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Endoplasmic reticulum (ER) stress exerts significant effects on cell growth, proliferation, migration, invasion, chemoresistance, and angiogenesis in various cancers. However, the impact of ER stress on the outcomes of osteosarcoma patients remains unclear. In this study, we established an ER stress risk model based on The Cancer Genome Atlas (TARGET) osteosarcoma dataset to reflect immune features and predict the prognosis of osteosarcoma patients. Survival analysis revealed significant differences in overall survival among osteosarcoma patients with different ER stress-related risk scores. Furthermore, ER stress-related risk features were significantly associated with the clinical pathological characteristics of osteosarcoma patients and could serve as independent prognostic indicators. Functional enrichment analysis indicated associations of the risk model with cell chemotaxis, leukocyte migration, and regulation of leukocyte migration. Additionally, the ER stress-related risk model suggested the presence of an immunosuppressive microenvironment and immune checkpoint responses. We validated the significance of 7 ER stress-related genes obtained from LASSO regression analysis through RT-qPCR testing on osteosarcoma samples from a local hospital, and inferred the importance of STC2 based on the literature. Subsequently, IHC experiments using samples from 70 osteosarcoma cases and 21 adjacent tissue samples confirmed differential expression of STC2 between cancer and normal tissues, and explored the gene's expression in pan-cancer and its association with clinical pathological parameters of osteosarcoma. In conclusion, we have proposed an ER stress risk model as an independent prognostic factor and identified STC2 as a novel risk indicator for disease progression, providing a promising direction for further research and treatment of osteosarcoma.
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Due to persistent inflammation and oxidative stress reactions, achieving drug absorption in diabetic wounds is challenging. To overcome this problem, our article presents a composite hydrogel, GelMA-GA/DMOG@GDNP, which consists of gelatin methacryloyl (GelMA) treated with gallic acid (GA) and encapsulating ginseng-derived nanoparticles (GDNPs) loaded with dimethyloxallyl glycine (DMOG). The composite hydrogel demonstrates excellent biocompatibility. In laboratory settings, the hydrogel inhibits the production of nitric oxide synthase 2 (iNOS) in mouse immune cells (RAW264.7 cells), enhances the growth and migration of mouse connective tissue cells (L929 cells) and human endothelial cells (HUVECs), and promotes tube formation in HUVECs. In a rat model of type 1 diabetes-induced wounds, the composite hydrogel attenuates inflammatory reactions, facilitates the formation of fibres and blood vessels, accelerates wound healing, and elucidates specific pathway mechanisms through transcriptome sequencing. Therefore, the GelMA-GA/DMOG@GDNP hydrogel can serve as a safe and efficient wound dressing to regulate the inflammatory response, promote collagen fiber and blood vessel formation, and accelerate wound healing. These findings suggest that utilizing this multifunctional engineered nanoparticle-loaded hydrogel in a clinical setting may be a promising strategy for diabetic wound healing.
