Asthma ; diagnosis ; therapy ; Child ; China ; Chronic Disease ; Cough ; diagnosis ; therapy ; Humans

Objective To investigate the clinical spectrum of respiratory viruses in infants and young children with acute lower respiratory infection(ALRI) in Chongqing area from 2003-2007.And to assess the clinical diagnostic value of virus detection in nasopharyngeal secretions(NPS) and serum viral antibody detection for ALRI.Methods Cases of 2 529 specimens of NPS in hospitalized children with ALRI from Apr.2003 to Oct.2007 were taken for detecting 7 common respiratory virus antigens by immunofluorescence assay including respiratory syncytial virus (RSV),adenovirus(ADV),influenza A(IA),influenza B (IB),parainfluenza virus1-3 (PIV1,PIV2,PIV3).Fifty-five thousand eight hundred and eighty-seven samples were tested for ADV-IgM by ELISA.Among those,45 159 cases were further tested for RSV-IgM by ELISA.Results Respiratory virus pathogens were detected in 778 samples out of 2 529(30.76%) including RSV positive in 668 samples (85.86%),PIV3 positive in 75 samples (9.64%),IA positive in 22 samples (2.57%),ADV positive in 15 samples ( 1.93%),only 1 sample ( 0.13%) positive for both PIV1 and RSV. And the positive rate of RSV-IgM was 0.9%-15.2%,and the positive rate for ADV-IgM was about 0.6%-10.6%.RSV infection occured mainly in winter and spring.Conclusions Respiratory virus is the most common pathogen in children with ALRI during the survey period in Chongqing area,especially for RSV infection.The pattern of RSV circulation varied every year with seasonality.It is suggest that this year is peak one for RSV infection from the monthly positive results,especially in Feburary(50%) in 2007.But the infection rate of PIV3,IA,ADV and PIV1 are lower,particularly IB and PIV2 infection have not been seen for the last 5 years.It is fast and accurate to detect RSV antigen and suit to clinical diagnosis by using immunofluorescence assay than other antibody detection.

To investigate the effect of Tanreqing injection on immune activity of peripheral blood lymphocytes of patients with lung cancer. The peripheral blood lymphocytes of patients with lung cancer and healthy persons were separated by the density gradient centrifugation method for subsequent experiments, with those from healthy persons as the positive control. The effect of Tanreqing injection on stimulating the proliferation of lymphocytes with phytohemagglutinin (PHA) was determined by MTT method. The effect of Tanreqing injection on the lymphocyte secretions of IFN-γ and TNF-α and the subset ratio of lymphocytes cultured separately or with Tanreqing injection of different concentrations were examined by ELISA and flow cytometry (FCM) respectively. In addition, the LDH release assay was used to detect the cytotoxicity of cytotoxic T cells (CTL) and natural killer cells (NK). According to the findings, all of immunological indexes of lymphocytes from patients with lung cancer were weaker than that of healthy persons, but with the obvious increases in proliferation activity and IFN-γ and TNF-α secretions of lymphocytes co-cultured with Tanreqing Injection (P < 0.05). Among lymphocyte subsets co-cultured with Tanreqing Injection, CD3+, CD3+ CD4+ and CD3- CD16 + 56+ cell ratios notably increased, whereas CD4+ CD25+ Treg cell ratio obviously decreased (P < 0.05). In the meantime, Tanreqing injection can markedly promote the cytotoxicities of CTL and NK (P < 0.05). In conclusion, Tanreqing injection shows a significant effect in promoting the immune activity of lymphocytes from patients with lung cancer and their anti-tumor immunity.
Cell Proliferation ; drug effects ; Cells, Cultured ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Interferon-gamma ; genetics ; immunology ; Killer Cells, Natural ; drug effects ; immunology ; Lung Neoplasms ; drug therapy ; genetics ; immunology ; physiopathology ; T-Lymphocytes, Cytotoxic ; drug effects ; immunology ; Tumor Necrosis Factor-alpha ; genetics ; immunology

OBJECTIVETo study the effects of extremely low frequency electromagnetic fields (ELF EMFs) on apoptosis and cell cycle of mouse brain and liver cells.

METHODSMice were exposed to 50 Hz, 0.2 mT or 6.0 mT electromagnetic fields for 2 weeks. TUNEL and flow cytometric methods were used to analyze apoptosis and cell cycle of brain and liver cells.

RESULTSAfter exposure to 0.2 mT and 6.0 mT ELF EMFs for 2 weeks, apoptosis rates of brain cells [(5.60 +/- 1.47)% and (4.73 +/- 0.48)% respectively] were higher than that of control [(2.90 +/- 0.75)%], and apoptosis rates of liver cells [(4.19 +/- 2.08)% and (3.38 +/- 0.65)% respectively] were higher than that of control [(1.84 +/- 0.76)%]. G0/G1 cell percentage of brain cells [(80.21 +/- 1.68)% and (79.54 +/- 0.56)% respectively] were higher than that of control [(76.85 +/- 0.83)%], and those of liver cells [(79.42 +/- 1.80)% and (80.47 +/- 1.79)% respectively] were higher than that of control [(73.36 +/- 3.10)%]. The above differences were all statistically significant as P < 0.05. At the same time S and G2 + M cell percentage of brain and liver cells were significantly decreased.

