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1.
Molecules ; 28(15)2023 Jul 25.
Artículo en Inglés | MEDLINE | ID: mdl-37570602

RESUMEN

Bupleurum chinense DC. and Bupleurum scorzonerifolium Willd. have different clinical efficacies, with the former typically used to treat typhoid fever and the latter mainly used to clear liver heat. The differences in their clinical efficacy are closely related to their complex chemical composition, especially the active components. In this study, the saponins and volatile oils in two varieties of Radix Bupleuri grown in different regions were extracted and analyzed using high-performance liquid chromatography (HPLC) and gas chromatography coupled with mass spectrometry (MS), and the absolute contents of five saikosaponins were accurately quantified using an established HPLC-MS method in the multiple reaction monitoring mode. Multivariate statistical analysis was performed to reveal the difference in the active components between the two varieties. The saikosaponin content was significantly affected by variety and growing region, with all five saikosaponins being significantly higher in Bupleurum chinense DC. than in Bupleurum scorzonerifolium Willd. The results of principal component analysis and hierarchical cluster analysis show a clear distinction between the two varieties in terms of both saponins and volatile oils. Twenty-one saponins, including saikosaponin b2 and b1, and fifty-two volatile oils, including 2-tetradecyloxirane and chloromethyl cyanide, were screened and identified as differential compounds contributing to the significant difference between the two varieties. These compounds may also be responsible for the difference in clinical efficacy between Bupleurum chinense DC. and Bupleurum scorzonerifolium Willd. All the results suggest that the accumulation and diversity of active components in Radix Bupleuri are significantly affected by the variety. In contrast to previous reports, this study provides the absolute contents of five saikosaponins in Radix Bupleuri of different varieties and reduces the influence of the growing region on the analytical results by collecting samples from different regions. The results of this study may provide a reference for the identification and quality evaluation of different varieties of Radix Bupleuri.


Asunto(s)
Bupleurum , Aceites Volátiles , Ácido Oleanólico , Saponinas , Bupleurum/química , Cromatografía Líquida de Alta Presión/métodos , Cromatografía de Gases y Espectrometría de Masas , Espectrometría de Masas , Saponinas/análisis , Ácido Oleanólico/análisis , Aceites Volátiles/análisis , Raíces de Plantas/química
2.
Virol J ; 19(1): 207, 2022 12 03.
Artículo en Inglés | MEDLINE | ID: mdl-36463170

RESUMEN

Foot-and-mouth disease (FMD) is a highly contagious viral disease that affects cloven-hoofed animals. Vaccination is the most effective measure to control FMD. However, FMDV particles are prone to dissociation, leading to insufficient potency of vaccine. Based on this characteristic, a combination of twenty percentage trehalose, 500 mM NaCl and 3 mM CuSO4·5H2O was developed to increase viral stability. Heating-resistance test showed that FMDV infectivity was maintained when formulated with formulation. Additionally, the half-life of FMDV inactivation was prolonged remarkably. Sequencing analysis demonstrated that viral genome could not be altered in serial passages. Vaccine stability was monitored for up to 1 year at 4 °C, with a higher level of 146S content remained. This study suggested that the formulation could protect FMDV against massive structural breakdown and extend the shelf life of vaccine. Our findings could provide strategy to develop more solutions for the stabilization of viral vaccine.


Asunto(s)
Boca , Vacunas Virales , Animales , Vacunación , Genoma Viral , Pase Seriado
3.
Virol J ; 19(1): 16, 2022 01 21.
Artículo en Inglés | MEDLINE | ID: mdl-35062983

RESUMEN

BACKGROUND: African swine fever (ASF) is a highly fatal swine disease, which threatens the global pig industry. There is no commercially available vaccine against ASF and effective subunit vaccines would represent a real breakthrough. METHODS: In this study, we expressed and purified two recombinant fusion proteins, OPM (OprI-p30-modified p54) and OPMT (OprI-p30-modified p54-T cell epitope), which combine the bacterial lipoprotein OprI with ASF virus proteins p30 and p54. Purified recombinant p30 and modified p54 expressed alone or fused served as controls. The activation of dendritic cells (DCs) by these proteins was first assessed. Then, humoral and cellular immunity induced by the proteins were evaluated in mice. RESULTS: Both OPM and OPMT activated DCs with elevated expression of relevant surface molecules and proinflammatory cytokines. Furthermore, OPMT elicited the highest levels of antigen-specific IgG responses, cytokines including interleukin-2, interferon-γ, and tumor necrosis factor-α, and proliferation of lymphocytes. Importantly, the sera from mice vaccinated with OPM or OPMT neutralized more than 86% of ASF virus in vitro. CONCLUSIONS: Our results suggest that OPMT has good immunostimulatory activities and immunogenicity in mice, and might be an appropriate candidate to elicit immune responses in swine. Our study provides valuable information on further development of a subunit vaccine against ASF.