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Diabetes Mellitus Experimental , Ácido Gálico , Gelatina , Nanopartículas , Panax , Cicatrización de Heridas , Animales , Gelatina/química , Cicatrización de Heridas/efectos de los fármacos , Ácido Gálico/química , Ácido Gálico/farmacología , Ratas , Nanopartículas/química , Diabetes Mellitus Experimental/tratamiento farmacológico , Humanos , Ratones , Panax/química , Células RAW 264.7 , Masculino , Células Endoteliales de la Vena Umbilical Humana/efectos de los fármacos , Hidrogeles/química , Hidrogeles/farmacología , Metacrilatos/química , Metacrilatos/farmacología , Ratas Sprague-DawleyRESUMEN
The regenerative microenvironment after peripheral nerve injury is imbalanced and difficult to rebalance, which is mainly affected by inflammation, oxidative stress, and inadequate blood supply. The difficulty in remodeling the nerve regeneration microenvironment is the main reason for slow nerve regeneration. Traditional drug treatments have certain limitations, such as difficulty in penetrating the blood-nerve barrier and lack of pleiotropic effects. Therefore, there is an urgent need to build multifunctional nerve grafts that can effectively regulate the regenerative microenvironment and promote nerve regeneration. Nitric oxide (NO), a highly effective gas transmitter with diatomic radicals, is an important regulator of axonal growth and migration, synaptic plasticity, proliferation of neural precursor cells, and neuronal survival. Moreover, NO provides potential anti-inflammation, anti-oxidation, and blood vessel promotion applications. However, excess NO may cause cell death and neuroinflammatory cell damage. The prerequisite for NO treatment of peripheral nerve injury is that it is gradually released over time. In this study, we constructed an injectable NO slow-release system with two main components, including macromolecular NO donor nanoparticles (mPEG-P(MSNO-EG) nanoparticles, NO-NPs) and a carrier for the nanoparticles, mPEG-PA-PP injectable temperature-sensitive hydrogel. Due to the multiple physiological regulation of NO and better physiological barrier penetration, the conduit effectively regulates the inflammatory response and oxidative stress of damaged peripheral nerves, promotes nerve vascularization, and nerve regeneration and docking, accelerating the nerve regeneration process. STATEMENT OF SIGNIFICANCE: The slow regeneration speed of peripheral nerves is mainly due to the destruction of the regeneration microenvironment. Neural conduits with drug delivery capabilities have the potential to improve the microenvironment of nerve regeneration. However, traditional drugs are hindered by the blood nerve barrier and cannot effectively target the injured area. NO, an endogenous gas signaling molecule, can freely cross the blood nerve barrier and act on target cells. However, excessive NO can lead to cell apoptosis. In this study, a NO sustained-release system was constructed to regulate the microenvironment of nerve regeneration through various pathways and promote nerve regeneration.
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Preparaciones de Acción Retardada , Regeneración Nerviosa , Óxido Nítrico , Animales , Óxido Nítrico/metabolismo , Preparaciones de Acción Retardada/farmacología , Preparaciones de Acción Retardada/química , Preparaciones de Acción Retardada/farmacocinética , Regeneración Nerviosa/efectos de los fármacos , Traumatismos de los Nervios Periféricos/tratamiento farmacológico , Traumatismos de los Nervios Periféricos/terapia , Traumatismos de los Nervios Periféricos/patología , Traumatismos de los Nervios Periféricos/metabolismo , Ratas Sprague-Dawley , Ratas , Nervios Periféricos/efectos de los fármacos , Nervios Periféricos/patología , Nanopartículas/química , Donantes de Óxido Nítrico/farmacología , Donantes de Óxido Nítrico/uso terapéutico , Masculino , Hidrogeles/química , Nervio Ciático/efectos de los fármacosRESUMEN
In this report, a multiplex PCR method was developed for the detection of three diarrhea-associated viruses in mink, including circovirus (MCV), bocavirus (MBoV), and enteritis virus (MEV). Three compatible sets of primers specific for each virus were designed respectively based on their conserved sequences. After optimization of the crucial factors such as primer concentration and annealing temperature in single and multiple amplification, three specific fragments were simultaneously amplified with the highest sensitivity and specificity in one PCR reaction. The fragments amplified were 259â¯bp (MCV)ï¼455â¯bp (MBoV) and 671â¯bp (MEV). The sensibility of this one-step multiplex PCR is about 10 times lower than that of regular singleplex PCR. There were no cross-reactions with some relevant pathogens like mink coronavirus, canine distemper virus, and aleutian mink disease virus. In our study we analyzed viral DNA in mink fecal samples by multiplex PCR assay from China, which revealed the occurrence of MCV, MBoV, and MEV as 3.1â¯%, 5.7â¯%, and 9.8â¯%, respectively. The testing results of multiplex PCR agreed with the singleplex PCR results with a coincidence rate of 100â¯%. These results indicated that the method could provide technical support for rapid detection of the three diarrhea-associated viruses, and epidemiological investigation of mink viral diarrhea.
