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The proportion of human isolates with reduced neuraminidase inhibitors (NAIs) susceptibility in highly pathogenic avian influenza (HPAI) H7N9 virus was high. These drug-resistant strains showed good replication capacity without serious loss of fitness. In the presence of oseltamivir, R229I substitution were found in HA1 region of the HPAI H7N9 virus before NA R292K appeared. HPAI H7N9 or H7N9/PR8 recombinant viruses were developed to study whether HA R229I could increase the fitness of the H7N9 virus bearing NA 292K. Replication efficiency was assessed in MDCK or A549 cells. Neuraminidase enzyme activity and receptor-binding ability were analyzed. Pathogenicity in C57 mice was evaluated. Antigenicity analysis was conducted through a two-way HI test, in which the antiserum was obtained from immunized ferrets. Transcriptomic analysis of MDCK infected with HPAI H7N9 24hpi was done. It turned out that HA R229I substitution from oseltamivir induction in HA1 region increased (1) replication ability in MDCK(P < 0.05) and A549(P < 0.05), (2) neuraminidase enzyme activity, (3) binding ability to both α2,3 and α2,6 receptor, (4) pathogenicity to mice(more weight loss; shorter mean survival day; viral titer in respiratory tract, P < 0.05; Pathological changes in pneumonia), (5) transcriptome response of MDCK, of the H7N9 virus bearing NA 292K. Besides, HA R229I substitution changed the antigenicity of H7N9/PR8 virus (>4-fold difference of HI titre). It indicated that through the fine-tuning of HA-NA balance, R229I increased the fitness and changed the antigenicity of H7N9 virus bearing NA 292K. Public health attention to this mechanism needs to be drawn.
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Antivirales , Subtipo H7N9 del Virus de la Influenza A , Neuraminidasa , Infecciones por Orthomyxoviridae , Oseltamivir , Replicación Viral , Animales , Oseltamivir/farmacología , Subtipo H7N9 del Virus de la Influenza A/genética , Subtipo H7N9 del Virus de la Influenza A/efectos de los fármacos , Subtipo H7N9 del Virus de la Influenza A/patogenicidad , Subtipo H7N9 del Virus de la Influenza A/inmunología , Subtipo H7N9 del Virus de la Influenza A/fisiología , Neuraminidasa/genética , Neuraminidasa/metabolismo , Perros , Replicación Viral/efectos de los fármacos , Antivirales/farmacología , Humanos , Ratones , Infecciones por Orthomyxoviridae/virología , Células de Riñón Canino Madin Darby , Células A549 , Ratones Endogámicos C57BL , Farmacorresistencia Viral/genética , Sustitución de Aminoácidos , Gripe Humana/virología , Hurones , Glicoproteínas Hemaglutininas del Virus de la Influenza/genética , Glicoproteínas Hemaglutininas del Virus de la Influenza/inmunología , Glicoproteínas Hemaglutininas del Virus de la Influenza/metabolismo , Femenino , Proteínas Virales/genética , Proteínas Virales/metabolismoRESUMEN
OBJECTIVE: To evaluate the effect of hydroxychloroquine on conjunctival and retinal microvascular density in rheumatoid arthritis (RA) patients. METHODS: Ten healthy controls, 10 RA patients who had not been treated with hydroxychloroquine, and 10 RA patients who had been treated with chloroquine for more than 5 years were recruited. Optical coherence tomography (OCTA) was used to examine the conjunctival and superficial and deep retinal microvascular density and compared the differences in microvascular density between the three groups. RESULTS: The vascular density in RA group in superficial microvascular was significantly lower than that in control group (p < 0.001). Compared with RA group, the chloroquine group showed statistically significantly lower microvascular (p < 0.001) and deep microvascular (p = 0.018). Superficial microvascular was positively correlated with conjunctival vessel density in RA patients (r = 0.868, p = 0.0048). CONCLUSIONS: The use of chloroquine could further reduce the vascular density in the absence of statistical difference in the course of the disease.
