RESUMEN
Surface-enhanced Raman spectroscopy (SERS) is a powerful technique for discrimination of bimolecules in complex systems. However, its practical applications face challenges such as complicated manufacturing procedures and limited scalability of SERS substrates, as well as poor reproducibility during detection which compromises the reliability of SERS-based analysis. In this study, we developed a convenient method for simultaneous fabrication of massive SERS substrates with an internal standard to eliminate the substrate-to-substrate differences. We first synthesized Au@CN@Au nanoparticles (NPs) which contain embedded internal standard molecules with a single characteristic peak in the Raman-silent region, and then deposited the NPs on 6 mm glass wafers in a 96-well plate simply by centrifugation for 3 min. The one-time obtained 96 SERS substrates have excellent intrasubstrate uniformity and intersubstrate repeatability for SERS detection by using the internal standard (relative standard deviation = 10.47%), and were able to detect both charged and neutral molecules (crystal violet and triphenylphosphine) at a concentration of 10-9 M. Importantly, cells can be directly cultured on glass wafers in the 96-well plate, enabling real time monitoring of the secretes and metabolism change in response to external stimulation. We found that the release of nucleic acids, amino acids and lipids by MDA-MB-231 cells significantly increased under hypoxic conditions. Overall, our approach enables fast and large-scale production of Au@CN@Au NPs-coated glass wafers as SERS substrates, which are homogeneous and highly sensitive for monitoring trace changes of biomolecules.
Asunto(s)
Vidrio , Oro , Nanopartículas del Metal , Espectrometría Raman , Oro/química , Espectrometría Raman/métodos , Nanopartículas del Metal/química , Humanos , Vidrio/química , Línea Celular TumoralRESUMEN
Recently, an increasing number of studies have shown that long noncoding RNAs (lncRNAs) are involved in host metabolism after infection with pseudorabies virus (PRV). In our study, via RNA sequencing analysis, a total of 418 mRNAs, 137 annotated lncRNAs, and 312 new lncRNAs were found to be differentially expressed. These lncRNAs were closely associated with metabolic regulation and immunity-related signalling pathways, including the T-cell receptor signalling pathway, chemokine signalling pathway, mitogen-activated protein kinase (MAPK) signalling pathway, TNF signalling pathway, Ras signalling pathway, calcium signalling pathway, and phosphatidylinositol signalling system. Real-time PCR indicated that several mRNAs and lncRNAs involved in the regulation of the immune effector process, T-cell receptor signalling pathway, TNF signalling pathway, MAPK signalling pathway, and chemokine signalling pathways were significantly expressed. These mRNAs and lncRNAs might play a role in PRV infection.
Asunto(s)
Herpesvirus Suido 1 , Seudorrabia , ARN Largo no Codificante , Animales , ARN Largo no Codificante/genética , Herpesvirus Suido 1/genética , Seudorrabia/genética , ARN Mensajero/genética , Receptores de Antígenos de Linfocitos T , QuimiocinasRESUMEN
The physicochemical properties of nanoparticles (NPs) significantly influence their deposition at the disease site, ultimately impacting the overall therapeutic efficacy; however, precisely assessing the effects of various factors on NP accumulation within a single cell/tumor tissue is challenging due to the lack of appropriate labeling techniques. Surface-enhanced Raman spectroscopy (SERS) tag is a powerful encoding method that has recently been intensively employed for immunodetection of biomarkers. Herein, we introduce a multiplexed SERS tracking approach for systematic investigation of size-dependent accumulation and distribution of NPs within the same tumor. Four-sized (34, 60, 108, and 147 nm) NPs encoded with different SERS "colors" were fabricated, mixed, and incubated with monolayer tumor cells, multicellular tumor spheroids, or injected into mouse models bearing xenograft solid tumors in a single dose. Multicolor SERS detection of the specimens revealed that NP accumulation in tumor cells, tumor spheroids, and solid tumors was in the order of 34 nm > 60 nm > 108 nm > 147 nm, 60 nm > 34 nm > 108 nm > 147 nm, and 34 nm > 147 nm > 108 nm > 60 nm, respectively. Inductively coupled plasma mass spectroscopy determination performed in parallel samples were in alignment with the four-color SERS probing results, demonstrating the effectiveness of this multiplexed evaluation assay. Furthermore, in combination with fluorescence labeling of specific biomolecules, this method can be applied for the colocalization of different NPs in various pathological structures and provide additional information for analysis of the possible mechanisms.
