ABSTRACT
BACKGROUND: A novel avian influenza A (H7N9) virus emerged in eastern China in February 2013. 413 confirmed human cases, including 157 deaths, have been recorded as of July 31, 2014. METHODS: Clinical specimens, including throat swabs, sputum or tracheal aspirates, etc., were obtained from patients exhibiting influenza-like illness (ILIs), especially from those having pneumonia and a history of occupational exposure to poultry and wild birds. RNA was extracted from these samples and a multiplex one-step real-time RT-PCR assay was developed to specifically detect the influenza A virus (FluA). PCR primers targeted the conserved M and Rnase P (RP) genes, as well as the hemagglutinin and neuraminidase genes of the H7N9 virus. RESULTS: The multiplex assay specifically detected the avian H7N9 virus, and no cross-reaction with other common respiratory pathogens was observed. The detection limit of the assay was approximately 0.05 50% tissue culture infective doses (TCID50), or 100 copies per reaction. Positive detection of the H7N9 virus in sputum/tracheal aspirates was higher than in throat swabs during the surveillance of patients with ILIs. Additionally, detection of the matrix (M) and Rnase P genes aided in the determination of the novel avian H7N9 virus and ensured the quality of the clinical samples. CONCLUSIONS: These results demonstrate that the multiplex assay detected the novel avian H7N9 virus with high specificity and sensitivity, which is essential for the early diagnosis and treatment of infected patients.
Subject(s)
Influenza A Virus, H7N9 Subtype/genetics , Influenza, Human/diagnosis , Animals , China/epidemiology , Dogs , Female , Humans , Influenza, Human/virology , Limit of Detection , Madin Darby Canine Kidney Cells , Male , Molecular Diagnostic Techniques , Multiplex Polymerase Chain Reaction , Real-Time Polymerase Chain Reaction , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
Enterovirus 71 (EV71) is a major causative agent of hand, foot, and mouth disease (HFMD) in children. The role of T follicular helper (TFH) cells in EV71-infected children remains unclear in regulating humoral immunity. The frequency of circulating ICOS(high)/PD-1(high)CXCR5(+)CD4(+) TFH cells in the children with mild and severe EV71 infection and healthy controls (HC) was detected by flow cytometry, respectively. IL-21 and IL-6 mRNA expression and their serum levels, Bcl-6 mRNA expression, and specific neutralizing antibodies against EV71 (NAb-EV71) were measured. In the acute stage of patients with EV71 infection, increased frequencies of circulating TFH cells with ICOS(high) and PD-1(high) expression in the mild and severe patients were observed, and the positive correlations among the frequencies of circulating TFH cells and the serum levels of IL-21, IL-6, and NAb-EV71 titres were detected, respectively. Moreover, the expressions of IL-6 and IL-21 mRNA in PBMCs from patients were also significantly higher than those of HC. However, further analysis did not reveal any significant differences between mild and severe patients. These data indicate a role of TFH cells and associated cytokines in modulating the humoral response during the pathogenesis of EV71 infection.
Subject(s)
Enterovirus A, Human/immunology , Hand, Foot and Mouth Disease/immunology , T-Lymphocytes, Helper-Inducer/immunology , Age Factors , Antibodies, Neutralizing/immunology , Antibodies, Viral/immunology , Antigens, Surface/metabolism , Case-Control Studies , Child , Child, Preschool , Cytokines/blood , Cytokines/genetics , Cytokines/metabolism , Female , Flow Cytometry , Gene Expression , Hand, Foot and Mouth Disease/blood , Hand, Foot and Mouth Disease/diagnosis , Humans , Immunophenotyping , Infant , Lymphocyte Count , Male , Receptors, CXCR5/metabolism , T-Lymphocyte Subsets/immunology , T-Lymphocyte Subsets/metabolism , T-Lymphocytes, Helper-Inducer/metabolismABSTRACT
Serological surveillance conducted in areas of an outbreak of influenza A(H7N9) infection in China found no seropositivity for antibodies specific for avian-origin influenza A(H7N9) among 1129 individuals of the general population, whereas >6% of 396 poultry workers were positive (on the basis of a hemagglutination inhibition titer of ≥ 80) for this subtype, confirming that infected poultry is the principal source of human infections and that subclinical infections are possible. Fourteen days after symptom onset, elevated levels of antibodies to A(H7N9) were found in 65.8% of patients (25/38) who survived but in only 28.6% of those (2/7) who died, suggesting that the presence of antibodies may improve clinical outcome in infected patients.
