| Objectives Pneumoconiosis is the systemic disease of pulmonary fibrosis arising from long-term inhalation of dust. On the cause of the illness is clear, but there is no effective treatment yet. Coal worker pneumoconiosis (CWP) is one of the most serious occupational lung diseases currently, which is due to long-term inhalation of dust containing some dissociative silica. In the course of coal mining, workers exposed to either coal dust or silica dust because of the different types of work thus giving rise to diffuse pulmonary fibrosis, which is collectively referred to as coal worker's pneumoconiosis. Coal worker's pneumoconiosis incidence was differences due to different ways of the exploitation. Pneumoconiosis prevalence rate among open-cut coal workers is very low, while the pneumoconiosis prevalence and morbidity rates of those working underground are both higher with the dust dispersion higher. With the Formation of China's vast complex structure, the prevalence of coal worker's pneumoconiosis have much difference between 0.92% and 24.1%, during which silicosis accounts for 11.4% of, coal silicosis 87.6%, coal lung 1.0%. The disease accounts for 80 percent of the total number of occupational ones, which diagnosis is mainly based on occupational history and chest X-ray. In recent years the research about mechanisms of pneumoconiosis induced by mineral dust, as one of the main research methods, achieves greater progress. By means of the mechanical erosion of normal saline and direct injection into alveolus, Lung lavage removes dust from lungs , with a view to achieve the treatment of pneumoconiosis. In the treatment of pneumoconiosis and its complications, Lung lavage can improve respiratory function, enhance oxygen saturation, and reduce the suffering of patients, which great significance makes the analysis of pneumoconiosis research lavage imperative. This study to investigate the total protein levels of coal worker's pneumoconiosis bronchoalveolar lavage fluid, Clara cell protein, SOD activity, as well as cell components. It also studies SDS polyacrylamide gel electrophoresis to the proteins and the 2-DE separation to explore the early prevention, diagnosis and treatment of coal worker's pneumoconiosis. Methods There are 22 hospitalized cases about workers who exposed to dust and pneumoconiosis patients proved with our country's pneumoconiosis GBZ70-2002 diagnostic criteria of the 0+,â… ,â…¡period and the patients of high-volume whole lung lavage without contraindication. The selection of indications: workers are with a variety of mixed and simple pneumoconiosis; pulmonary alveolar proteinosis; aspiration pneumonia, bronchial asthma, lung infections and so on. background of all subject: Age is from38 to 58 years old, with an average of 52.1±5.6 and all is male the types of dust contactedincluding coal dust, rock dust or mixed dust; the mean of exposure history is 26.8±4.3 years; 7 cases were 0+ and 11 Periodâ… , 4cases with Phaseâ…¡bronchoalveolar lavage fluid were obtained from t Chinese coal patients in the Beidaihe Sanatorium. The samples were concentration by using of low-voltage low-temperature fluid mixing Miriam 10ml concentrated lavage fluid is stored under -70℃to perform protein analysis. The CC-10, superoxide dismutase SOD will be determined by using the method of double-antibody sandwich ELISA, and then protein in fluid will be determined by the means of polyacrylamide gel electrophoresis and 2-DE. Centrifuged cells should be observed by Wright's staining method, and macrophages should be observed by electron microscopy techniques.Results1 Cytological examination1. According to the number of the cells in term of the three-stage patients of 0+,â… ,â…¡, the cells number of lavage fluid in total 24 bottles are characterized with skewness of the distribution, of which the peak is in the second and third bottle liquid. It is the second bottle liquid that has the most pneumoconiosis fluid cells among the 24 bottles of 0+ period patients', around 1.55×1011/ L with the total number of cells around 1.00×1012 / L. However among 1 period patient's, it is the third bottle that has the most pneumoconiosis