Berberine interferes with RSV infection of HEp-2 cells by inducing mitophagy autophagy / 中国药理学通报

Aim To explore the effect of berberine (B E) on RSV infected HEp-2 cells and the related mechanism. Methods HEp-2 cells were infected with RSV and treated with BE. Cell viability was assessed using the CCK-8 assay. Protein expression levels of NLRP3, ASC, caspase-1, PINK1, Parkin, Beclinl, p62, LC3 I,LC3 II,and BNIP3 in HEp-2 cells were detected by Western blot. The secretion level of IL-1 p in HEp-2 cells was measured using ELISA. Apoptosis rate and mitochondrial membrane potential of HEp-2 cells were examined by flow cytometry. Mitochondrial ROS (mtROS) in HEp-2 cells was detected through MitoSOX staining. Colocalization of mitochondria and autophagosomes in HEp-2 cells was investigated using immunofluorescence staining. Cyclosporin A was used for validation experiments. Results BE could significantly improve the activity of RSV-infected HEp-2 cells,reduce the apoptosis rate (P < 0. 05), and decrease the activation level of NLRP3 inflammasomes and IL-lp level (P <0. 05); BE improved mitochondrial function by increasing mitochondrial membrane potential and ATP levels,and reduced mtROS. BE significantly promoted the colocalization of mitochondria-autophagosome in RSV infected cells, inducing PINK1/ Parkin and BNIP3 to mediate mitochondrial autophagy; cyclosporine A aggravated RSV infection. Conclusions BE has protective effects on HEp-2 cells infected by RSV. The mechanism may be related to the inhibitory effect of BE on the production of mtROS and the activation of NLRP3 inflammasomes by inducing PINK1/ Parkin and BNIP3-mediated mitochondrial autophagy.

Preparation and immunization effect evaluation of recombinant F protein vaccine of respiratory syncytial virus / 中国生物制品学杂志

@#Objective To prepare recombinant F protein vaccine of respiratory syncytial virus(RSV) and evaluate its immunization effect.Methods Two RSV vaccines based on RSV F protein were prepared:one was a mucosal vaccine with bacterial like particle(BLP)as adjuvant and the other was an injectable vaccine with aluminium hydroxide as adjuvant.Forty female BALB/c mice were randomly divided into four groups:BLP-F,BLP control,AL-F and AL control group,with 10mice in each group.BLP-F and BLP control group were inoculated intranasally,and AL-F and AL control group were inoculated subcutaneously.The mice were immunized once each at day 0,14 and 28,respectively.Two weeks after the last immunization,the titers of serum IgG antibody and IgA antibody in nasal lotion were detected by ELISA,and the titers of neutralizing antibody were detected by plaque test.Results Both vaccines induced high levels of serum binding antibodies and neutralizing antibodies,and the induction capacity of injected vaccine was stronger than that of mucosal vaccine.The injected vaccine induced the increase of IgG in serum,which was about 10 times higher than the mucosal immune response,but could not induce the increase of IgA.However,the mucosal vaccine induced the high level of mucosal IgA,but the serum IgG antibody was relatively low.Conclusion Both vaccines based on RSV F protein are promising candidates,and each vaccine has its own advantages.Follow-up studies will evaluate the feasibility of these two vaccines as immunogens using a combination immunization approach to simultaneously enhance systemic and mucosal immune responses against RSV.

Development progress and application of monoclonal antibody drugs against respiratory virus / 中国生物制品学杂志

@#Abstract：In recent years，the outbreak and prevalence of respiratory infectious diseases in the world seriously endanger human health，among which the respiratory infectious diseases caused by viral infection account for a large proportion. The use of vaccines and common antiviral drugs is an effective way to fight viral infection，but there are also problems such as lag and drug resistance. Monoclonal antibodies against respiratory viral infections provide a new strategy for clinical treatment. This paper reviews the development of monoclonal antibody against respiratory virus and its application in respiratory viral infectious diseases. Keywords：Respiratory viral infectious diseases；Respiratory syncytial virus（RSV）；Influenza virus（IFV）；Coronavirus （CoV）；Monoclonal antibody

Modified Wuhutang Alleviates Airway Inflammation and Down-regulates Expression of Muc5AC， STAT3， NF-κB， and NLRP3 in Lung Tissues of RSV-infected Asthmatic Mice / 中国实验方剂学杂志

