This study was to establish a model to explore anti- RSV effect of different administration method of Chinese medicine realgar on respiratory syncytial virus type A (RSV-A) replication in Hep-2 cells. Using high-energy ball milling with distilled water to prepare realgar nanoparticles,the concentration of nanometer realgar was tested by molybdenum blue staining method and the size of realgar nanoparticles was tested on Nano Series. Cell culture with ribavirin as a positive control was applied to observe the effect of anti-respiratory syncytial virus type A replication through prevention, treatment or direct inactivation of three different drug administration methods. Realgar nano-particles was found to be a potential inhibitor of RSV-A in a concentration-dependent manner with the median toxic concentration(TC50) of 0.649 microg/mL in Hep-2 cell culture. The median inhibition concentration (IC50) was 0.20 microg/mL when drug was added before virus infection. The IC50 was 0.13 microg/mL when drug was added after virus infection,and it was 0.16 microg/mL when the drug was mixed with virus and added. The therapeutic index (TI) was 3.18, 4.99 and 4.11, respectively. The results showed realgar nanoparticles could inhibit the replication of the RSV and inactivate the RSV in vitro.
Arsenicals ; pharmacology ; Nanoparticles ; Respiratory Syncytial Viruses ; drug effects ; physiology ; Sulfides ; pharmacology ; Virus Replication ; drug effects

OBJECTIVETo evaluate possible inactivating effect of recombined decoction in on mumps virus.

METHODSBy adopting tissue cell culturing technology, a group of viruses including the mumps virus, herpes simplex virus (type I, II), rubella virus, cytomegalovirus (CMV), herpes zoster virus, influenza virus, parainfluenza virus, adeno viruses, respiratory syneytial virus (RSV) were cultured. The cells infected with the viruses were treated with the decoction.

RESULTSThe decoction showed remarkable inhibitory and killing effects on the mumps virus while had no obvious inhibitory and killing effects on host's cells, herpes simplex virus (type I, II), rubella virus, cytomegalovirus (CMV), herpes zoster virus, influenza virus, parainfluenza virus, adeno viruses, respiratory syneytial virus (RSV).

CONCLUSIONSThe decoction had obvious inhibitory and killing effects on mumps virus during single layer cells culture.

Cells, Cultured ; Cytomegalovirus ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Mumps virus ; drug effects ; Respiratory Syncytial Viruses ; drug effects ; Respirovirus ; drug effects ; Rubella virus ; drug effects ; Simplexvirus ; drug effects

OBJECTIVETo study the effect on anti-respiratory syncytial virus of an active compound (AP3) from a Chinese medicinal herb-Herba patriniae in vitro.

METHODSActive component of herba patriniae (AP3) was extracted and its anti-respiratory syncytial virus (RSV) effect was tested. A water soluble substance (AP3) was isolated from a Chinese herb Herba patriniae, by hot water extraction, ethol precipitation and gel permeation column chromatography. The cytotoxicity of AP3 was tested by adding it to HeLa cells directly. Its effect against RSV was estimated by CPEI assay while ribavirin was used as positive control.

RESULTSChemical test showed that the nature of substance AP3 was polysaccharide. The median cytotoxic concentration (TC(50)) of AP3 was 11.45 mg/ml by morphological observation and the median effective concentration (50% effective concentration, EC(50)) of it against replication of the long strain of RSV in HeLa cells was 0.0986 mg/ml. The Therapeutic index (TI = TC(50)/EC(50)) of AP3 was 116.12, much higher than the TI of herba patriniae (AP1) (TI = 59.26) and ribavirin (TI = 53.45). Moreover, AP3 gave a dose-dependent response in inhibiting RSV. In the assay, the effect of AP3 against RSV growth was also tested. In addition, the effect of AP3 on virus growth, AP3 inhibited replication of RSV in HeLa cells, when added at 0 h, 2 h, 4 h after virus infection, were also tested.

CONCLUSIONThis study suggested that the AP3 exerted an obvious inhibitory effect to RSV in HeLa cell culture. This study furnished a reliable evidence for development of a new antiviral drug.

Antiviral Agents ; pharmacology ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; pharmacology ; HeLa Cells ; Humans ; Respiratory Syncytial Viruses ; drug effects ; Time Factors

OBJECTIVETo understand the clinical characteristics and outcome associated with respiratory syncytial virus (RSV) infection in hematopoietic stem cell transplantation (HSCT) recipients with primary immunodeficiencies (PIDs).