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Cartilla de ADN , Diarrea , Heces , Visón , Reacción en Cadena de la Polimerasa Multiplex , Sensibilidad y Especificidad , Animales , Visón/virología , Reacción en Cadena de la Polimerasa Multiplex/métodos , Reacción en Cadena de la Polimerasa Multiplex/veterinaria , China , Diarrea/virología , Diarrea/veterinaria , Diarrea/diagnóstico , Cartilla de ADN/genética , Heces/virología , Circovirus/genética , Circovirus/aislamiento & purificación , Bocavirus/genética , Bocavirus/aislamiento & purificación , Virus de la Enteritis del Visón/genética , Virus de la Enteritis del Visón/aislamiento & purificación , Técnicas de Diagnóstico Molecular/métodos , Técnicas de Diagnóstico Molecular/veterinariaRESUMEN
Sophoridine, which is derived from the Leguminous plant Sophora alopecuroides L., has certain pharmacological activity as a new anticancer drug. Herein, a series of novel N-substituted sophoridine derivatives was designed, synthesized and evaluated with anticancer activity. Through QSAR prediction models, it was discovered that the introduction of a benzene ring as a main pharmacophore and reintroduced into a benzene in para position on the phenyl ring in the novel sophoridine derivatives improved the anticancer activity effectively. In vitro, 28 novel compounds were evaluated for anticancer activity against four human tumor cell lines (A549, CNE-2, HepG-2, and HEC-1-B). In particular, Compound 26 exhibited remarkable inhibitory effects, with an IC50 value of 15.6 µM against HepG-2 cells, surpassing cis-Dichlorodiamineplatinum (II). Molecular docking studies verified that the derivatives exhibit stronger binding affinity with DNA topoisomerase I compared to sophoridine. In addition, 26 demonstrated significant inhibition of DNA Topoisomerase I and could arrest cells in G0/G1 phase. This study provides valuable insights into the design and synthesis of N-substituted sophoridine derivatives with anticancer activity.
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Alcaloides , Matrinas , Simulación del Acoplamiento Molecular , Relación Estructura-Actividad Cuantitativa , Quinolizinas , Sophora , Inhibidores de Topoisomerasa I , Humanos , Inhibidores de Topoisomerasa I/farmacología , Inhibidores de Topoisomerasa I/síntesis química , Quinolizinas/farmacología , Quinolizinas/síntesis química , Quinolizinas/química , Estructura Molecular , Sophora/química , Alcaloides/farmacología , Alcaloides/síntesis química , Alcaloides/química , Línea Celular Tumoral , Antineoplásicos Fitogénicos/farmacología , Antineoplásicos Fitogénicos/síntesis química , Indolizinas/farmacología , Indolizinas/química , Indolizinas/síntesis química , ADN-Topoisomerasas de Tipo I/metabolismo , Fitoquímicos/farmacología , Fitoquímicos/síntesis químicaRESUMEN
Inflammation stands as a pivotal factor in the pathogenesis of glucocorticoid-associated osteonecrosis of the femoral head (GA-ONFH). However, the vital role played by M1 macrophages, the principal constituents of the inflammatory process, remains largely underexplored. In this study, we employed reverse transcription-quantitative polymerase chain Reaction (RT-PCR), western blot, and flow cytometry to assess the impact of M1-conditioned medium on cultures of mouse bone marrow-derived mesenchymal stem cells (BMSCs) and Murine Long bone Osteocyte-Y4 (MLO-Y4) in vitro. Moreover, we quantified the levels of inflammatory cytokines in the M1-conditioned medium through the employment of an enzyme-linked immunosorbent assay (ELISA). For in vivo analysis, we examined M1 macrophages and investigated the NF-kB signaling pathway in specimens obtained from the femoral heads of animals and humans. We found that the number of M1 macrophages in the femoral head of GA-ONFH patients grew significantly, and in the mice remarkably increase, maintaining high levels in the intramedullary. In vitro, the M1 macrophage-conditioned medium elicited apoptosis in BMSCs and MLO-Y4 cells, shedding light on the intricate interplay between macrophages and these cell types. The presence of TNF-α within the M1-conditioned medium activated the NF-κB pathway, providing mechanistic insight into the apoptotic induction. Moreover, employing a robust rat macrophage clearance model and GA-ONFH model, we demonstrated a remarkable attenuation in TNF-α expression and NF-kB signaling subsequent to macrophage clearance. This pronounced reduction engenders diminished cellular apoptosis and engenders a decelerated trajectory of GA-ONFH progression. In conclusion, our study reveals the crucial involvement of M1 macrophages in the pathogenesis of GA-ONFH, highlighting their indispensable role in disease progression. Furthermore, early clearance emerges as a promising strategy for impeding the development of GA-ONFH.