ObjectiveTo explore effect of modified Wuhutang on airway inflammation and expression of mucin （Muc） 5AC， signal transducer and activator of transcription 3 （STAT3）， nuclear factor kappa B （NF-κB）， and NOD-like receptor thermal protein domain associated protein 3 （NLRP3） in respiratory syncytial virus （RSV）-infected asthmatic mice. MethodSeventy male BALB/c mice of 6-8 weeks old were randomized into normal control （CON）， asthma （ovalbumin， OVA）， RSV infection-induced asthma （OVA+RSV）， high-， medium-， and low-dose （4.08， 2.04， 1.02 g·kg-1·d-1， respectively） modified Wuhutang， and dexamethasone （Dxms， 0.1 g·kg-1d-1） groups （n=10）. The model of asthma was established by sensitization and atomization inhalation with OVA. The RSV infection-induced asthma model was established by three consecutive RSV nasal infusions （1.0 × 106 PFU·mL-1， 50 μL）. Wuhutang was administrated by gavage， and Dxms by intraperitoneal injection. The CON group was given the same amount of normal saline by gavage. The mice were anesthetized with 2.5% pentobarbital sodium 24 h after the last administration， and then the lung tissue was stained by hematoxylin-eosin （HE） and Van Gieson （VG） for observation of airway inflammation. The immunohistochemical assay was employed to detect the expression of Muc5AC. Western blot was employed to determine the protein levels of phosphorylated （p）-STAT3， STAT3， p-NF-κB， NF-κB， and NLRP3. ResultCompared with the CON group， the OVA group presented airway inflammatory cell infiltration， tissue hyperemia and edema， and collagen fiber deposition. The OVA+RSV group showed severer airway inflammatory cell infiltration and tissue hyperemia and edema than the OVA group. Compared with the OVA+RSV group， modified Wuhutang alleviated the airway inflammatory cell infiltration， tissue hyperemia and edema， and collagen fiber deposition， and the high-dose group had the best performance. Compared with the CON group， the OVA group and the OVA+RSV group showed increased expression level of Muc5AC （P<0.01）. Compared with the OVA+RSV group， modified Wuhutang reduced the expression level of Muc5AC， and the reduction was significant in the high-dose group （P<0.05）. Compared with the high-dose modified Wuhutang group， Dxms lowered the expression level of Muc5AC （P<0.05）. Compared with the CON group， the OVA and OVA+RSV groups showed up-regulated protein levels of p-STAT3， p-NF-κB， and NLRP3 （P<0.05， P<0.01）. Compared with the OVA+RSV group， modified Wuhutang down-regulated the protein levels of p-STAT3， p-NF-κB， and NLRP3 （P<0.01）. Compared with the high-dose modified Wuhutang group， the Dxms group showed up-regulated levels of p-STAT3， p-NF-κB proteins （P<0.01）. ConclusionModified Wuhutang can reduce airway inflammation and down-regulate the expression of Muc5AC， p-STAT3， p-NF-κB， and NLRP3 in RSV-infected asthmatic mice， which suggests that Wuhutang reduces airway inflammation in RSV-infected asthma by regulating the STAT3/NF-κB signaling pathway.

Mechanism of Resveratrol in Treatment of Lung Adenocarcinoma： An Exploration Based on Database Mining and Molecular Biology / 中国实验方剂学杂志

Objective:To explore the mechanism of resveratrol （RSV） in the treatment of lung adenocarcinoma （LUAD） based on bioinformatics and molecular biology. Method:The targets of RSV were retrieved from DrugBank and then imported into STRING for constructing a protein-protein interaction （PPI） network.TCGA database was utilized to analyze the expression of target genes in tumor and normal tissues， followed by the prediction of their impacts on tumor occurrence and development and the screening of target genes using random forest and univariate Cox regression models.With the results of bioinformatics taken into consideration， the mechanism of RSV in inhibiting LUAD was further explored by molecular biology. Result:Ten Hub genes were screened out from the PPI network of RSV targets.Among them， solute carrier family 2 member 1 （SLC2A1）， arachidonate 5-lipoxygenase （ALOX5）， peroxisome proliferative activated receptor gamma （PPARG）， and arachidonate 15-lipoxygenase （ALOX15） differed significantly in their expression in tumor and normal tissues.As revealed by random forest and univariate COX regression analysis， SLC2A1 was of great significance to the survival and prognosis of patients with LUAD.The survival analysis through Kaplan-Meier （KM） plotter indicated that the SLC2A1 expression was closely related to the overall survival （OS）， first progression （FP）， and post-progression survival （PPS） of LUAD patients.The molecular biological experiments further proved that RSV inhibited the proliferation and migration of LUAD cells by reducing the expression of SLC2A1.As verified by immunohistochemical scoring， SLC2A1 protein expression in tumor tissue was significantly different from that in normal tissue. Conclusion:RSV inhibits the proliferation and migration of LUAD cells by reducing the expression of SLC2A1， which has far-reaching significance in the clinical treatment of LUAD.

