RESUMEN
BACKGROUND: Esophageal atresia (EA) is a life-threatening congenital malformation in newborns, and the traditional repair approaches pose technical challenges and are extremely invasive. Therefore, surgeons have been actively investigating new minimally invasive techniques to address this issue. Magnetic compression anastomosis has been reported in several studies for its potential in repairing EA. In this paper, the primary repair of EA with magnetic compression anastomosis under thoracoscopy was reported. CASE SUMMARY: A full-term male weighing 3500 g was diagnosed with EA gross type C. The magnetic devices used in this procedure consisted of two magnetic rings and several catheters. Tracheoesophageal fistula ligation and two purse strings were performed. The magnetic compression anastomosis was then completed thoracoscopically. After the primary repair, no additional operation was conducted. A patent anastomosis was observed on the 15th day postoperatively, and the magnets were removed on the 23rd day. No leakage existed when the transoral feeding started. CONCLUSION: Thoracoscopic magnetic compression anastomosis may be a promising minimally invasive approach for repairing EA.
RESUMEN
Following the publication of this paper, it was drawn to the Editors' attention by a concerned reader that certain of the western blot data shown in Figs. 4B and 7, the stratchwound assay data shown in Figs. 2B and E and Fig. 6B, and the cellular images shown in Fig. 3 were strikingly similar to data appearing in different form in other articles written by different authors in different research institutions. Owing to the fact that the contentious data in the above article had already been published elsewhere prior to its submission to Oncology Reports, the Editor has decided that this paper should be retracted from the Journal. The authors were asked for an explanation to account for these concerns, but the Editorial Office did not receive a reply. The Editor apologizes to the readership for any inconvenience caused. [Oncology Reports 38: 10751082, 2017; DOI: 10.3892/or.2017.5781].
RESUMEN
Interferon (IFN)-stimulated gene 15 (ISG15) encodes a ubiquitin-like protein that is heavily involved in immune response elicitation. As an important member of interferon regulatory factor (IRF) family, IRF1 can activate the expression of multiple genes, including the human optineurin gene (Sudhakar et al., 2013). In this study, a sequence in the promoter region of the optineurin gene was compared to the 5' flanking region of the porcine isg15 gene. Porcine IRF1 also possesses antiviral activity against several swine viruses (Li et al., 2015), but the mechanism is not well understood. Herein, we report that porcine IRF1 and ISG15 were up-regulated in porcine kidney (PK-15) cells following stimulation with double-stranded RNA (dsRNA) or classical swine fever virus (CSFV) infection. We also found that siRNA-mediated knockdown of IRF1 expression resulted in lower ISG15 expression in response to polyinosinic:polycytidylic acid [poly(I:C)] or CSFV infection. The overexpression of IRF1 resulted in ISG15 up-regulation. IRF1 was shown to translocate to the nucleus in response to dsRNA stimulation. To further identify the functional domain of the isg15 gene that promotes IRF1 transcriptional activity, firefly luciferase and ISG15 reporter systems were constructed. The results of the firefly luciferase and ISG15 reporter assay suggested that IRF1 mediates the up-regulation of ISG15. Nucleotides -487 to -325, located in the 5' flanking region of the isg15 gene, constituted the promoter region of IRF1. ChIP assay indicated that IRF1 protein was able to interact with the DNA in the 5'fr of isg15 gene in cells. As an innate immune response protein with broad-spectrum antiviral activity, the up-regulation of ISG15 mediated by IRF1 in porcine cells is reported for the first time. These results warrant further investigation into the antiviral activity of porcine IRF1 against reported swine viruses.
Asunto(s)
Región de Flanqueo 5'/genética , Peste Porcina Clásica/genética , Factor 1 Regulador del Interferón/fisiología , ARN Bicatenario/fisiología , Ubiquitinas/genética , Animales , Células Cultivadas , Peste Porcina Clásica/inmunología , Virus de la Fiebre Porcina Clásica/fisiología , Cricetinae , Citocinas/genética , Expresión Génica , Interacciones Huésped-Patógeno/genética , Inmunidad Innata/genética , Porcinos , Regulación hacia Arriba/genéticaRESUMEN
Understanding the molecular basis of the neutralizing antibody response to dengue virus (DENV) is an essential component in the design and development of effective vaccines and immunotherapeutics. Here we present the structure of a cross-reactive, neutralizing antibody, 3E31, in complex with domain III (DIII) of the DENV envelope (E) protein and reveal a conserved, temperature-sensitive, cryptic epitope on DIII that is not available in any of the known conformations of E on the dengue virion. We observed that 3E31 inhibits E-mediated membrane fusion, suggesting that the antibody is able to neutralize virus through binding an as-yet uncharacterized intermediate conformation of DENV E and sterically block trimer formation. Finally, we show that, unlike cross-reactive fusion peptide-specific antibodies, 3E31 does not promote antibody-dependent enhancement of infection at sub-neutralizing concentrations. Our results highlight the 3E31 epitope on the E protein DIII as a promising target for immunotherapeutics or vaccine design.