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Artritis Reumatoide , Hidroxicloroquina , Humanos , Hidroxicloroquina/uso terapéutico , Hidroxicloroquina/farmacología , Vasos Retinianos/diagnóstico por imagen , Cloroquina/uso terapéutico , Cloroquina/farmacología , Angiografía con Fluoresceína/métodos , Artritis Reumatoide/diagnóstico por imagen , Artritis Reumatoide/tratamiento farmacológico , Tomografía de Coherencia Óptica/métodosRESUMEN
[This corrects the article DOI: 10.3389/fnagi.2022.877281.].
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PURPOSE: To investigate the altered functional connectivity (FC) of the cerebral hemispheres in patients with morbid obesity (MO) with meibomian gland dysfunction (MGD) by voxel-mirrored homotopic connectivity (VMHC). METHODS: Patients and matched healthy controls (HCs) were recruited, and all subjects underwent functional resonance magnetic imaging (fMRI), and VMHC results were processed statistically to assess the differences in FC in different brain regions between the two groups. We further used ROC curves to evaluate the diagnostic value of these differences. We also used Pearson's correlation analysis to explore the relationship between changes in VMHC values in specific brain regions, visual acuity, and Mini-Mental State Examination (MMSE) score. CONCLUSIONS: Patients with morbid obesity and MGD had abnormal FC in the cerebral hemispheres in several specific brain areas, which were mainly concentrated in pathways related to vision and perception and may correlate to some extent with the clinical presentations of the patients.
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Purpose: To investigate spontaneous brain activity in patients with dry eye (DE) and healthy control (HC) using the fractional amplitude of low frequency fluctuation (fALFF) technique with the aim of elucidating the relationship between the clinical symptoms of DE and changes in brain function. Material and Methods: A total of 28 patients with DE and 28 matched healthy volunteers (10 males and 18 females in each group) were enrolled. Resting-state functional magnetic resonance imaging scans were performed in both groups. Then all subjects were required to complete a comprehensive Hospital Anxiety and Depression Scale (HADS). Receiver operating characteristic (ROC) curve analysis was used to evaluate the differences in fALFF values between the two groups and their diagnostic value. Linear correlations between HADS and fALFF values in different brain regions of DE patients were analyzed using the Pearson correlation coefficient. Results: Patients with DE had significantly higher fALFF values in the left calcarine sulcus (CS) than the HC group, while fALFF values in the bilateral middle frontal gyrus (MFG) and right MFG/right inferior frontal gyrus (IFG) were significantly lower in DE patients than in HC group. fALFF values had a high diagnostic value for differentiating patients with DE from the HC group (P < 0.001). Right MFG and right MFG/IFG were significantly correlated with HADS values. Conclusion: Our study found that DE mainly involved functional disorders in the brain areas of the left CS, bilateral MFG and right MFG/right IFG, which helped us to find possible clinical features of DE disease and reflected the potential pathological mechanism of DE.
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Purpose: To explore alterations in macular retinal thickness (RT) and analyze correlation between macular RT and pterygium area, length in pterygium patients. Methods: Totally 13 patients with pterygium (left eye) and 13 healthy controls (left eye) were recruited. OCTA was applied to scan each eye to generate three-dimensional images. Based on the Early Treatment Diabetic Retinopathy Study (ETDRS) method, each image was divided into nine subregions for the ETDRS: central (C); inner superior (IS); outer superior (OS); inner nasal (IN); outer nasal (ON); inner inferior (II); outer inferior (OI); inner temporal (IT); and outer temporal (OT). The macular RT in each subregion was measured. Furthermore, the correlation between RT and the area, length of pterygium was analyzed. Results: The visual acuity of pterygium patient was different from that of the control (P < 0.05). Besides, decreased intraretinal thickness of the IN and ON, increased intraretinal thickness of OT, decreased extraretinal thickness of OT, IN, ON, OS, and decreased retinal full layer thickness of medial superior, OS, IN, ON, and II subregions in pterygium group were observed. There was a negative correlation between RT of the IN and ON subregions and the length of pterygium (r = -0.5803 and r = -0.6013, P = 0.0376 and P = 0.0297). The RT of IN subregion was negatively correlated with pterygium area (r = -0.5844, P = 0.0359). According to the receiver operating characteristic analysis, in the ON subregion, the areas under the curve of the inner retinal thickness, outer retinal thickness and the whole retinal thickness were 1.0 (95% CI: 1.0), 0.882 (95% CI: 0.715 and 0.963), and 1.0 (95% CI: 1.0). The smallest area under the curve of retinal thickness in OT subregion was 0.018 (95% CI: 0-0.059). Conclusion: RT of pterygium patients was significantly decreased, and the main alterations occurred in the temporal side suggesting there might exist retinal structural alterations in pterygium.