Asunto(s)
Virus de la Fiebre Porcina Africana , Fiebre Porcina Africana , Vacunas Virales , Virus de la Fiebre Porcina Africana/genética , Animales , Anticuerpos Antivirales , Lipoproteínas/genética , Ratones , Proteínas Recombinantes de Fusión/genética , Porcinos , Proteínas Virales/metabolismo , Vacunas Virales/genética
4.
Entropy (Basel) ; 24(3)2022 Mar 18.
Artículo en Inglés | MEDLINE | ID: mdl-35327934

RESUMEN

A novel time-varying channel adaptive low-complexity chase (LCC) algorithm with low redundancy is proposed, where only the necessary number of test vectors (TVs) are generated and key equations are calculated according to the channel evaluation to reduce the decoding complexity. The algorithm evaluates the error symbol numbers by counting the number of unreliable bits of the received code sequence and dynamically adjusts the decoding parameters, which can reduce a large number of redundant calculations in the decoding process. We provide a simplified multiplicity assignment (MA) scheme and its architecture. Moreover, a multi-functional block that can implement polynomial selection, Chien search and the Forney algorithm (PCF) is provided. On this basis, a high-efficiency LCC decoder with adaptive error-correcting capability is proposed. Compared with the state-of-the-art LCC (TV = 16) decoding, the number of TVs of our decoder was reduced by 50.4% without loss of the frame error rate (FER) performance. The hardware implementation results show that the proposed decoder achieved 81.6% reduced average latency and 150% increased throughput compared to the state-of-the-art LCC decoder.

5.
Entropy (Basel) ; 24(3)2022 Mar 19.
Artículo en Inglés | MEDLINE | ID: mdl-35327937

RESUMEN

An end-to-end joint source-channel (JSC) encoding matrix and a JSC decoding scheme using the proposed bit flipping check (BFC) algorithm and controversial variable node selection-based adaptive belief propagation (CVNS-ABP) decoding algorithm are presented to improve the efficiency and reliability of the joint source-channel coding (JSCC) scheme based on double Reed-Solomon (RS) codes. The constructed coding matrix can realize source compression and channel coding of multiple sets of information data simultaneously, which significantly improves the coding efficiency. The proposed BFC algorithm uses channel soft information to select and flip the unreliable bits and then uses the redundancy of the source block to realize the error verification and error correction. The proposed CVNS-ABP algorithm reduces the influence of error bits on decoding by selecting error variable nodes (VNs) from controversial VNs and adding them to the sparsity of the parity-check matrix. In addition, the proposed JSC decoding scheme based on the BFC algorithm and CVNS-ABP algorithm can realize the connection of source and channel to improve the performance of JSC decoding. Simulation results show that the proposed BFC-based hard-decision decoding (BFC-HDD) algorithm (ζ = 1) and BFC-based low-complexity chase (BFC-LCC) algorithm (ζ = 1, η = 3) can achieve about 0.23 dB and 0.46 dB of signal-to-noise ratio (SNR) defined gain over the prior-art decoding algorithm at a frame error rate (FER) = 10-1. Compared with the ABP algorithm, the proposed CVNS-ABP algorithm and BFC-CVNS-ABP algorithm achieve performance gains of 0.18 dB and 0.23 dB, respectively, at FER = 10-3.

6.
J Clin Microbiol ; 59(3)2021 02 18.
Artículo en Inglés | MEDLINE | ID: mdl-33328177

RESUMEN

Foot-and-mouth disease virus (FMDV) has led to serious losses in animal husbandry worldwide. Seromonitoring of FMDV postvaccination is important for the control and eradication of foot-and-mouth disease (FMD) in regions and countries where vaccination is widespread. However, many commercial kits present high false-positive rates. In this study, a multiepitope-based indirect chemiluminescence immunoassay (ME-CLIA) was developed for specifically detecting antibodies against FMDV serotype O in swine sera. The developed method presented high diagnostic sensitivity and excellent diagnostic specificity, and it could detect a broad spectrum of antibodies against FMDV serotype O. The diagnostic performance, accuracy rate, and analytical sensitivity of ME-CLIA were compared with those of three commercial kits. The immune protection value of multiple-epitope recombinant vaccine detected using ME-CLIA was preliminarily determined by observation of clinical symptoms postimmunization challenge, the results of which indicated that the ME-CLIA can be employed as a matching detection method for evaluating multiple-epitope recombinant vaccine. The percent positive values of ME-CLIA determined using swine vaccinated with inactivated vaccine were significantly positively correlated with the titers of liquid-phase-blocking enzyme-linked immunosorbent assay (ELISA) (LBPE) (r = 0.8361; P < 0.0001). These results indicated that ME-CLIA is suitable for detection of antibodies against FMDV serotype O in swine and for potency evaluation of multiple-epitope and inactivated vaccines.