Asunto(s)
Anticuerpos Monoclonales/química , Anticuerpos Neutralizantes/química , Anticuerpos Antivirales/química , Virus del Dengue/inmunología , Epítopos/química , Proteínas del Envoltorio Viral/inmunología , Secuencia de Aminoácidos , Animales , Anticuerpos Monoclonales/biosíntesis , Anticuerpos Monoclonales/inmunología , Anticuerpos Monoclonales/farmacología , Anticuerpos Neutralizantes/biosíntesis , Anticuerpos Neutralizantes/inmunología , Anticuerpos Neutralizantes/farmacología , Anticuerpos Antivirales/biosíntesis , Anticuerpos Antivirales/inmunología , Anticuerpos Antivirales/farmacología , Especificidad de Anticuerpos , Sitios de Unión , Chlorocebus aethiops , Reacciones Cruzadas , Dengue/prevención & control , Dengue/virología , Vacunas contra el Dengue/biosíntesis , Virus del Dengue/química , Virus del Dengue/efectos de los fármacos , Mapeo Epitopo/métodos , Epítopos/inmunología , Humanos , Hibridomas/inmunología , Fusión de Membrana/efectos de los fármacos , Ratones , Ratones Endogámicos BALB C , Unión Proteica , Dominios y Motivos de Interacción de Proteínas , Estructura Secundaria de Proteína , Alineación de Secuencia , Homología de Secuencia de Aminoácido , Bazo/citología , Bazo/inmunología , Células Vero , Proteínas del Envoltorio Viral/químicaRESUMEN
Pancreatic ductal adenocarcinoma (PDAC) is the most common type of malignant pancreatic tumor. MicroRNAs (miRNAs) are a group of small, non-protein coding, endogenous RNAs that play critical roles in tumorigenesis and progression of PDAC. In the present study, we demonstrated that miR-448 expression was downregulated in PDAC tissues and cell lines. Clinical association analysis indicated that low expression of miR-448 was associated with poor prognostic features and conferred a significant reduced survival of PDAC patients. Overexpression of miR-448 suppressed PDAC cell migration and invasion, while its loss showed the opposite effects on these cellular processes. In vivo experiments revealed that miR-488 restoration prohibited liver metastasis of PDAC in nude mice. Moreover, we found that Janus kinase 1 (JAK1) was a direct target gene of miR-448 in PDAC cells. We further demonstrated that the expression of JAK1 mRNA was upregulated in PDAC tissues. Notably, the expression of JAK1 mRNA was inversely correlated with the level of miR-448 in PDAC tissues. In addition, JAK1 knockdown showed similar effects of miR-448 on the metastasis of PDAC cells. JAK1/STAT3 pathway may be involved in the function of miR-448 in PDAC cells. Taken together, these findings suggest that miR-448 functions as a tumor suppressor in the development of PDAC through targeting the JAK1/STAT3 pathway.
