Humans ; Proteomics ; methods ; Silicosis ; metabolism ; Specimen Handling ; methods

OBJECTIVETo observe the effects of complement fragment C3f on expression and secretion of collagen I, III and transforming growth factor( TGF)-beta1 in human embryonic lung fibroblast (MRC-5) cells.

METHODSMRC-5 cells were cultured with C3f (the synthetic 17 peptides fragments of complement C3). The extracellular and intracellular expression levels of type I, III collagens and TGF-beta1 in MRC-5 cultures were detected by ELISA and immunohistochemistry, respectively.

RESULTSThe expression levels of type I, III collagen and TGF-beta1 in the supernatant of MRC-5 cultures decreased significantly with the concentrations of C3f as compared with controls (P < 0.05). Also the expression level of TGF-beta1 in MRC-5 cytoplasm reduced significantly as compared with controls (P < 0.05).

CONCLUSIONThe results of present in vitro study showed that the complement fragment C3f could reduce the formation of TGF-beta1 and type I, III collagens in MRC-5 cells, and inhibit the lung tissue fibrosis.

Cell Line ; Collagen Type I ; metabolism ; Collagen Type III ; metabolism ; Complement C3b ; pharmacology ; Fibroblasts ; drug effects ; metabolism ; Humans ; Lung ; cytology ; drug effects ; embryology ; Transforming Growth Factor beta1 ; metabolism

OBJECTIVETo establish 2-dimensional gel electrophoresis (2-DE) images and seek differentially expressed serum proteins for understanding the pathogenesis of silicosis.

METHODS2-DE and matrix-assisted laser desorption/ionization time of flight tandem mass spectrometry (MALDI-TOF-MS/MS) were used to screen differentially expressed serum proteins among silica-exposed population, suspect of silicosis (0+), phase one (I) group with silicosis and control group(non silica exposure).

RESULTSComplement C4, leucine-rich alpha-2-glycoprotein and alpha-1-antitrypsin were significantly highly expressed in suspect of silicosis (0+) group(P < 0.01), but lowly in other groups. Inversely, serotransferrin was significantly down-regulated only in suspect of silicosis (0+) group(P < 0.01). Plasma glutathione peroxidase, tetranectin, apolipoprotein A-I and transthyretin were equally expressed in the serum of control group and silica-exposed population group, but decreased in the suspect of silicosis (0+) and phase (I) group.

CONCLUSIONComplement C4, leucine-rich alpha-2-glycoprotein, alpha-1-antitrypsin, serotransferrin, plasma glutathione peroxidase, tetranectin, apolipoprotein A-I and transthyretin are differentially expressed in the silica-exposed group and phase (I) group with silicosis, and the result should be validated by other biochemical technologies.

Adult ; Aged ; Aged, 80 and over ; Blood Proteins ; analysis ; Electrophoresis, Gel, Two-Dimensional ; Humans ; Middle Aged ; Proteomics ; methods ; Silicosis ; blood ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

OBJECTIVETo explore changes of Clara cell protein (CC16) and surfactant protein-D (SP-D) in the serum of patients with silicosis.

METHODThe concentrations of CC16 and SP-D were measured in the serum by sandwich enzyme-linked immunosorbent assays. The subjects consisted of 30 healthy volunteers and 90 silica-exposed workers including silica-exposed group, the silicosis of suspects group (0(+)) and the silicosis phase I group, 30 subjects each groups.

RESULTSThe concentrations of CC16 in the serum was significantly decreased in silica-exposed workers compared to controls (P < 0.01); The concentrations of CC16 in the serum were higher in lifelong nonsmokers than the current smokers in control subjects (P < 0.05), but they were no differences between lifelong nonsmokers and current smokers of 90 silica-exposed workers. Compared with control subjects, the levels of SP-D in the serum of silicosis suspects (0(+)) and silicosis phase I groups were significantly elevated (P < 0.01, respectively), which were also higher than silica-exposed group (P < 0.05 and P < 0.01, respectively), Discriminant equations set by CC16 and SP-D were used in diagnosis of silicosis, and the rate of accuracy in healthy volunteers, the silica-exposed group and the silicosis phase I group were 86.7%, 86.7% and 76.7%, respectively, The total rate of correct classification hit 84.2%.

CONCLUSIONThe serum CC16 of long-term silica-exposed workers is decreased, and SP-D is increased gradually.