Asunto(s)
Virus de la Fiebre Aftosa , Fiebre Aftosa , Enfermedades de los Porcinos , Vacunas Virales , Animales , Anticuerpos Antivirales , Ensayo de Inmunoadsorción Enzimática , Fiebre Aftosa/diagnóstico , Fiebre Aftosa/prevención & control , Virus de la Fiebre Aftosa/genética , Luminiscencia , Proteínas Recombinantes , Serogrupo , Porcinos , Enfermedades de los Porcinos/diagnóstico , Enfermedades de los Porcinos/prevención & control
7.
Exp Physiol ; 106(4): 1061-1071, 2021 04.
Artículo en Inglés | MEDLINE | ID: mdl-33527539

RESUMEN

NEW FINDINGS: What is the central question of this study? What is the mechanism of miR-211 in an Alzheimer's disease cell model? What is the main finding and its importance? miR-211 was upregulated in an Alzheimer's disease cell model. It targeted neurogenin 2, reduced the activation of the phosphoinositide 3-kinase-Akt signalling pathway, inhibited the proliferation of the Alzheimer's disease cell model and promoted apoptosis. ABSTRACT: MicroRNAs (miRs) are aberrantly expressed in Alzheimer's disease (AD) patients. This study was intended to investigate the effect of miR-211 on an AD cell model and the involvement of neurogenin 2 (Ngn2). The appropriate dose and time for the effect of Aß1-42 on PC12 cells were determined to establish an AD cell model. An effect of miR-211 expression on cell viability, proliferation and apoptosis was detected after cell transfection. Online prediction and a dual luciferase reporter gene assay were utilized to confirm the binding sequence of miR-211 and Ngn2. qRT-PCR and western blot analysis were applied to measure Ngn2 expression. A gain and loss of function assay of miR-211 and Ngn2 was performed, and activation of the phosphoinositide 3-kinase (PI3K)-Akt signaling pathway was detected. The AD cell model was induced by Aß1-42 treatment. miR-211 expression was significantly enhanced after miR-211 transfection, leading to suppressed proliferation and promotion of apoptosis in Aß1-42 -treated PC12 cells. In addition, miR-211 could downregulate Ngn2 mRNA and protein expression, while overexpression of Ngn2 could reverse the effects of miR-211 on Aß1-42 -treated PC12 cells and significantly enhance the phosphorylated Akt and PI3K protein levels. miR-211 could inhibit growth of PC12 cells by suppressing Ngn2 expression and inactivating the PI3K-Akt signalling pathway.


Asunto(s)
Enfermedad de Alzheimer , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/metabolismo , MicroARNs , Proteínas del Tejido Nervioso/metabolismo , Enfermedad de Alzheimer/genética , Péptidos beta-Amiloides/genética , Péptidos beta-Amiloides/metabolismo , Animales , Apoptosis , Proliferación Celular/genética , MicroARNs/genética , MicroARNs/metabolismo , Células PC12 , Fosfatidilinositol 3-Quinasas/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Ratas
8.
Virol J ; 18(1): 97, 2021 05 05.
Artículo en Inglés | MEDLINE | ID: mdl-33952293

RESUMEN

BACKGROUND: African swine fever (ASF), characterized by acute, severe, and fast-spreading, is a highly lethal swine infectious disease caused by the African swine fever virus (ASFV), which has caused substantial economic losses to the pig industry worldwide in the past 100 years. METHODS: This study started with bioinformatics methods and verified the epitope fusion protein method's reliability that does not rely on traditional epitope identification. Meanwhile, it will also express and purify the constructed genes through prokaryotic expression and establish antibody detection methods. RESULTS: The results indicated that the protein had good reactivity and did not cross-react with other swine diseases. The receiver-operating characteristic analysis was performed to verify the determination. The area under the receiver-operating characteristic curve was 0.9991 (95% confidence interval 0.9973 to 1.001). CONCLUSIONS: It was proved that the recombinant protein is feasible as a diagnostic antigen to distinguish ASFV and provides a new idea for ASFV antibody detection.


