao Ruanjiaonang; what's more, it could also provide a reference for the quality control of traditional Chinese medicine and its preparations at the same time.
Asunto(s)
Medicamentos Herbarios Chinos/normas , Control de Calidad , Cromatografía Líquida de Alta Presión , Medicamentos Herbarios Chinos/química , Análisis de Componente PrincipalRESUMEN
Autophagy is an evolutionarily conserved degradative process that allows cells to maintain homoeostasis in numerous physiological situations. This process also functions as an essential protective response to endoplasmic reticulum (ER) stress, which promotes the removal and degradation of unfolded proteins. However, little is known regarding the mechanism by which autophagy is initiated and regulated in response to ER stress. In this study, different types of autophagy were identified in human gastric cancer MKN45 cells in response to the stress induced by nutrient starvation or lipotoxicity in which the regulation of these pathways is mammalian target of rapamycin (mTOR)-dependent or -independent, respectively. Interestingly, we found that p8, a stress-inducible transcription factor, was enhanced in MKN45 cells treated with palmitic acid to induce lipotoxicity. Furthermore, an increase in autophagy was observed in MKN45 cells stably overexpressing p8 using a lentivirus system, and autophagy induced by palmitic acid was blocked by p8 RNAi compared with the control. Western blotting analyses showed that autophagy was regulated by p8 or mTOR in response to the protein kinase-like endoplasmic reticulum kinase/activating transcription factor 6-mediated ER stress of lipotoxicity or the parkin-mediated mitochondrial stress of nutrient starvation, respectively. Furthermore, our results indicated that autophagy induced by palmitic acid is mTOR-independent, but this autophagy pathway was regulated by p8 via p53- and PKCα-mediated signaling in MKN45 cells. Our findings provide insights into the role of p8 in regulating autophagy induced by the lipotoxic effects of excess fat accumulation in cells.
Asunto(s)
Autofagia , Proteínas de Unión al ADN/metabolismo , Estrés del Retículo Endoplásmico , Ácidos Grasos no Esterificados/efectos adversos , Transducción de Señal , Factores de Transcripción/metabolismo , Regulación hacia Arriba , Autofagia/efectos de los fármacos , Línea Celular Tumoral , Proteínas de Unión al ADN/antagonistas & inhibidores , Proteínas de Unión al ADN/genética , Estrés del Retículo Endoplásmico/efectos de los fármacos , Proteínas Fluorescentes Verdes/genética , Proteínas Fluorescentes Verdes/metabolismo , Células HEK293 , Humanos , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Microscopía Fluorescente , Ácido Palmítico/efectos adversos , Inhibidores de Proteínas Quinasas/farmacología , Interferencia de ARN , Proteínas Recombinantes de Fusión/química , Proteínas Recombinantes de Fusión/metabolismo , Transducción de Señal/efectos de los fármacos , Serina-Treonina Quinasas TOR/metabolismo , Factores de Transcripción/antagonistas & inhibidores , Factores de Transcripción/genética , Regulación hacia Arriba/efectos de los fármacosRESUMEN
A gas chromatography-mass spectrometry(GC-MS)method was established for the analysis of volatile components in Mentha haplocalyx, and seven principal components were quantified by gas chromatography(GC). Based on these analyses, the differences of volatile components in M. haplocalyx from Jiangsu, Anhui and other regions were compared. The results showed that the volatile oil of M. haplocalyx was divided into four chemical typesï¼menthol-menthone type, pulegone-menthone type, piperitone-menthol type, piperitone epoxide type, and menthol-menthone type was the principal type. Menthol was the highest and pulegone was the lowest. The differences of M. haplocalyx from Anhui and other regions were obvious. The major volatile components and the differences of M. haplocalyx from different regions were confirmed and a quantitative method was established for the determination of volatile components, which provided the basis for improving the quality standard of M. haplocalyx.
