Objective To evaluate the efficacy and feasibility of the coil-and-glue method for the reduction of lung volume in rabbit emphysema model.Methods Sixteen rabbits of emphysema model were divided into the occlusion group(n=10),in which both anterior bronchi were occluded using the coil-and- glue method,and the control group(n=6).The maximal static pressure of airway(P_(max)),peak expiratory flow(PEF),end-expiratory volume(EEV)and pressure of oxygen(PO_2)were measured at ante- emphysema,post-emphysema,1 week and 4 week after occlusion respectively.The expectoration(or migration)of coil and collapse of lung were also investigated.Results P_(max)was(20.0?1.3)and(17.1? 1.4)cm H_2O(1 cm H_2O=0.098 kPa)in the occlusion group at ante-emphysema and post-emphysema respectively.P_(max)was(19.2?1.4)cm H_2O in the occlusion group in the 1 week after the occlusion,while (17.1?1.5)cm H_2O in the control group(F=6.68,P

OBJECTIVETo evaluate the authenticity of the parents' memory on their children's immunization status.

METHODSTwo counties and 1 district in each of the 18 prefectures were selected, and parents of the children 1 - 2 years old, residents in counties or floating in district, were studied on the authenticity of their memory regarding their children's immunization status.

RESULTSThe rates of inoculation with all the four expanded programme on immunization (EPI) vaccines were 89.7% in the whole province, and 77.9% among floating children. The authenticity of the reply from parents on their children, inoculation status with vaccines was above 96%. However, less than 50% of the parents could remember what specific vaccines that their children had received. The authenticity of parents' memory was higher in the parents with high school or college education than those who were illiterates or only having had elementary school education. Mothers had better memory than the fathers. Of the children whose parents could not remember the vaccination status, 97% of them had been inoculated.

CONCLUSIONThe definite answer of parents to children's immunization status had high creditability, especially when the mothers having had more schooling. Those children whose parents failed to remember whether vaccination had been received should not be ranked as unimmuned.

Cross-Sectional Studies ; Humans ; Immunization ; statistics & numerical data ; Immunization Schedule ; Infant ; Memory ; Parent-Child Relations ; Parenting ; psychology ; Social Class

Objective To study the function of interleukin (IL)-25 for rheumatoid arthritis (RA) fibroblast-like synoviocytes (FLS) differentiation as well as on the expression of extracellular regulating protein kinase (ERK) and matrix metalloproteinases-3 (MMP-3).Methods The differences on ERK1/2 and MMP-3 protein levels were tested in RA-FLS of RA patients and healthy controls,then IL-17A (10 ng/ml) was tested when the RA-FLS were co-stimulated with different concentrations of IL-25 (0.01,0.1,1 and 10 ng/ml) and IL-17A(10 ng/ml) for 24 hours respectively.The expression of ERK1/2 and MMP-3 protein was detected by the Western blot.T test was used for the comparison between different groups.Results The expression of ERK1/2 (1.71±0.17) and MMP-3 (0.50±0.13) proteins in RA-FLS was higher than the healthy controls (0.50±0.15,0.17±0.05) (t=-9.13,P＜0.01 and t=-4.10,P＜0.05),after stimulated with IL-17A,the expression of ERK1/2 (0.77±0.22) and MMP-3 (0.59±0.13) proteins in RA-FLS were increased compared with the untreated groups (0.18±0.35,0.04±0.03) (t=-4.69,P＜0.01 and t=-7.47,P＜0.01).With increase of the concentration on IL-25,the level of ERK1/2 (0.54±0.26,0.48±0.18,0.48±0.23,0.23±0.06) and MMP-3 (0.58±0.09,0.59±0.14,0.21±0.04,0.04±0.02) in RA-FLS which were stimulated by IL-17A was decreased slowly (t=4.22,P＜0.05 and t=4.95,P＜0.01 and t=7.47,P＜0.01).Conclusion IL-25 can inhibit the stimulation of IL-17A on ERK1/2 and MMP-3 fractionally,which implies that it may take part in the development of RA through this pathway and may be a target for the RA treatment.

OBJECTIVETo examine the effect of Buyanghuanwu decoction (BYHWD) in inducing nerve proliferation in rats with sequelae of ischemic stroke.