Asunto(s)
Adenocarcinoma/prevención & control , Carcinoma Ductal Pancreático/prevención & control , Janus Quinasa 1/antagonistas & inhibidores , Neoplasias Hepáticas/prevención & control , MicroARNs/genética , Neoplasias Pancreáticas/prevención & control , Factor de Transcripción STAT3/antagonistas & inhibidores , Adenocarcinoma/genética , Adenocarcinoma/metabolismo , Adenocarcinoma/secundario , Animales , Apoptosis , Biomarcadores de Tumor , Carcinoma Ductal Pancreático/genética , Carcinoma Ductal Pancreático/metabolismo , Carcinoma Ductal Pancreático/secundario , Estudios de Casos y Controles , Movimiento Celular , Proliferación Celular , Femenino , Estudios de Seguimiento , Regulación Neoplásica de la Expresión Génica , Humanos , Janus Quinasa 1/genética , Janus Quinasa 1/metabolismo , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/secundario , Metástasis Linfática , Masculino , Ratones , Ratones Desnudos , Persona de Mediana Edad , Invasividad Neoplásica , Recurrencia Local de Neoplasia/genética , Recurrencia Local de Neoplasia/metabolismo , Recurrencia Local de Neoplasia/patología , Recurrencia Local de Neoplasia/prevención & control , Neoplasias Pancreáticas/genética , Neoplasias Pancreáticas/metabolismo , Neoplasias Pancreáticas/patología , Pronóstico , Factor de Transcripción STAT3/genética , Factor de Transcripción STAT3/metabolismo , Tasa de Supervivencia , Células Tumorales Cultivadas , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
Glutamatergic retinal waves, the spontaneous patterned neural activities propagating among developing retinal ganglion cells (RGCs), instruct the activity-dependent refinement of visuotopic maps. However, its initiation and underlying mechanism remain largely elusive. Here using larval zebrafish and multiple in vivo approaches, we discover that bipolar cells (BCs) are responsible for the generation of glutamatergic retinal waves. The wave originates from BC axon terminals (ATs) and propagates laterally to nearby BCs and vertically to downstream RGCs and the optic tectum. Its initiation is triggered by the activation of and consequent glutamate release from BC ATs, and is mediated by the N-methyl-D-aspartate subtype of glutamate receptors (NMDARs) expressed at these ATs. Intercellular asymmetry of NMDAR expression at BC ATs enables the preferential initiation of waves at the temporal retina, where BC ATs express more NMDARs. Thus, our findings indicate that glutamatergic retinal waves are initiated by BCs through a presynaptic NMDA autoreceptor-dependent process.
Asunto(s)
Receptores de N-Metil-D-Aspartato/metabolismo , Células Bipolares de la Retina/metabolismo , Células Ganglionares de la Retina/metabolismo , Transmisión Sináptica/fisiología , Vías Visuales/fisiología , Pez Cebra/embriología , Animales , Sinapsis Eléctricas/fisiología , Ácido Glutámico/metabolismo , Larva/crecimiento & desarrollo , Terminales Presinápticos/metabolismo , Pez Cebra/genéticaRESUMEN
The envelope domain III (EDIII) of the dengue virus (DENV) has been confirmed to be involved in receptor binding. It is the target of specific neutralizing antibodies, and is considered to be a promising subunit dengue vaccine candidate. However, several recent studies have shown that antiEDIII antibodies contribute little to the neutralizing or enhancing ability of human DENVinfected serum. The present study involved an analysis of the neutralization and antibodydependent enhancement (ADE) activities of EDIIIreactive antibodies in human convalescent sera from patients with primary DENV1 infection and rabbit antiserum immunized with recombinant DENV1 EDIII protein. The results indicated that serum neutralization was not associated with titres of EDIIIbinding antibodies in the human DENV1infected sera. The depletion of antiEDIII antibodies from these serum samples revealed that the antiEDIII antibodies of the patients contributed little to neutralization and ADE. However, the EDIIIreactive antibodies from the rabbit antiserum exhibited protective abilities of neutralization at a high dilution (~1:50,000) and ADE at a low dilution (~1:5,000) for the homotypic DENV infection. Notably, the rabbit antiserum displayed ADE activity only at a dilution of 1:40 for the heterotypic virus infection, which suggests that EDIIIreactive antibodies may be safe in secondary infection with heterotypic viruses. These results suggest that DENV EDIII is not the predominant antigen of the DENV infection process; however, purified or recombinant DENV EDIII may be used as a subunit vaccine to provoke an effective and safe antibody response.