Asunto(s)
Medicamentos Herbarios Chinos/normas , Mentha/química , Aceites Volátiles/química , Cromatografía de Gases y Espectrometría de Masas , Aceites de Plantas/química , Plantas Medicinales/químicaRESUMEN
C1q is a key subcomponent of the complement C1 complex. This subcomponent contains a globular C1q (gC1q) domain with remarkable ligand binding properties. C1q domain-containing (C1qDC) proteins are composed of all proteins with a gC1q domain. C1qDC proteins exist in many invertebrates and recognize non-self-ligands. In our study, four C1qDC genes, namely, HcC1qDC1-HcC1qDC4, were identified from Hyriopsis cumingii. HcC1qDC1-HcC1qDC4 encode a protein of 224, 204, 305, and 332 amino acids, respectively. All C1qDC proteins consist of a gC1q domain at the C terminal. In addition to the gC1q domain, a coiled-coil region is found in HcC1qDC4. Multiple alignments and phylogenetic tree analysis revealed that the C1qDC proteins highly differ from one another. Tissue distribution analysis demonstrated that HcC1qDC1-HcC1qDC4 are widely distributed in hemocytes, hepatopancreas, gills, mantle, and foot. These C1qDC genes are regulated by bacteria to varying degrees. These recombinant HcC1qDC proteins exhibit a binding activity against different bacterial species. Our results may suggest the roles of HcC1qDC genes in anti-bacterial immune defense.
Asunto(s)
Complemento C1q/genética , Expresión Génica , Inmunidad Innata/genética , Unionidae/genética , Aeromonas hydrophila/fisiología , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Complemento C1q/metabolismo , Hepatopáncreas/inmunología , Hepatopáncreas/microbiología , Especificidad de Órganos , Filogenia , Alineación de Secuencia , Staphylococcus aureus/fisiología , Unionidae/inmunología , Unionidae/microbiologíaRESUMEN
As a type of pattern-recognition proteins, lectins perform important functions in the innate immunity of crustaceans, including prawns. Although several reports showed that C-type lectin domain family (CLEC) importantly functions in host-pathogen interactions, limited research has focused on CLEC in Macrobrachium rosenbergii. In the present study, a new single CRD containing CLEC (designated as MrLec) was reported in freshwater prawns, M. rosenbergii. The full-length cDNA of MrLec consisted of 1027 bp with an open reading frame of 801 bp, which encoded a peptide of 266 amino acid residues. Genomic sequence for MrLec was also obtained from the M. rosenbergii, which contain 4 exons and 3 introns. MrLec was found to contain a single carbohydrate-recognition domain with an EPN motif. MrLec was ubiquitously distributed in various tissues of a normal prawn, particularly in the hepatopancreas and gills. MrLec expression in the gills was significantly upregulated after a challenge with Vibrio parahaemolyticus and downregulated at 24 h after MrLec RNA interference (MrLec-RNAi). The expression levels of some AMPs, including antilipopolysaccharide factor 1 (Alf1) and lysozyme 2 (Lyso2), also markedly decreased after MrLec-RNAi. Recombinant MrLec can agglutinate (Ca(2+)-dependent) and bind both Gram-negative and Gram-positive bacteria. Results suggested that MrLec participates in the recognition of invading pathogens and functions in the immune response of prawn against pathogen infections.
Asunto(s)
Proteínas de Artrópodos/inmunología , Lectinas Tipo C/inmunología , Palaemonidae/inmunología , Vibriosis/inmunología , Pruebas de Aglutinación , Secuencia de Aminoácidos , Animales , Proteínas de Artrópodos/genética , Secuencia de Bases , ADN Complementario/genética , Branquias/metabolismo , Bacterias Gramnegativas/efectos de los fármacos , Bacterias Grampositivas/efectos de los fármacos , Hepatopáncreas/metabolismo , Inmunidad Innata , Lectinas Tipo C/genética , Palaemonidae/genética , ARN Mensajero/metabolismo , Vibriosis/veterinariaRESUMEN
C-type lectins (CTLs) are found in a wide number of invertebrates, and have been reported to participate in immune responses, such as the activation of prophenoloxidase, cell adhesion, bacterial clearance and phagocytosis. Previous studies on CTLs focused on the function of their carbohydrate recognition domains (CRDs). Currently, studies on lectins with multi-CRDs are limited. In this study, a lectin with four CRDs was cloned from Hyriopsis cumingii, and called HcLec4. HcLec4 was widely distributed in several tissues and was significantly down-regulated at the early stage (2 h) of bacterial infection. We further analyzed the bacteria and carbohydrate binding activities of HcLec4. The results showed that HcLec4 could bind to several bacteria, lipopolysaccharide (LPS) and peptidoglycan (PGN). In HcLec4 knockdown mussels, the bacterial clearance rate was increased, and the expression level of antimicrobial peptides (AMPs) was up-regulated. This study reveals that HcLec4 exerts its antibacterial effect by regulating the expression of AMPs at the early stage of bacterial infection.