METHODNasopharyngeal aspirate samples were collected consecutively before and after HSCT from 9 recipients from Apr. 2009 to Sep. 2010 and analyzed for the presence of RSV using real-time polymerase chain reaction assay. To further verify the presence of the virus, positive samples for PCR were isolated for RSV. RSV G gene was amplified, sequenced and used for phylogenetic analysis.

RESULTThe presence of RSV was detected in 3 out of 9 children. The viral replication in all the patients was prolonged for months. All the 3 patients with RSV infection were treated with intravenous immune globulin (IVIG) and one was treated with antiviral medication. All patients survived and achieved successful immune reconstitution.

CONCLUSIONThis study indicates that the HSCT recipients with PID are at increased risk for RSV infection. RSV can shed for months after the initial infection and the patients recover with the course of immune reconstitution.

Child, Preschool ; Female ; Hematopoietic Stem Cell Transplantation ; adverse effects ; Humans ; Immunologic Deficiency Syndromes ; surgery ; virology ; Infant ; Prognosis ; Respiratory Syncytial Virus Infections ; diagnosis ; drug therapy ; Respiratory Syncytial Viruses ; genetics ; isolation & purification ; physiology ; Virus Replication ; Virus Shedding

OBJECTIVE: To explore the mechanisms for an increase in susceptibility of asthma induced by respiratory syncytial virus (RSV), to observe the expression of interleukin-8 (IL-8) in human bronchial epithelial cells (HBECs) after RSV infection and to invesigate the regulatory effect of IL-8 on Th17/Treg differentiation.
 METHODS: HBECs were divided into a control group and a RSV infected group. The RSVE-infected model of HBECs was established and examined. The expression of IL-8 mRNA was detected by real-time PCR, and the levels of IL-8 were measured by ELISA. Peripheral blood lymphocytes in healthy people were extracted and divided into a control group and an IL-8 treatment group. Based on concentration of IL-8 in RSV-infected HBECs, lymphocytes were treated by a matched concentration of human recombinant IL-8 for 24 h. The distribution of Th17 and Treg subsets in lymphocytes were examined by flow cytometry.
 RESULTS: The RSV-infected HBECs model was successfully established. The infected HBECs were still able to split and passage. The RSV could be detected in every passage in the infected cells. Virus particles indicated by bright yellow green fluorescence were seen under fluorescence microscope. Edema of mitochondrias, expansion of endoplasmic reticulum, fissure around nucleus and intracellular virus particles were all observed under electron microscope. The expression IL-8 mRNA were significantly enhanced in the RSV-infected group, and the level of IL-8 in the RSV-infected group was higher than that in the control group (P<0.05). After IL-8 treatment for 24 h, the ratio of Th17 subsets in lymphocytes were dramatically increased compared to the control group (P<0.05), but there was no difference in the ratio of Treg subsets between the 2 groups (P>0.05).
 CONCLUSION Over-secretion of IL-8 by the RSV-infected HBECs may promote the differentiation of Th17 subsets and maintain the Th17/Tred imbalance.
Cell Differentiation ; Cells, Cultured ; Epithelial Cells ; drug effects ; virology ; Flow Cytometry ; Humans ; Interleukin-8 ; immunology ; pharmacology ; Real-Time Polymerase Chain Reaction ; Recombinant Proteins ; pharmacology ; Respiratory Syncytial Virus Infections ; immunology ; Respiratory Syncytial Viruses ; T-Lymphocytes, Regulatory ; cytology ; Th17 Cells ; cytology

OBJECTIVEDNAzyme/Deoxyribozyme is another novel molecular biological tool following the ribozyme. DNAzyme consists of a 15-nucleotide (nt) internal loop as its catalytic domain and two flanking substrate-recognition domains of 7 to 8 nt each which is complementary to substrate. The RNA substrate is cleaved at a particular phosphodiester located between an unpaired purine and a paired pyrimidine residue. DNAzyme has been applied in fields such as viral infectious disease, tumor, cardiovascular disease and genetic disease. But there is no report about using DNAzyme for anti-respiratory syncytial virus purpose. To observe the inhibitory effects of RNA-cleaving DNAzymes on respiratory syncytial virus (RSV) replication in cultured cells.

METHODSAnti-RSV RNA-cleaving DNAzyme DZ604 was designed to target the RSV genome at the start of the NS2 gene in an effort to inhibit the RNA replication. Microscope and electron microscope were used to observe the effects of anti-RSV genomic RNA DNAzyme on cytopathogenic effect (CPE) and ultrastructural change of 9HTE cell induced by RSV infection. Viral plaque forming reduction assay and MTT assay were used to detect the anti-RSV activity and protective function for RSV infected 9HTE cells of DNAzyme.