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Necrosis de la Cabeza Femoral , Glucocorticoides , Humanos , Ratas , Ratones , Animales , Cabeza Femoral , Necrosis de la Cabeza Femoral/inducido químicamente , Necrosis de la Cabeza Femoral/metabolismo , Factor de Necrosis Tumoral alfa , FN-kappa B , Medios de Cultivo Condicionados , Macrófagos/metabolismoRESUMEN
In this study, the diversity and regularity of two new feline calicivirus (FCV) isolates, QD-7 and QD-164, were investigated. The genomes of these new strains were compared with 39 strains from the NCBI database including isolates from China, United States, Germany, South Korea, the United Kingdom and Japan. The nucleotide sequence identities ranged from 75-88%, indicating a high degree of variability. These variations were not related to distributions of the virus by time of isolation and geographical location. Cats that were experimentally infected with the new isolate QD-164 showed typical clinical symptoms of sneezing, fever and conjunctivitis and all recovered within 30 days. In contrast, QD-7 infections were asymptomatic and the virus was cleared within 16 days. These results indicate that QD-7 and QD-164 were naturally attenuated strains. NNS mutations characteristic of highly virulent strains at positions 441-443 were absent in QD-7 while QD-164 possessed an N at position 442. This indicated that mutations in regions 441-443 may be linked to disease severity.
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Infecciones por Caliciviridae , Calicivirus Felino , Enfermedades de los Gatos , Gatos , Animales , Calicivirus Felino/genética , Virulencia/genética , Infecciones por Caliciviridae/veterinaria , Secuencia de Bases , ChinaRESUMEN
Most recently, the path integral molecular dynamics has been successfully used to consider the thermodynamics of single-component identical bosons and fermions. In this work, the path integral molecular dynamics is developed to simulate thermodynamics, Green's function, and momentum distribution of two-component bosons in three dimensions. As an example of our general method, we consider the thermodynamics of up to 16 bosons in a three-dimensional harmonic trap. For noninteracting spinor bosons, our simulation shows a bump in the heat capacity. As the repulsive interaction strength increases, however, we find the gradual disappearance of the bump in the heat capacity. We believe that this simulation result can be tested by ultracold spinor bosons with optical lattices and magnetic-field Feshbach resonance to tune the inter-particle interaction. We also calculate Green's function and momentum distribution of spinor bosons. Our work facilitates the exact numerical simulation of spinor bosons, whose property is one of the major problems in ultracold Bose gases.
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Sarcopenia is an age-related disease in which muscle mass, strength and function may decline with age or can be secondary to cachexia or malnutrition and can lead to weakness, falls and even death. With the increase in life expectancy, sarcopenia has become a major threat to the health of the elderly. Currently, our understanding of bone-muscle interactions is not limited to their mechanical coupling. Bone and muscle have been identified as secretory endocrine organs, and their interaction may affect the function of each. Both muscle-derived factors and osteokines can play a role in regulating muscle and bone metabolism via autocrine, paracrine and endocrine mechanisms. Herein, we comprehensively summarize the latest research progress on the effects of the osteokines FGF-23, IGF-1, RANKL and osteocalcin on muscle to explore whether these cytokines can be utilized to treat and prevent sarcopenia.
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Sarcopenia is an aged-related syndrome that is progressive and can be accelerated by other concomitant disease states. Sarcopenia, characterized by loss of skeletal muscle mass, reduced muscle strength, and/or reduced physical performance, is one of the main reasons for limitation of daily activities in the elderly. It is associated with an increased incidence of many adverse events, such as dysfunction, falls, weakness, hospitalization, disability and even death. Sarcopenia justifies one of the most widely accepted theories that low-grade chronic inflammation associated with aging, known as inflammatory aging, is important to the pathogenesis of many age-related diseases. Currently, the diagnosis of sarcopenia is based on a comprehensive assessment of three aspects: muscle mass, muscle strength and physical performance. The measurement of muscle mass is complicated, as the measurement of muscle strength and gait speed is easily affected by the physical conditions of the subjects. This makes the measurements inaccurate and prospective, and it is difficult to achieve continuous, purposeful monitoring. In addition, serum levels of inflammatory cytokines change as inflammatory states develop in the elderly population. This manuscript focuses on the correlation between serum inflammatory cytokines and sarcopenia in recent years, plus the possible underlying mechanisms.