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Objective: To analyze the potential changes in brain neural networks in resting state functional magnetic resonance imaging (rs-fMRI) scans by regional homogeneity (ReHo) in patients with mild cognitive impairment (MCI). Methods: We recruited and selected 24 volunteers, including 12 patients (6 men and 6 women) with MCI and 12 healthy controls matched by age, sex, and lifestyle. All subjects were examined with rs-fMRI to evaluate changes in neural network connectivity, and the data were analyzed by ReHo method. Correlation analysis was used to investigate the relationship between ReHo values and clinical features in different brain regions of MCI patients. The severity of MCI was determined by the Mini-Mental State Examination (MMSE) scale. Results: The signals of the right cerebellum areas 4 and 5, left superior temporal, right superior temporal, left fusiform, and left orbital middle frontal gyri in the patient group were significantly higher than those in the normal group (P < 0.01 by t-test of paired samples). The signal intensity of the right inferior temporal and left inferior temporal gyri was significantly lower than that of the normal group (P < 0.01). The ReHO value for the left inferior temporal gyrus correlated negatively with disease duration, and the value for the right inferior temporal gyrus correlated positively with MMSE scores. Conclusion: Mild cognitive impairment in patients with pre- Alzheimer's disease may be related to the excitation and inhibition of neural networks in these regions. This may have a certain guiding significance for clinical diagnosis.
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Functional connectivity of the primary visual cortex was explored with resting functional magnetic resonance imaging among adults with strabismus and amblyopia and healthy controls. We used the two-sample test and receiver operating characteristic curves to investigate the differences in mean functional connectivity values between the groups with strabismus and amblyopia and healthy controls. Compared with healthy controls, functional connectivity values in the left Brodmann areas 17, including bilateral lingual/angular gyri, were reduced in groups with strabismus and amblyopia. Moreover, functional connectivity values in the right Brodmann area 17, including left cuneus, right inferior occipital gyrus, and left inferior parietal lobule, were reduced in adults with strabismus and amblyopia. Our findings indicate that functional connectivity abnormalities exist between the primary visual cortex and other regions. This may be the basis of the pathological mechanism of visual dysfunction and stereovision disorders in adults with strabismus and amblyopia.
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Ambliopía/fisiopatología , Conectoma , Corteza Visual Primaria/fisiopatología , Estrabismo/fisiopatología , Adulto , Ambliopía/diagnóstico por imagen , Femenino , Humanos , Imagen por Resonancia Magnética , Masculino , Corteza Visual Primaria/diagnóstico por imagen , Estrabismo/diagnóstico por imagen , Adulto JovenRESUMEN
MicroRNA (miRNA) has emerged as a promising genetic marker for cancer diagnosis and therapy because its expression level is closely related to the progression of malignant diseases. Herein, a label-free and selective fluorescence platform was proposed for miRNA based on light-up "G-quadruplex nanostring" via duplex-specific nuclease (DSN) mediated tandem rolling circle amplification (RCA). First, a long DNA generated from upstream RCA was designed with the antisense sequences for miR-21 and downstream RCA primer. Upon recognizing miR-21, the resulting DNA-RNA permitted DSN digestion and triggered downstream two-way RCA, and generation of abundant "G-quadruplex nanostring" binding with ZnPPIX for label-free fluorescent responses. In our strategy, the strong preference of DSN for perfectly matched DNA/RNA ensures its excellent selectivity. The developed method generated wide linear response with LOD of 1.019 fM. Additionally, the miR-21 levels in cell extracts have been evaluated, revealing the utility of this tool for biomedical research and clinical diagnosis.