Asunto(s)
Nanopartículas , Neoplasias , Animales , Humanos , Ratones , Modelos Animales de Enfermedad , Espectrometría RamanRESUMEN
Hematopoietic stem and progenitor cell (HSPC) research will help elucidate the pathogenesis of hematologic diseases. The present study aimed to establish an isolation method and culture system for chicken bone marrow (BM)-derived HSPCs and test their proliferation and differentiation abilities. Mononuclear cells were collected from chicken BM, and CD34+ HSPCs were isolated. Then, the cells were cultured in media with different cytokine compositions, and the growth status, cell phenotype, and morphological appearance of the cells were analyzed at different time points. Our results showed that Iscove's Modified Dulbecco's Medium supplemented with 50 ng/mL stem cell factor, 30 ng/mL Flt-3 ligand, 10 µg/mL interleukin 3, 50 ng/mL interleukin 6%, and 10% chicken serum supported chicken CD34+ HSPC survival ex vivo for approximately 10 d. Further, 80 ng/mL granulocyte-colony stimulating factor and 30 ng/mL granulocyte macrophage-colony stimulating factor were added into the above culture system to form a myeloid cell differentiation induction culture system. After culturing in this system for 72 h, approximately 66% of chicken CD34+ HSPCs exhibited a CD11b+ phenotype, indicating that HSPCs differentiated into myeloid cells. In conclusion, chicken BM-derived CD34+ cells possess HSPC characteristics that can self-renew and differentiate into myeloid cells in a culture medium containing growth factors.
Asunto(s)
Médula Ósea , Pollos , Animales , Antígenos CD34 , Células Madre Hematopoyéticas , Diferenciación Celular , Células Mieloides , Células de la Médula Ósea , Células CultivadasRESUMEN
Logic-in-memory architecture holds great promise to meet the high-performance and energy-efficient requirements of data-intensive scenarios. Two-dimensional compacted transistors embedded with logic functions are expected to extend Moore's law toward advanced nodes. Here we demonstrate that a WSe2/h-BN/graphene based middle-floating-gate field-effect transistor can perform under diverse current levels due to the controllable polarity by the control gate, floating gate, and drain voltages. Such electrical tunable characteristics are employed for logic-in-memory architectures and can behave as reconfigurable logic functions of AND/XNOR within a single device. Compared to the conventional devices like floating-gate field-effect transistors, our design can greatly decrease the consumption of transistors. For AND/NAND, it can save 75% transistors by reducing the transistor number from 4 to 1; for XNOR/XOR, it is even up to 87.5% with the number being reduced from 8 to 1.
RESUMEN
Seneca Valley virus (SVV) is an oncolytic virus, which belongs to the Picornaviridae family, that causes blisters on the nose and hooves, affecting the production performance of pigs. However, the function of proinflammatory cytokines and chemokines in SVV infection is still unclear. In our study, SVV infection could induce a high expression of proinflammatory cytokines interleukin (IL)-1α, IL-1ß, and tumor necrosis factor α (TNF-α) and chemokines, including chemokine (C-C motif) ligand 2 (CCL2), chemokine (C-C motif) ligand 5 (CCL5), and chemokine (C-X-C motif) ligand 10 (CXCL10). Interfered genes of IL-1α, IL-1ß, and TNF-α inhibited virus replication, but interfered genes of CCL2, CCL5, and CXCL10 promoted virus replication. These results indicate that proinflammatory cytokines and chemokines are involved in SVV infection; this will be beneficial to explore the pathogenesis and cytokine therapy of SVV.
Le virus de la Vallée de Seneca (SVV) est un virus oncolytique, qui appartient à la famille des Picornaviridae, qui provoque des cloques sur le nez et les sabots, affectant les performances de production des porcs. Cependant, la fonction des cytokines pro-inflammatoires et des chimiokines dans l'infection par le SVV n'est toujours pas claire. Dans notre étude, l'infection par le SVV pourrait induire une forte expression des cytokines pro-inflammatoires interleukine (IL)-1α, IL-1ß, et du facteur de nécrose tumorale α (TNF-α) et des chimiokines, y compris la chimiokine (motif C-C) ligand 2 (CCL2), chimiokine (motif C-C) ligand 5 (CCL5) et chimiokine (motif C-X-C) ligand 10 (CXCL10). Les gènes interférés d'IL-1α, IL-1ß et TNF-α inhibent la réplication virale, mais les gènes interférents de CCL2, CCL5 et CXCL10 favorisent la réplication virale. Ces résultats indiquent que les cytokines pro-inflammatoires et les chimiokines sont impliquées dans l'infection par le SVV; cela sera bénéfique pour explorer la pathogenèse et la thérapie par cytokines du SVV.(Traduit par Docteur Serge Messier).