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Sarcopenia , Anciano , Citocinas , Fuerza de la Mano , Humanos , Inflamación/patología , Fuerza Muscular , Músculo Esquelético/patología , Estudios Prospectivos , Sarcopenia/patologíaRESUMEN
Canine parvovirus type 2 (CPV-2) is currently circulating in domestic and wild animals, but our knowledge about CPV-2 infections in raccoon dogs is limited. In this study, VP2 gene sequences of CPV-2 were amplified from rectal swabs of 14 diarrhetic raccoon dogs (Nyctereutes procyonoides) in Hebei province, China, in 2016 and 2017. Phylogenetic analysis of the VP2 gene sequences revealed that most of these sequences (11 of 14) belonged to the same subclade as raccoon dog strain CPV-2/Raccoon_Dog/China/DP-1/16 isolated from Shandong province in 2016. A comparison of deduced amino acid sequences revealed presence of the substitutions S297A and S27T in 11 of those 14 sequences. I418T was observed in a minority of the sequences (4 of 14). In addition, A300D and T301I, P13S and I219V, and N419K were found in three of the sequences. This study shows that CPV-2 strains with different substitutions in their VP2 amino acid sequences were spreading among raccoon dogs in Hebei during 2016 and 2017 and suggests that further studies are needed to monitor the distribution of these strains in China.
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Infecciones por Parvoviridae/veterinaria , Parvovirus Canino/clasificación , Perros Mapache/virología , Secuencia de Aminoácidos , Animales , Proteínas de la Cápside/genética , China/epidemiología , Infecciones por Parvoviridae/epidemiología , Infecciones por Parvoviridae/virología , Parvovirus Canino/aislamiento & purificaciónRESUMEN
Ostrich diseases characterized by paralysis have been breaking out in broad areas of China since 2015, causing major damage to the ostrich breeding industry in China. This report describes a parvovirus detected in ostriches from four different regions. The entire genomes of four parvovirus strains were sequenced following amplification by PCR, and we conducted comprehensive analysis of the ostrich parvovirus genome. Results showed that the length genomes of the parvovirus contained two open reading frames. Ostrich parvovirus (OsPV) is a branch of goose parvovirus (GPV). Genetic distance analysis revealed a close relationship between the parvovirus and goose parvovirus strains from China, with the closest being the 2016 goose parvovirus RC16 strain from Chongqing. This is the first report of a parvovirus in ostriches. However, whether OsPV is the pathogen of ostrich paralysis remains uncertain. This study contributes new information about the evolution and epidemiology of parvovirus in China, which provides a new way for the study of paralysis in ostriches.
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Evolución Molecular , Genoma Viral , Infecciones por Parvoviridae/virología , Parvovirus/fisiología , Struthioniformes/virología , Animales , Secuencia de Bases , Pruebas Genéticas , Genómica/métodos , Infecciones por Parvoviridae/diagnóstico , Filogenia , Reacción en Cadena de la PolimerasaRESUMEN
In this study, we describe a novel and rapid method for the construction of a full-length infectious clone (pPPV). The constructed clone contained an engineered EcoRv site that served as a genetic marker and was shown to be infectious when transfected into a monolayer of PK-15 cells. The rescued virus (rPPV) of the infectious clone was found to be indistinguishable from wild-type virus BQ in terms of its biological properties. The generation of this PPV infectious clone provides a potentially powerful tool with which to elucidate the molecular pathogenesis of PPV.