CONCLUSIONExposure to 50 Hz EMFs may alter cell cycle and induce apoptosis of mouse brain and liver cells.

Animals ; Apoptosis ; radiation effects ; Brain ; cytology ; radiation effects ; Cell Cycle ; radiation effects ; Electromagnetic Fields ; Flow Cytometry ; In Situ Nick-End Labeling ; Liver ; cytology ; radiation effects ; Male ; Mice

OBJECTIVETo study the effects of extremely low frequency electromagnetic fields (ELF EMFs) on c-fos gene expression in mouse brain and liver tissues.

METHODSMice were exposed to 50 Hz sinusoidal 0.2 mT or 6.0 mT electromagnetic field for 2 weeks or 4 weeks. Competitive RT-PCR method was used to measure c-fos mRNA level.

RESULTSAfter exposure to 0.2 mT or 6.0 mT field for 2 weeks, c-fos mRNA levels in brain tissue [(0.0178 +/- 0.0076) amol/120 ng cDNA and (0.0092 +/- 0.0042) amol/120 ng cDNA respectively] were higher than that of control level [(0.0012 +/- 0.0005) amol/120 ng cDNA] (P < 0.05). In liver tissue c-fos mRNA levels [(0.0117 +/- 0.0055) amol/120 ng cDNA and (0.0148 +/- 0.0162) amol/120 ng cDNA respectively] were also higher than that of control level [(0.0005 +/- 0.0005) amol/120 ng cDNA] (P < 0.05). After exposure to 0.2 mT or 6.0 mT field for 4 weeks, c-fos mRNA levels in brain tissue [(0.0100 +/- 0.0054) amol/120 ng cDNA and (0.0198 +/- 0.0079) amol/120 ng cDNA respectively] were higher than that of control level [(0.0015 +/- 0.0008) amol/120 ng cDNA] (P < 0.05). In liver tissue the exposure induced much higher expression level [(0.0173 +/- 0.0122) amol/120 ng cDNA and (0.0133 +/- 0.0090) amol/120 ng cDNA respectively] while no expression was found in the control.

CONCLUSIONExposure to 50 Hz electromagnetic fields may induce up-regulation of c-fos transcription in mouse brain and liver tissue.

Animals ; Brain ; metabolism ; radiation effects ; Dose-Response Relationship, Radiation ; Electromagnetic Fields ; Gene Expression Regulation ; radiation effects ; Genes, fos ; genetics ; Liver ; metabolism ; radiation effects ; Male ; Mice ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Time Factors

AIM: The study was undertaken to investigate the effort of Dexamethasone (DEX) on cultured rabbit corneal epithelial (RCE) cells and rabbit corneal epithelial wound healing.METHODS: For the in vitro experiments, primary cultures of RCE cells were used. DEX in different concentrations was added to cultured RCE cells. The effects were measured with tetrazolium salt (MTT)method and flow cytometry. For the in vivo wound-healing experiments, a central corneal deepithelialization was created and were treated with 0.1g/L DEX eyedrop randomly explain how randomly. Epithelial wound healing was evaluated clinically and analyzed histopathologically using light microscopy along with immunohistochemical staning and electronic microscopy.RESULTS: Less than 0.1g/L DEX didn't influence survival rate in cell culture conditions by MTT assay. Flow cytometric studies revealed that 0.1g/L DFX had no effect on cellular growth phase in cultured rabbit corneal epithelial cells. The mean time of the epithelial healing was significantly shorter in the DEX-treated group than in the control group at 24h. There were strong proliferative-cell-nuclear-antigen(PCNA) expressions in newly generated epithelial cells of both groups. The Dex-treated group had a more regular architecture of stromal lamella and significantly less inflammatory response than the control group under electronic microscopy.CONCLUSION: Less than 0.1g/L DEX had no inhibiting effect on cultured rabbit corneal epithelial cell growth.0.1g/L DEX eye drops can effectively promote epithelial growth and reduce inflammatory response, which may have useful clinical application at the early stage of corneal wound healing process.

OBJECTIVEThe impact of dendritic cells (DCs) and regulatory T cells (Tr) on the pathogenesis of asthma have been investigated over the past decades. As the professional antigen presenting cells, DCs not only prime immune response directing Th0 cells toward different T subtypes but also induce immune tolerance. As an immunoregulator, Bacillus Calmette-Guerin (BCG) has potential to be applied in allergic diseases such as asthma for prevention. Previous study showed that neonatal BCG vaccination could induce Th1/Tr1 development in mice in vivo. To further identify the mechanism of neonatal BCG vaccination on T cell subsets differentiation, the present study was designed to investigate the impact of BCG vaccination on splenic DCs development in neonatal mice.