Asunto(s)
Citocinas , Picornaviridae , Animales , Porcinos , Citocinas/genética , Factor de Necrosis Tumoral alfa/metabolismo , Quimiocinas/genética , Quimiocinas/metabolismoRESUMEN
Two-dimensional (2D) van der Waals heterostructures based on transition metal dichalcogenides are expected to be unique building blocks for next-generation nanoscale electronics and optoelectronics. The ability to control the properties of 2D heterostructures is the key for practical applications. Here, we report a simple way to fabricate a high-performance self-driven photodetector based on the MoTe2/MoSe2 p-n heterojunction, in which the hole-dominated transport polarity of MoTe2 is easily achieved via a straightforward thermal annealing treatment in air without any chemical dopants or special gases needed. A high photoresponsivity of 0.72 A W-1, an external quantum efficiency up to 41.3%, a detectivity of 7 × 1011 Jones, and a response speed of 120 µs are obtained at zero bias voltage. Additionally, this doping method is also utilized to realize a complementary inverter with a voltage gain of 24. By configuring 2D p-MoTe2 and n-MoSe2 on demand, logic functions of NAND and NOR gates are also accomplished successfully. These results present a significant potential toward future larger-scale heterogeneously integrated 2D electronics and optoelectronics.
RESUMEN
Vesicular disease caused by Seneca Valley virus (SVV) has recently emerged throughout China and caused certain industry losses. We used immunofluorescence and western blotting to confirm that 3 new SVV strains (CH-GDSG-2018-1, CH-GDSG-2018-2, and CH-GDSG-2018-3) were from 1 pig farm. Phylogenetic analysis revealed the following: i) all 3 strains belong to USA-GBI29-2015-like clades, ii) CH-GDSG-2018-3 might have diverged from CH-GDSG-2018-1 and CH-GDSG-2018-2, and iii) CH-GDSG-2018-3 is a recombinant of the CHhb17 and HeNKF-1 strains. Virus growth curves showed that CH-GDSG-2018-3 had stronger proliferation ability in vitro. Seneca Valley virus has evolved extensively within China and this study has furthered our understanding of SVV epidemiology.
La maladie vésiculeuse causée par le virus de la vallée de Seneca (SVV) est récemment apparue dans toute la Chine et a causé certaines pertes dans l'industrie. Nous avons utilisé l'immunofluorescence et l'immunobuvardage pour confirmer que trois nouvelles souches de SVV (CH-GDSG-2018-1, CH-GDSG-2018-2 et CH-GDSG-2018-3) provenaient d'un seul élevage de porcs. L'analyse phylogénétique a révélé ce qui suit : i) les trois souches appartiennent à des clades de type USA-GBI29-2015, ii) CH-GDSG-2018-3 pourrait avoir divergé de CH-GDSG-2018-1 et CH-GDSG-2018-2, et iii) CH-GDSG-2018-3 est un recombinant des souches CHhb17 et HeNKF-1. Les courbes de croissance virale ont montré que CH-GDSG-2018-3 avait une capacité de prolifération in vitro plus forte. Le virus SVV a considérablement évolué en Chine et cette étude a approfondi notre compréhension de l'épidémiologie de ce virus.(Traduit par Docteur Serge Messier).
Asunto(s)
Infecciones por Picornaviridae , Enfermedades de los Porcinos , Animales , Granjas , Filogenia , Picornaviridae , Infecciones por Picornaviridae/epidemiología , Infecciones por Picornaviridae/veterinaria , Porcinos , Enfermedades de los Porcinos/epidemiologíaRESUMEN
Long noncoding RNAs (lncRNAs) have been demonstrated to play key roles in various biological processes. However, the contributions of lncRNAs to Seneca Valley virus (SVV) infection and host defense remain largely unknown. In this study, differentially expressed lncRNAs and mRNAs in SVV-infected PK15 cells were detected by genome-wide analysis. A total of 14,127 lncRNAs and 63,562 mRNAs were identified, and 1,780 lncRNAs were differentially expressed. The functional prediction of SVV-induced lncRNAs showed high associations with biological regulation and many immunity-related signaling pathways, including the B-cell receptor pathway, RIG-I-like receptor signaling pathway, and NF-kappa B (NF-κB) signaling pathway. We next screened lncRNAs and target genes related to immune response pathways and further demonstrated their differential expression in SVV-infected PK15 cells. Our study investigated the function of lncRNAs involved in SVV infection and provided new insight into the pathogenic mechanisms of SVV.
