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PURPOSE: Research has examined peer mentorship to understand how it may help people with spinal cord injury (SCI) adapt and thrive. We still lack an in-depth understanding of the perspectives of SCI peer mentors and mentees on their dyadic relationship. This study was to explore the dyadic interactions and relationships between SCI peer mentors and mentees in a peer mentorship program delivered at a rehabilitation center. RESEARCH METHOD: Between 2016 and 2017, we recruited two dyads of peer mentor and mentee with SCI (N = 4). Each participant completed three one-on-one interviews (N = 12). Data were analyzed using a creative nonfiction approach. RESULTS: Three unique dialogical stories were developed. Story 1 (A slow and steady start) described how mentors took a mentee-centered approach in building the relationship. Story 2 (Mentorship and friendship: negotiating the "grey zone") highlighted how mentees and mentors experienced challenges in navigating the boundaries between mentorship and friendship. Story 3 (The "endless" job for mentor) showcased how the relationship could enter a phase in which it could affect mentors' well-being. CONCLUSIONS: The stories highlighted important attributes to the relationships between SCI mentors and mentees. Considerations were suggested for community-based SCI organizations to integrate peer mentorship into rehabilitation settings, including optimizing mentorship introductions and matching, defining mentors' role explicitly, and building support systems for mentors. (PsycInfo Database Record (c) 2024 APA, all rights reserved).
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Mentores , Grupo Associado , Traumatismos da Medula Espinal , Humanos , Traumatismos da Medula Espinal/reabilitação , Traumatismos da Medula Espinal/psicologia , Masculino , Feminino , Mentores/psicologia , Adulto , Pessoa de Meia-Idade , Relações Interpessoais , Tutoria/métodos , Pesquisa QualitativaRESUMO
Introduction: Community-based spinal cord injury (SCI) organizations deliver peer mentorship programs in rehabilitation settings. Little is known on how these programs are delivered through the collaboration between community-based SCI organizations and rehabilitation institutions. This study aimed to identify barriers, facilitators, and collaboration processes within a SCI peer mentorship program provided by a community-based organization at a rehabilitation center. Methods: A qualitative case study design was applied. Seven participants were recruited, including two mentees, two mentors, one program director of the community-based SCI organization, and two healthcare professionals of the rehabilitation center. Each participant completed a one-on-one interview. Data were analyzed inductively and deductively based on the Consolidated Framework for Implementation Research (CFIR). Results: Ten factors were identified to influence the delivery of the peer mentorship program, including nine CFIR constructs. Successful delivery of the program required strong, collaborative inter-professional relationships between health professionals and community organizational staff (e.g., peer mentors) as facilitators; whereas potential cost, minimal patient needs, and limited mentor resources were found to be barriers. Engaging health professionals by initiating communications, reflecting and evaluating the program collectively with health professionals were important collaboration processes for the community-based organization to maintain effective partnership with the rehabilitation center. Discussion: The collaboration processes and strategies to addressing/leveraging the barriers and facilitators may inform evidence-based practice to establish and optimize the delivery of SCI peer mentorship programs in various rehabilitation settings.
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Tutoria , Traumatismos da Medula Espinal , Humanos , Mentores , Técnica Delphi , Grupo AssociadoRESUMO
OBJECTIVE: Peer mentorship is a flagship program utilized by Canadian community-based spinal cord injury (SCI) organizations. Through connecting trained SCI peer mentors with fellow adults with SCI, these programs help adults adapt and thrive following their injury. The objective of this meta-synthesis was to work with SCI community organizations and to identify outcomes of participating in community- or rehabilitation-based peer mentorship programs using an integrated knowledge translation approach. DESIGN: A meta-synthesis of 21 qualitative peer-reviewed studies and 66 community documents was conducted. MAIN OUTCOME MEASURES: A total of 87 outcomes of peer mentorship were identified. RESULTS: The outcomes of peer mentorship were grouped according to six higher-order themes: 1) Independence: enhanced self-sufficiency; 2) Personal growth: positive psychological changes; 3) Activities and participation: greater participation in activities and events; 4) Adaptation: adapting to life with disability; 5) Knowledge: obtaining new information, resources, and opportunities; and 6) Connection: developing and maintaining social relationship. CONCLUSION: The positive nature of the identified outcomes suggests that participating in peer mentorship can promote improved health and quality of life for adults with SCI. Furthermore, the integrated knowledge translation approach helped identify outcomes that were previously not examined within SCI peer mentorship research, thus providing important insight for future research.