Subject(s)
Influenza A Virus, H7N9 Subtype/isolation & purification , Influenza, Human/epidemiology , Influenza, Human/virology , Occupational Exposure , Adolescent , Adult , Aged , Aged, 80 and over , Antibodies, Viral/blood , Child , Child, Preschool , China/epidemiology , Female , Hemagglutination Inhibition Tests , Humans , Infant , Influenza, Human/immunology , Male , Middle Aged , Seroepidemiologic Studies , Survival Analysis , Young AdultABSTRACT
Rituximab is the first line drug to treat non Hodgkin's lymphoma (B-NHL) alone or in combination with chemotherapy. However, 30-40% of B-NHL patients are unresponsive to rituximab or resistant after therapy. Human phosphatidylethanolamine-binding protein 4 (hPEBP4) is a novel member of PEBP family and functions as an anti-apoptotic molecule. In this study, we found hPEBP4 to be expressed in up to 90% of B-cell lymphoma patients, but in only 16.7% of normal lymph nodes. Interestingly, hPEBP4 overexpression inhibited rituximab-mediated complement dependent cytotoxicity (R-CDC) and antibody-dependent cell-mediated cytotoxicity (ADCC) in B-NHL cells while downregulation of hPEBP4 augmented the therapeutic efficacy of rituximab both in vitro and in vivo. Furthermore, hPEBP4 silencing sensitized the primary B-acute lymphocytic leukemia (B-ALL) cells to R-CDC. During rituximab-mediated complement dependent cytotoxicity, hPEBP4 was recruited to the cell membrane in a PE-binding domain dependent manner and inhibited R-CDC induced calcium flux and reactive oxygen species (ROS) generation. These events contributed to the decrease of cell death induced by R-CDC in B-cell lymphomas. Meanwhile, hPEBP4 knockdown potentiated the chemosensitization of the rituximab in B-cell lymphoma cells by regulating the expression of Bcl-xl, Cycline E, p21(waf/cip1) and p53 and the activation of caspase-3 and caspase-9. Considering that hPEBP4 conferred cellular resistance to rituximab treatment and was preferentially expressed in lymphoma tissue, it could be a potential valuable target for adjuvant therapy for B-cell lymphoma.
Subject(s)
Antibodies, Monoclonal, Murine-Derived/pharmacology , Antibodies, Monoclonal, Murine-Derived/therapeutic use , Lymphoma, B-Cell/drug therapy , Phosphatidylethanolamine Binding Protein/metabolism , Animals , Apoptosis/drug effects , Apoptosis/genetics , Cell Line, Tumor , Drug Resistance, Neoplasm/genetics , Female , Gene Silencing , Humans , Immunohistochemistry , In Vitro Techniques , Lymphoma, B-Cell/genetics , Lymphoma, B-Cell/metabolism , Mice , Mice, Inbred BALB C , Mice, Nude , Microscopy, Confocal , Phosphatidylethanolamine Binding Protein/genetics , Reactive Oxygen Species/metabolism , RituximabABSTRACT
OBJECTIVE: Use proteomic technique to research the effects of Qishenyiqi formula in acute myocardial infarcted rats, and further explore its protein-level mechanism. METHOD: The rat hearts of sham, myocardial infarcted and Qishenyiqi formula treated groups were collected. Analyzing and comparing the differences of protein expressions in each groups with two-dimensional electrophoresis, computer-assisted image analysis and matrix-assisted laser desorption/ionization mass spectrometry. RESULT: Compared with myocardial infarcted group, there were 14 protein expressions changed in Qishenyiqi formula treated group. The result showed that 9 protein expressions were up-regulated, and 5 ones were down-regulated significantly. These proteins are associated with energy metabolism, mitochondrial function, oxidative stress, cytoskeleton and others. CONCLUSION: Myocardial protective effects of Qishenyiqi formula in myocardial infarcted procedure might be closely related to the recovery of energy supply, the reduction of oxidative stress, as well as the promotion of cell recovery.