RESUMEN
Current liquid biopsy assays lack sufficient sensitivity to detect copy number loss, which limits the interrogation of critical tumor suppressor gene deletions during cancer progression and treatment. Here we describe a liquid biopsy assay with improved sensitivity for detection of copy number loss in blood samples with low levels of circulating tumor DNA, and demonstrate its utility by profiling PTEN, RB1, and TP53 genetic loss in metastatic prostate cancer patients.
RESUMEN
Congenital avian leukosis virus subgroup J (ALV-J) infection can induce persistent immunotolerance in chicken, however, the underlying mechanism remains unclear. Here, we demonstrate that congenital ALV-J infection induces the production of high-frequency and activated CD4+CD25+ Tregs that maintain persistent immunotolerance. A model of congenital infection by ALV-J was established in fertilized eggs, and hatched chicks showed persistent immunotolerance characterized by persistent viremia, immune organ dysplasia, severe imbalance of the ratio of CD4+/CD8+ T cells in blood and immune organs, and significant decrease in CD3+ T cells and Bu-1+ B cells in the spleen. Concurrently, the mRNA levels of IL-2, IL-10, and IFN-γ showed significant fluctuations in immune organs. Moreover, the frequency of CD4+CD25+ Tregs in blood and immune organs significantly increased, and the frequency of CD4+CD25+ Tregs was positively correlated with changes in ALV-J load in immune organs. Interestingly, CD4+CD25+ Tregs increased in the marginal zone of splenic nodules in ALV-J-infected chickens and dispersed to the germinal center. In addition, the proliferation and activation of B cells in splenic nodules was inhibited, and the number of IgM+ and IgG+ cells in the marginal zone significantly decreased. We further found that the mRNA levels of TGF- ß and CTLA-4 in CD4+CD25+ Tregs of ALV-J-infected chickens significantly increased. Together, high-frequency and activated CD4+CD25+ Tregs inhibited B cells functions by expressing the inhibitory cytokine TGF-ß and inhibitory surface receptor CTLA-4, thereby maintaining persistent immunotolerance in congenital ALV-J-infected chickens.
Asunto(s)
Virus de la Leucosis Aviar/inmunología , Leucosis Aviar/inmunología , Pollos , Tolerancia Inmunológica , Enfermedades de las Aves de Corral/inmunología , Linfocitos T/inmunología , Animales , Linfocitos T CD4-Positivos , Embrión de Pollo , Organismos Libres de Patógenos EspecíficosRESUMEN
Porcine reproductive and respiratory syndrome (PRRS) is a highly contagious infectious disease caused by porcine reproductive and respiratory syndrome virus (PRRSV), which inflicts major economic losses on the global pig farming industry. Based on its similarity to highly pathogenic strains, the GDzj strain isolated in this study was predicted to be highly pathogenic. We therefore analyzed the pathogenicity of this strain experimentally in piglets. All piglets challenged with this virus experienced fever or high fever, loss of appetite, decreased food intake, daily weight loss, shortness of breath, and listlessness, and the necropsy results showed that they had experienced severe interstitial pneumonia. We then used the BAC system to construct a full-length cDNA infectious clone of GDzj, and the rescued virus displayed in vitro proliferation characteristics similar to those of the parental PRRSV strain. In summary, we successfully isolated a highly pathogenic PRRSV strain and constructed a full-length infectious cDNA clone from it, thereby providing an effective reverse genetics platform for further study of viral pathogenesis.