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Clonación Molecular/métodos , Genoma Viral/genética , Parvovirus Porcino/genética , Secuencias Repetidas Terminales/genética , Animales , Línea Celular , Técnica del Anticuerpo Fluorescente Indirecta , Marcadores Genéticos/genética , Técnicas de Amplificación de Ácido Nucleico , PorcinosRESUMEN
Canine distemper virus (CDV), bearing a close resemblance to measles virus, represents a promising candidate for oncolytic therapy; however, its application and underlying oncolytic mechanisms in canine mammary carcinoma cells remain to be explored. Here, we found that an attenuated canine distemper vaccine strain, CDV-L, efficiently infected and inhibited the growth of canine mammary tubular adenocarcinoma CIPp cells but not MDCK cells in vitro. Transcriptomic analysis of CDV-L-infected CIPp cells revealed substantially differentially expressed genes in apoptotic and NF-κB signalling pathways. Subsequent validations confirmed that CDV-L-induced apoptosis of CIPp cells through the caspase-8 and caspase-3 pathway. Identification of phosphorylated-IκBα, phosphorylated-p65 and the nuclear translocation of p65 confirmed the activation of the NF-κB signalling pathway. Inhibition of the NF-κB pathway abrogated CDV-L-induced cleaved-caspase-3 and cleaved-PARP. In a CIPp subcutaneous xenograft mouse model, intratumoural injections of CDV-L significantly restricted tumour growth without apparent pathology, and virus remained localized within the tumour. Taken altogether, these findings indicate that CDV-L exerts an antitumour effect in CIPp cells, and that apoptosis and the NF-κB pathway play essential roles in this process.
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Adenocarcinoma/veterinaria , Neoplasias de la Mama/veterinaria , Línea Celular Tumoral/efectos de los fármacos , Virus del Moquillo Canino/patogenicidad , Enfermedades de los Perros/virología , Virus Oncolíticos/patogenicidad , Adenocarcinoma/virología , Animales , Apoptosis/efectos de los fármacos , Neoplasias de la Mama/virología , Inhibidores de Caspasas , Perros , Femenino , FN-kappa B/genética , Análisis de Secuencia de ARN/veterinariaRESUMEN
Feline panleukopenia virus (FPV) infects cats and can be fatal to kittens. There is evidence that canine parvovirus originated from FPV, which makes FPV important in studies of the family Parvoviridae. In the present study, the entire genome of FPV strain HH-1/86 was converted into a full-length infectious clone (pFPV). The FPV strain HH-1/86 has a 5123-nt single stranded DNA genome with a Y-shaped inverted 3' terminal repeat (ITR) and a U-shaped inverted 5' ITR. Feline kidney cells (F81) were transfected with the pFPV clone which contained a genetic marker, and a rescued virus was obtained (rFPV). The rFPV was identified by its cytopathic effects, indirect immunofluorescence, growth curve analysis, western blot assay and hemagglutination, and was indistinguishable from the parent virus. The FPV infectious clone will facilitate the study of pathogenicity and viral replication of FPV and the inter-species transmission of parvoviruses.
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Virus de la Panleucopenia Felina/genética , Panleucopenia Felina/virología , Genética Inversa , Animales , Gatos , Clonación Molecular , ADN Viral , Marcadores Genéticos , Genoma Viral , Genómica/métodos , Hemaglutinación , Hemaglutininas Virales/metabolismo , Genética Inversa/métodos , Secuenciación Completa del GenomaRESUMEN
We have isolated 4 naturally-occurring strains of CPV in mainland China and have identified them as CPV-2, 2a, 2b and 2c genotypes according to their VP2 sequences which also revealed substitutions within their right terminal regions. To determine if these substitutions affected the growth characteristics of the 4 strains, we constructed plasmids based on their genomic sequences minus their right terminal sequences, with the latter replaced by a single right terminal region. Analysis of rescued recombinants showed that the substitutions within their natural right termini had no significant effect on their growth characteristics.