Asunto(s)
Fiebre Porcina Africana , Anticuerpos Antivirales/aislamiento & purificación , Ensayo de Inmunoadsorción Enzimática , Fiebre Porcina Africana/diagnóstico , Virus de la Fiebre Porcina Africana/inmunología , Animales , Biología Computacional , Epítopos , Proteínas Recombinantes , Reproducibilidad de los Resultados , Porcinos
9.
Appl Opt ; 59(22): 6751-6759, 2020 Aug 01.
Artículo en Inglés | MEDLINE | ID: mdl-32749381

RESUMEN

Visible light communication (VLC) offers a unique advantage of electromagnetic interference immunity and wide bandwidth. It has gradually become the main candidate in marine communication with great potential. As a green communication link, VLC requires reliable beam tracking as a prerequisite. Therefore, based on the received signal strength of a single-input-multiple-output system in the VLC scenario, this paper first proposes a geometrical algorithm for transmitter localization. On this basis, a linear iterative algorithm using Taylor expansion, implicit function theorem, and time-domain expansion is presented to realize online beam tracking. Under the marine VLC system, an iterative denoising algorithm based on the hidden Markov model and principal component analysis is proposed for denoising. Simulation results show that the proposed algorithms have predominance in high tracking accuracy (within 10 cm) and desirable real-time performance.

10.
Metab Brain Dis ; 35(6): 959-970, 2020 08.
Artículo en Inglés | MEDLINE | ID: mdl-32246322

RESUMEN

Paeoniflorin is a natural monoterpene glucoside from Paeoniae Radix with neuroprotective properties. However, it is still unclear whether paeoniflorin has neuroprotective effects on subarachnoid hemorrhage (SAH). This study explores the effect of paeoniflorin on early brain injury (EBI) using rat SAH model. We found that paeoniflorin significantly improves neurological deficits, attenuates brain water content and Evans blue extravasation at 72 h after SAH. Paeoniflorin attenuates the oxidative stress following SAH as evidenced by decrease of reactive oxygen species (ROS), malondialdehyde (MDA), 3-Nitrotyrosine, and 8-Hydroxy-2-deoxy guanosine (8-OHDG) level, increase of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase activity, and up-regulates the nuclear factor erythroid­related factor 2 (Nrf2)/heme oxygenase­1 (HO-1) pathway. Inhibition of microglia activation and neuro-inflammatory response both contributed to paeoniflorin's protective effects. Moreover, paeoniflorin treatment significantly reduces the ratio of Bax/Bcl-2, active caspase-3/ neuronal nuclei (NeuN) and TUNEL/DAPI positive cells at 72 h following SAH. Our results indicate that paeoniflorin may attenuate early brain injury after experimental SAH.


Asunto(s)
Antiinflamatorios no Esteroideos/administración & dosificación , Apoptosis/efectos de los fármacos , Lesiones Encefálicas/prevención & control , Glucósidos/administración & dosificación , Monoterpenos/administración & dosificación , Estrés Oxidativo/efectos de los fármacos , Hemorragia Subaracnoidea/tratamiento farmacológico , Animales , Apoptosis/fisiología , Lesiones Encefálicas/metabolismo , Lesiones Encefálicas/patología , Células Cultivadas , Inyecciones Intraperitoneales , Masculino , Neuronas/efectos de los fármacos , Neuronas/metabolismo , Neuronas/patología , Estrés Oxidativo/fisiología , Ratas , Ratas Sprague-Dawley , Hemorragia Subaracnoidea/metabolismo , Hemorragia Subaracnoidea/patología
11.
Biotechnol Lett ; 42(8): 1317-1325, 2020 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-32185620

RESUMEN

OBJECTIVES: Marc-145 cells (monkey embryonic kidney epithelial cells) play a critical role in the biotechnology industry as certain virus host cells. To investigate the expression of enhanced green fluorescent protein (eGFP) gene as a foreign gene in Marc-145 cells, which we developed an approach of foreign gene site-specific knock-in into Marc-145 cells by clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated 9 (Cas9) and putatively explored appropriate genomic recombination sites in Marc-145 cells. RESULTS: Our study demonstrated that the specific homologous recombination (HR) site between the Rac GTPase activating protein 1 (RACGAP1) and the acid-sensing ion channel subunit 1 (ASIC1) genes of the 11th chromosome could be used as the target site of Cas9 for the generation of target gene knock-in into Marc-145 cells, by the insertion of the eGFP cassette into the specific HR site and subsequent expression. CONCLUSIONS: Junction PCR, sequencing, Southern blot and fluorescence assay determined eGFP gene-specific knock-in HR site between the RACGAP1 and ASIC1 genes of the 11th chromosome, which was identified by the genomic safe harbours in Marc-145 cells. Our study encouraged a broader range of applications, such as Marc-145 cells development and engineering for virus adaption and yield increase in the vaccine biotechnology industry.