Asunto(s)
Síndrome Respiratorio y de la Reproducción Porcina/etiología , Virus del Síndrome Respiratorio y Reproductivo Porcino/genética , Virus del Síndrome Respiratorio y Reproductivo Porcino/patogenicidad , Animales , Cromosomas Artificiales Bacterianos , ADN Complementario/genética , Genoma Viral , Pulmón/virología , Ganglios Linfáticos/patología , Ganglios Linfáticos/virología , Filogenia , Neumonía Viral/patología , Neumonía Viral/virología , Síndrome Respiratorio y de la Reproducción Porcina/virología , Virus del Síndrome Respiratorio y Reproductivo Porcino/crecimiento & desarrollo , Virus del Síndrome Respiratorio y Reproductivo Porcino/aislamiento & purificación , PorcinosRESUMEN
Since June 2017, several outbreaks of a Seneca Valley virus (SVV) USA/GBI29/2015-like strain have emerged in pigs in China. In our study, we successfully isolated the SVV strain CH-GDZQ-2018, confirmed by immunofluorescence and Western blot assays. Phylogenetic and recombinant analyses showed that the USA/GBI29/2015-like CH-GDZQ-2018 strain was the result of recombination between epidemic strains local to Guangdong, showing that SVV has undergone evolution in China.
Depuis juin 2017, plusieurs foyers d'une souche apparentée au virus de la vallée de Seneca (SVV) USA/GBI29/2015 sont apparus chez des porcs en Chine. Dans la présente étude, nous avons isolé avec succès la souche SVV CH-GDZQ-2018, confirmée par des tests d'immunofluorescence et d'immunobuvardage. Des analyses phylogénétiques et recombinantes ont montré que la souche CH-GDZQ-2018 de type USA/GBI29/2015 était le résultat d'une recombinaison entre des souches épidémiques locales au Guangdong, indiquant une évolution du SVV en Chine.(Traduit par Docteur Serge Messier).
Asunto(s)
Infecciones por Picornaviridae/veterinaria , Picornaviridae/genética , Enfermedades de los Porcinos/virología , Animales , China/epidemiología , Filogenia , Picornaviridae/clasificación , Infecciones por Picornaviridae/epidemiología , Infecciones por Picornaviridae/virología , Virus Reordenados , Porcinos , Enfermedades de los Porcinos/epidemiologíaRESUMEN
Porcine reproductive and respiratory syndrome virus (PRRSV) is a notable threat to the pig industry. Long-term epidemiological investigations and genetic variation analyses of PRRSV isolates benefit PRRSV prevention and control. In our study, 43 PRRSV strains were successfully isolated from the lungs of sick pigs, and the genetic variations of these isolates were analyzed. Phylogenetic analysis showed that the isolates belonged to PRRSV2 and that lineage 8 (8.7) subgroup III strains remained the dominant type circulating in South China. In addition, sequence alignment analysis identified many novel deletions and mutations in the Nsp2 and GP5 genes. Furthermore, phylogenetic analysis showed that highly frequent recombination events of PRRSV between different lineages might occur in Guangdong Province. These results may help to elucidate the epidemiology and genetic variation of PRRSV isolates in Guangdong Province. Keywords: GP5; Nsp2; phylogenetic analysis; sequence alignment; porcine reproductive and respiratory syndrome virus (PRRSV).
Asunto(s)
Síndrome Respiratorio y de la Reproducción Porcina , Virus del Síndrome Respiratorio y Reproductivo Porcino , Animales , China/epidemiología , Variación Genética , Filogenia , Síndrome Respiratorio y de la Reproducción Porcina/epidemiología , Virus del Síndrome Respiratorio y Reproductivo Porcino/genética , PorcinosRESUMEN
Porcine circovirus type 3 (PCV3) has been widely detected throughout the world since it was first discovered on pig farms in 2015. PCV3 is closely associated with cardiac and multisystem inflammation, respiratory disease, congenital tremors, myocarditis, diarrhea, encephalitis and neurologic disease, and periarteritis. However, there have been few reports on the relationship between PCV3 and inflammatory pathways. The NF-κB signaling pathway plays an important role in the defense against viral infection. Here, we demonstrate that the capsid protein (Cap) of PCV3 plays a key role in the activation of NF-κB signaling in HEK-293T cells. Furthermore, PCV3 Cap promotes the mRNA expression of the pro-inflammatory cytokines IL6 and TNFα. In addition, PCV3 Cap promotes RIG-I and MDA5 mRNA expression in RIG-like receptor (RLR) signaling and MyD88 mRNA expression in Toll-like receptor (TLR) signaling but does not influence TRIF mRNA expression in TLR signaling. These results show that PCV3 Cap activates NF-κB signaling, possibly through the RLR and the TLR signaling pathways. This work illustrates that PCV3 Cap activates NF-κB signaling and thus may provide a basis for the pathogenesis of PCV3 and the innate immunity of the host.