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Mentores , Traumatismos da Medula Espinal , Adulto , Canadá , Humanos , Mentores/psicologia , Grupo Associado , Qualidade de VidaRESUMO
STUDY DESIGN: A generic qualitative design. OBJECTIVES: To obtain a deeper understanding of the outcomes of spinal cord injury (SCI) peer mentorship programs delivered by community-based organizations. SETTING: Peer mentorship programs of community-based SCI organizations METHODS: We interviewed 36 individuals who shared their experiences of SCI peer mentorship from the perspective of a peer mentee, peer mentor, or family member of a peer mentee/mentor, or staff of SCI community-based organizations. Interview data were analyzed using an inductive thematic analysis approach. RESULTS: Four overarching themes with sub-themes were identified. (1) Positive outcomes for mentees such as understanding, emotional outlet/psychological support, inspiration/hope, and belonging. (2) Positive outcomes for mentors such as gaining gratitude, confidence, pride, and personal growth. (3) Reciprocity in positive/negative outcomes for mentors and mentees, such as shared learning and a lack of connection. (4) Negative outcomes for mentors such as impact of negativity, emotional toll, and time/energy demands. CONCLUSIONS: Peer mentorship programs delivered by community-based SCI organizations are important, impactful resources for individuals with SCI who engage in these programs. These results provide insights into the variety of positive and negative outcomes linked with these programs.
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Mentores , Traumatismos da Medula Espinal , Canadá , Humanos , Mentores/psicologia , Grupo Associado , Traumatismos da Medula Espinal/psicologiaRESUMO
OBJECTIVE: First, to examine whether participants reported changes in (1) leisure-time physical activity (LTPA) participation and social inclusion variables and (2) well-being outcomes before and after joining a community-based LTPA program for adults with physical disabilities. Second, to explore the longitudinal relationship between LTPA and the other aforementioned outcomes. DESIGN: A double baseline longitudinal design with measurements at 4-6 weeks (baseline 1) and immediately (baseline 2) before and 2 and 4 months after joining the community-based LTPA program. SETTING: Community. PARTICIPANTS: Adults (N=43) with a physical disability who reported no cognitive impairment, were new members of the community-based LTPA program, and spoke English or French. INTERVENTIONS: A community-based physical activity program for adults with physical disabilities. Participants were provided an individualized exercise program and accessed the program at designated times during the week. MAIN OUTCOME MEASURES: Primary: LTPA (LTPA Questionnaire for People with Spinal Cord Injury), participation (Patient-Perceived Participation in Daily Activities Questionnaire), and social inclusion. Secondary: depression severity, self-esteem, resilience, and life satisfaction. RESULTS: After joining the program, participants reported an increase in total LTPA (meanbaseline2, 177.80±211.32; mean2months, 299.31±298.70; mean4months, 288.14±292.14), moderate-to-vigorous LTPA (meanbaseline2, 83.95±123.95; mean2months, 142.00±198.38; mean4months, 163.23±182.08), and participation in health (meanbaseline2, 6.24±1.16; mean2months, 6.58±1.25; mean4months, 6.97±0.82) and family-related activities (meanbaseline2, 12.18±2.43; mean2months, 12.60±2.30; mean4months, 13.47±2.01). A significant increase (ß=3.46, P<.001) in social inclusion before joining the program was followed by a decrease (ß=-1.09, P<.05) 4 months later. Improvements related to depression severity were noted (ßbaseline1-baseline2=-1.51, P<.05; ßbaseline2-4 months=-0.28, P>.05). CONCLUSIONS: The results support the role of a community-based LTPA program in increasing LTPA levels and enhancing participation in some activities among adults with physical disabilities.