Asunto(s)
Sistemas CRISPR-Cas/genética , Técnicas de Sustitución del Gen/métodos , Genes Reporteros/genética , Recombinación Homóloga/genética , Animales , Línea Celular , Chlorocebus aethiops , Proteínas Fluorescentes Verdes/genética , Proteínas Fluorescentes Verdes/metabolismo , Reacción en Cadena de la Polimerasa , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo
12.
Microb Pathog ; 135: 103638, 2019 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-31326561

RESUMEN

Recently, amiloride was shown to potently suppress Coxsackievirus B3 (CVB3) replication. In the current study, we investigated whether amiloride could also exhibit antiviral activity against foot-and-mouth disease virus (FMDV), which belongs to the same family (Picornaviridae) as CVB3. We found that amiloride exerted antiviral activity in a dose-dependent manner against two strains of FMDV in IBRS-2 cells, with slight cytotoxicity at 1000 µM. Besides, amiloride did not inhibit the attachment and entry of FMDV in IBRS-2 cells, but prevented early viral replication. These data implied that amiloride could be a promising candidate for further research as a potential antiviral drug against FMDV infection.


Asunto(s)
Amilorida/farmacología , Antivirales/farmacología , Virus de la Fiebre Aftosa/efectos de los fármacos , Replicación Viral/efectos de los fármacos , Animales , Técnicas de Cultivo de Célula , Línea Celular , Supervivencia Celular , Replicación del ADN/efectos de los fármacos , Fiebre Aftosa/virología , Humanos , ARN Mensajero/metabolismo , Proteínas Virales
13.
Appl Microbiol Biotechnol ; 103(19): 8075-8086, 2019 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-31463546

RESUMEN

Foot-and-mouth disease virus (FMDV) has led to serious losses in the farming industry worldwide, particularly in cattle and swine. In developing countries, the control and eradication of FMD rely upon vaccination, in which the inactivated vaccine is predominant. In the preparation of inactivated vaccine, a series of purification methods were used to remove non-structural proteins (NSPs). It is necessary to develop a quantitative detection method of residual NSP and confirm a threshold value for the evaluation of the vaccine. Meanwhile, it is also important to develop a sensitive and rapid diagnostic method to distinguish infected animals from vaccinated animals (DIVA). In this study, three monoclonal antibodies (MAbs) against NSP 3ABC, designated 2G5, 9E2, and 1E10, were used. Subsequently, a series of overlapping peptides were expressed using a prokaryotic expression system to determine the minimal epitopes identified by the MAbs. Three linear B cell epitopes (BCEs), "92EYIEKA97" "23EGPYAGPLE31" and "209EPHH212", were identified by MAbs 2G5, 9E2, and 1E10, respectively. Alanine-scanning mutagenesis analysis confirmed the critical amino acid in these epitopes. The epitope "92EYIEKA97" is located in 3A, which is deleted in some natural deletion mutants that result in a change in virus tropism. MAb 9E2 that identified the epitope "23EGPYAGPLE31" reacted with 3B1 and 3B2, but did not react with 3B3. In combination with sequence alignment analysis, the epitope "23EGPYAGPLE31" is highly conserved among different FMDV isolates. Preliminary screening using the known positive and negative sera indicated the MAb 9E2 has the potential for the development of a diagnostic method for DIVA. The residual NSP in inactivated vaccines can be detected using 9E2-HRP, which indicated the MAb 9E2 is able to evaluate inactivated vaccines. The four-amino acid epitope is the first reported to date that is recognized by 1E10. These results provide valuable insight into the diagnosis of DIVA and the NSP residual evaluation in inactivated vaccines.


Asunto(s)
Anticuerpos Monoclonales/inmunología , Anticuerpos Antivirales/inmunología , Mapeo Epitopo , Epítopos de Linfocito B/inmunología , Virus de la Fiebre Aftosa/inmunología , Proteínas no Estructurales Virales/inmunología , Animales , Anticuerpos Monoclonales/aislamiento & purificación , Anticuerpos Antivirales/aislamiento & purificación , Ratones
14.
Biochem Biophys Res Commun ; 506(3): 543-547, 2018 11 30.
Artículo en Inglés | MEDLINE | ID: mdl-30366671

RESUMEN

Adenylate cyclase 7 (AC7) has been reported to participate in various biological processes during cancer progression. However, the roles of AC7 in all-trans retinoic acid (ATRA)-induced differentiation of acute promyelocytic leukemia (APL) cells are still unknown. In this study, firstly, our results showed that AC7 affected intracellular cAMP level and influenced ATRA-induced differentiation of APL cells. Secondly, we revealed that miR-192 could directly target AC7 expression and knockdown of miR-192 promoted ATRA-induced APL cell differentiation by regulating AC7 expression. Furthermore, we found that AC7 expression was lower in patients with relapsed APL than that in patients with newly diagnosed APL, while miR-192 expression was relatively higher in patients with relapsed APL. Taken together, our results show that miR-192-mediated AC7 could play important roles in differentiation of APL cells, AC7 and miR-192 might be new biomarkers and therapeutic targets for patients with relapsed APL.