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Endonucleasas/metabolismo , G-Cuádruplex , MicroARNs/genética , Técnicas de Amplificación de Ácido Nucleico/métodos , Estudios de Factibilidad , Células HeLa , Humanos , MicroARNs/metabolismo , Espectrometría de FluorescenciaRESUMEN
OBJECTIVE: In China, 24 cases of human infection with highly pathogenic avian influenza (HPAI) H5N6 virus have been confirmed since the first confirmed case in 2014. Therefore, we developed and assessed two H5N6 candidate vaccine viruses (CVVs). METHODS: In accordance with the World Health Organization (WHO) recommendations, we constructed two reassortant viruses using reverse genetics (RG) technology to match the two different epidemic H5N6 viruses. We performed complete genome sequencing to determine the genetic stability. We assessed the growth ability of the studied viruses in MDCK cells and conducted a hemagglutination inhibition assay to analyze their antigenicity. Pathogenicity attenuation was also evaluated in vitro and in vivo. RESULTS: The results showed that no mutations occurred in hemagglutinin or neuraminidase, and both CVVs retained their original antigenicity. The replication capacity of the two CVVs reached a level similar to that of A/Puerto Rico/8/34 in MDCK cells. The two CVVs showed low pathogenicity in vitro and in vivo, which are in line with the WHO requirements for CVVs. CONCLUSION: We obtained two genetically stable CVVs of HPAI H5N6 with high growth characteristics, which may aid in our preparedness for a potential H5N6 pandemic.
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Virus de la Influenza A/inmunología , Vacunas contra la Influenza/inmunología , Gripe Aviar/epidemiología , Gripe Aviar/prevención & control , Gripe Humana/epidemiología , Gripe Humana/prevención & control , Pandemias/prevención & control , Animales , Aves , China , HumanosRESUMEN
OBJECTIVE: Interferon-induced transmembrane protein 3 (IFITM3) is an important member of the IFITM family. However, the molecular mechanisms underlying its antiviral action have not been completely elucidated. Recent studies on IFITM3, particularly those focused on innate antiviral defense mechanisms, have shown that IFITM3 affects the body's adaptive immune response. The aim of this study was to determine the contribution of IFITM3 proteins to immune control of influenza infection in vivo. METHODS: We performed proteomics, flow cytometry, and immunohistochemistry analysis and used bioinformatics tools to systematically compare and analyze the differences in natural killer (NK) cell numbers, their activation, and their immune function in the lungs of Ifitm3-/- and wild-type mice. RESULTS: Ifitm3-/- mice developed more severe inflammation and apoptotic responses compared to wild-type mice. Moreover, the NK cell activation was higher in the lungs of Ifitm3-/- mice during acute influenza infection. CONCLUSIONS: Based on our results, we speculate that the NK cells are more readily activated in the absence of IFITM3, increasing mortality in Ifitm3-/- mice.