Asunto(s)
Péptidos Catiónicos Antimicrobianos/genética , Complemento C1q/genética , Regulación de la Expresión Génica/genética , Inmunidad Innata/genética , Unionidae/genética , Secuencia de Aminoácidos , Animales , Péptidos Catiónicos Antimicrobianos/química , Péptidos Catiónicos Antimicrobianos/metabolismo , Fenómenos Fisiológicos Bacterianos , Complemento C1q/química , Complemento C1q/metabolismo , Lipopolisacáridos/farmacología , Especificidad de Órganos , Peptidoglicano/farmacología , Filogenia , Alineación de Secuencia , Unionidae/inmunología , Unionidae/microbiologíaRESUMEN
Regulation of the cell cycle is complex but critical for proper development, reproduction and stress resistance. To survive unfavourable environmental conditions, the crustacean Artemia produces diapause embryos whose metabolism is maintained at extremely low levels. In the present study, the expression profiles of miRNAs during Artemia diapause entry and termination were characterized using high-throughput sequencing. A total of 13 unclassified miRNAs and 370 miRNAs belonging to 87 families were identified; among them, 107 were differentially expressed during diapause entry and termination. We focused on the roles of two of these miRNAs, miR-100 and miR-34, in regulating cell cycle progression; during the various stages of diapause entry, these miRNAs displayed opposing patterns of expression. A functional analysis revealed that miR-100 and miR-34 regulate the cell cycle during diapause entry by targeting polo-like kinase 1 (PLK1), leading to activation of the mitogen-activated protein kinase kinase-extracellular signal-regulated kinase-ribosomal S6 kinase 2 (MEK-ERK-RSK2) pathway and cyclin K, leading to suppression of RNA polymerase II (RNAP II) activity respectively. The findings presented in the present study provide insights into the functions of miR-100 and miR-34 and suggest that the expression profiles of miRNAs in Artemia can be used to characterize their functions in cell cycle regulation.
Asunto(s)
Artemia/metabolismo , MicroARNs/metabolismo , Animales , Artemia/citología , Artemia/embriología , Artemia/crecimiento & desarrollo , Puntos de Control del Ciclo Celular , Proteínas de Ciclo Celular/metabolismo , Ciclinas/metabolismo , Oviparidad , Ovoviviparidad , Proteínas Serina-Treonina Quinasas/metabolismo , Proteínas Proto-Oncogénicas/metabolismo , ARN Polimerasa II/metabolismo , Transducción de Señal , Quinasa Tipo Polo 1RESUMEN
The toll-like receptor/NF-κB signaling pathways play an important role in the innate immune system. In the present study, one Pelle gene (named EsPelle) was identified for the first time from the Chinese mitten crab Eriocheir sinensis. The full-length cDNA of EsPelle is 3797 bp with a 3156 bp-long open reading frame that encodes a 1051 amino acid protein. EsPelle protein contains a death domain at the N-terminal and a serine/threonine kinase domain at the C-terminal. A neighbor joining phylogenetic tree showed that the EsPelle protein, which is closest to those of Scylla paramamosain Pelle and Litopenaeus vannamei Pelle, was clustered to a group of crustacean Pelle proteins. EsPelle was expressed in all tested tissues of normal crabs, and its expression was regulated in hemocytes and hepatopancreas of crabs challenged with lipopolysaccharide, peptidoglycan, Staphyloccocus aureus, Vibrio parahaemolyticus, and Aeromonas hydrophila. Overexpression of EsPelle in Drosophila Schneider 2 cells could upregulate the expression of Drosophila antimicrobial peptides, namely, metchnikowin (Mtk), attacinA (Atta), drosomycin (Drs), and cecropinA (CecA). Moreover, EsPelle silencing by siRNA reduced the transcription of anti-lipopolysaccharide factor 1 and 2, crustin 2, and lysozyme in crabs challenged with V. parahaemolyticus. From the results, we speculated that EsPelle was involved in innate immune defense against V. parahaemolyticus in E. sinensis.
