| There have all along been a lot of toxic chemicals in circumstance, including physical, chemic and biologic factors, which could damage genetic matter in human (chromosome, gene and DNA) and induce mutation. They could do harm to our health and children. Previous studies indicated that many medical difficult problem have to do with mutation, for example cancer, genetic disease and caducity. HPRT gene upright mutation test is a kind of methods determining mutagens, which has been adopted all over the world. In the past, there were only two kinds of methods determining mutation at the HPRT locus, that is, autoradiography assay and cyt-B incorporation assay. Though these two assays are simple and relatively inexpensive, there are some fatal weakness of not allowing recovery and further analysis of gene component or structure of HPRT- mutation (location and qualitation of gene mutation). This is always an obstacle to limit the further development of studies on HPRT gene mutation. The mutant cloning assay has the advantage in the two respects. It could be the most valuable methods in studies on HPRT gene mutation in human. So, it will be one of pressing assignment in mutagenesis study to confirm data of human HPRT gene mutation and to obtain molecular spectra of mutation. A great deal of HPRT gene mutation datum have been accumulated for constituting data-base by various methods about gene mutation detecting. The international organization proposed in many times that various suspected mutagens should be examined in different cell lines in vivo and in vitro to get more HPRT gene mutation datum. These datum will lay a foundation for researching mutation mechanism, hot spot, mutation patterm comparation and so on.On the other hand, 1,3-Butadiene (BD) is a volatile organic compound that is widely used in the production of resins and plastics. Since it has been detected in cigarette smoke, gasoline vapour and lampblack of cooking oil, BD is thought to be a public circumstance contamination. It was concluded that there is "sufficient evidence" for the carcinogenicity of BD in experimental animals, but "inadequate evidence" for the carcinogenicity in human. But there is a few appropriate BD-exposed locale being found, it is difficult to get direct evidence about mutagenesis of BD to human. In order to get details about HPRT gene plating efficiency, cloning efficiency and mutation frequency of lymphocytes in vivo, and to understand the spectra and mechanism of HPRT gene BD-induced mutation, lymphocytes from BD-exposed workers (74) and control (157) were tested. Some classical and new techniques such as epidemiology research, microscopic observation, single cell clone culturing (231), two-way screening count, multiplex PCR (783), RT-PCR, double electrophoresis and sequence analysis (146), were used in our studies. This experimental study was composed of three parts: (1) The HPRT gene mutation assay was established in our lab with several chemical and physical factors as models. (2) The studies on the occupational exposure and physical check-up in workers of BD monomer plant. (3) The laboratory analysis of HPRT gene mutation frequency and molecular spectrum in lymphocytes from BD-exposed workers. The main results are as follows:1. We have sumed up 4 standards and 2 reference factors about choosing BD-exposed occupational locale. They were obtained from our field survey and correlative datum. We choosed the BD workshop of olefin plant in Nanking Yangtse Petrifaction Corporation as BD-exposed occupational locale, which was up to the mustard and representational. We investigated the BD producing technics, the workers' labor flow, the general health and living habit background datum, which made it to be confirmed to choose BD-exposed group, control group and air sampling spot. The BD concentration was 20.79±34.95mg/m3 in the workaround of BD-exposed group, and couldn't be examined in the workaround of control group. This kind of methods can be used for reference in researching other occupational contacting compounds.
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