fluid cells among the 24 bottles of which are around 1.56×1011/L with the total number of cells around 1.16×1012/L. It is the third bottle that has the most pneumoconiosis fluid cells among the 24 bottles of II period patients', around 1.65×1011/L with the total number of cells around 1.18×1012/L. Compared with the total number of cells between these fluids, the conclusion can be drawn as: 0+<â… <â…¡1.2 A smear of centrifuge tube fluid that holds the most cells of Phaseâ…¡patients should be taken, and then the macrophages are observed by Wright staining. There are more black particles within macrophages with a clearer outline of cells and other obvious cells. 1.3 Under a Microscope, macrophage and their nuclei are more complete, the edge of the nucleus ambiguous and sometimes shrinkage phenomenon can be seen with some increasing. Within macrophage cells there are a lot of dust particles with its edge vague and nucleus clear.2 protein measurement in lavage fluid2.1 Total protein content was measured in BALF. 0+ total protein content 0.28±0.07 mg/ml,â… total protein content 0.51±0.09 mg/ml,â…¡total protein content 0.82±0.07 mg/ml,The total protein content in 0+,â… ,â…¡is significantly higher, and there is significant difference (F=60.29, P <0.001).2.2 Detection of SOD vitality in BALF. 0+ SOD vitality 306.04±6.12μg/ml;â… SOD vitality 313.20±7.52μg/ml;â…¡SOD vitality 356.83±7.56μg/ml。In the three stages the difference in the vitality of SOD has much statistical significance (F=71.83, P <0.001).2.3 The total content of CC-10 in BALF. 0+ CC-10 content 8.23±0.62 ng/ml;â… CC-10 content 9.53±0.81 ng/ml;â…¡CC-10 content 6.48±0.45 ng/ml.In 0+,â… ,â…¡has significantly decreased. The difference has statistical significance (F=28.54, P <0.001). But the content in Period I is more than those in the period of 0+ andâ…¡. 2.4 Pneumoconiosis fluid polyacrylamide gel electrophoresis and two-dimensional gel electrophoresisFrom the map of polyacrylamide gel electrophoresis, the good separation of protein has a clear strip which shows the types of protein in coal worker's pneumoconiosis fluid is relatively constant. Therefore, two-dimensional gel electrophoresis is used to separate protein groups and then the first isoelectric focusing is done by the 17cm long linear IPG strips with 3-10 pH, and then 12% SDS-PAGE is used as the second phase separation so as to obtain a more clear BALF protein patterns of two-dimensional gel electrophoresis of 0+dust-exposed workers and Phaseâ…¡pneumoconiosis of coal workers. Isoelectric point like protein increases from left to right, and from top to bottom, the molecular weight decreases.Conclusions1 After classification of Lotion cell, Can see macrophages, lymphocytes,neutrophil cells, eosinophil apparent from the number on the rise(P<0.05). Under the microscopy macrophages can reflect its change of the morphology of macrophages in the process of pneumoconiosis pathogenesis, which lays a good foundation on better understanding of the pneumoconiosis mechanism.2 CWP in bronchoalveolar lavage fluid total protein content increased with a view to significantly higher level, the vitality of SOD increased with higher-level view. Therefore, through analysis of its mechanism, Able to clarify the pathogenesis of pneumoconiosis must play the role of the reference.3 The CWP was found in bronchoalveolar lavage fluid of CC10 with the dust content is the amount of particles entering the body to change, the general trend of changes in order to reduc(e0+<â… ,â… >â…¡,0+>â…¡). CC10 may therefore reflect the early detection of Clara cell damage, Judge for the early emergence of lung injury and provide the basis for recovery.4 CWP by bronchoalveolar lavage fluid proteome of SDS polyacrylamide gel electrophoresis and two-dimensional gel electrophoresis analysis, Of BALF in the soluble protein components of the separation, To obtain a clear view of the 0+ dust-exposed workers and pneumoconiosis of stageâ… BALF protein patterns of two-dimensional gel electrophoresis, Contrast through the image reveals the protein composition of BALF in the quality and quantity of change... |
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