Asunto(s)
Adenilil Ciclasas/metabolismo , Diferenciación Celular/efectos de los fármacos , Leucemia Promielocítica Aguda/genética , Leucemia Promielocítica Aguda/patología , MicroARNs/metabolismo , Tretinoina/farmacología , Adenilil Ciclasas/genética , Secuencia de Bases , Línea Celular Tumoral , AMP Cíclico/biosíntesis , Regulación hacia Abajo/efectos de los fármacos , Regulación hacia Abajo/genética , Regulación Leucémica de la Expresión Génica/efectos de los fármacos , Técnicas de Silenciamiento del Gen , Humanos , Leucemia Promielocítica Aguda/diagnóstico , MicroARNs/genética , ARN Mensajero/genética , ARN Mensajero/metabolismo , Recurrencia , Regulación hacia Arriba/efectos de los fármacos
15.
BMC Vet Res ; 14(1): 240, 2018 Aug 17.
Artículo en Inglés | MEDLINE | ID: mdl-30119669

RESUMEN

BACKGROUND: Porcine reproductive and respiratory syndrome (PRRS) is a severe disease, causing great economic losses to the pig industry. The causative agent, porcine reproductive and respiratory syndrome virus (PRRSV) is highly variable. Since the emergence of highly pathogenic PRRSV (HP-PRRSV) in China in 2006, this virus strain has undergone extensive variation. To investigate the genetic variation and pathogenicity of currently isolated PRRSV GSWW/2015 strain, its whole genome was sequenced and analyzed for the specific variation in NSP2, GP3 and GP5 regions. Pigs were challenged with the isolated virus to investigate its pathogenicity. METHODS: The PRRSV GSWW/2015 strain was isolated by seeding the viral material in Marc-145 cells. The virus specific cytopathic effect (CPE) was confirmed by indirect immunofluorescent assay (IFA) and PCR to detect the virus protein and RNA. Nine pairs of primers were designed to obtain the complete genome by PCR. All PCR fragments were cloned into T-vector for sequencing. The genetic variation of GSWW/2015 strain was analyzed by multiple sequence alignments. Nineteen PRRSV-free piglets were intranasally challenged with 108 copies of GSWW virus, while seven piglets were housed together as contact-infected control. Clinical signs were recorded daily after challenge. Blood samples were obtained every week and the viral titer was detected by quantitative real-time PCR (qRT-PCR). The PRRSV specific antibody was detected by LSI ELISA kit. RESULTS: The complete genome of PRRSV GSWW/2015 strain (GenBank accession number KX767091) was obtained. The whole genome of this strain shares 88.5 and 60.6% identity with VR-2332 and LV respectively, indicating that it belongs to the North American type (NA-type). Sequence alignments revealed that GSWW/2015 strain has a discontinuous deletion of 30 amino acids in NSP2, which is similar with HP-PRRSV. Some amino acids mutations can be observed in antigenic epitope regions of GP3 and GP5 compared with earlier strains of HP-PRRSV. Some piglets showed typical clinical signs of PRRSV after challenge. Only four pigs showed viremia within 3 days after challenge, most pigs showed peaked viremia after 21-28 days including 7 contact-infected pigs. Two pigs were detected to be positive for antibody to PRRSV at 14 days post infection (DPI), and 11 pigs (11/26) show seroconversion for PRRSV at 49 DPI. Twelve piglets died of PRRSV infection within two months. CONCLUSIONS: The genome of PRRSV GSWW/2015 strain shows the features of HP-PRRSV with 30 discontinuous amino acids deletion in NSP2 and some new amino acid mutations in epitope regions of GP5 and GP3, which might alter the antigenicity of the virus. Furthermore, the virus showed high virulence to piglets as reported in HP-PRRSV, and induced long-lasting viremia and low level of antibody responses. This work further enriched our knowledge on PRRSV evolution and pathogenicity.


Asunto(s)
Variación Genética , Síndrome Respiratorio y de la Reproducción Porcina/patología , Virus del Síndrome Respiratorio y Reproductivo Porcino/genética , Virus del Síndrome Respiratorio y Reproductivo Porcino/patogenicidad , Secuencia de Aminoácidos , Animales , Anticuerpos Antivirales/sangre , Genoma Viral/genética , Filogenia , Síndrome Respiratorio y de la Reproducción Porcina/inmunología , Síndrome Respiratorio y de la Reproducción Porcina/virología , Alineación de Secuencia , Eliminación de Secuencia , Porcinos , Viremia/veterinaria , Virulencia/genética , Secuenciación Completa del Genoma
16.
Can J Microbiol ; 64(2): 155-166, 2018 02.
Artículo en Inglés | MEDLINE | ID: mdl-29253356