Asunto(s)
Anticuerpos Antivirales/inmunología , Virus del Dengue/inmunología , Dengue/inmunología , Sueros Inmunes/inmunología , Dominios Proteicos/inmunología , Proteínas del Envoltorio Viral/inmunología , Animales , Anticuerpos Neutralizantes/inmunología , Anticuerpos Antivirales/sangre , Dengue/sangre , Virus del Dengue/clasificación , Ensayo de Inmunoadsorción Enzimática , Humanos , Pruebas de Neutralización , Unión Proteica/inmunología , Conejos , Serogrupo , Proteínas del Envoltorio Viral/químicaRESUMEN
Dopamine plays crucial roles in a broad spectrum of brain functions, and neural circuit mechanisms underlying dopaminergic regulation have been intensively studied in the past decade. As larval zebrafish have relatively simple and highly conserved dopaminergic systems, it can serve as an ideal vertebrate animal model to tackle this issue at a whole-brain scale. For this purpose, it is important to develop methods for monitoring endogenous dopamine release in intact larval zebrafish. Here, we developed a real-time method to monitor dopamine release at high spatiotemporal resolution in the brain of awake larval zebrafish using carbon fiber microelectrodes. As an example for application, we combined this method with genetic tools and in vivo calcium imaging and found that food extract can activate pretectal dopaminergic neurons, which in turn release dopamine at the visual center through their projection, providing a dopaminergic circuit mechanism for olfactory modulation of visual functions. Thus, our study demonstrates, for the first time, the utility of carbon fiber microelectrodes for monitoring sensory-evoked dopamine release in the brain of an awake small organism. SIGNIFICANCE STATEMENT: With carbon fiber microelectrodes, we have succeeded in monitoring sensory-evoked dopamine release in the brain of an awake small organism for the first time. By elucidating the circuitry origin of the dopamine release, we illustrated the potential application of this method in dissection of the neural circuitry mechanisms underlying dopaminergic neuromodulation.
Asunto(s)
Dopamina/metabolismo , Olfato/fisiología , Vigilia/fisiología , Animales , Animales Modificados Genéticamente , Calcio/metabolismo , Proteínas de Transporte de Dopamina a través de la Membrana Plasmática/genética , Proteínas de Transporte de Dopamina a través de la Membrana Plasmática/metabolismo , Neuronas Dopaminérgicas/fisiología , Electroquímica , Equidae , Proteínas Fluorescentes Verdes/genética , Proteínas Fluorescentes Verdes/metabolismo , Larva , Terapia por Láser , Microelectrodos , OptogenéticaRESUMEN
Dengue virus (DENV) is a mosquito-borne virus that causes severe health problems. An effective tetravalent dengue vaccine candidate that can provide life-long protection simultaneously against all four DENV serotypes is highly anticipated. A better understanding of the antibody response to DENV envelope protein domain III (EDIII) may offer insights into vaccine development. Here, we identified 25 DENV cross-reactive mAbs from immunization with Pichia pastoris-expressed EDIII of a single or all four serotype(s) using a prime-boost protocol, and through pepscan analysis found that 60â% of them (15/25) specifically recognized the same highly conserved linear epitope aa 309-320 of EDIII. All 15 complex-reactive mAbs exhibited significant cross-reactivity with recombinant EDIII from all DENV serotypes and also with C6/36 cells infected with DENV-1, -2, -3 and -4. However, neutralization assays indicated that the majority of these 15 mAbs were either moderately or weakly neutralizing. Through further epitope mapping by yeast surface display, two residues in the AB loop, Q316 and H317, were discovered to be critical. Three-dimensional modelling analysis suggests that this epitope is surface exposed on EDIII but less accessible on the surface of the E protein dimer and trimer, especially on the surface of the mature virion. It is concluded that EDIII as an immunogen may elicit cross-reactive mAbs toward an epitope that is not exposed on the virion surface, therefore contributing inefficiently to the mAbs neutralization potency. Therefore, the prime-boost strategy of EDIII from a single serotype or four serotypes mainly elicited a poorly neutralizing, cross-reactive antibody response to the conserved AB loop of EDIII.
Asunto(s)
Anticuerpos Neutralizantes/inmunología , Anticuerpos Antivirales/inmunología , Vacunas contra el Dengue/inmunología , Virus del Dengue/inmunología , Proteínas del Envoltorio Viral/inmunología , Secuencia de Aminoácidos , Sustitución de Aminoácidos , Anticuerpos Monoclonales/inmunología , Reacciones Cruzadas , Vacunas contra el Dengue/química , Virus del Dengue/metabolismo , Mapeo Epitopo , Epítopos/química , Epítopos/inmunología , Modelos Moleculares , Pichia/metabolismo , Estructura Terciaria de Proteína , Proteínas del Envoltorio Viral/química , Proteínas del Envoltorio Viral/metabolismoRESUMEN
The risk of antibody-dependent enhancement (ADE) of dengue virus (DENV) infection is a major obstacle for the development of dengue vaccine candidates. Here, we described a novel approach for assessment of ADE by measuring DENV nonstructural protein 1 (NS1) production in culture supernatants with Fcγ receptor-expressing K562 cells in ELISA format (ELISA-ADE). Enhancing activities quantified by measurement of kinetics of NS1 production were in a good agreement with the results of the virus titration assay. In conjunction with the previously established enzyme-linked immunospot-based micro-neutralization test (ELISPOT-MNT) in 96-well format, the observable dose-response profiles of enhancing and neutralizing activities against all four DENV serotypes were produced with two flaviviral envelope cross-reactive monoclonal antibodies and four primary DENV-1-infected human sera. The simple high-throughput ELISA-ADE assay offers advantages for quantitative measurement of infection enhancement that can potentially be applied to large-scale seroepidemiological studies of DENV infection and vaccination.