METHODSA rat model of ischemic stroke sequelae was established by means of craniectomy in which the right common carotid artery was ligated with 4-0 silk thread followed by cauterization of the right middle cerebral artery. Programmed electric shock was administered 24 h after the onset of ischemic stroke for 2 h daily for 20 consecutive days. The rats in sham operation group were not subjected to ligation of the right common carotid artery or right middle cerebral artery occlusion. The rats in the treatment groups were given oral BYHWD for 15 consecutive days. All the rats received repeated intraperitoneal injections of the cell proliferation-specific marker 5-bromodeoxyuridine (BrdU), and the intake of BrdU in the cerebral tissues was determined by immunohistochemistry.

RESULTSThe number of BrdU-immunoreactive cells in the cerebral tissues of BYHWD-treated rats was significantly greater than that in the untreated model group.

CONCLUSIONBYHWD can promote nerve proliferation in rats with ischemic stroke sequelae.

Administration, Oral ; Animals ; Bromodeoxyuridine ; metabolism ; pharmacokinetics ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; administration & dosage ; therapeutic use ; Female ; Hippocampus ; drug effects ; metabolism ; pathology ; Immunohistochemistry ; Infarction, Middle Cerebral Artery ; drug therapy ; physiopathology ; Male ; Neurons ; drug effects ; metabolism ; pathology ; Neuroprotective Agents ; administration & dosage ; therapeutic use ; Phytotherapy ; Random Allocation ; Rats ; Rats, Wistar

AIMTo construct recombinant retroviruses expressing anti-CD20 scFv/CD80/CD28/zeta gene and detect its expression in Jurkat cells.

METHODCD28-zetacDNA were amplified from plasmids pBULLET and inserted into pLNCX vector that contained anti-CD20 scFv/CD80 gene. The recombinant plasmids were transfected into PA 317 cells. Retroviruses were harvested from culture medium of PA 317 cells. Then NIH 3T3 were transfected with retroviruses. Objective gene expression was determined by PCR and FACS. Jurkat cells were transfected with high titer of retroviruses and resistant clones were obtained by G418 selection. Objective mRNA was determined by RT- PCR.

RESULTSThe recombinant eukaryotic vector was constructed successfully by PCR and enzyme digestion analysis and objective gene was amplified from NIH 3T3 cells transfected with retroviruses by PCR; FACS showed that objective protein could be expressed in NIH 3T3 cells. Objective gene was amplified from Jurkat cells transfected with retroviruses by RT-PCR.

CONCLUSIONRecombinant retrovirus expressing anti-CD20 scFv/CD80/CD28/zeta gene was successfully constructed and objective protein could be expressed in Jurkat cells.

Antigens, CD20 ; genetics ; metabolism ; B7-1 Antigen ; genetics ; metabolism ; CD28 Antigens ; genetics ; metabolism ; Genetic Vectors ; Humans ; Immunoglobulin Fab Fragments ; genetics ; metabolism ; Immunoglobulin Variable Region ; genetics ; metabolism ; Jurkat Cells ; Recombinant Fusion Proteins ; genetics ; metabolism ; Retroviridae ; genetics ; T-Lymphocytes ; metabolism ; Transfection

OBJECTIVETo construct anti-CD20scFv/CD80/CD28/zeta recombinant gene modified T cells, test its effectiveness of eradicating CD20+ lymphoma cells and provide a probably new approach to tumor adoptive immunotherapy.

METHODSCD28-zeta cDNA were amplified from vector pBULLET and inserted into pLNCX vector that contained anti-CD20scFv/CD80 gene. The recombinant vectors were transduced into PA317 cells and high titer retroviruses were obtained to infect human peripheral blood T lymphocytes. Resistant T cells were obtained by G418 selection at one week. Then transduced T lymphocytes and lymphoma cell lines Daudi Raji were cocultured. The cytotoxicity and cytokine production of transduced T cells were determined by non-radio-activation cytotoxicity assay and ELISA respectively.

RESULTSThe recombinant eukaryotic vector was constructed successfully as proved by enzyme digestion analysis and sequencing. These T cells were able to lyse CD20+ target cells and secrete high levels of IL-2 and IFN-gamma in vitro.

CONCLUSIONRecombinant gene modified T cells can be constructed successfully. It can specially kill CD20 positive lymphoma cells in vitro.