Asunto(s)
Proteínas de la Cápside/inmunología , Circovirus/metabolismo , Citocinas/genética , Transducción de Señal , Circovirus/inmunología , Proteína 58 DEAD Box/genética , Células HEK293 , Humanos , Helicasa Inducida por Interferón IFIH1/genética , Interleucina-6/genética , Factor 88 de Diferenciación Mieloide/genética , FN-kappa B/metabolismo , Factor de Necrosis Tumoral alfa/genéticaRESUMEN
Immune tolerance induced by avian leukosis virus subgroup J (ALV-J) is a prerequisite for tumorigenesis. Although we had reported that B cell anergy induced by ALV-J was the main reason for immune tolerance, the molecular mechanism still remains unclear. Here, we found SU protein of ALV-J interacted with tyrosine kinase Lyn (a key protein in BCR signaling pathway) by confocal laser scanning microscopy and co-immunoprecipitation test, which suggested that Lyn might play an important role in B cell anergy induced by ALV-J. Correspondingly, the mRNA and protein level of Lyn was significantly up-regulated in B cells after ALV-J infection. Subsequently, the phosphorylated protein levels of Lyn at Tyr507 site were significantly up-regulated in ALV-J-infected B cells after BCR signal activation, but the phosphorylated protein level of Syk (a direct substrate of Lyn) at Tyr525/526 site, Ca2+ flux, and NF-κB p65 protein level were significantly down-regulated. Interestingly, the phosphorylated protein level of Syk at Tyr525/526 site, Ca2+ flux, and NF-κB p65 protein level were both significantly retrieved after the shLyn treatment in B cells infected by ALV-J. In summary, these results indicated that ALV-J activated the negative regulatory effect of phosphorylated Lyn protein at 507 site in BCR signal transduction pathway and then mediated B cell anergy, which will provide a new insight for revealing the pathogenesis of immune tolerance induced by ALV-J.
Asunto(s)
Virus de la Leucosis Aviar/inmunología , Linfocitos B/inmunología , Anergia Clonal , Transducción de Señal/inmunología , Familia-src Quinasas/genética , Animales , Leucosis Aviar/inmunología , Leucosis Aviar/virología , Virus de la Leucosis Aviar/clasificación , Linfocitos B/virología , Pollos/inmunología , Pollos/virología , Regulación de la Expresión Génica , Fosforilación , Enfermedades de las Aves de Corral/virología , Organismos Libres de Patógenos Específicos , Regulación hacia ArribaRESUMEN
Porcine epidemic diarrhea virus (PEDV) belongs to the Alphacoronavirus genus in the Coronaviridae family. Similar to other coronaviruses, PEDV encodes two papain-like proteases. Papain-like protease (PLP)2 has been proposed to play a key role in antagonizing host innate immunity. However, the function of PLP1 remains unclear. In this study, we found that overexpression of PLP1 significantly promoted PEDV replication and inhibited production of interferon-ß. Immunoprecipitation and mass spectrometry were used to identify cellular interaction partners of PLP1. Host cell poly(C) binding protein 2 (PCBP2) was determined to bind and interact with PLP1. Both endogenous and overexpressed PCBP2 co-localized with PLP1 in the cytoplasm. Overexpression of PLP1 upregulated expression of PCBP2. Furthermore, overexpression of PCBP2 promoted PEDV replication. Silencing of endogenous PCBP2 using small interfering RNAs attenuated PEDV replication. Taken together, these data demonstrated that PLP1 negatively regulated the production of type 1 interferon by interacting with PCBP2 and promoted PEDV replication.