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Pessoas com Deficiência/psicologia , Pessoas com Deficiência/reabilitação , Exercício Físico , Atividades de Lazer , Inclusão Social , Adulto , Idoso , Feminino , Humanos , Estudos Longitudinais , Masculino , Pessoa de Meia-Idade , Inquéritos e QuestionáriosRESUMO
STUDY DESIGN: Generic qualitative design. OBJECTIVES: To explore how Chinese adults living with spinal cord injury (SCI) viewed the prospect of inpatient peer support programs within a rehabilitation setting. SETTING: Hospital in China. METHODS: A purposive sample of adult inpatients with SCI (N = 6) currently undergoing rehabilitation was recruited. Each participant was interviewed twice. Twelve interview transcripts were analyzed using a thematic method. RESULTS: Five higher-order themes were developed. First, participants had unique backgrounds and personal lives before and after their SCI and reported frustrations about their lives resulting from their SCI. Second, participants reported varying degrees of satisfaction with their rehabilitation and identified the facilitators and barriers to their rehabilitation. Third, their perspectives on peer support were shaped by their rehabilitation goals. For example, participants who solely focused on the recovery of physical functioning noted that peers could help to supplement existing rehabilitation programming by guiding their rehabilitation exercises. Participants who concentrated on their future lives believed peers could teach them new skills to facilitate their integration in the community. However, some participants felt they could not trust peers' advice because peers are not healthcare providers. Fourth, peer support delivery options varied from online chat groups (i.e., WeChat), in-person conversations, and mentoring lectures. Finally, anticipated outcomes were related to obtaining practical and emotional support from peers, being motivated, and feeling understood. CONCLUSIONS: Participants harbored mixed views on potential use-value and necessity of hospital-based peer support programs, which could inform future utilization of SCI peer support within Chinese hospitals.
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Pacientes Internados , Traumatismos da Medula Espinal , Adulto , China , Hospitais , Humanos , Grupo AssociadoRESUMO
Human infection with H7N9 virus has provoked global public health concern due to the substantial morbidity and mortality. Neuraminidase inhibitors (NAIs) are used as first-line drugs to treat the infection. However, virus quasispecies can evolve rapidly under drug pressure, which may alter various biological characteristics of virus. Using an in vitro evolution platform and next-generation sequencing, we found the presence of peramivir led to changes to the dominant populations of the virus. Two important amino acid substitutions were identified in NA, I222T and H274Y, which caused reduced susceptibilities to oseltamivir or both oseltamivir and peramivir as confirmed by enzyme- and cell-based assays. The NA-H274Y variant showed decreased replicative fitness at the early stage of infection accompanied with impaired NA function. The quasispecies evolution of H7N9 virus and the potential emergence of these two variants should be closely monitored, which may guide the adjustment of antiviral strategies.
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Antivirais/farmacologia , Ciclopentanos/farmacologia , Farmacorresistência Viral/genética , Guanidinas/farmacologia , Subtipo H7N9 do Vírus da Influenza A/efeitos dos fármacos , Neuraminidase/genética , Proteínas Virais/genética , Ácidos Carbocíclicos , Substituição de Aminoácidos , Animais , Cães , Evolução Molecular , Expressão Gênica , Humanos , Subtipo H7N9 do Vírus da Influenza A/enzimologia , Subtipo H7N9 do Vírus da Influenza A/genética , Subtipo H7N9 do Vírus da Influenza A/crescimento & desenvolvimento , Influenza Humana/virologia , Células Madin Darby de Rim Canino , Neuraminidase/metabolismo , Oseltamivir/farmacologia , Carga Viral/efeitos dos fármacos , Proteínas Virais/metabolismo , Replicação Viral/efeitos dos fármacosRESUMO
Metagenomic analysis through high-throughput sequencing is a tool for detecting both known and novel viruses. Using this technique, a novel circular single-stranded DNA (ssDNA) virus genome was discovered in respiratory secretions from a febrile traveler. The virus, named human respiratory-associated PSCV-5-like virus (HRAPLV), has a genome comprising 3,018 bases, with two major putative ORFs inversely encoding capsid (Cap) and replicase (Rep) protein and separated by two intergenic regions. One stem-loop structure was predicted in the larger intergenic region (LIR). The predicted amino acid sequences of the Cap and Rep proteins of HRAPLV showed highest identity to those of porcine stool-associated circular virus 5 isolate CP3 (PoSCV 5) (53.0% and 48.9%, respectively). The host tropism of the virus is unknown, and further study is warranted to determine whether this novel virus is associated with human disease.