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Gripe Humana/virología , Proteínas de la Membrana/genética , Infecciones por Orthomyxoviridae/veterinaria , Enfermedades de los Roedores/virología , Enfermedad Aguda , Animales , Modelos Animales de Enfermedad , Femenino , Humanos , Masculino , Proteínas de la Membrana/metabolismo , Ratones , Ratones Endogámicos C57BL , Infecciones por Orthomyxoviridae/virologíaRESUMEN
OBJECTIVE: To recover broad-neutralizing monoclonal antibodies (BnAbs) from avian influenza A (H5N1) virus infection cases and investigate their genetic and functional features. METHODS: We screened the Abs repertoires of expanded B cells circulating in the peripheral blood of H5N1 patients. The genetic basis, biological functions, and epitopes of the obtained BnAbs were assessed and modeled. RESULTS: Two BnAbs, 2-12D5, and 3-37G7.1, were respectively obtained from two human H5N1 cases on days 12 and 21 after disease onset. Both Abs demonstrated cross-neutralizing and Ab-dependent cellular cytotoxicity (ADCC) activity. Albeit derived from distinct Ab lineages, i.e., V H1-69-D2-15-J H4 (2-12D5) and V H1-2-D3-9-J H5 (3-32G7.1), the BnAbs were directed toward CR6261-like epitopes in the HA stem, and HA 2 I45 in the hydrophobic pocket was the critical residue for their binding. Signature motifs for binding with the HA stem, namely, IFY in V H1-69-encoded Abs and LXYFXW in D3-9-encoded Abs, were also observed in 2-12D5 and 3-32G7.1, respectively. CONCLUSIONS: Cross-reactive B cells of different germline origins could be activated and re-circulated by avian influenza virus. The HA stem epitopes targeted by the BnAbs, and the two Ab-encoding genes usage implied the VH1-69 and D3-9 are the ideal candidates triggered by influenza virus for vaccine development.
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Anticuerpos Antivirales/sangre , Anticuerpos ampliamente neutralizantes/inmunología , Hemaglutininas/inmunología , Subtipo H5N1 del Virus de la Influenza A/inmunología , Gripe Humana/inmunología , Adulto , Reacciones Cruzadas , Epítopos/inmunología , Femenino , Humanos , Gripe Humana/virología , Masculino , Adulto JovenRESUMEN
OBJECTIVE: To determine whether it could protect mice from challenge of lethal influenza virus which group prior infected A(H1N1) pdm09 and H9N2 virus respectively. METHODS: 150 BALB/c mice are divided into three groups. Mice are infected A(H1N1) pdm09 virus (pCA07) and poultry H9N2 virus (GZ333) respectively. Infected mice are challenged with 10 times of lethal dose virus (PR8) then compare the viral load, antibody and survival of the two group mice before and after challenged. RESULTS: Both experimental group mice survived after challenge of lethal influenza virus and lung viral load are lower than that of the first infection. Antibodies derived from the infective virus and challenge virus. CONCLUSION: Prior infected A(H1N1) pdm09 and H9N2 virus could protect mice from challenge of lethal influenza virus.
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Subtipo H1N1 del Virus de la Influenza A/fisiología , Subtipo H9N2 del Virus de la Influenza A/fisiología , Gripe Humana/prevención & control , Gripe Humana/virología , Animales , Anticuerpos Antivirales/inmunología , Femenino , Humanos , Gripe Humana/inmunología , Pulmón/inmunología , Pulmón/virología , Ratones , Ratones Endogámicos BALB C , Carga ViralRESUMEN
Electrospinning technique is the main method of preparing polymer nanofiber simply, directly and continuously at present. In this work, electrospinning blend solution was prepared by in-situ polymerization using acid-modified multi-walled carbon nanotubes (MWNTs), m-phenylenediamine (MPD) and isophthaloyl chloride (IPC). And then composite nanofibers were prepared by electrospinning. MWNTs played an important role in nanofiber's properties. The effects of MWNTs on the morphology and characterization of the MWNTs/PMIA composite nanofibers were investigated. Scanning electron microscopy (SEM), thermal gravimetric analyzer (TGA), and X-ray diffraction (XRD) were utilized to characterize the MWNTs/PMIA nanofibers morphology and properties. The experimental results indicated that the nanofibers diameter decreased and solution dynamic viscosity increased with increasing MWNTs contents. XRD data demonstrated that PMIA composite nanofibers had the same crystal type as the pure PMIA nanofiber, and crystallinity was improved with increasing MWNTs loading. Transmission electron microscopy (TEM) was used to confirm MWNTs aligned along the axis of composite nanofibers.