RESUMEN

The innate immune system acts as the first line of defense against invasion by bacterial and viral pathogens. The role of macrophages in innate immune responses to foot-and-mouth disease virus (FMDV) is poorly understood. To determine the mechanism underlying activation of innate immunity after FMDV infection in macrophages, we performed FMDV infection in mouse macrophage RAW 264.7 cells and found that FMDV serotype O infection induced a cytopathic effect. We then evaluated the gene expression profile in macrophage RAW 264.7 cells after FMDV infection using systematic microarray analysis. Gene ontology annotation and enrichment analysis revealed that FMDV promoted expression in a group of genes that are enriched in innate immune response and inflammatory response processes. Further research demonstrated that FMDV serotype O infection enhanced NF-κB, Toll-like, and RIG-I-like receptor signaling pathways and proteins expression and increased transcription and expression of a series of cytokines and interferons, as proved by qRT-PCR, Western blot, ELISA, and dual-luciferase reporter assay. Our study concluded that FMDV infection triggers the innate immune response in macrophages after activation of multiple innate immune pathway receptors and proteins by FMDV serotype O, resulting in activation and secretion of a series of cytokines and interferons.


Asunto(s)
Virus de la Fiebre Aftosa/inmunología , Fiebre Aftosa/inmunología , Macrófagos/inmunología , Transducción de Señal/inmunología , Animales , Línea Celular , Citocinas/genética , Citocinas/inmunología , Regulación de la Expresión Génica/inmunología , Inmunidad Innata/genética , Interferones/genética , Interferones/inmunología , Ratones , Células RAW 264.7 , Transducción de Señal/genética , Transcriptoma
17.
J Gen Virol ; 98(11): 2725-2730, 2017 11.
Artículo en Inglés | MEDLINE | ID: mdl-29068273

RESUMEN

CD59 protein functions as a negative regulator of the terminal pathway of the complement system by binding to the C8/C9 factors. To date, little is known about the role of CD59 in coronavirus infectious bronchitis virus (IBV) infection. In this study, we discovered that CD59 was downregulated in IBV-infected cells and was associated with IBV virions. This association protected IBV particles from antibody-dependent complement-mediated lysis. IBV titres in the supernatant were significantly increased when CD59 proteins were overexpressed in cells followed by IBV infection, and this observation was further supported by knockdown or cleavage of CD59. Because no considerable change in IBV N protein and viral RNA levels was detected in total cell lysates prepared from the overexpression, knockdown or cleavage of CD59 groups, our data indicated that CD59 was involved in IBV particle release and that IBV had evolved a mechanism to utilize CD59 to evade complement-mediated destruction.


Asunto(s)
Anticuerpos/metabolismo , Antígenos CD59/metabolismo , Proteínas del Sistema Complemento/metabolismo , Interacciones Huésped-Patógeno , Evasión Inmune , Factores Inmunológicos/metabolismo , Virus de la Bronquitis Infecciosa/inmunología , Animales , Línea Celular , Humanos , Unión Proteica
18.
Virol J ; 11: 1, 2014 Jan 06.
Artículo en Inglés | MEDLINE | ID: mdl-24393133

RESUMEN

BACKGROUND: Foot-and-mouth disease virus (FMDV) causes a severe vesicular disease in domestic and wild cloven-hoofed animals. Because of the limited early protection induced by current vaccines, emergency antiviral strategies to control the rapid spread of FMD outbreaks are needed.Here we constructed multiple microRNAs (miRNAs) targeting the internal ribosome entry site (IRES) element of FMDV and investigated the effect of IRES-specific miRNAs on FMDV replication in baby hamster kidney (BHK-21) cells and suckling mice. RESULTS: Four IRES-specific miRNAs significantly reduced enhanced green fluorescent protein (EGFP) expression from IRES-EGFP reporter plasmids, which were used with each miRNA expression plasmid in co-transfection of BHK-21 cells. Furthermore, treatment of BHK-21 cells with Bi-miRNA (a mixture of two miRNA expression plasmids) and Dual-miRNA (a co-cistronic expression plasmid containing two miRNA hairpin structures) induced more efficient and greater inhibition of EGFP expression than did plasmids carrying single miRNA sequences.Stably transformed BHK-21 cells and goat fibroblasts with an integrating IRES-specific Dual-miRNA were generated, and real-time quantitative RT-PCR showed that the Dual-miRNA was able to effectively inhibit the replication of FMDV (except for the Mya98 strain) in the stably transformed BHK-21 cells.The Dual-miRNA plasmid significantly delayed the deaths of suckling mice challenged with 50× and 100× the 50% lethal dose (LD50) of FMDV vaccine strains of three serotypes (O, A and Asia 1), and induced partial/complete protection against the prevalent PanAsia-1 and Mya98 strains of FMDV serotype O. CONCLUSION: These data demonstrate that IRES-specific miRNAs can significantly inhibit FMDV infection in vitro and in vivo.