Asunto(s)
Acrecentamiento Dependiente de Anticuerpo , Virus del Dengue/fisiología , Dengue/inmunología , Dengue/virología , Ensayo de Inmunoadsorción Enzimática/métodos , Ensayos Analíticos de Alto Rendimiento/métodos , Anticuerpos Neutralizantes/inmunología , Anticuerpos Antivirales/inmunología , Dengue/diagnóstico , Virus del Dengue/clasificación , Virus del Dengue/inmunología , Humanos , Proteínas no Estructurales Virales/inmunologíaRESUMEN
Studying neural circuits is a crucial step for understanding neural mechanisms underlying animal behaviors. Larval zebrafish is a low vertebrate animal model with incomparable advantages in neural circuit study. In this review, we describe the zebrafish visual system and its downstream targets, with special emphasis on their possible roles in prey capture behavior. Prey capture is executed mainly through the visual system and its downstream circuits, including reticulospinal commanding neurons, motor-controlling circuits within the spinal cord, and other un-identified functional units. With the development of approaches in monitoring and manipulating neuronal activity and behavioral assays, we will get deep insights about neural basis for prey capture in near future, which will shed light on elucidating neural circuit mechanisms of behavior.
Asunto(s)
Sistema Nervioso Central/fisiología , Conducta Predatoria/fisiología , Percepción Visual/fisiología , Pez Cebra/fisiología , Animales , Larva/fisiología , Movimiento/fisiologíaRESUMEN
Porcine reproductive and respiratory syndrome (PRRS) is considered to be one of the most important infectious diseases affecting livestock. This study used gene sequence analysis of ORF5 and Nsp2 to determine the molecular epidemiology of PRRSV in different parts of the Guangxi province of China. These genes were selected due to their extensive variation within the genome. Out of 189 samples from animals suspected to have PRRS, 145 were PRRSV RNA positive. ORF5 and Nsp2 gene sequence analysis of 31 of these samples showed that all of the Guangxi isolates were of type 2. A phylogenetic tree analysis based on ORF5 showed that the Guangxi isolates were divided into two groups. Most of these were closely related to highly pathogenic strains, showing a 30 amino acid deletion at positions 481 and 533-561 of Nsp2, but an additional unique isolate (GXNN06) possessed a further four amino acid deletion at positions 485-488 of Nsp2.
Asunto(s)
Síndrome Respiratorio y de la Reproducción Porcina/epidemiología , Virus del Síndrome Respiratorio y Reproductivo Porcino/clasificación , Virus del Síndrome Respiratorio y Reproductivo Porcino/genética , Animales , China/epidemiología , Análisis por Conglomerados , Genotipo , Epidemiología Molecular , Datos de Secuencia Molecular , Filogenia , Polimorfismo Genético , Síndrome Respiratorio y de la Reproducción Porcina/virología , Virus del Síndrome Respiratorio y Reproductivo Porcino/aislamiento & purificación , ARN Viral/genética , Análisis de Secuencia de ADN , Eliminación de Secuencia , Porcinos , Proteínas Virales/genéticaRESUMEN
Visual cues often modulate auditory signal processing, leading to improved sound detection. However, the synaptic and circuit mechanism underlying this cross-modal modulation remains poorly understood. Using larval zebrafish, we first established a cross-modal behavioral paradigm in which a preceding flash enhances sound-evoked escape behavior, which is known to be executed through auditory afferents (VIII(th) nerves) and command-like neurons (Mauthner cells). In vivo recording revealed that the visual enhancement of auditory escape is achieved by increasing sound-evoked Mauthner cell responses. This increase in Mauthner cell responses is accounted for by the increase in the signal-to-noise ratio of sound-evoked VIII(th) nerve spiking and efficacy of VIII(th) nerve-Mauthner cell synapses. Furthermore, the visual enhancement of Mauthner cell response and escape behavior requires light-responsive dopaminergic neurons in the caudal hypothalamus and D1 dopamine receptor activation. Our findings illustrate a cooperative neural mechanism for visual modulation of audiomotor processing that involves dopaminergic neuromodulation.