Antigens, CD20 ; genetics ; immunology ; B7-1 Antigen ; genetics ; immunology ; CD28 Antigens ; genetics ; immunology ; Cell Line ; Cytotoxicity, Immunologic ; Genetic Vectors ; Humans ; Immunotherapy, Adoptive ; Plasmids ; genetics ; T-Lymphocytes ; immunology ; metabolism ; Transfection

OBJECTIVEPseudomonas aeruginosa is a ubiquitous and opportunistic pathogen that uses the type III secretion system (TTSS) to inject effector proteins directly into the cytosol of target cells to subvert the host cell's functions. Specialized bacterial chaperones are required for effective secretion of some effectors. To identify the chaperone of ExoS, the representative effector secreted by the TTSS of P. aeruginosa, we analyzed the role of a postulated chaperone termed Orf1.

METHODSBy allelic exchange, we constructed the mutant with the deletion of gene Orf1. Analysis of secreted and cell-associated fractions was performed by SDS-PAGE and Western blotting. Using strain expressing in trans Orf1, tagged by V5 polypeptide and histidine, protein-protein interaction was determined by affinity resin pull-down assay in combination with MALDI-TOF. The role of Orf1 in the expression of exoS was evaluated by gene reporter analysis.

RESULTSPull-down assay showed that Orf1 binds to ExoS and ExoT. Secretion profile analysis showed that Orf1 was necessary for the optimal secretion of ExoS and ExoT. However, Orf1 had no effect on the expression of exoS.

CONCLUSIONOrf1 is important for the secretion of ExoS probably by maintaining ExoS in a secretion-competent conformation. We propose to name Orf1 as SpcS for "specific Pseudomonas chaperone for ExoS".

ADP Ribose Transferases ; genetics ; metabolism ; Bacterial Toxins ; genetics ; metabolism ; Base Sequence ; Blotting, Western ; DNA Primers ; Electrophoresis, Polyacrylamide Gel ; Kinetics ; Molecular Chaperones ; genetics ; metabolism ; Protein Binding ; Pseudomonas aeruginosa ; metabolism ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

BACKGROUNDChronic obstructive pulmonary disease (COPD) has a variable natural history and not all individuals follow the same course. This study aimed to identify the prevalence and characteristics of asymptomatic COPD patients from a population-based survey in China.

METHODSA multistage cluster sampling strategy was used in a population from seven different provinces/cities. All residents (over 40 years old) were interviewed with a standardized questionnaire and spirometry. Post-bronchodilator forced expiratory volume in 1 second (FEV(1))/forced vital capacity (FVC) of less than 70% was defined as the diagnostic criterion of COPD. All COPD patients screened were divided into symptomatic group and asymptomatic group according to the presence or absence of chronic respiratory symptoms. Socio-demographic, personal and exposure variables were collected and analyzed.

RESULTSAmong the 1668 patients who were diagnosed with COPD from the 25 627 sampling subjects, 589 (35.3%) were asymptomatic. The age, sex, body mass index (BMI), rural and urban distributions, smoking habit and education levels were similar in the two groups. A total of 64.7% of the asymptomatic patients had no comorbidities. Cardiovascular diseases and lung cancer were more common among symptomatic COPD patients than asymptomatic group. Asymptomatic COPD group were less likely to present with poor ventilation in the kitchen, a family history of respiratory disease and recurrent childhood cough. Asymptomatic COPD patients had significantly higher FEV(1) (73.1% vs. 61.0%), FVC (91.9% vs. 82.0%), and a higher ratio of FEV(1)/FVC (62.9% vs. 58.7%) (all P < 0.001) than symptomatic group. More asymptomatic patients were underdiagnosed (91.9% vs. 54.3%, P < 0.001) than symptomatic patients.

CONCLUSIONSThis large population-based survey confirmed a high prevalence of asymptomatic COPD patients in China. More use of spirometry screening test may be important to the early detection of COPD.

Adult ; Aged ; Aged, 80 and over ; China ; epidemiology ; Educational Status ; Female ; Humans ; Male ; Middle Aged ; Pulmonary Disease, Chronic Obstructive ; diagnosis ; epidemiology ; Risk Factors ; Smoking ; Spirometry ; Surveys and Questionnaires