Granulocyte-Macrophage colony stimulating factor in asthmatic patients infected with respiratory syncytial virus

@#Introduction: It is estimated that at least 30 to 40% of asthma attacks in adults are related to respiratory infections with viruses. The majority of asthma-related viruses include respiratory syncytial virus (RSV), rhinovirus, and parainfluenza. Inflammatory cytokines are supposed to play a vital role in causing inflammation of the respiratory tract as regulators of proliferation, chemotaxis, and activation of inflammatory cells. Objectives: The aim of this study is to assess the role of Granulocyte Macrophage-Colony Stimulating Factor (GMCSF) in asthmatic airway hyper-responsiveness associated with RSV infections. Materials and Methods: Forty five asthmatic cases and 45 healthy individuals were studied in a cross-sectional design. All asthmatics underwent symptom score assessment.GMCSF concentrations in sputum and RSV-IgM/IgG in serum samples were measured for all participants by Enzyme Linked Immuno-Sorbent Assay (ELISA). Results: The GM-CSF concentration level was significantly higher in asthmatics (270.27± 194.87pg/mL) especially among moderate and severe disease with mean concentration of 197.33±98.47 and 521.08± 310.04 respectively, compared to healthy controls (22.20±21.27 pg/ mL) (p=0.0001). The sputum level of GM-CSF in asthmatics is highly significant associated with positive anti-RSV IgG sera which represents 35/45(77.8%) with mean GM-CSF concentration of (276.99± 86.42) compared with controls at about 31/45 (68.9%) with GM-CSF mean concentration of (22.84±23.47). On the other hand, positive anti-RSV IgM in asthma cases was 8 out of 45(17.8 %) with GM-CSF mean concentration of (307.25± 306.65). Furthermore, GM-CSF sputum level was significantly correlated with eosinophil count especially in moderate and severe asthma. Conclusions: This study revealed that GM-CSF level is associated with eosinophilia and indicates asthma severity that might be evident during RSV infection .The distinctive GM-CSF features observed in the sputum from asthmatics with RSV may be useful as a diagnostic methods to help match patients with antibody therapy.

Factores de riesgo de Bronquiolitis en pacientes menores de 2 años

Resumen Introducción: Se estima que a nivel mundial, cada año alrededor del 10% de los lactantes tienen bronquiolitis. Se produce un pico entre los 2 y los 6 meses de edad. Objetivo: Determinar los factores de riesgo de bronquiolitis en menores de dos años que consultaron al servicio de Pediatría del Instituto de Medicina Tropical entre el 2018 y 2019. Método: Estudio de tipo descriptivo, transversal en el que se incluyeron pacientes menores de 2 años internados en el Servicio de Pediatría del Instituto de Medicina Tropical de enero de 2018 a febrero de 2019 con diagnóstico de bronquiolitis en el que se analizaron los factores de riesgo de la enfermedad. Resultados: Se estudió una población de 38 pacientes que presentaron bronquiolitis antes de los 2 años, 19 fueron niños (50%) La edad media al ingreso fue de 3,5 ± 2,8 meses (límites, 1-10 meses). No se han encontrado datos de tabaco en el embarazo. Se detectó VRS en el aspirado nasofaríngeo de 8 niños (21%), la detección viral fue negativa en 14 niños (37%) y no se realizó la determinación en 18 casos (47%). Los siguientes factores de riesgo se asociaron de forma independiente con la bronquiolitis: Asma de los padres, infección por VSR, sibilancia, cianosis y hacinamiento. Los factores de riesgo estudiado, exposición al tabaco, sexo, edad en el momento de la bronquiolitis o lactancia materna exclusiva, no se asociaron con el ingreso a UTI de los pacientes con bronquiolitis. Conclusión: Hemos encontrado factores de riesgo asociado relacionados al huésped como asma de los padres, y otros como sibilancia, cianosis, hacinamiento e infección por VRS.

Abstract Introduction: It is estimated that globally, each year around 10% of infants have bronchiolitis. A peak occurs between 2 and 6 months of age Objective: To determine the risk factors of bronchiolitis in children less than two years of age who admitted at the pediatric service of the Institute of Tropical Medicine between 2017 and 2019 Method: A descriptive, cross-sectional study in which patients under 2 years of age admitted to the Pediatric Service of the Institute of Tropical Medicine from January 2018 to February 2019 were included with a diagnosis of bronchiolitis in which the risk factors of the disease were analyzed. Results: We studied a population of 38 patients who presented bronchiolitis before 2 years, 19 were children (50%) The average age at admission was 3.5 ± 2.8 months (range, 1-10 months). No tobacco data were found in pregnancy. RSV was detected in the nasopharyngeal aspirate of 8 children (21%), viral detection was negative in 14 children (37%) and the determination was not made in 18 cases (47%). The following risk factors were independently associated with bronchiolitis: Asthma of the parents, RSV infection, wheezing, cyanosis and overcrowding. The risk factors studied exposure to tobacco, sex, age at the time of bronchiolitis or exclusive breastfeeding were not associated with admission to the ICU of patients with bronchiolitis. Conclusion: We have found associated risk factors related to the host such as asthma of the parents, and others such as wheezing, cyanosis, overcrowding and RSV infection.