Asunto(s)
Comunicación Celular/fisiología , Neuronas Dopaminérgicas/fisiología , Reacción de Fuga/fisiología , Hipotálamo/citología , Locomoción/fisiología , Vías Visuales/fisiología , 6-Ciano 7-nitroquinoxalina 2,3-diona/farmacología , Estimulación Acústica/efectos adversos , Potenciales de Acción/efectos de los fármacos , Potenciales de Acción/fisiología , Animales , Apomorfina/farmacología , Conducta Animal , Benzazepinas/farmacología , Biotina/análogos & derivados , Biotina/metabolismo , Comunicación Celular/efectos de los fármacos , Agonistas de Dopamina/farmacología , Antagonistas de Dopamina/farmacología , Neuronas Dopaminérgicas/efectos de los fármacos , Reacción de Fuga/efectos de los fármacos , Agonistas de Aminoácidos Excitadores/farmacología , Potenciales Postsinápticos Excitadores/efectos de los fármacos , Ácido Flufenámico/farmacología , Lateralidad Funcional , Ácido Glicirretínico/farmacología , Técnicas In Vitro , Larva , Luz , Microscopía Confocal , Morfolinos/farmacología , Técnicas de Placa-Clamp , Estimulación Luminosa/métodos , Psicoacústica , Receptores de Dopamina D1/fisiología , Relación Señal-Ruido , Factores de Tiempo , Valina/análogos & derivados , Valina/farmacología , Vías Visuales/efectos de los fármacos , Pez CebraRESUMEN
BACKGROUND: Classical swine fever virus (CSFV) can evade the immune response and establish chronic infection under natural and experimental conditions. Some genes related to antigen processing and presentation and to cytokine regulation are known to be involved in this response, but the precise mechanism through which each gene responds to CSFV infection remains unclear. RESULTS: In this study, the amplification standard curve and corresponding linear regression equations for the genes SLA-2, TAP1, SLA-DR, Ii, CD40, CD80, CD86, IFN-α, and IFN-ß were established successfully. Real-time RT-PCR was used to quantify the immune response gene transcription in PK-15 cells post CSFV infection. Results showed that: (1) immune response genes were generally down-regulated as a result of CSFV infection, and (2) the expression of SLA-2, SLA-DR, Ii and CD80 was significantly decreased (p < 0.001). CONCLUSION: We conclude that in vitro infection with CSFV inhibits the transcription of host immune response genes. These findings may facilitate the development of effective strategies for controlling CSF.
Asunto(s)
Virus de la Fiebre Porcina Clásica/inmunología , Genes MHC Clase II , Interacciones Huésped-Patógeno , Transcripción Genética , Animales , Línea Celular , Perfilación de la Expresión Génica , Evasión Inmune , Tolerancia Inmunológica , Reacción en Cadena en Tiempo Real de la Polimerasa , PorcinosRESUMEN
AIM OF THE STUDY: Reperfusion following cerebral ischemia leads to excessive production of reactive oxygen species (ROS) and consumption of endogenous antioxidants. Antioxidant enzymes are considered to have beneficial effects against various diseases mediated by ROS. Copper, zinc-superoxide dismutase (SOD1) is one of the major defensive mechanisms by which cells counteract the deleterious effects of ROS after ischemia. However, exogenous SOD1 can not be delivered into living cells because of the poor permeability and selectivity of the cell membrane, thus its application for protecting cells/tissues from oxidative stress damage is greatly limited. METHODS: The purified SOD1 or PEP-1-SOD1 fusion proteins were injected into rats via their tail veins, the transduction ability of PEP-1-SOD1 was examined with immunofluorescent staining and SOD1 activity was measured. Moreover, we determined whether or not PEP-1-SOD1 can protect brain from ischemic injury in an experimental asphyxial cardiac arrest rat model through histopathologic analysis, evaluating the levels of malondialdehyde (MDA), S100ß and neuron specific enolase (NSE). RESULTS: SOD1 protein was observed in PEP-1-SOD1-treated animals and SOD1 activity was significantly increased. However, SOD1 protein was not detected in SOD1-treated animals. The transduced PEP-1-SOD1 significantly attenuated cerebral ischemia-reperfusion damage, inhibited ischemia-induced lipid peroxidation, and protected neurons in hippocampus from the damage induced by transient global ischemic insults. CONCLUSIONS: PEP-1-SOD1 fusion protein can be transduced into the neurons in vivo and protect the neurons from the transient global ischemia-induced damage, suggesting PEP-1-SOD1 may be used for the treatment of oxidative stress-associated disorders such as transient global cerebral ischemia.