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Circovirus/genética , Circovirus/isolamento & purificação , DNA Circular/genética , DNA de Cadeia Simples/genética , DNA Viral/genética , Faringe/virologia , Genoma Viral , Humanos , Masculino , Pessoa de Meia-Idade , FilogeniaRESUMO
Objective To express and purify two kinds of antigens of Coxiella burnetii (C. burnetii), the main outer membrane protein Com1 and the acute disease antigen A (adaA), in prokaryotic expression system and to validate the two recombinant antigens by mass spectrometry and identify their antigenicity. Methods The gene sequences encoding Com1 and adaA were separately synthesized and constructed into the prokaryotic expression vector pET-20b(+). The constructed vectors were transformed into E.coli BL21(DE3), and the recombinant proteins were induced by IPTG. The recombinant Com1 and adaA were purified by His affinity chromatography and identified by mass spectrometry. The immunoreactivity of the two antigens was identified by Western blot analysis using Q fever positive bovine serum. Results The expression vectors pET-20b(+)-Com1 and pET-20b(+)-adaA were constructed and the recombinant Com1 and adaA were expressed and purified in a soluble form. High-purity recombinant Com1 and adaA were obtained after purification, and the SDS-PAGE showed that their relative molecular masses were Mr 27 000 and 25 000, respectively. The mass spectrometry confirmed the recombinant proteins were Com1 and adaA of C. burnetii. Both of the recombinant Com1 and adaA were able to react with the Q fever positive bovine serum in Western blotting, and the corresponding bands were in accordance with the SDS-PAGE. Conclusion We obtained high-purity Com1 and adaA in a soluble form and confirmed their immunoreactivity.
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Antígenos de Bactérias/isolamento & purificação , Proteínas da Membrana Bacteriana Externa/isolamento & purificação , Coxiella burnetii/imunologia , Animais , Antígenos de Bactérias/genética , Antígenos de Bactérias/imunologia , Proteínas da Membrana Bacteriana Externa/genética , Proteínas da Membrana Bacteriana Externa/imunologia , Bovinos , Escherichia coli/genética , Peso Molecular , Proteínas Recombinantes/isolamento & purificaçãoRESUMO
Influenza virus infections represent a worldwide public health and economic problem due to the significant morbidity and mortality caused by seasonal epidemics and pandemics. Sensitive and convenient methodologies for detection of influenza viruses are essential for further disease control. Loop-mediated isothermal amplification (LAMP) is the most commonly used method of nucleic acid isothermal amplification. However, with regard to multiplex LAMP, differentiating the ladder-like LAMP products derived from multiple targets is still challenging today. The requirement of specialized instruments has further hindered the on-site application of multiplex LAMP. We have developed an integrated assay coupling multiplex reverse transcription LAMP with cascade invasive reaction using nanoparticles (mRT-LAMP-CIRN) as a sensor for the detection of three subtypes of influenza viruses: A/H1N1pdm09, A/H3 and influenza B. The analytic sensitivities of the mRT-LAMP-CIRN assay were 101 copies of RNA for both A/H1N1pdm09 and A/H3, and 102 copies of RNA for influenza B. This assay demonstrated highly specific detection of target viruses and could differentiate them from other genetically or clinically related viruses. Clinical specimen analysis showed the mRT-LAMP-CIRN assay had an overall sensitivity and specificity of 98.3% and 100%, respectively. In summary, the mRT-LAMP-CIRN assay is highly sensitive and specific, and can be used as a cost-saving and instrument-free method for the detection of influenza viruses, especially for on-site use.