Effect of resveratrol treatment on apoptosis and apoptotic pathways during boar semen freezing / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Resveratrol (3,5,4'-trihydroxystilbene, RSV) has been widely used in mammalian cells, but whether it can be used during freezing boar semen is still unknown. The effects of RSV treatment during boar semen freezing on its anti-freezing ability, apoptosis, and possible apoptotic pathways were observed in this study. Sperm motility, mitochondrial membrane potential (ΔΨm), adenosine triphosphate (ATP) content, terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick-end labeling (TUNEL)-positive apoptotic state, and messenger RNA (mRNA) expression levels of apoptotic genes involved in different apoptotic pathways after freezing with or without RSV treatment were tested. The results showed that: (1) Compared with fresh sperm, the motility, normal acrosome rate, and plasma membrane integrity rate of frozen boar sperm decreased significantly (P0.05), but it did significantly improve the normal acrosome rate (57.65% vs. 47.00%, P<0.05) and plasma membrane integrity rate (46.67% vs. 38.85%, P<0.05). (2) After freezing, most boar sperm showed low mitochondrial ΔΨm. RSV treatment could increase the rate of high mitochondrial ΔΨm of boar sperm. (3) RSV treatment significantly decreased reactive oxygen species (ROS) levels (58.65% vs. 88.41%, P<0.05) and increased the ATP content (0.49 μmol/L vs. 0.25 μmol/L, P<0.05) of boar sperm during freezing. (4) The apoptotic rate of the freezing group (80.41%) with TUNEL detection increased significantly compared to the fresh group (9.70%, P<0.05), and RSV treatment greatly decreased the apoptotic rate (68.32%, P<0.05). (5) Real-time polymerase chain reaction (RT-PCR) showed that not only the genes from the death receptor-mediated apoptotic pathway (tumor necrosis factor-α (TNF-α), Fas ligand (FasL), and Caspase-8), but also the genes from the mitochondria-mediated apoptotic pathway (manganese superoxide dismutase (MnSOD), B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax), and Caspase-9) were both significantly changed after freezing. RSV treatment during freezing greatly changed their expression levels. Although RSV treatment during boar semen freezing did not significantly increase motility after thawing, it still played an efficient antioxidant role, which could enhance the mitochondrial function and decrease the apoptotic level induced by both the death receptor- and mitochondria-mediated apoptotic pathways.

Effect of resveratrol treatment on apoptosis and apoptotic pathways during boar semen freezing / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Resveratrol (3,5,4'-trihydroxystilbene, RSV) has been widely used in mammalian cells, but whether it can be used during freezing boar semen is still unknown. The effects of RSV treatment during boar semen freezing on its anti-freezing ability, apoptosis, and possible apoptotic pathways were observed in this study. Sperm motility, mitochondrial membrane potential (ΔΨ), adenosine triphosphate (ATP) content, terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick-end labeling (TUNEL)-positive apoptotic state, and messenger RNA (mRNA) expression levels of apoptotic genes involved in different apoptotic pathways after freezing with or without RSV treatment were tested. The results showed that: (1) Compared with fresh sperm, the motility, normal acrosome rate, and plasma membrane integrity rate of frozen boar sperm decreased significantly (P0.05), but it did significantly improve the normal acrosome rate (57.65% vs. 47.00%, P<0.05) and plasma membrane integrity rate (46.67% vs. 38.85%, P<0.05). (2) After freezing, most boar sperm showed low mitochondrial ΔΨ. RSV treatment could increase the rate of high mitochondrial ΔΨ of boar sperm. (3) RSV treatment significantly decreased reactive oxygen species (ROS) levels (58.65% vs. 88.41%, P<0.05) and increased the ATP content (0.49 μmol/L vs. 0.25 μmol/L, P<0.05) of boar sperm during freezing. (4) The apoptotic rate of the freezing group (80.41%) with TUNEL detection increased significantly compared to the fresh group (9.70%, P<0.05), and RSV treatment greatly decreased the apoptotic rate (68.32%, P<0.05). (5) Real-time polymerase chain reaction (RT-PCR) showed that not only the genes from the death receptor-mediated apoptotic pathway (tumor necrosis factor-α (TNF-α), Fas ligand (FasL), and Caspase-8), but also the genes from the mitochondria-mediated apoptotic pathway (manganese superoxide dismutase (MnSOD), B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax), and Caspase-9) were both significantly changed after freezing. RSV treatment during freezing greatly changed their expression levels. Although RSV treatment during boar semen freezing did not significantly increase motility after thawing, it still played an efficient antioxidant role, which could enhance the mitochondrial function and decrease the apoptotic level induced by both the death receptor- and mitochondria-mediated apoptotic pathways.

Caracterización de una cohorte durante cinco años después de un episodio de bronquiolitis que requiere hospitalización en una clínica de tercer nivel de Bogotá, Colombia / Characterization of a cohort five years after an episode of bronchiolitis that required hospitalization in a third level clinic of Bogotá, Colombia

