| At present, the pneumoconiosis is the most serious occupational diseases in china. Both prevalence rate and incidence rate are very high. Therefore, the prevention and therapy of pneumoconiosis are very important. But the mechanism of it is complicated and unclear. So the key to control is prevention. It is an important precautionary measure to detect the high-risk group.Some studies showed that the heat shock protein70 (HSP70) may be related to pneumoconiosis. There are some polymorphism sites in the gene of HSP70. And it is reported that gene polymorphism of encoding chaperone cohort is closely related to lung cancer susceptibility. So 100 0+ Sternum cases and 100 contacts of coal-silica mixed dust were selected based on GBZ70-2002 and investigated by questionnaires. Extracting DNA from 3ml peripheral vein blood cells of all subjects. Polymerse chain reaction-restriction fragment length polymorphisms (PCR-RFLP)techniques was used, the polymorphisms of HSP70-1 +190 site, HSP70-2 +1267 site and HSP70-hom +2437 site in the HSP70 gene of all the objects are detected. Then the relationship between the HSP70 gene polymorphism and genetic susceptibility in dust-exposed workers was studied.Materials and methods:Sample Collection: In the same mine area, subjects esposed to coal-silica mixed dust (n=100) and the 0+ Sternum cases by chest radiographic determining (n=100) were selected(GBZ70-2002 was the standard of Chest radiographic determination). All subjects were male workers of Han race and without other pulmonary or autoirnmune diseases. Three milliliter peripheral vein blood was drawn from all subjects.HSP70 gene polymorphism detecting: DNA was extracted from 3ml peripheral vein blood. Genotype of the HSP70 target gene were analyzed by polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLP) techniques. The sequence of primer is as following: HSP70-1 : 5'-TCC GGC GTC CGG AAG GAC C-3', 5'-TGC GGC CAA TCA GGC GCT T-3'; HSP70-2 : 5'-CAT CGA CTT CTA CAC GTC CA-3', 5'-CAA AGT CCT TGA GTC CCA AC-3'; HSP70-hom: 5'-GGA CAA GTC TGA GAA GGT ACA G-3', 5'-GTA ACT TAG ATT CAG GTC TGG-3'. PCR reaction system was 10×Buffer 2.5μl, DNTP 2.0μl, upper primer 1μl, lower primer 1μl, Taq Enzyme 0.25μl, H2O 15.25μl, DNA 3μl. PCR Reaction conditions were:HSP70-1:94℃5minâ†'94℃30sâ†'54.5℃30sâ†'72℃45sâ†'35 recyclesâ†'72℃10min; HSP70-2: 94℃3minâ†'94℃1minâ†'61℃1minâ†'72℃2minâ†'35 recyclesâ†'72℃10min; HSP70-hom: 94℃2minâ†'94℃45sâ†'57℃45sâ†'72℃1minâ†'35 recyclesâ†'72℃5min. Restriction enzymes were BsrBI, PstI, and NcoI.The interactions were analyzed by logistic regression methods. Process the data by meanes of SPSS13.0 software. The significant level was set atα=0.05.Results:1 General condition of the investigated subjectsAll subjects were mining workers. The type of dust was mixed dust. There was no statistics significance(P>0.05)for the subjects in the two groups of age, working age, height, weight, physical condition, education degree and the history of smoking and drinking.2 Comparative in the different distribution of genotypes among the three groups There was significant difference between 0+ group and dust exposed group in distribution frequencies of HSP70-1 gene +190 b2/b2 (OR = 2.357). There was no significant difference between 0+ group and dust exposed group in distribution frequencies of HSP70-2 and HSP70-hom.3 The Results of logistic regression Analysis.Analyzed by logistic regression model, the index showed that there was significant difference between 0+ group and dust exposed group in distribution frequencies of HSP70-1. There was no statistic significance between 0+ group and dust exposed group in the smoking indexes and working age.Conclusion:The genotype distribution of HSP70-1 gene b2/b2 is not balance. And it may be one of dust-exposed workers susceptibility genes. |
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