Asunto(s)
Antivirales/metabolismo , Productos Biológicos/metabolismo , Virus de la Fiebre Aftosa/fisiología , MicroARNs/metabolismo , Biosíntesis de Proteínas , Ribosomas/metabolismo , Replicación Viral , Animales , Fusión Artificial Génica , Sitios de Unión , Células Cultivadas , Cricetinae , Modelos Animales de Enfermedad , Fiebre Aftosa/terapia , Virus de la Fiebre Aftosa/genética , Genes Reporteros , Cabras , Proteínas Fluorescentes Verdes/análisis , Proteínas Fluorescentes Verdes/genética , Ratones , MicroARNs/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Ribosomas/genética , Análisis de Supervivencia , Resultado del Tratamiento
19.
Arch Esp Urol ; 77(1): 98-103, 2024 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-38374019

RESUMEN

OBJECTIVE: The current status and influencing factors of sleep quality in chronic nephritis patients (CNPs) were explored to provide clinical basis for improving the sleep quality of these patients. METHODS: A total of 197 CNPs admitted to our hospital from June 2021 to June 2023 were retrospectively analysed. The sleep status of patients was evaluated by the Pittsburgh sleep quality index (PSQI). According to the PSQI scores, patients were divided into good sleep quality (n = 93) and poor sleep quality (n = 104) groups. The clinical indicators between the two groups were detected. The influencing factors of sleep quality in CNP were explored by univariate and multivariate logistic regression analysis. RESULTS: Statistical differences existed in age, gender, course of disease, hypertension, neutrophilic granulocyte (NEUT) percentage (NEUT %), haemoglobin (Hb), urea, total carbon dioxide (TCO2), and serum phosphorus (P) between both groups (p < 0.05). Multivariate logistic regression analysis showed that age, course of disease, hypertension, NEUT %, Hb, and TCO2 were independent influencing factors for poor sleep quality in CNPs (p < 0.05). CONCLUSIONS: Older age, longer course of disease, hypertension, higher NEUT %, lower Hb, and higher TCO2 are associated with poorer sleep quality in CNPs. Therefore, targeted interventions for sleep quality should be given priority in clinical practice.


Asunto(s)
Hipertensión , Nefritis , Humanos , Calidad del Sueño , Estudios Retrospectivos , Hospitales
20.
J Orthop Surg Res ; 19(1): 164, 2024 Mar 04.
Artículo en Inglés | MEDLINE | ID: mdl-38439085

RESUMEN

BACKGROUND: Previous studies evaluating the influence of diabetes on the risk of deep vein thrombosis (DVT) after total knee arthroplasty (TKA) showed inconsistent results. The aim of the study was to systematically evaluate the association between diabetes and DVT after TKA in a meta-analysis. METHODS: An extensive search was conducted in PubMed, Embase, and Web of Science to identify relevant cohort studies. Random-effects models were employed to pool the results after taking account of the potential influence of heterogeneity. RESULTS: Thirteen cohort studies involving 546,156 patients receiving TKA were included, with 71,110 (13.0%) diabetic patients before surgery and 1479 (2.1%) patients diagnosed as DVT after surgery. Overall, diabetes was associated with an increased risk of DVT after TKA (risk ratio [RR]: 1.43, 95% confidence interval [CI]: 1.12-1.84, p = 0.004; I2 = 44%). Sensitivity analysis limited to studies with chemoprophylaxis (RR: 1.96, 95% CI: 1.50-2.54), and studies with multivariate analysis (RR: 1.54, 95% CI: 1.12-2.11) showed consistent results. Subgroup analysis showed that diabetes was associated with higher risk of postoperative DVT in Asian countries (RR: 1.93, 95% CI: 1.49-2.52, p < 0.001; I2 = 1%) but not in Western countries (RR: 1.07, 95% CI: 0.86-1.34, p = 0.52; I2 = 0%; p for subgroup difference < 0.001). CONCLUSION: Diabetes may be a risk factor for DVT after TKA, even with the chemoprophylaxis of anticoagulants. The association between diabetes and DVT after TKA may be more remarkable in patients from Asian countries.


Asunto(s)
Artroplastia de Reemplazo de Rodilla , Diabetes Mellitus , Trombosis de la Vena , Humanos , Anticoagulantes , Artroplastia de Reemplazo de Rodilla/efectos adversos , Quimioprevención , Diabetes Mellitus/epidemiología , Factores de Riesgo , Trombosis de la Vena/etiología
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