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Técnicas de Amplificação de Ácido Nucleico/métodos , Infecções por Orthomyxoviridae/virologia , Orthomyxoviridae/genética , Humanos , Influenza Humana/virologia , Nanopartículas , Técnicas de Amplificação de Ácido Nucleico/instrumentação , Orthomyxoviridae/isolamento & purificação , Transcrição Reversa , Sensibilidade e EspecificidadeRESUMO
Considering the fatal human victims and economic loss caused by influenza virus infection every year, methodologies for rapid and on-site detection of influenza viruses are urgently needed. LAMP is the most commonly used nucleic acid isothermal amplification technology suitable for on-site use. However, for multiplex LAMP, differentiation of the amplicons derived from multiple targets is still challengeable currently. Here we developed a multiplex RT-LAMP assay for simultaneous amplification of three prominent subtypes of influenza viruses (A/H5, A/H7 and 2009A/H1). The amplicons were further identified by cascade invasive reaction and nanoparticle hybridization in separate target-specific detection tubes (referred to as mRT-LAMP-IRNH). The analytic sensitivities of the assay are 10 copies of RNA for all the three HA subtypes, and the specificity reached 100%. Clinical specimen analysis showed this assay had a combined sensitivity and specificity of 98.1% and 100%, respectively. Overall, the mRT-LAMP-IRNH assay can be used as a cost-saving method that utilizes a simple instrument to detect A/H5, A/H7, and 2009A/H1 influenza viruses, especially in resource-limited settings.
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Vírus da Influenza A/classificação , Vírus da Influenza A/genética , Influenza Humana/virologia , Nanopartículas , Técnicas de Amplificação de Ácido Nucleico , Hibridização de Ácido Nucleico , Transcrição Reversa , Ouro , Humanos , Influenza Humana/diagnóstico , Técnicas de Amplificação de Ácido Nucleico/métodos , RNA Viral , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
Hand, foot, and mouth disease (HFMD), mainly caused by coxsackievirus A16 (CVA16) and enterovirus 71 (EV71) infections, remains a serious public health issue with thousands of newly diagnostic cases each year since 2008 in China. The mechanisms underlying viral infection, however, are elusive to date. In the present study, we systematically investigated the host cellular microRNA (miRNA) expression patterns in response to CVA16 and EV71 infections. Through microarray examination, 27 miRNAs (15 upregulated and 12 downregulated) were found to be coassociated with the replication process of two viruses, while the expression levels of 15 and 5 miRNAs were significantly changed in CVA16- and EV71-infected cells, respectively. A great number of target genes of 27 common differentially expressed miRNAs were predicted by combined use of two computational target prediction algorithms, TargetScan and MiRanda. Comprehensive bioinformatic analysis of target genes in GO categories and KEGG pathways indicated the involvement of diverse biological functions and signaling pathways during viral infection. These results provide an overview of the roles of miRNAs in virus-host interaction, which will contribute to further understanding of HFMD pathological mechanisms.