Asunto(s)
Papaína/metabolismo , Virus de la Diarrea Epidémica Porcina/metabolismo , Proteínas no Estructurales Virales/metabolismo , Replicación Viral/fisiología , Animales , Chlorocebus aethiops , Proteasas Similares a la Papaína de Coronavirus , Regulación de la Expresión Génica/efectos de los fármacos , Regulación de la Expresión Génica/fisiología , Células HEK293 , Humanos , Interferón beta/genética , Interferón beta/metabolismo , Proteína Proteolipídica de la Mielina/metabolismo , Papaína/genética , Virus de la Diarrea Epidémica Porcina/genética , Virus de la Diarrea Epidémica Porcina/fisiología , Interferencia de ARN , Proteínas de Unión al ARN , Factor de Necrosis Tumoral alfa/farmacología , Células Vero , Proteínas no Estructurales Virales/genéticaRESUMEN
Porcine epidemic diarrhea (PED) is an acute enteric disease caused by porcine epidemic diarrhea virus (PEDV). In China, variant PEDV causes severe watery diarrhea, vomiting, and dehydration in piglets, leading to very high morbidity and mortality. However, the pathogenesis of PEDV is still not fully understood. In our study, we analyzed the long noncoding RNA (lncRNA) and mRNA expression profiles of PEDV GDgh16 in infected Vero cells at 60 h postinfection. A total of 61,790 annotated mRNAs, 14,247 annotated lncRNAs and 1290 novel lncRNAs were identified. A total of 227 annotated lncRNAs and 13 novel lncRNAs were significantly and differentially expressed after viral infection. The Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) databases were used to identify genes adjacent to the lncRNAs, and it was found that these lncRNAs were enriched in pathways related to immune and antiviral responses. Next, we selected candidate lncRNAs and their predicted target genes for study. RT-qPCR demonstrated that these lncRNAs and genes were differentially expressed after PEDV infection. Our study investigated the function of lncRNAs involved in PEDV infection, providing new insight into the pathogenic mechanisms of PEDV.
Asunto(s)
Infecciones por Coronavirus/veterinaria , Virus de la Diarrea Epidémica Porcina/genética , ARN Largo no Codificante/genética , Enfermedades de los Porcinos/genética , Animales , China , Chlorocebus aethiops , Infecciones por Coronavirus/genética , Genoma , Virus de la Diarrea Epidémica Porcina/fisiología , Porcinos , Enfermedades de los Porcinos/virología , Células VeroRESUMEN
In this study, we aimed to test the effects of treadmill running on depression induced olfactory functions and OB neurogenesis in depression model. Depression model was created with chronic unpredictable mild stress (CUMS) and treadmill running was performed as the antidepressant treatment. Behavioral results showed that treadmill running not only attenuated the depression mood but also improved the olfactory discrimination and sensitivity in CUMS depression model. Immune-staining further indicates treadmill running promoted neurogenesis in hippocampal OB region. Moreover, treadmill running prevented the loss of DAergic neurons in glomerular layer of OB region, indicating the critical role of DAergic neuronal functions in regulating treadmill running mediated olfactory functions. In depression model, inhibiting DAergic neurons by intra-OB injection of 6-OHDA resulted in the compromised improving effects of treadmill running olfactory discrimination. In conclusion, treadmill running could attenuate depression associated olfactory deficits by promoting olfactory neurogenesis and improve DAergic neural functions.
Asunto(s)
Depresión/terapia , Terapia por Ejercicio , Neurogénesis , Bulbo Olfatorio/fisiopatología , Animales , Depresión/fisiopatología , Modelos Animales de Enfermedad , Neuronas Dopaminérgicas/citología , Neuronas Dopaminérgicas/patología , Hipocampo/citología , Hipocampo/fisiopatología , Masculino , Ratones Endogámicos C57BL , Bulbo Olfatorio/citología , CarreraRESUMEN
Since 2010, Porcine epidemic diarrhea virus (PEDV) has caused severe diarrhea disease in piglets in China, resulting in large economic losses. To understand the genetic characteristics of the PEDV strains that circulated in some provinces of China between 2015 and 2018, 375 samples of feces and small intestine were collected from pigs and tested. One hundred seventy-seven samples tested positive and the PEDV-positive rate was 47.20%. A phylogenetic tree analysis based on the entire S gene showed that these strains clustered into four subgroups, GI-a, GI-b, GII-a, and GII-b, and that the GII-b strains have become dominant in recent years. Compared with previous strains, these strains have multiple variations in the SP and S1-NTD domains and in the neutralizing epitopes of the S protein. We also successfully isolated and identified a new virulent GII-b strain, GDgh16, which is well-adapted to Vero cells and caused a high mortality rate in piglets in challenge experiments. Our study clarifies the genetic characteristics of the prevalent PEDV strains in parts of China, and suggests that the development of effective novel vaccines is both necessary and urgent.