Introducción. La Bronquiolitis secundaria al Virus Sincitial Respiratorio, se han asociado con episodios sibilantes recurrentes y desarrollo de asma, incluso en la adultez. Sin embargo, la relación entre estas patologías es controvertida, y aun no se conoce el comportamiento de este fenómeno en Colombia. El objetivo de este estudio fue describir la evolución clínica a cinco años, de los niños con antecedente de bronquiolitis que requirieron hospitalización Materiales y métodos. Estudio descriptivo de cohorte retrospectiva de menores de dos años, con diagnóstico de Bronquiolitis atendidos en la Clínica Universitaria Colombia en los años 2008 a 2011 con seguimiento de hospitalizaciones por patologías respiratorias hasta el año 2016. Se revisaron 306 historias clínicas de pacientes y se analizaron características socio- demográficas, aislamientos virales y manejo farmacológico. Resultados. Los años con mayor número de hospitalizaciones por episodios sibilantes posterior al episodio bronquiolitis fueron el 2009 y 2011 con una incidencia acumulada de 15,6% y 9,9%. La edad promedio de hospitalización fue 6 meses y más frecuente en hombres. El virus sincital fue aislado con mayor frecuencia en los años de seguimiento, y la mayoría de los casos requirió manejo antibiótico; la ampicilina sulbactam (28,5%) y la ampicilina (22,6%).

Introduction. Bronchiolitis secondary to Respiratory Syncytial Virus, have been associated with recurrent wheezing episodes and development of asthma, even in adulthood. However, the relationship between these pathologies is controversial, and the behavior of this phenomenon in Colombia is not yet known. The aim of this study was to describe the five-year clinical course of children with a history of bronchiolitis who required hospitalization Materials and methods. Descriptive study of a retrospective cohort of children under two years of age, with a diagnosis of Bronchiolitis attended at the University Clinic Colombia in the years 2008 to 2011 with follow-up of hospitalizations for respiratory pathologies up to 2016. 306 patient clinical histories were reviewed and characteristics were analyzed. socio-demographic, viral isolates and pharmacological management. Results. The years with the highest number of hospitalizations for wheezing episodes after the bronchiolitis episode were 2009 and 2011 with an accumulated incidence of 15.6% and 9.9%. The average age of hospitalization was 6 months and more frequent in men. The syncytial virus was isolated more frequently in the years of follow-up, and most of the cases required antibiotic management; ampicillin sulbactam (28.5%) and ampicillin (22.6%).

Detection of respiratory syncytial virus & Mycoplasma pneumoniae in paediatric lower respiratory tract infections

Background & objectives: Respiratory syncytial virus (RSV) and Mycoplasma pneumoniae are considered common cause of lower respiratory tract infections (LRTIs) in children. The present study was conducted to detect M. pneumoniae and RSV in paediatric LRTIs employing serology, polymerase chain reaction (PCR) and reverse transcriptase PCR (RT-PCR) analysis. Methods: Seventy five children aged one month to five years with acute LRTIs were investigated for M. pneumoniae antibodies and RSV antigen using immunochromatographic test, RT-PCR for RSV and M. pneumoniae by PCR on nasopharyngeal aspirates. Results: RSV infection was observed in 33 (44%) and M. pneumoniae was positive in 26 (35%) children. No significant difference in infection was noted between male and female children. Clinical and radiological features among RSV and M. pneumoniae positive and negative cases were similar. Considering RT-PCR for RSV as gold standard, RSV antigen immunochromatography was 90.90 per cent sensitive and 100 per cent specific. Interpretation & conclusions: Our study showed the presence of RSV and M. pneumoniae infection in 44 and 35 per cent children, respectively with community-acquired LRTIs and aged less than five years.

IL-17A involved in respiratory syncytial virus-associated pathogenesis by promoting IFN-γand inhib-iting viral clearance in mice / 中华微生物学和免疫学杂志

Objective To identify the role of IL-17A during respiratory syncytial virus (RSV) in-fection in a mouse model. Methods Female wild-type C57BL/6 mice and IL-17A knockout ( IL-17A-/-) mice at the age of 6 to 8 weeks were both randomly divided into two groups:control and RSV groups. Mice in the control groups were given the supernatant of Hep-2 cell culture, while those in the RSV groups were treated with RSV A2 through intranasal administration. Leukocytes in bronchoalveolar lavage fluid ( BALF) samples were counted. Left lung tissues were stained with hematoxylin and eosin ( HE ) to evaluate his-topathological scores. Airway hyperresponsiveness ( AHR) was measured by whole-body plethysmography. The concentrations of IFN-γ were determined with ELISA. RSV titers were measured by plaque assay. To assess the effects of IL-17A on IFN-γproduction and its role in RSV infection, IL-17A-/- mice were treated with exogenous recombinant murine IFN-γ or IL-17A, while wild-type mice were given IFN-γ neutralizing antibody intervention. Results The counts of inflammatory cells and neutrophils in BALF, lung tissue his-topathological scores, AHR, IFN-γlevels and virus titers of the wild-type group were higher than those of the IL-17A-/-group after RSV infection. IFN-γlevels, inflammatory cell counts in BALF, AHR and lung tissue histopathological scores were significantly increased in RSV-infected IL-17A-/- mice after the intervention of recombinant murine IL-17A or IFN-γ. RSV titers were much higher in the recombinant murine IL-17A-trea-ted group, but not affected by the recombinant murine IFN-γ intervention. Inflammatory cell counts in BALF, AHR and lung tissue histopathological scores were significantly decreased in RSV-infected wild-type mice following IFN-γ neutralizing antibody treatment, but no significant changes were found in RSV titers. Conclusions IL-17A might be involved in the pathogenesis of pulmonary diseases during RSV infection through promoting IFN-γ production and inhibiting viral clearance in mice.