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Enterovirus Humano A/patogenicidade , Enterovirus/patogenicidade , Doença de Mão, Pé e Boca/genética , Doença de Mão, Pé e Boca/virologia , MicroRNAs/genética , Algoritmos , Linhagem Celular , China , Biologia Computacional , Enterovirus/fisiologia , Enterovirus Humano A/fisiologia , Ontologia Genética , Interações Hospedeiro-Patógeno/genética , Humanos , Reação em Cadeia da Polimerase em Tempo Real , Transcriptoma , Replicação Viral/genéticaRESUMO
We develop color code-based pyrosequencing with di-base addition for analysis of single nucleotide polymorphisms (SNPs). When a di-base is added into the polymerization, one or several two-color code(s) containing the type and the number of incorporated nucleotides will be produced. The code information obtained in a single run is useful to genotype SNPs as each allelic variant will give a specific pattern compared to the two other variants. Special care has to be taken while designing the di-base dispensation order. Here, we present a detailed protocol for establishing sequence-specific di-base addition to avoid nonsynchronous extension at the SNP sites. By using this technology, as few as 50 copies of DNA templates were accurately sequenced. Higher signals were produced and thus a relatively lower sample amount was required. Furthermore, the read length of per flow was increased, making simultaneous identification of multiple SNPs in a single sequencing run possible. Validation of the method was performed by using templates with two SNPs covering 37 bp and with three SNPs covering 58 bp as well as 82 bp. These SNPs were successfully genotyped by using only a sequencing primer in a single PCR/sequencing run. Our results demonstrated that this technology could be potentially developed into a powerful methodology to accurately determine SNPs so as to diagnose clinical settings.
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Genótipo , Polimorfismo de Nucleotídeo Único , Análise de Sequência de DNA/métodos , HumanosRESUMO
BACKGROUND: Sarcopenia is a common condition in older people. The aim of the present study was to examine the prevalence and factors associated with sarcopenia in an elderly Chinese suburb-dwelling population. METHODS: This study was conducted on 1,069 Chinese suburb-dwelling participants aged ≥60 years to evaluate sarcopenia using the Asian Working Group for Sarcopenia criteria. Sociodemographic and behavioral characteristics, as well as medical conditions, were considered independent variables to determine factors associated with sarcopenia using a logistic regression model. RESULTS: The prevalence of sarcopenia was 6.4% in men and 11.5% in women. Age was a significant factor in both sexes. In addition, presence of sarcopenia was inversely associated with BMI for both sexes. The odds ration and 95% confidence interval for factors statistically significantly associated with sarcopenia were 5.04 (1.70-14.89) and 2.36 (1.06-5.25) for diabetes in males and females, respectively; 10.60 (1.75-64.24) for daily consumption of alcohol (daily drinkers), 5.58 (2.13-14.59) for peptic ulcer in female (not statistically significant in males). CONCLUSIONS: The Asian Working Group for Sarcopenia criterion is useful for defining sarcopenia, and our data suggest that the prevalence of sarcopenia in the general elderly suburb-dwelling Chinese population is high. Moreover, we find that high body mass index is inversely associated with the likelihood of being sarcopenic and that several others factors such as diabetes, peptic ulcer, and drinking habits increase the prevalence of sarcopenia.
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Sarcopenia/epidemiologia , Fatores Etários , Idoso , Idoso de 80 Anos ou mais , Índice de Massa Corporal , China/epidemiologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Prevalência , Fatores de Risco , Fatores Sexuais , População SuburbanaRESUMO
BACKGROUND: Severe fever with thrombocytopenia syndrome virus (SFTSV), the causative agent for the fatal life-threatening infectious disease, severe fever with thrombocytopenia syndrome (SFTS), was first identified in the central and eastern regions of China. Although the viral RNA was detected in free-living and parasitic ticks, the vector for SFTSV remains unsettled. METHODOLOGY/PRINCIPAL FINDINGS: Firstly, an experimental infection study in goats was conducted in a bio-safety level-2 (BSL-2) facility to investigate virus transmission between animals. The results showed that infected animals did not shed virus to the outside through respiratory or digestive tract route, and the control animals did not get infected. Then, a natural infection study was carried out in the SFTSV endemic region. A cohort of naïve goats was used as sentinel animals in the study site. A variety of daily samples including goat sera, ticks and mosquitoes were collected for viral RNA and antibody (from serum only) detection, and virus isolation. We detected viral RNA from free-living and parasitic ticks rather than mosquitoes, and from goats after ticks' infestation. We also observed sero-conversion in all members of the animal cohort subsequently. The S segment sequences of the two recovered viral isolates from one infected goat and its parasitic ticks showed a 100% homology at the nucleic acid level. CONCLUSIONS/SIGNIFICANCE: In our natural infection study, close contact between goats does not appear to transmit SFTSV, however, the naïve animals were infected after ticks' infestation and two viral isolates derived from an infected goat and its parasitic ticks shared 100% of sequence identity. These data demonstrate that the etiologic agent for goat cohort's natural infection comes from environmental factors. Of these, ticks, especially the predominant species Haemaphysalis longicornis, probably act as vector for this pathogen. The findings in this study may help local health authorities formulate and focus preventive measures to contain this infection.