Asunto(s)
Isquemia Encefálica/prevención & control , Cisteamina/análogos & derivados , Paro Cardíaco/complicaciones , Péptidos/farmacología , Daño por Reperfusión/prevención & control , Superóxido Dismutasa/farmacología , Análisis de Varianza , Animales , Isquemia Encefálica/etiología , Isquemia Encefálica/fisiopatología , Distribución de Chi-Cuadrado , Cisteamina/administración & dosificación , Cisteamina/metabolismo , Cisteamina/farmacología , Paro Cardíaco/fisiopatología , Peroxidación de Lípido , Masculino , Factores de Crecimiento Nervioso/metabolismo , Estrés Oxidativo , Péptidos/administración & dosificación , Péptidos/metabolismo , Fosfopiruvato Hidratasa/metabolismo , Distribución Aleatoria , Ratas , Ratas Sprague-Dawley , Daño por Reperfusión/etiología , Daño por Reperfusión/fisiopatología , Subunidad beta de la Proteína de Unión al Calcio S100 , Proteínas S100/metabolismo , Superóxido Dismutasa/administración & dosificación , Superóxido Dismutasa/metabolismo , Superóxido Dismutasa-1RESUMEN
OBJECTIVE: To study the effects of CpG oligodeoxyribonucleotide (ODN) as adjuvant on the immune responses in PBMC and CD4+ T cell with chronic hepatitis B virus. METHODS: The selected 20 infections were averagely divided two groups. The frequency of IFN-gamma secreting PBMC and CD4+ T cell in immune tolerant phase and in the immune clearance phase that had stimulated by CpG ODN, HBsAg and Mixture [CpG ODN + HBsAg] were analyzed by enzyme linked immune spot (ELISOT). RESULTS: The PBMC and CD4+ T cell were differently incubated by CpG ODN, HBsAg and M [CpG ODN + HBsAg]. The number of IFN-gamma spot differently are 3 +/- 8, 339 +/- 429, 375 +/- 496, 1 +/- 4, 5 +/- 16 and 5 +/- 12; the results of immume tolerance are 3 +/- 8, 361 +/- 153, 375 +/- 276, 0 +/- 2, 2 +/- 2 and 4 +/- 4; but the results of immune clearance are 3 +/- 21, 289 +/- 345, 405 +/- 656, 2 +/- 14, 8 +/- 40 and 7 +/- 30. The IFN-gamma spots statistical analysis of PBMC were differently incubated by HBsAg and M, the total is P = 0.720, The IFN-gamma spots statistical analysis of CD4+ T cell were differently incubated by HBsAg and M, the total is P = 0.890, The IFN-gamma spots statistical analysis of PBMC and CD4+ T cell were differently incubated by M, the total is P = 0.000. CONCLUSIONS: The ability that CpG ODN can not significantly increase the IFN-gamma secreting of PBMC and CD4+ T cell that were incubated by HBsAg to the infection in immune tolerant phase and in the immune clearance phase, but the PBMC outweighed The CD4 T cell.
Asunto(s)
Linfocitos T CD4-Positivos/inmunología , Virus de la Hepatitis B/inmunología , Hepatitis B Crónica/inmunología , Leucocitos Mononucleares/inmunología , Oligodesoxirribonucleótidos/inmunología , Adulto , Células Cultivadas , Femenino , Antígenos de la Hepatitis B/inmunología , Hepatitis B Crónica/virología , Humanos , Interferón gamma/inmunología , Masculino , Persona de Mediana Edad , Adulto JovenRESUMEN
OBJECTIVE: To apply pulse-field gel electrophoresis analysis(PFGE) in analysing a case of food poisoning caused by Vibrio parahaemolyticus. METHODS: PFGE using restriction enzyme Not I was employed in molecular subtyping of thirty strains of V. parahaemolyticus isolated from a case of food poisoning in Guangzhou city and PFGE patterns were analyzed by using BioNumerics Version 4.0 software to perform cluster analysis. Pattern profiles were compared by using the Dice coefficient and unweighted pair group method with arithmetic averages (UPGMA). RESULTS: Thirty strains were of the same type of pulsotype. CONCLUSIONS: Molecular subtyping by PFGE might disclose the epidemiological relationships of the strains from humans, food and the environment, giving a strong molecular epidemiological evidence and a support for the source-tracking of outbreak events.