Asunto(s)
Proteínas de Artrópodos , Braquiuros , Proteínas Serina-Treonina Quinasas , Aeromonas hydrophila , Secuencia de Aminoácidos , Animales , Proteínas de Artrópodos/genética , Proteínas de Artrópodos/inmunología , Proteínas de Artrópodos/metabolismo , Secuencia de Bases , Braquiuros/genética , Braquiuros/inmunología , Braquiuros/metabolismo , Línea Celular , Drosophila , Proteínas de Drosophila/genética , Branquias/inmunología , Branquias/metabolismo , Infecciones por Bacterias Gramnegativas/inmunología , Infecciones por Bacterias Gramnegativas/veterinaria , Hemocitos/inmunología , Hemocitos/metabolismo , Hepatopáncreas/inmunología , Hepatopáncreas/metabolismo , Lipopolisacáridos/farmacología , Datos de Secuencia Molecular , Peptidoglicano/farmacología , Proteínas Serina-Treonina Quinasas/genética , Proteínas Serina-Treonina Quinasas/inmunología , Proteínas Serina-Treonina Quinasas/metabolismo , Infecciones Estafilocócicas/inmunología , Infecciones Estafilocócicas/veterinaria , Staphylococcus aureus , Vibrio parahaemolyticusRESUMEN
Temperature is a very important parameter in scientific research, production and life. Almost all the properties of materials are related to temperature. The precise measurement of the temperature is a very important task, so the temperature sensor is widely used as a core part in the temperature measuring instrument. A novel surface plasmon micro-ring sensor suitable for humidity sensing is presented in this paper. The sensor uses a multi-layered surface plasmon waveguide structure and choosing Polyimide (Polyimide, PI) as the moisture material. We get the transfer function of surface plasmon micro-ring sensor by using transfer matrix method. Refractive indexes of Polyimide and the multilayer waveguide structure change as environment relative humidity changes, thus leading to an obvious peak drift of output spectrum. The paper mainly discusses the influence of the changes of the refractive index of humidity-sensing parts on the output spectrum, and the transmission characteristics of multilayer waveguide structure. Through the finite element method and the theoretical simulation of Matlab, We can draw: When the length between the two coupling points of the U-shaped waveguide is an integer multiple of circumference of the micro-ring, an obvious drift in the horizontal direction appears, the free spectral range (FSR) doubled and the sensitivity is 0.0005 µm/%RH; When the external environment relative humidity RH changes from 10% to 100% RH, scatter is change between including (including 0.005 m to 0.005 m, compared to other humidity sensor, the Sensitivity of sensor improves 10~50 times and the transmission is very stable. Results show that the design of surface plasma micro ring sensors has better sensitivity, stable performance and can be used in the humidity measurement, achieving a high sensitivity in the sense of humidity when the wide range of filter frequency selection is taken into account, and providing a theoretical basis for the preparation of micro-optics.
RESUMEN
Although numerous miRNAs have been already isolated from fruit trees, knowledge about miRNA biogenesis is largely unknown in fruit trees. Double-strand RNA-binding (DRB) protein plays an important role in miRNA processing and maturation; however, its role in the regulation of economically important traits is not clear yet in fruit trees. EST blast and RACE amplification were performed to isolate apple MdDRB1 gene. Following expression analysis, RNA binding and protein interaction assays, MdDRB1 was transformed into apple callus and in vitro tissue cultures to characterize the functions of MdDRB1 in miRNA biogenesis, adventitious rooting, leaf development and tree growth habit. MdDRB1 contained two highly conserved DRB domains. Its transcripts existed in all tissues tested and are induced by hormones. It bound to double-strand RNAs and interacted with AtDCL1 (Dicer-Like 1) and MdDCL1. Chip assay indicated its role in miRNA biogenesis. Transgenic analysis showed that MdDRB1 controls adventitious rooting, leaf curvature and tree architecture by modulating the accumulation of miRNAs and the transcript levels of miRNA target genes. Our results demonstrated that MdDRB1 functions in the miRNA biogenesis in a conserved way and that it is a master regulator in the formation of economically important traits in fruit trees.