Respiratory Syncytial Virus Fusion Protein-encoding DNA Vaccine Is Less Effective in Conferring Protection against Inflammatory Disease than a Virus-like Particle Platform

Formalin-inactivated respiratory syncytial virus (RSV) vaccination causes vaccine-enhanced disease (VED) after RSV infection. It is considered that vaccine platforms enabling endogenous synthesis of RSV immunogens would induce favorable immune responses than non-replicating subunit vaccines in avoiding VED. Here, we investigated the immunogenicity, protection, and disease in mice after vaccination with RSV fusion protein (F) encoding plasmid DNA (F-DNA) or virus-like particles presenting RSV F (F-VLP). F-DNA vaccination induced CD8 T cells and RSV neutralizing Abs, whereas F-VLP elicited higher levels of IgG2a isotype and neutralizing Abs, and germinal center B cells, contributing to protection by controlling lung viral loads after RSV challenge. However, mice that were immunized with F-DNA displayed weight loss and pulmonary histopathology, and induced F specific CD8 T cell responses and recruitment of monocytes and plasmacytoid dendritic cells into the lungs. These innate immune parameters, RSV disease, and pulmonary histopathology were lower in mice that were immunized with F-VLP after challenge. This study provides important insight into developing effective and safe RSV vaccines.

Low-dose Sekikaic Acid Modulates Host Immunity and Protects Cells from Respiratory Syncytial Virus Infection

Introduction: Investigations of Ramalina farinaceae and its isolated compounds had reportedly shown appreciable anti-RSV activity albeit within a slim therapeutic window. Sekikaic acid represent one of such compounds. In this present study, sekikaic acid in very low dose was evaluated as a cell restorative and immunomodulatory factor. Methodology: The effect of sekikaic acid on the cell viability of RSV-infected HEp2 cells was investigated. Further assay for cellular immune response in primed mouse splenocytes was established by cell culture and flow cytometry. Splenocytes were treated with graded concentrations of sekikaic acid (6.25-25 µg/ml) and screened for their effect on the expression of IFNγ and IL-2 and T-lymphocytes markers CD4+ and CD8+ using intracellular cytokine staining and Fluorescence-activated cell sorting (FACS) analysis. Results: Results reveal that sekikaic acid protects RSV-infected Hep 2 cells from infection-induced cytopathology. Moreover, sekikaic acid displayed some degree of immune-regulatory activity in the primed mouse splenocytes increasing the proportion of CD8+/IFNγ+ (70.95-73.8%) and CD4+/ IFNγ+ (3.44-4.13%) T lymphocytes when compared to the cells in untreated controls 2.75% (CD4+) and 69.35% (CD8+) respectively. Interleukin-2 (IL-2) expression and signalling by T-Lymphocytes was selective but pronounced for CD4+ cells activation (sekikaic acid 2.67-3.6%, control 0.44%). Similar scenario was recorded for Intracellular IL-2 secretion by the T-lymphocytes. Conclusion: Low-dose sekikaic acid protects RSV infected cells and lead to immune lymphocytes proliferation. This recorded T-lymphocyte-specific immune-modulatory property may contribute to explain in part the dynamics associated with the overall antiviral effect of Sekikaic acid, and may also find relevance as a necessary cellular immune response precursor to infection-associated disease management.

Heliox delivered by high flow nasal cannula improves oxygenation in infants with respiratory syncytial virus acute bronchiolitis / Heliox administrado por cânula nasal de alto fluxo melhora a oxigenação em crianças com bronquiolite aguda por vírus sincicial respiratório

Abstract Objective: The objective of this study is to evaluate the hypothesis that use of heliox would result in improvement of gas exchange when used with high flow nasal cannula in infants with RSV acute bronchiolitis. Methods: All patients that met the inclusion criteria were randomized to either heliox (70:30) or air-oxygen mixture 30% via high flow nasal cannula at 8 L/min for a continuous 24 h. Measurements were taken at baseline, after 2 h, and at the end of the 24 h. Results: This prospective study included 48 patients. After 2 h of treatment with heliox, the oxygen saturation and PaO2 significantly improved when compared with the air-oxygen group, 98.3% vs. 92.9%, 62.0 mmHg vs. 43.6 mmHg (p = 0.04 and 0.01), respectively. Furthermore, PaO2/FiO2 ratio was significantly higher in the heliox group when compared with the air-oxygen group, 206.7 vs. 145.3. Nevertheless, CO2 showed better elimination when heliox was used, without significance. MWCA score dropped significantly in the heliox group, 2.2 points vs. 4.0 points in air-oxygen (p = 0.04), 2 h after starting the therapy. Conclusion: Transient improvement of oxygenation in infants with RSV acute bronchiolitis during the initial phase of the therapy is associated with heliox when provided with HFNC, may provide a precious time for other therapeutic agents to work or for the disease to resolve naturally, avoiding other aggressive interventions.