Asunto(s)
Técnicas de Tipificación Bacteriana/métodos , Electroforesis en Gel de Campo Pulsado/métodos , Enfermedades Transmitidas por los Alimentos/microbiología , Vibrio parahaemolyticus/clasificación , China , Humanos , Vibrio parahaemolyticus/genética , Vibrio parahaemolyticus/aislamiento & purificaciónRESUMEN
OBJECTIVE: To apply pulse-field gel electrophoresis analysis(PFGE) in the analysis of cholera outbreak events and to determine the molecular epidemiological characteristics of Vibrio cholerae ( V. cholerae) isolates. METHODS: PFGE using restriction enzyme Not I was employed in the molecular subtyping of forty-one strains of V. cholerae isolated in cholera outbreak events from 2003 to 2005 in Guangzhou area and PFGE patterns were analyzed by BioNumerics Version 4.0 software to perform cluster analysis. Pattern profiles were compared by utilizing of Dice coefficient and UPGMA(unweighted pair group method with arithmetic averages). Comparison of PFGE typing results was performed with phage-biological typing and pathogenicity-associated genes typing. RESULTS: In cholera outbreak events, PFGE could discriminate epidemiologically related and unrelated strains, having more discriminatory power than phage-biological typing and pathogenicity-associated genes-typing. CONCLUSIONS: Molecular sub-typing by PFGE could disclose the epidemiological relationships of strains from humans and the environment, providing molecular epidemiological evidence and support for the source-tracking of cholera outbreak events.
Asunto(s)
Técnicas de Tipificación Bacteriana/métodos , Cólera/epidemiología , Cólera/microbiología , Brotes de Enfermedades , Vibrio cholerae/genética , Vibrio cholerae/aislamiento & purificación , Electroforesis en Gel de Campo Pulsado , Humanos , Epidemiología Molecular , Vibrio cholerae/clasificaciónRESUMEN
OBJECTIVE: To apply multiplex polymerase chain reaction (MPCR) assay and sequencing in study of the carrying status of four pathogenicity-related genes of Vibrio cholerae (V.cholerae) and the variation of ctxA. METHODS: Primers targeting cholera toxin sub-unit A gene (ctxA), toxin-coregulated pilus gene (tcpA), accessory cholera enterotoxin gene (ace), zonula occludens toxin gene (zot) were designed and the MPCR method was applied to detect the pathogenicity-related genes of 276 strains of V.cholerae isolates. The amplified fragments of ctxA gene were sequenced and the genetic homology of the amplified fragments of ctxA was analyzed. RESULTS: Of the 276 strains of V.cholerae, 93.9% strains from human sources belong to the pathogenicity-related genes type A (ctxA(+)tcpA(+)ace(+)zot(+) type) and 6.1% belong to pathogenicity-related genes type C (ctxA(-)tcpA(-)ace(-)zot(-) type). Type A strains from clinical sources were isolated from patients with mild to severe symptom and carriers, among which 68.5% were isolated from patients with mild symptom and 21.9% from carriers. All 63.6% of type C strains from clinical sources were isolated from patients with mild symptom and 36.4% from carriers. The proportion of type C strains that caused mild symptom was higher than that of type A strains. Of the 78 strains isolated from the environment, 9.0% strains belong to pathogenicity-related type A and 35.9% belong to the pathogenicity-related genes type B (ctxA(-)tcpA(-)ace(+)zot(+) type), while 55.1% belong to pathogenicity-related genes type C. The sequencing results showed little genetic variation among the amplified fragments for ctxA. CONCLUSION: MPCR disclosed the polymorphic status of pathogenicity-related gene patterns in V.cholerae isolates of Guangzhou, providing effective means for further study on evolution of pathogenicity-related genes among V.cholerae isolates from human and environmental sources. This study also offers significant guidance for effective prevention, control and warning against cholera epidemic in local area.