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Vetores Aracnídeos , Infecções por Bunyaviridae/veterinária , Doenças das Cabras/etiologia , Phlebovirus/isolamento & purificação , Carrapatos/virologia , Animais , Cabras , RNA Viral/análiseRESUMO
In April 2013, human infections with a novel avian influenza (H7N9) virus emerged in China. It has caused serious concerns for public health throughout the world. However, there is presently no effective treatment, and an A (H7N9) H7 subtype influenza vaccine is not available. Vaccination with virus-like particles (VLPs) has showed considerable promise for many other subtype influenza viruses. To produce H7N9 VLPs, full length, unmodified hemagglutinin (HA), neuraminidase (NA), and matrix1 (M1) genes from the A/Wuxi/1/2013(H7N9) were cloned into a pCDNA5.1 FRT vector. By co-transfection, VLPs containing HA, NA, and M1 were secreted by 293T cells. VLPs were purified by ultracentrifugation and injected into mice by the intramuscular route. In animal experiments, humoral and cellular immunoresponse were all triggered by H7N9 VLPs. High levels of specific antibodies and the isotypes of IgG were detected by ELISA. Anamnestic cellular immune responses were examined by detecting specific cytotoxic T cell for IFN-Υ production in ELISPOT assay. The hemagglutination-inhibition (HAI) against the homologous virus was more than 1:64, and cross-reactive HAI titers against the heterologous virus (H1N1 and H3N2) were more than 1:16. Moreover, VLPs immunized mice showed a rapid increase of neutralizing antibodies, with neutralizing antibody titers more than 1:8, which increased four-fold against PBS immunized mice in week four. By week six, the mice had high neutralization ability against the given strain and held a potent homologous virus neutralizing capacity. Thus, VLPs represent a potential strategy for the development of a safe and effective vaccine against novel avian influenza (H7N9) virus.
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Anticorpos Antivirais/sangue , Subtipo H7N9 do Vírus da Influenza A/imunologia , Vacinas contra Influenza/imunologia , Vacinas de Partículas Semelhantes a Vírus/imunologia , Animais , Anticorpos Neutralizantes/sangue , Linhagem Celular , Ensaio de Imunoadsorção Enzimática , Feminino , Testes de Inibição da Hemaglutinação , Glicoproteínas de Hemaglutininação de Vírus da Influenza/genética , Glicoproteínas de Hemaglutininação de Vírus da Influenza/imunologia , Humanos , Imunoglobulina G/sangue , Subtipo H7N9 do Vírus da Influenza A/genética , Vacinas contra Influenza/administração & dosagem , Vacinas contra Influenza/genética , Vacinas contra Influenza/isolamento & purificação , Injeções Intramusculares , Interferon gama/metabolismo , Camundongos Endogâmicos BALB C , Neuraminidase/genética , Neuraminidase/imunologia , Testes de Neutralização , Linfócitos T Citotóxicos/imunologia , Vacinas de Partículas Semelhantes a Vírus/administração & dosagem , Vacinas de Partículas Semelhantes a Vírus/genética , Vacinas de Partículas Semelhantes a Vírus/isolamento & purificação , Proteínas da Matriz Viral/genética , Proteínas da Matriz Viral/imunologia , Proteínas Virais/genética , Proteínas Virais/imunologiaRESUMO
The genome sequence of the strain A/chicken/Changzhou/C08/2013 (H9N9) shows that the hemagglutinin (HA) genes of this strain are closely related to those of the strain A/chicken/Shanghai/1107/2013 (H9N2) and share 99.2% nucleotide homology, while the other seven genes had the greatest sequence identities with the novel H7N9 virus. We speculate that this strain may be a novel natural reassortant avian influenza virus (AIV).