Resumo Objetivo: Avaliar a hipótese de que o uso da mistura heliox resultaria em melhoria da troca gasosa quando usado com cânula nasal de alto fluxo em crianças com bronquiolite aguda por VSR. Métodos: Todos os pacientes que atenderam aos critérios de inclusão foram randomizados para receber a mistura heliox (70:30) ou a mistura ar/oxigênio a 30% por meio da cânula nasal de alto fluxo a 8 L/min por 24 horas contínuas. As medições foram feitas no início, depois de duas horas e ao fim de 24 horas. Resultados: Fizemos um estudo prospectivo em que foram incluídos 48 pacientes. Após duas horas de tratamento com a mistura heliox, a saturação de oxigênio e a PaO2 apresentaram melhoria significativa em comparação com o grupo da mistura ar/oxigênio: 98,3% em comparação com 92,9%, 62,0 mmHg em comparação com 43,6 mmHg (p = 0,04 e 0,01), respectivamente. Além disso, a relação PaO2/FiO2 era significativamente mais alta no grupo da mistura heliox do que no grupo da mistura ar/oxigênio, 2.067 em comparação com 1.453. Contudo, o CO2 apresentou melhor eliminação quando a mistura heliox foi usada, sem relevância. O Escore MWCA caiu significativamente no grupo da mistura heliox, 2,2 pontos em comparação com 4,0 pontos da mistura ar/oxigênio (p = 0,04) duas horas após o início da terapia. Conclusão: A breve melhoria da oxigenação em crianças com bronquiolite aguda por VSR na fase inicial da terapia está associada à mistura heliox quando administrada pela CNAF e poderá fornecer um tempo precioso para outros agentes terapêuticos funcionarem ou para a própria doença se curar naturalmente e evitar outras intervenções agressivas.

In vitro Antiviral Activity of Nauclea latifolia Root Bark Extract Against the Respiratory Syncytial Virus

Extract of Nauclea latifolia (NL) root bark collected from the Nigerian flora was examined for anti-RSV activity. Preliminary data showed anti-RSV activities with IC50 =75.62 µg/ml when tested against the recombinant strain rgRSV expressing the green fluorescent protein. Corresponding assays for the cytotoxic effect of the extract against utilized cell lines gave TC50 = 333.82 µg/ml. Further screening of against the circulating RSV A2 strain established their promising anti-RSV utility. Time of additional studies for the elucidation of the possible mechanism of action gave 74.38, 69.42, and 71.90% reduction of RSV plaque forming units at the respective 0, 2, and 4 hours post-infection addition times.

Label-free quantitative proteomics reveals fibrinopeptide B and heparin cofactor II as potential serum biomarkers in respiratory syncytial virus-infected mice treated with Qingfei oral liquid formula / 中国天然药物

Respiratory syncytial virus (RSV) is a leading cause of acute lower respiratory tract infections. Qingfei oral liquid (QFOL), a traditional Chinese medicine, is widely used in clinical treatment for RSV-induced pneumonia. The present study was designed to reveal the potential targets and mechanism of action for QFOL by exploring its influence on the host cellular network following RSV infection. We investigated the serum proteomic changes and potential biomarkers in an RSV-infected mouse pneumonia model treated with QFOL. Eighteen BALB/c mice were randomly divided into three groups: RSV pneumonia model group (M), QFOL-treated group (Q) and the control group (C). Serum proteomes were analyzed and compared using a label-free quantitative LC-MS/MS approach. A total of 172 protein groups, 1009 proteins, and 1073 unique peptides were successfully identified. 51 differentially expressed proteins (DEPs) were identified (15 DEPs when M/C and 43 DEPs when Q/M; 7 DEPs in common). Classification and interaction network showed that these proteins participated in various biological processes including immune response, blood coagulation, complement activation, and so forth. Particularly, fibrinopeptide B (FpB) and heparin cofactor II (HCII) were evaluated as important nodes in the interaction network, which was closely involved in coagulation and inflammation. Further, the FpB level was increased in Group M but decreased in Group Q, while the HCII level exhibited the opposite trend. These findings not only indicated FpB and HCII as potential biomarkers and targets of QFOL in the treatment of RSV pneumonia, but also suggested a regulatory role of QFOL in the RSV-induced disturbance of coagulation and inflammation-coagulation interactions.

