生物硝化池污水中硝化细菌的快速定量研究

实验采用聚合酶链式反应（PCR）技术与最大几率数法（MPN）相结合的MPN－PCR法对生物硝化池污水中的硝化细菌进行快速定量。所用的一对PCR引物是在对硝化细菌的16SrRNA基因进行系统比较的基础上设计合成的，可以扩增出大小为388bp的DNA片段。以从生物硝化池污水中抽提的含硝化细菌DNA的混合DNA为模板，进行PCR扩增并确定合适的扩增条件。运用MPN－PCR法进行定量检测的整个过程可在几小时之内完成。     

Prenatal diagnosis of medium-chain acyl-CoA dehydrogenase (MCAD) deficiency in a family with a previous fatal case of sudden unexpected death in childhood

Medium-chain acyl-CoA dehydrogenase (MCAD) deficiency is a potentially fatal inherited disease with a carrier frequency of approximately 1:100 in most Caucasian populations. The disease is implicated in sudden unexpected death in childhood. A prevalent disease-causing point mutation (A985G) in the MCAD gene has been characterized, thus rendering diagnosis easy in the majority of cases. Since the clinical spectrum of MCAD deficiency ranges from death in the first days of life to an asymptomatic life, there are probably other genetic factors—in addition to MCAD mutations—involved in the expression of the disease. Thus, families who have experienced the death of a child from MCAD deficiency might have an increased risk of a seriously affected subsequent child. In such a family we have therefore performed a prenatal diagnosis on a chorionic villus sample by a highly specific and sensitive polymerase chain reaction (PCR) assay for the G985 mutation. The analysis was positive and resulted in abortion. We verified the diagnosis by direct analysis on blood spots and other tissue material from the aborted fetus and from family members.     

Oxidative stress and DNA damages induced by cadmium accumulation

Experimental evidence shows that cadmium （Cd） could induce oxidative stress and then causes DNA damage in animal cells, however, whether such effect exists in plants is still unclear. In the present study, Vicia faba plants was exposed to 5 and 10 mg/L Cd for 4 d to investigate the distribution of Cd in plant, the metal effects on the cell lipids, antioxidative enzymes and DNA damages in leaves. Cd induced an increase in Cd concentrations in plants. An enhanced level of lipid peroxidation in leaves and an enhanced concentration of H2O2 in root tissues suggested that Cd caused oxidative stress in Vicia faba. Compared with control, Cd-induced enhancement in superoxide dismutase activity was significant at 5 mg/L than at 10 mg/kg in leaves, by contrast, catalase and peroxidaseactivities were significantly suppressed by Cd addition. DNA damage was detected by neutral/neutral, alkaline/neutral and alkaline/alkaline Comet assay. Increased levels of DNA damages induced by Cd occurred with reference to oxidative stress in leaves, therefore, oxidative stress induced by Cd accumulation in plants contributed to DNA damages and was possibly an important mechanism of Cd-phytotoxicity in Vicia faba plants.     

含铅,无铅汽油车尾气排出物遗传毒性比较

利用台架试验模拟汽车行驶工况，收集了发动机使用无铅汽油，含铅汽油燃烧后的尾气颗粒物，对其中吸附的有机物进行了提取，并以Ａｍｅｓ试验，中国仓鼠肺细胞体外微核实验等方法，从基因，染色体水平检测了两种颗粒提取的遗传毒性。     

Determination of alkali-labile phosphoprotein phosphorus from fish plasma using the Tb^3＋-tiron complex as a fluorescence probe

A sensitive method based on the fluorescence quenching effect of the Tb^3＋-Tiron complex is proposed for the determination of alkali-labile phosphoprotein phosphorus （ALP） released from fish plasma. The detection limit was 5.4 ng/ml （S/N=2）, and the relative standard deviation of the quenching effect （6 replicates） was 4.6%. The results obtained by the proposed method were in good agreement with those obtained by the colorimetric assay. The advantages of the present method are its relatively simple detection procedure, the lack of toxic organic solvents, and high sensitivity.     