METHODSNeonatal specific pathogen free (SPF) BALB/c mice (2-3 days) were divided into intraperitoneal BCG-treated group, subcutaneous BCG-treated group and control group; simultaneously adult SPF BALB/c mice (6-8 weeks) were divided into intraperitoneal BCG-treated and control group. The BCG-treated mice were inoculated with 1 x 10(5) CFU BCG, the mice in control group were not inoculated with any vaccine. Four weeks post BCG vaccination, spleen cells were isolated. With flow cytometry, subtype and maturity of splenic DCs were analysed. Moreover, cells were further separated into mononuclear cell by Ficoll solution. The mononuclear cells were stimulated by 1 microg/ml lipopolysaccharide (LPS) for 18 or 10(5) CFU /ml BCG for 48 hours at 37 degrees C in a humidified atmosphere containing 5% CO2 and cytokines concentration was detected by ELISA.

RESULTS(1) CD11c(+) CD8alpha(+) and CD11c(+) CD8alpha(-) DCs were found in spleen cells of the BALB/c mice. In comparison with the control group, the percent of CD11c(+) CD8alpha(-) DCs in intraperitoneal BCG group significantly declined (P < 0.01) and that of CD11c(+) CD8alpha(+) DCs significantly increased (P < 0.01), there were no significant difference in DC subtypes between intraperitoneal and subcutaneous BCG-vaccinated mice. In contrast, the percent of CD11c(+) CD8alpha(-)DCs markedly increased (P < 0.01) and that of CD11c(+) CD8alpha(+)DCs noticeably reduced (P < 0.01) in adult BCG-vaccinated mice. The percent of CD11c(+)CD8alpha(-)DC was significantly higher and that of CD11c(+) CD8alpha(+)DC was significantly lower in adult-vaccinated BALB/c mice than that of neonatal-vaccinated ones. (2) The expression of costimulatory molecules CD40 on CD11c(+) CD8alpha(-) DCs and CD86 on CD11c(+) CD8alpha(+) DCs in neonatal BCG-treated BALB/c mice was higher than the controls. There were no significant difference in expression of costimulatory molecules on DC between neonatal BCG-vaccinated mice. Compared with the control group, expression of CD40 and MHC-II molecules on CD11c(+) CD8alpha(-) and CD11c(+) CD8alpha(+)DC was significantly higher and that of CD86 was significantly lower in adult BCG-vaccinated mice. The expression of costimulatory molecules on DC had no significant difference between neonatal and adult BCG vaccinated BALB/c mice. (3) As compared with the control mice, concentration of IL-12p70 induced by LPS and IL-10 induced by BCG in vitro from spleen cells culture supernatant was noticeably elevated (P < 0.05) in neonatal BCG-treated BALB/c mice, but that of IL-6 did not change by LPS or BCG stimulation.

CONCLUSION(1) By up-regulating splenic CD8alpha(+)DCs and inducing IL-12p70 and IL-10 production in BALB/c mice, neonatal BCG vaccination promoted Th1/Tr1 development. (2) The effect of BCG vaccination on DC was different between neonates and adult BALB/c mice.

Animals ; Animals, Newborn ; BCG Vaccine ; immunology ; Cell Differentiation ; Cells, Cultured ; Dendritic Cells ; cytology ; immunology ; metabolism ; Immunophenotyping ; Mice ; Mice, Inbred BALB C ; Spleen ; cytology ; immunology ; metabolism

OBJECTIVETo study the impact of neonatal bacillus Calmette-Guerin(BCG) vaccination on lung Th17 cells and IL-17 in murine asthma model.

METHODSNeonatal BALB/c mice were divided into three groups: control, OVA and BCG/OVA groups. BCG was administerd in the BCG/OVA group on postnatal day 2 or 3. Except the control group, the mice in the other two groups were sensitized and undergone OVA challenge. Inflammatory cell numbers and morphological identification of leucocytes in bronchoalveolar lavage fluid (BALF) were measured by light microscopy. Cytokine IFN-gamma and IL-17 levels in BALF were measured using ELISA. The percentage of lung Th17 cells were assayed by flow cytometry.

RESULTSThere was significantly larger number of total cells, lymphocytes, eosinophils and neutrophils in BALF in the OVA and BCG/OVA groups compared with the control group. The number or percentage of those cells in the BCG/OVA group was lower than that in the OVA group. The level of IL-17 in BALF was significantly higher in the OVA and the BCG/OVA groups compared with the control group, while the level of IFN-gamma was lower. The OVA group had higher level of IL-17 than the BCG/OVA group. The mice in the OVA and the BCG/OVA groups had a higher percentage of Th17 cells in lungs compared with the control group, but there were no significant differences in the percentage of Th17 cells between the OVA and the BCG/OVA groups.

CONCLUSIONSTh17 cells and IL-17 play roles in the pathogenesis of asthma. BCG vaccination can reduce the level of IL-17 in BALF and the reduced IL-17 may be mainly from other IL-17-producing cells in the lungs, not Th17 cells.

Animals ; Animals, Newborn ; Asthma ; immunology ; BCG Vaccine ; immunology ; Disease Models, Animal ; Interferon-gamma ; analysis ; Interleukin-17 ; analysis ; Lung ; immunology ; Mice ; Mice, Inbred BALB C ; Ovalbumin ; immunology ; T-Lymphocytes, Helper-Inducer ; immunology ; Vaccination