Adult ; Case-Control Studies ; Epithelial Cells ; metabolism ; Humans ; Male ; Middle Aged ; Pulmonary Surfactant-Associated Protein D ; blood ; Silicosis ; blood ; Uteroglobin ; blood

Objective:To evaluate the immune response efficiency of the recombinant abrin B chain protein (rATB) in BALB/c mice.Methods:BALB/c mice were vaccinated intraperitoneally (i.p.) with the purified rATB protein.ELISA was used to detect the IgG,IgG1 and IgG2a.Meanwhile,the expression of IL-4 and IFN-γwere detected by flow cytometry.Then the protein neutralization assay and efficacy of passive protection were done .Results:The anti-rATB antibody was detected by ELISA after vaccinated the rATB in the mice and results showed that the highest titer reached 106 ,which was acquired after the last vaccination .Meanwhile,a strong secondary response was triggered in mice when challenged with toxin .In contrast ,the antibody response of the PBS-vaccinated mice was less than 1:10 and P<0.05 indicated obvious difference between test group and control group .The result of IgG 1 was the same as the IgG ,while the result of IgG2a had not changed.There was significant difference for IL-4 between two groups (P<0.05),while no significant difference for IFN-γwas observed.All BALB/c mice survived after challenged with 5×LD50 of AT.Transfusion of sera from immunized mice provided passive protection in naive mice.Furthermore, the rATB could induce specific neutralizing antibodies against abrin toxin.Conclusion: The recombinant protein can induce the Th 2-type immune response and trigger a good immune response and protective efficacy,which means it may be a promising vaccine candidate against human exposure to AT .

OBJECTIVETo study the factors in relation to severe acute respiratory syndromes (SARS) among health care workers and to develop related protective measures.

METHODSCase-control study was applied. A standardized questionnaire was used to collect SARS related information for health care workers who had contacted or treated SARS patients. Univariate analysis was conducted using SPSS 10.0 software package and multivariate logistic regression analysis was conducted using SAS 6.12.

RESULTSTwenty-seven of the 49 factors under study were significantly associated with SARS infection, in which 22 factors were protective, and the other 5 were risk factors. 27 factors were included for multivariate logistic regression analysis. Results showed that six factors as wearing eye glasses, wearing protection gowns, exposure to secrets/mode of contact with SARS patients, types of mask and the working years atc, remained significant association with hospital infection of SARS.

CONCLUSIONSARS infection in heath care workers was related to many factors during the process of diagnoses and/or treatment. It is recommended that adequate masks, eye-protection and protective gowns should be adopted for heath care workers during the process of clinical diagnoses and treatment of SARS patients.

Case-Control Studies ; China ; epidemiology ; Cross Infection ; prevention & control ; Female ; Health Personnel ; Humans ; Infectious Disease Transmission, Patient-to-Professional ; prevention & control ; Logistic Models ; Male ; Risk Factors ; Severe Acute Respiratory Syndrome ; epidemiology ; transmission ; Surveys and Questionnaires

OBJECTIVESTo study the correlation between positive rates of RNA in clinical confirmed severe acute respiratory syndrome (SARS) patients and its appearance in relation to the development of the disease in order to provide scientific basis for early diagnosis, effective prevention and treatment of the disease.

METHODSOne-step reverse transcription-polymerase chain reaction (RT-PCR) was used to amplify the SARS RNA in the clinical specimens from different courses of the disease. The representative amplicons were then sequenced. Chi-square for trend test was performed to study the correlation between positive rates of RT-PCR and at different periods after the onset of the disease.

RESULTSThe fragments amplified from the sputum specimens of SARS patients were shown to share 100% homology with the published SARS-associated coronavirus. Of the different clinical specimens, positive rate in the stools appeared to be the highest (21.55%). Chi-square for trend test revealed that the positive rates of stools and sputa of SARS patients decreased with the development of the disease (chi(2) for trend = 12.55 and 16.408, P = 0.0004 and P = 0.000 05 respectively).

CONCLUSIONOne-step RT-PCR proved to be an effective method for the detection of SARS-associated coronavirus from clinical specimens. Data as indicated that the positive rates of SARS coronavirus were decreasing in SARS patients along with the disease progression.

Adolescent ; Adult ; Aged ; Chi-Square Distribution ; China ; Disease Progression ; Feces ; virology ; Female ; Humans ; Male ; Middle Aged ; Mucus ; virology ; RNA, Viral ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; SARS Virus ; genetics ; isolation & purification ; Severe Acute Respiratory Syndrome ; pathology ; virology

OBJECTIVETo analyze the differences of lung tissue proteins in rats exposed to silica early by using comparative proteomics method and investigate the related mechanism with the occurrence and development of silicosis.