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To establish the mouse-lethal model for pandemic H1N1 influenza virus, provide an animal model for studying the pathogenicity and host adaptation of 2009 pandemic H1N1 influenza virus, and find out the key amino acid mutations which may affect viral virulence and replication. A pandemic H1N1 influenza virus strain, A/Sichuan/SWL1/2009 (H1N1, SC/1) was passaged in mouse lung by 15 cycles with intranasal infection. The passaged viruses were all propagated in MDCK cells and sequenced. Based on the sequencing results, four mice in each group were inoculated with 6 selected viruses and their weight and survival rate were monitored during the following 14 days after infection. Additionally, SC/1-MA P14 and P15 viruses were sequenced after purification by Plague Assay. Viral virulence was increased after serial passages and the mortality of 100% was detected after 7 passages. Several amino acid residue mutations of passaged viruses which may contribute to the enhanced virulence were observed. The increased virulence of passaged viruses and mammalian host adaptation maybe associated with amino acid mutations in viral functional proteins. Finally, we established a mouse-lethal model.
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Modelos Animales de Enfermedad , Subtipo H1N1 del Virus de la Influenza A/fisiología , Subtipo H1N1 del Virus de la Influenza A/patogenicidad , Gripe Humana/epidemiología , Gripe Humana/virología , Pandemias , Sustitución de Aminoácidos , Animales , Secuencia de Bases , Línea Celular , China/epidemiología , Perros , Femenino , Humanos , Subtipo H1N1 del Virus de la Influenza A/genética , Subtipo H1N1 del Virus de la Influenza A/crecimiento & desarrollo , Ratones , Ratones Endogámicos BALB C , Análisis de Supervivencia , Ensayo de Placa Viral , Virulencia , Replicación ViralRESUMEN
A randomized clinical trial was conducted to assess whether the immunogenicity of seasonal and pandemic (H1N1/09) influenza vaccines is affected by the order of vaccine administration. 151 healthy adult volunteers were randomized into three groups. All groups received one dose (15 µg haemagglutinin) each of a pandemic H1N1 vaccine and a seasonal trivalent vaccine. Group 1 received the pandemic H1N1 vaccine first, followed by the seasonal vaccine 21 days later. Group 2 received vaccinations in vice versa and Group 3 received both vaccines simultaneously. Post-vaccination blood samples were collected to determine the immunogenicity by hemagglutination-inhibition (HI), microneutralization (MN), and B cell ELISPOT assays. All three vaccination strategies were well-tolerated and generated specific immune responses. However, we found a significant difference in magnitude of antibody responses to pandemic H1N1 between the three groups. Pre- or co-vaccination with the seasonal flu vaccine led to a significant reduction by 50% in HI titre to pandemic H1N1 virus after pandemic vaccination. Pre- or co-vaccination of pandemic H1N1 vaccine had no effect on seasonal flu vaccination. MN and ELISPOT assays showed a similar effect. Vaccination with pandemic H1N1 vaccine first is recommended to avoid an associated inhibitory effect by the seasonal trivalent flu vaccine.
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Vacunas contra la Influenza/administración & dosificación , Vacunas contra la Influenza/inmunología , Gripe Humana/prevención & control , Vacunación/métodos , Adolescente , Adulto , Anticuerpos Neutralizantes/sangre , Anticuerpos Antivirales/sangre , Linfocitos B/inmunología , Ensayo de Immunospot Ligado a Enzimas , Femenino , Pruebas de Inhibición de Hemaglutinación , Humanos , Vacunas contra la Influenza/efectos adversos , Masculino , Persona de Mediana Edad , Pruebas de Neutralización , Adulto JovenRESUMEN
M2 protein of type A influenza virus is a good candidate for universal influenza vaccine, exotoxin A of Pseudomonas aeruginosa may facilitate the immunogenicity of M2 protein. We constructed and expressed a prokaryotic expression plasmid containing a chimeric gene of M2 extracellular coding region and a partial PEA gene, and observed the immunoprotection in BALB/c mice vaccinated with the fusion protein. The fusion protein (ntPE-M2e) was generated by inserting the coding sequence of the M2e in place of Ib loop in PEA. This fusion protein was used to immunize BALB/c mice by subcutaneously injection with incomplete Freund's adjuvant and boost at weeks 3 and 7. The immunized mice were challenged with influenza virus strain A/PR/34/8. The fusion protein (ntPE-M2e) immunization protected mice against lethal viral challenge. ELISA and ELISPOT results demonstrated that the fusion protein could induce a strong systemic immune response against synthetic M2e peptide, and virus replication in the lungs of mice was inhibited in comparison with the control. This study provides foundation for developing broad-spectrum vaccines against type A influenza viruses.