RESULTSAnti-RSV genomic RNA DNAzyme (DZ604) significantly improved CPE of RSV-infected 9HTE cells. The time to appearance of CPE and of total CPE was delayed by using DZ604 in a dose-dependent manner. At a 5 micro mol/L concentration of DZ604, CPE of 9HTE cells induced by RSV infection at 10 and 1 multiplicity of infection (MOI) was not improved. At smaller MOI (0.1, 0.01, 0.001, 0.0001) of RSV infection, CPE of 9HTE cells was significantly lightened by DZ604 at the same concentration. DZ604 also significantly improved ultrastructural change of 9HTE cells at early stage of RSV infection. Reduction in RSV yield was 85.56% and 8.33% at concentrations of 5 micro mol/L and 0.25 micro mol/L of DZ604. DZ604 inhibited RSV yield in a dose-dependent manner (P < 0.05). Non-specific DNAzyme did not have anti-RSV activity (P > 0.05).

CONCLUSIONAnti-RSV genomic RNA DNAzyme designed and synthesized in our laboratory was capable of inhibiting respiratory syncytial virus replication specifically in cultured cells. Our data indicated that DNAzymes could be useful for the prevention against respiratory syncytial virus infection.

Animals ; Cell Line ; Cercopithecus aethiops ; DNA, Catalytic ; pharmacology ; Dose-Response Relationship, Drug ; Humans ; Microscopy, Electron ; Respiratory Mucosa ; cytology ; ultrastructure ; virology ; Respiratory Syncytial Viruses ; drug effects ; genetics ; physiology ; Vero Cells ; Virus Replication ; drug effects

OBJECTIVETo investigate the effects of leukotriene receptor antagonist on the levels of Th1 and Th2 cytokines and the serum cysteinyl leukotrenes (CysLTs) in infants and young children with respiratory syncytial virus (RSV) pneumonia.

METHODSThirty-seven infants and young children with RSV pneumonia were divided into two groups after discharge. The cases in group 1 (n=24) were treated with a leukotriene receptor antagonist, Singulair 4 mg once daily for 12 weeks; the cases in group 2 (n=13) were treated with budesonide aerosol 200 ug once or twice daily for 12 weeks. The serum CysLTs, IFN-gamma and IL-4 were detected with enzyme_linked immunosorbent assays (ELISA) for all the 37 cases, and 10 healthy infants of the same age served as controls.

RESULTSThe serum CysLTs level in the cases with RSV pneumonia was significantly higher than that in controls (P<0.05). There was an imbalance in expression of Th1 and Th2 cytokines (IFN-gamma and IL-4 ) in these cases. Both Singulair and budesonide aerosol could correct the imbalance of Th1 and Th2 cytokines. The serum CysLTs level declined after treatment with Singulair in 24 cases, but no significant change occurred after treatment with budesonide aerosol in the remaining 13 cases.

CONCLUSIONSThe serum CysLTs level in children with RSV pneumonia was higher than that in healthy children, and there was an imbalance of Th1 and Th2 cytokines in these infants, which was similar to those with asthma. Leukotriene receptor antagonist may be effective in preventing children with RSV pneumonia from evolving into asthma.

Child, Preschool ; Cytokines ; blood ; Female ; Humans ; Infant ; Leukotriene Antagonists ; administration & dosage ; pharmacology ; Male ; Pneumonia, Viral ; drug therapy ; immunology ; virology ; Respiratory Syncytial Virus Infections ; drug therapy ; immunology ; virology ; Respiratory Syncytial Viruses ; drug effects ; immunology ; Th1 Cells ; drug effects ; immunology ; Th2 Cells ; drug effects ; immunology

OBJECTIVEThe etiology of acute lower respiratory tract infection (LRTI) in children in Wenzhou City remains poorly defined. This study investigated the etiological agents responsible for acute LRTI and patterns of the antibiotic resistant bacterial pathogens in children with acute LRTI from Wenzhou City.

METHODSLower respiratory tract secretions were obtained from 454 children with acute LRTI (aged 1 month to 10 years, median age 6 months) within 24 hrs after admission for bacterial culture. Meanwhile respiratory viruses were detected by the Direct immunofluorescence (DIF) assay. The K-B method was applied for the drug susceptibility test.