METHODSAdult male Wistar rats were randomly divided into control group and silica-treated group. The animal model was established by intratracheal (IT) instillation with silica suspension. On the 14th day after establishment of animal model, rats were sacrificed and lung tissues were collected. The total proteins were separated by means of two-dimensional gel electrophoresis (2-DE) and the differentially expressed proteins were identified by using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). In addition, Western blotting was performed to verify the expression of certain candidate protein.

RESULTSEleven differential expression protein spots were tested by MALDI-TOF-MS, and six proteins were identified. The levels of cathepsin D precursor, peroxiredoxin-1 (Prx-1), heat shock cognate 71 000 protein (HSP7C), heterogeneous nuclear ribonucleoprotein A3 (hnRNPA3) and fatty acid-binding protein (epidermal, E-FABP) were up-regulated in silica-treated group with the optical density (A) values. These values were 116.50+/-12.56, 148.75+/-22.40; 40.00+/-1.63, 66.00+/-13.93; 51.25+/-7.37, 92.75+/-8.69; 83.00+/-6.48, 122.75+/-24.62; 50.75+/-6.50, 93.50+/-23.10 and 100.25+/-19.99, 142.50+/-21.21 respectively. The statistical difference was observed as compared with control group (t=-2.51, -3.71, -7.28, -3.12, -3.56 and -2.90, P<0.05). However, SEC14-like protein 3 with the A values 153.00+/-11.28, 109.75+/-18.32 was down-regulated (t=4.02, P<0.01). Western blotting showed that in the expression of Prx-1 was higher in silica-treated group (0.61+/-0.05) than that in the control (0.35+/-0.05) (t=-7.24, P<0.01) when calculating the semi-quantification of this protein using ratio of optical density.

CONCLUSION2-DE pattern of lung tissue from rats exposed to silica has been established and six differentially expressed proteins have been identified. Our study is of help for further research of the mechanisms of silicosis.

Animals ; Disease Models, Animal ; Electrophoresis, Gel, Two-Dimensional ; Environmental Exposure ; Lung ; metabolism ; pathology ; Male ; Proteomics ; Rats ; Rats, Wistar ; Silicon Dioxide ; toxicity ; Silicosis ; metabolism

OBJECTIVETo explore the correlation between the exposure levels and serum protein fingerprints in population exposed to silica.

METHODSLiquid chip time-of-flight mass spectrometry technology was used to investigate the serum profiles in control group (30 cases), group exposed to silica (30 cases), silicosis group (I stage, 25 cases) and suspected silicosis group (30 cases), and screen the differential expression proteins. The correlation between the levels of the differential expression proteins and the exposure levels was performed.

RESULTSFive differential expression proteins were found among 4 groups, the expression of 5081 and 5066 proteins was upregulated, and the expression of 3954, 2021 and 1777 proteins was downregulated. There was no the correlation between the exposure levels and the peak with M/Z among those proteins.

CONCLUSIONthe results of present investigation indicated there was no correlation between the exposure levels and protein/peptide peak.

Adult ; Blood Proteins ; analysis ; Case-Control Studies ; Dust ; analysis ; Female ; Humans ; Male ; Mass Spectrometry ; Middle Aged ; Occupational Exposure ; analysis ; Peptide Mapping ; Proteomics ; Silicon Dioxide ; toxicity ; Silicosis ; blood

OBJECTIVETo study the differential gene expression profiling of rats exposed to silica using the normal rats as control.

METHODSAnimal models were established using intratracheal injection of the lung and 22 107 genes were screened in the differential expression profiling of silicosis by using oligonucleotide bead array. Differential expression profiling data were analyzed by using DAVID bioinformation software.

RESULTSTotally 1567 differentially expressed genes were identified in lungs of silica exposed rats including 765 up-regulated genes and 802 down-regulated genes as compared to the normal controls. Among 406 annotated genes in KEGG pathways, 204 genes and 11 pathways were up-regulated as well as 202 genes and 3 pathways were down-regulated in silica exposed rats.

CONCLUSIONAll 1567 genes are involved in the formation of silicosis. The differential gene expression profile of silicosis describes the general changes in the gene expressions in silicosis at transcriptional level. Further analysis of the identified genes might help reveal the molecular mechanism of pulmonary fibrosis induced by silica.

Animals ; Disease Models, Animal ; Gene Expression Profiling ; Lung ; metabolism ; pathology ; Male ; Oligonucleotide Array Sequence Analysis ; Pulmonary Fibrosis ; genetics ; metabolism ; Rats ; Rats, Wistar ; Silicosis ; genetics ; metabolism ; pathology