Correlation Research of Serum Specific Antibody Expression of IgM, IgG and IgA in Children with Respiratory Syncytial Virus Infection / 现代检验医学杂志

Objective To explore the correlation between the expression of serum specific antibody IgM,IgG and IgA in the respiratory syncytial virus (RSV) in hospitalized children,and the antibody indexes with early auxiliary diagnostic signifi cance were screened.Methods Using fluorescent quantitative polymerase chain reaction (FQ-PCR) to screen 50 cases of throat swabs which RSV RNA were positive in hospitalized children from 2015 to 2017 and using enzyme linked immunosorbent assay (ELISA) detected specific antibodies IgM,IgG and lgA in the children's serum.Meanwhile 95 cases of children's serum specimens without respiratory symptoms were taken as the control group.The results were analyzed by chi-square test.Results In the 50 cases of serum from children who's RSV RNA were positive from throat swabs,the positive rates of IgM,IgG,IgA and the three forms coming together were 24.00％,60.00％00,22.00％ and 16.00％ respectively.And the difference was statistically significant (x2 =28.19,P＜0.01).About the serums which were the same gender child patient in the experimental group and the control group,the positive rates of IgM,IgG and IgA had significant differences (x2 =9.16,P＜0.01).There was no significant difference in the positive rate of each antibody in the same experimental group or control group or control group (x2 =0.10,P＞0.05).IgM,IgG and IgA were not detected in acute laryngotracheal bronchitis.Only 1 case with IgG was detected in the acute upper respiratory infection.It was the highest that the detection rates of IgG in bronchial pneumonia and acute bronchitis were 41.38％ and 23.53％,and IgA was not been detected alone.In the group of ＜6 months,IgM and IgA were not detected within 7 days and 21 days.In the group of 1～5 years old,the positive rate of IgM within 7 days was 50％,which was the highest in all groups.And IgM,IgG and IgA could be detected in 21 days also.The positive rates of IgG and IgA in the age group of 5～10 years old were 100％ and 66.67％ respectively.Conclusion Specific antibodies of RSV-IgM,IgG and IgA cannot be used as a diadynamic criteria alone in early RSV infection.The spe cific antibody was not affected by gender.Upper respiratory tract infection RSV was difficult to produce IgM,IgG and IgA.The younger the child,was the slower the IgM and IgA were produced.At the same time,IgG was able to vertical transmis sion and it had no protective effect on body.That RSV could infect the body and cause clinical respiratory symptoms would be related with immunity.Finally,IgA was the earliest specific antibody and could not be alone.

Identification of Respiratory Syncytial Virus A and B Subtypesby Tube Double Nested Polymerase Chain Reaction and Gene Sequencing Technology / 现代检验医学杂志

Objective Objective to establish a method for identification of respiratory syncytial virus (RSV) A and B subtypes for clinical research and application.Methods Using fluorescent quantitative polymerase chain reaction (FQ-PCR) screened 50 cases of throat swab that RSV were positive in hospitalized children from 2015 to 2017.The genotyping was performed according to the nucleotide sequence of G protein coding gene,and a single tube double nested PCR primers was designed for it.A and B subgroup by sequencing to conduct comparative analysis with nucleotide sequence in the Genebank.The results were analyzed by chi-square test.Results In the 50 cases of throat swab RSV positive children,respiratory syncytial virus A and B subtype and mixed infection rates were 82.00％,14.00％ and 4.00％,respectively.The difference was statistically significant (x2=81.06,P＜0.01).The RSV fractal results were consistent with the gene sequencing results.Conclusion The eastern part of shenzhen was dominated by respiratory syncytial virus A subtype epidemic and mixed infection.Single tube double nested polymerase chain reaction (PCR) and gene sequencing technique is suitable for the identification of A and B subtypes of RSV.It is characterized by high sensitivity,specificity and high accuracy.

Label-free quantitative proteomics reveals fibrinopeptide B and heparin cofactor II as potential serum biomarkers in respiratory syncytial virus-infected mice treated with Qingfei oral liquid formula / 中国天然药物

Respiratory syncytial virus (RSV) is a leading cause of acute lower respiratory tract infections. Qingfei oral liquid (QFOL), a traditional Chinese medicine, is widely used in clinical treatment for RSV-induced pneumonia. The present study was designed to reveal the potential targets and mechanism of action for QFOL by exploring its influence on the host cellular network following RSV infection. We investigated the serum proteomic changes and potential biomarkers in an RSV-infected mouse pneumonia model treated with QFOL. Eighteen BALB/c mice were randomly divided into three groups: RSV pneumonia model group (M), QFOL-treated group (Q) and the control group (C). Serum proteomes were analyzed and compared using a label-free quantitative LC-MS/MS approach. A total of 172 protein groups, 1009 proteins, and 1073 unique peptides were successfully identified. 51 differentially expressed proteins (DEPs) were identified (15 DEPs when M/C and 43 DEPs when Q/M; 7 DEPs in common). Classification and interaction network showed that these proteins participated in various biological processes including immune response, blood coagulation, complement activation, and so forth. Particularly, fibrinopeptide B (FpB) and heparin cofactor II (HCII) were evaluated as important nodes in the interaction network, which was closely involved in coagulation and inflammation. Further, the FpB level was increased in Group M but decreased in Group Q, while the HCII level exhibited the opposite trend. These findings not only indicated FpB and HCII as potential biomarkers and targets of QFOL in the treatment of RSV pneumonia, but also suggested a regulatory role of QFOL in the RSV-induced disturbance of coagulation and inflammation-coagulation interactions.