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ADN Viral/genética , Mutación , Parvovirus Canino/crecimiento & desarrollo , Parvovirus Canino/genética , Animales , Enfermedades de los Gatos/virología , Gatos , Línea Celular , China , Células Epiteliales/virología , Genotipo , Infecciones por Parvoviridae/veterinaria , Infecciones por Parvoviridae/virología , Parvovirus Canino/clasificación , Parvovirus Canino/aislamiento & purificación , Genética Inversa , Análisis de Secuencia de ADN , Proteínas Estructurales Virales/genética , VirulenciaRESUMEN
Pathogenic strains of Aleutian mink disease virus (AMDV) such as Utah-1 do not replicate in cell culture (e.g., Crandell Rees feline kidney cells) while the in vitro-adapted AMDV strain ADV-Gorham (ADV-G) is not pathogenic. Here, we constructed a full-length infectious clone (pADV-G). Alignment of the VP2 gene of ADV-G with that of other AMDV strains revealed many amino acid (a.a.) residues conserved among pathogenic isolates that differed in ADV-G. Four virulence-associated, conserved residues of pADV-G VP2 were studied by site-directed mutagenesis (H92A, Q94S, Y115F, and I116L). Mutation of residue 92 or 94 decreased viral-transcription and viral-infectivity levels, whereas mutation of residue 115 or 116 did not affect viral-infectivity in CRFK cells. These results indicated that VP2 residues 92 and 94, both located on the surface of the viral capsid, are critical for AMDV infectivity in vitro.
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Virus de la Enfermedad Aleutiana del Visón/fisiología , Proteínas de la Cápside/metabolismo , Internalización del Virus , Replicación Viral , Virus de la Enfermedad Aleutiana del Visón/genética , Animales , Proteínas de la Cápside/genética , Gatos , Técnicas de Cultivo de Célula , Células Cultivadas , Análisis Mutacional de ADN , Mutagénesis Sitio-Dirigida , Mutación Missense , Genética InversaRESUMEN
Canine parvovirus (CPV) is an important and highly prevalent pathogen of dogs that causes acute hemorrhagic enteritis disease. Here, we describe a rapid method for the construction and characterization of a full-length infectious clone (rCPV) of CPV. Feline kidney (F81) cells were transfected with rCPV incorporating an engineered EcoR I site that served as a genetic marker. The rescued virus was indistinguishable from that of wild-type virus in its biological properties.
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Enfermedades de los Perros/virología , Infecciones por Parvoviridae/virología , Parvovirus Canino/genética , Animales , Gatos , Línea Celular , ADN Viral/genética , Perros , Filogenia , Genética Inversa/mortalidad , Análisis de Secuencia de ADN/métodosRESUMEN
Mink enteritis virus (MEV) is one of the most important viral pathogens causing serious disease in mink. Type I interferon (IFN) plays a critical role in antiviral innate immunity and, for successful infection, many viruses have evolved evasive strategies against it. Here, we show that MEV infection does not evoke IFN or interferon-stimulated genes (ISGs) responses in feline kidney (CRFK) cells, and that MEV suppresses IFN production in both poly I:C-stimulated and untreated cells. In CRFK cells pre-exposure to IFN, show that infection with, and replication of, MEV remain unaffected. This inhibition appears to be mediated by the MEV nonstructural protein (NS1) with its ORI-binding domain playing a major role.
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Panleucopenia Felina/inmunología , Interferón Tipo I/inmunología , Virus de la Enteritis del Visón/fisiología , Animales , Gatos , Línea Celular , Regulación de la Expresión Génica/efectos de los fármacos , Inductores de Interferón/farmacología , Poli I-C/farmacología , Proteínas no Estructurales Virales/metabolismo , Replicación ViralRESUMEN
The genome of a highly pathogenic strain of Aleutian disease mink virus (AMDV-BJ) isolated from a domestic farm in North China has been determined and compared with other strains. Alignment analysis of the major structural protein VP2 revealed that AMDV-BJ is unique among 17 other AMDV strains. Compared with the nonpathogenic strain ADV-G, the 3' end Y-shaped hairpin was highly conserved, while a 4-base deletion in the 5' U-shaped terminal palindrome resulted in a different unpaired "bubble" group near the NS1-binding region of the 5' end hairpin which may affect replication efficiency in vivo. We also performed a protein analysis of the NS1, NS2, and new-confirmed NS3 of AMDV-BJ with some related AMDV DNA sequence published, providing information on evolution of AMDV genes. This study shows a useful method to obtain the full-length genome of AMDV and some other parvoviruses.