Immunotoxicity of bisphenol a to Carassius auratus Lymphocytes and macrophages following in vitro exposure

Bisphenol A （BPA） is the monomer component of polycarbonate plastics and classified as an endocrine disrupting chemical （EDC）. The reproductive toxicity of BPA has been extensively studied in mammals; however, relatively little information is available on the immunotoxic responses of fish to BPA. In this study, we investigated the effects of BPA on the immune functions of lymphocytes and macrophages in Carassius auratus. The effects of BPA were compared with those of two natural steroid hormones, estradiol and hydrocortisone. Proliferation of the two types of cells in response to PHA was measured using colorimetric MTT assay. Macrophage respiratory burst stimulated by Con A was measured using chemiluminescence assay. Results showed that BPA （0.054-5.4 mg/L）, estradiol （0.0002-2.0 mg/L） and hydrocortisone （5-50 mg/L） significantly induced Carassius auratus lymphocyte proliferation while higher doses of hydrocortisone （500-5000 mg/L） appeared to be inhibitory. BPA （0.005-50 mg/L）, estradiol （0.005-800 mg/L） and hydrocortisone （0.005-500 mg/L） markedly enhanced macrophage proliferation, whereas higher doses of BPA （500-1000 mg/L） appeared to inhibit cell proliferation. Furthermore, higher dosage of BPA （50 mg/L） and hydrocortisone （50 and 500 mg/L） suppressed the macrophages respiratory burst while estradiol is stimulative all the doses tested （0.05-500 mg/L）. In conclusion, BPA could have immunotoxicity to Carassius auratus and functional changes of lymphocyte and macrophage in Carassius auratus may be different between low and high dosages.     

用改良彗星试验检测工业废水的DNA损伤作用

采用改良彗星试验直接检测了多种工业废水不所引起的Ｖ７９细胞ＤＮＡ损伤。结果表明,所测的种各工业废话水均含有ＤＮＡ损伤剂,能够诱发培养的Ｖ７９细胞ＤＮＡ链断裂,其中以制革厂不的ＤＮＡ损伤作用最强。本研究显示出改良彗星试验在综合评价各种废水遗传毒性方面具有广阔的应用前景。     

Response of the Ascorbate-Peroxidase of Selenastrum capricornutum to Copper and Lead in Stormwaters

The green alga Selenastrum capricornutum expresses a uniqueascorbate peroxidase, that responds to copper and lead. Attemptswere made to test if this peroxidase could be used to monitor thelevels of copper and lead in natural waters. When S.capricornutum was exposed to a stormwater sample, the specificactivity of the peroxidase in the cell extract was commensuratewith the combined copper and lead contents in the sample. Theperoxidase responses were also correlated with the 96 hr biomasstoxicity assay of S. capricornutum. However, unlike thebiomass toxicity assay, the peroxidase activity was not affectedby the anions in the samples. The use of this peroxidase can beused as a marker for testing heavy metal toxicity in the water.     

东亚钳蝎抑制型神经毒素基因在昆虫细胞中的表达及活性鉴定

基因BmKIT3R 是根据东亚钳蝎 (ButhusmartensiiKarsch)抑制型神经毒素基因BmKIT3 优化而得的 .将优化过的蝎毒素基因BmKIT3R 通过Bac to Bac操作技术 ,使重组杆状病毒基因组DNA转染到草地粘虫sf2 1细胞中对IT3R 基因进行表达 ,经SDS PAGE检测在 8.7× 10 3 处有一表达条带 .表达产物经生物鉴定 ,证明具有很高的活性 ,它对昆虫具有非常明显的致死作用 .图 3参 5     

酚类化合物对不同组织细胞DNA损伤的研究

应用单细胞凝胶电泳技术 ,研究了酚类化合物对不同组织细胞DNA的损伤 .试验结果表明 ,酚类化合物均能引起不同组织细胞不同程度的DNA损伤 ,并呈现剂量效应关系 .其高剂量组与对照组相比 ,均有极显著性差异 (P <0 0 1 ) .不同组织细胞对同一种药物呈现出不同的敏感性 .酚类化合物遗传毒性效应与其结构密切相关