RESUMO
OBJECTIVE: To establish a rapid eukaryotic expression system of hemagglutinin (HA) gene of novel avian influenza H7N9 using lentiviral vector, express the recombinant protein and study its functions in human embryonic kidney HEK293T cells. METHODS: The full-length HA gene was amplified from H7N9 genomic RNA by reverse transcription PCR (RT-PCR) and linked with pMD18-T vector to generate pMD18-T-HA plasmid. Blunt-end HA gene with Kozak sequence was amplified from pMD18-T-HA vector, and then pLenti-HA-V5 expression vector was constructed by Topo cloning for transient expression in HEK293T cells. Expression of HA-V5 recombinant protein was confirmed by immunofluorescence assay (IFA) and Western blotting. Hemagglutination test was performed to evaluate the biological activity of the recombinant protein. RESULTS: The full-length HA gene (1 683 bp) was obtained and eukaryotic expression plasmid was constructed successfully. A recombinant protein with relative molecular mass (Mr) 70 000 was expressed and the antigenicity and binding specificity to positive serum were demonstrated by IFA and Western blotting. The hemagglutination activity was proved by hemagglutination test. IFA and Western blotting showed that the Mr 70 000 recombinant protein had an immuoreactivity to positive serum. The hemagglutination activity was confirmed by hemagglutination test. CONCLUSION: The rapid eukaryotic expression system of HA gene was successfully constructed, which laid a solid foundation for further research on subunit vaccine development, neutralizing epitope mapping and packaging pseudovirus.
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Glicoproteínas de Hemaglutininação de Vírus da Influenza/genética , Subtipo H7N9 do Vírus da Influenza A/genética , Influenza Aviária/virologia , Influenza Humana/virologia , Lentivirus/genética , Animais , Aves , Western Blotting , Técnica Indireta de Fluorescência para Anticorpo , Expressão Gênica , Vetores Genéticos/genética , Células HEK293 , Testes de Hemaglutinação , Glicoproteínas de Hemaglutininação de Vírus da Influenza/imunologia , Glicoproteínas de Hemaglutininação de Vírus da Influenza/metabolismo , Humanos , Subtipo H7N9 do Vírus da Influenza A/imunologia , Subtipo H7N9 do Vírus da Influenza A/metabolismo , Peso Molecular , Proteínas Recombinantes/química , Proteínas Recombinantes/imunologia , Proteínas Recombinantes/metabolismoRESUMO
Circulating microRNAs (miRNAs) may play an important role in pathogen-host interactions and can serve as molecular markers for the detection of infectious diseases. To date, the relationship between circulating miRNAs and varicella-zoster virus (VZV) caused varicella has not been reported. Using TaqMan Low-Density Array (TLDA) analysis, expression levels of miRNAs in serum samples from 29 patients with varicella and 60 patients with Bordetella pertussis (BP), measles virus (MEV) and enterovirus (EV) were analyzed. The array results showed that 247 miRNAs were differentially expressed in sera of the varicella patients compared with healthy controls (215 up-regulated and 32 down-regulated). Through the following qRT-PCR confirmation and receiver operational characteristic (ROC) curve analysis, five miRNAs (miR-197, miR-629, miR-363, miR-132 and miR-122) were shown to distinguish varicella patients from healthy controls and other microbial infections with moderate sensitivity and specificity. A number of significantly enriched pathways regulated by these circulating miRNAs were predicted, and some of them were involved in inflammatory response, nervous system and respiratory system development. Our results, for the first time, revealed that a number of miRNAs were differentially expressed during VZV infection, and these five serum miRNAs have great potential to serve as biomarkers for the diagnosis of VZV infection in varicella patients.