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ADP Ribosa Transferasas/genética , Toxinas Bacterianas/genética , Exotoxinas/genética , Virus de la Influenza A , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/inmunología , Proteínas de la Matriz Viral/genética , Proteínas de la Matriz Viral/inmunología , Factores de Virulencia/genética , Animales , Ensayo de Inmunoadsorción Enzimática , Escherichia coli/genética , Femenino , Expresión Génica , Inmunización , Virus de la Influenza A/inmunología , Virus de la Influenza A/fisiología , Pulmón/inmunología , Pulmón/virología , Ratones , Ratones Endogámicos BALB C , Proteínas Recombinantes de Fusión/biosíntesis , Proteínas Recombinantes de Fusión/aislamiento & purificación , Proteínas de la Matriz Viral/biosíntesis , Proteínas de la Matriz Viral/aislamiento & purificación , Exotoxina A de Pseudomonas aeruginosaRESUMEN
OBJECTIVE: To provide a technology platform for vaccine development as well as the research on transmission and pathogenesis, the reverse genetic system for H9N2 avian influenza virus was established. METHODS: Eight full-length cDNAs of avian influenza virus A/Guangzhou/333/99 (H9N2) were amplified by RT-PCR and separately cloned into the transcription/expression vector, pCI-polI. The 8 plasmids DNA was cotransfected into 293T cell, the cell supernatant was collected and inoculated into embryonated eggs, the rescued virus from the allantoic fluid was identified by hemagglutinination assay. RESULTS: The avian influenza H9N2 virus was successfully rescued by 8 plasmids co-transfection in 293T cells. The hemagglutinination titer of the rescued virus is up to 2(9)/50 microl and its growth curve remained relatively as to the wild-type virus. CONCLUSION: The reverse genetic for avian influenza H9N2 subtype virus has been established successfully.
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Ingeniería Genética/métodos , Subtipo H9N2 del Virus de la Influenza A/genética , Animales , Línea Celular , Embrión de Pollo , Femenino , Vectores Genéticos/genética , Humanos , Lactante , Subtipo H9N2 del Virus de la Influenza A/crecimiento & desarrollo , Subtipo H9N2 del Virus de la Influenza A/fisiología , Gripe Humana/virología , Plásmidos/genéticaAsunto(s)
Gripe Humana/virología , Infecciones por Orthomyxoviridae/veterinaria , Infecciones por Orthomyxoviridae/virología , Orthomyxoviridae/aislamiento & purificación , Enfermedades de los Porcinos/virología , Animales , Humanos , Orthomyxoviridae/genética , Infecciones por Orthomyxoviridae/epidemiología , Porcinos , Proteínas Virales/genéticaRESUMEN
OBJECTIVE: To evaluate the efficacy of the interferon alpha-2b nasal spray in prevention of rubella and measles virus infections. METHODS: The properly selected volunteer groups have been divided into interferon alpha-2b experimental and control group. The experimental group received interferon alpha-2b treatment by nasal spray for 2 days before the immunization, then both groups were challenged with rubella and measles attenuated live vaccine respectively through nasal spray. The sera from pre-immunization and 21 and 28 days after immunization were collected to test the IgG antibody titers. The influence on the viral antibody titer reflects the viral preventive effect by interferon alpha-2b. RESULTS: The antibody titer difference of measles virus between experimental and control group was 1.26 (21 day) and 2.96 (28 day), there were statistically difference between them; the difference of rubella virus was 0.95 (21 day) and 0.37 (28 day), but there were no statistically differences found. CONCLUSION: The preliminary results showed that the interferon alpha-2b can be used as prevention method for measles and rubella viral infections.