RESULTSEtiological agents were identified in 297 cases out of 454 patients (65.4%. Viral pathogens were identified in 229 cases (50.4%), bacteria in 135 cases (29.7%) and mixed viral-bacterial infections in 67 cases (14.8%). The isolating rate of Respiratory syncytial virus (RSV) was the highest (180 cases, 39.6%) in all of the samples. The isolating rates of other viral pathogens were as follows: Parainfluenza virus 3 type (PIV3) (6.6%), Adenovirus (2.2%), Influenza A (0.9%) and Influenza B (0.7%). Of the 135 strains of bacterial pathogens, 19 kinds of bacterial pathogens were isolated. The predominant isolate was Klebsiella pneumoniae (K. pneumoniae) (9.9%), followed by Escherichia coli (E.coli) (4.4%), Streptococcus pneumoniae (S. pneumoniae) (4.2%) and Staphylococcus aureus (S. aureus) (4.2%). The isolating rates of K. pneumoniae and E.coli with extended-spectrum beta-lactamases strains (ESBLs) positive were 42.2% and 65.0%, respectively. The pathogens isolated of the first 5 places in children with acute LRTI under six months were RSV, K. pneumoniae, PIV3, E.coli and S. aureus in turn. RSV, PIV3, S. pneumoniae, K. pneumoniae and E.coli were found to be the pathogens of the first 5 places in children with acute LRTI between six months and three years. The resistant rates of K. pneumoniae and E.coli to ampicillin were 97.8% and 75.0%, respectively. K. pneumoniae and E.coli with positive ESBLs were resistant to cephalosporin. The resistant rates of S. pneumoniae to erythromycin and penicilin were 100% and 68.4%, respectively. The resistant rates of S. aureus to erythromycin and penicillin were 94.7% and 89.5%, respectively.

CONCLUSIONSRSV is the most common pathogen responsible for acute LRTI in children in Wenzhou City, followed by K. pneumoniae and PIV3. The rate of antibiotic resistance of common bacteria and the isolating rate of Gram-negative bacillus with ESBLs positive are high.

Acute Disease ; Bacteria ; drug effects ; Child ; Child, Preschool ; Drug Resistance, Bacterial ; Female ; Humans ; Infant ; Klebsiella pneumoniae ; isolation & purification ; Male ; Microbial Sensitivity Tests ; Respiratory Syncytial Viruses ; isolation & purification ; Respiratory Tract Infections ; etiology

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.
Animals ; Biomarkers ; blood ; Chromatography, Liquid ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Fibrinopeptide B ; analysis ; genetics ; Gene Expression Regulation ; drug effects ; Heparin Cofactor II ; analysis ; genetics ; Lung ; pathology ; Mice, Inbred BALB C ; Proteome ; drug effects ; Proteomics ; Respiratory Syncytial Virus Infections ; blood ; drug therapy ; Respiratory Syncytial Viruses ; drug effects ; Tandem Mass Spectrometry

Viral infections are the major causes of morbidity and mortality in elderly people and young children throughout the world. The most common pathogens include coxsackie virus (CV) and respiratory syncytial virus (RSV). However, no antiviral agents with low toxicity and drug resistance are currently available in clinic therapy. The present study aimed to examine the antiviral activities of emodin (an ingredient of Rheum palmatum) against CVB5 and RSV infections, in an attempt to discover new antiviral agents for virus infection. The monomer emodin was extracted and isolated from Rheum palmatum. The antiviral activities of emodin on HEp-2 cells were evaluated, including virus replication inhibition, virucidal and anti-absorption effects, by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tet-razolium bromide (MTT) assay and plaque reduction assay (PRA). The kinetics of virus inhibition by emodin in a period of 14 h was further determined by plaque assay and quantitative real time PCR (qPCR). Cytokine (IFN-γ, TNF-α) mRNA expressions after emodin treatment (7.5, 15, 30 μmol/L) were also assessed by qPCR post-infection. The results showed that emodin had potent inhibitory activities against CVB5 and RSV, with the 50% effective concentration (EC50) ranging from 13.06 to 14.27 μmol/L and selectivity index (SI) being 5.38-6.41 μmol/L. However, emodin couldn't directly inactivate the viruses or block their absorption to cells. It acted as a biological synthesis inhibitor against CVB4 and RSV in a concentration- and time-dependent manner, especially during the first 0-4 h post-infection. Moreover, emodin could decrease the mRNA expression of IFN-α but enhance TNF-γ expression significantly compared to the viral controls in vitro. Our results provide a molecular basis for development of emodin as a novel and safe antiviral agent for human enterovirus and respiratory virus infection in the clinical therapy.
Antiviral Agents ; pharmacology ; Cell Line ; Cell Line, Tumor ; Emodin ; pharmacology ; Enterovirus B, Human ; drug effects ; physiology ; Humans ; In Vitro Techniques ; Respiratory Syncytial Viruses ; drug effects ; physiology ; Rheum ; chemistry ; Virus Replication