不同端粒酶抑制剂对肝癌细胞株蛋白表达的影响

目的以表面增强激光解吸及电离飞行时间质谱(SELDI-TOF-MS)技术检测针对人类端粒酶RNA的反义寡核苷酸(hTR-ASODN)、针对hTR的正义寡核苷酸(hTR-SODN)、针对人类端粒酶活性催化亚单位的ASODN(hTERT-ASODN)、针对hTERT的SODN(hTERT-SODN)、没食子酰表没食子儿茶素(EGCG)、3′-叠氮-3′-脱氧胸苷(AZT)、全反式维甲酸(ATRA)和盐酸阿酶素(ADM)8种端粒酶抑制剂分别作用人肝癌细胞SMMC-7721后差异蛋白表达的情况。方法运用SELDI-TOF-MS技术结合配套的弱阳离子交换型芯片(WCX-2)检测8种端粒酶抑制剂分别作用于SMMC-7721细胞后分泌蛋白和胞浆蛋白表达的变化。对照组为未加任何处理因素的SMMC-7721细胞。结果WCX-2蛋白芯片检测发现,8种端粒酶抑制剂作用后细胞株的分泌蛋白和胞浆蛋白分别存在不同程度的表达差异及共同低表达或高表达的差异蛋白。所有差异蛋白分子量均小于10000Da。结论8种端粒酶抑制剂作用后SMMC-7721细胞存在特异性差异蛋白和共性变化的蛋白,这些蛋白可能与端粒酶活性的调控密切相关。     

BED捕获酶联免疫法检测广西某吸毒监测哨点HIV新近感染率

目的 了解广西某吸毒监测哨点注射吸毒人群(injection drug users,IDU)的HIV新近感染情况,估算HIV新近感染率.方法 对广西某吸毒监测哨点2005-2007年787份血清样本进行监测,以酶联免疫吸附试验(ELISA)筛检出168份HIV-1阳性样本,再经BED捕获酶联免疫法(BED capture enzyme immunoassay,BED-CEIA)检测出其中新近感染的样本共17份,获得该人群HIV-1的新近感染率.结果 787份监测样本中共筛检出HIV-1确认阳性样本168份,新近感染样本17份.该哨点2005-2007年HIV感染率分别为23.06%(92/399)、21.90%(46/210)、16.85%(30/178),新近感染率分别为7.41%、6.94%、3.12%.结论 在广西某吸毒监测哨点IDU中,2005-2007年HIV新近感染率绝对数值有所降低,已发现的HIV感染者可能部分为既往感染者.     

大鼠胚胎脑新皮质神经干细胞的体外分离培养方法研究

目的建立大鼠胚胎脑新皮质神经干细胞的体外分离培养方法。方法从孕14天SD大鼠胚胎脑分离新皮质,通过机械吹打改善细胞分散状态、优化无血清培养基条件,从而改进神经干细胞的培养方法。利用神经干细胞标志蛋白(nestin和SOX2)、增殖和克隆成球能力以及多向分化能力测试三方面对神经干细胞进行鉴定。结果以该法培养的神经干细胞nestin蛋白呈强阳性表达,SOX2阳性表达率达99%以上,BrdU掺入阳性,培养3天后细胞量为接种量的10.55倍,6日克隆成球率为(33.00±4.40)%。经相应特异性标志蛋白检测证实神经干细胞经诱导分化后可形成神经元(MAP2)、星形胶质细胞(GFAP)和少突胶质细胞(O4)。结论获得的神经干细胞纯度和细胞产出率高,具有强自我更新和多向分化能力。     

食品中典型有机污染物和重金属的毒性通路研究进展

食品污染是影响食品安全和人群健康的重要风险因素。《21世纪毒性测试愿景与战略》提出未来的毒性测试应充分考虑人群暴露可能，基于体外人源性细胞系、高通量筛选和毒性通路的毒性测试战略，试图解决传统毒性测试成本高昂、耗时费力、种属差异方面的挑战。本综述围绕我国食品污染问题突出的典型重金属和有机污染物，分析归纳了有关毒性通路研究现况，并展望未来的研究方向。     

饮用水未受控消毒副产物的遗传毒性和致癌性研究进展

饮用水消毒副产物(disinfection by-products,DBPs)是饮用水加消毒剂消毒过程必然形成的污染物,目前已发现的DBPs约600～700种[1].     

高效液相色谱法检测上海市两水厂原水和出厂水中神经类毒素-a污染水平

目的建立高效液相色谱法(HPLC)检测饮用水中神经类毒素-a的方法。方法采集以青草沙水库为水源的A、B两水厂的原水和出厂水各1L,经GF/C滤膜过滤,弱阳离子交换(WCX)填料分别富集胞内和溶解性神经类毒素-a,同时,确定最佳的三氟乙酸(TFA)甲醇配比洗脱条件。采用HPLC定性定量分析以青草沙为水源的水厂原水和出厂水神经类毒素-a含量。结果 0.2%TFA甲醇洗脱效果最好。本研究建立的HPLC法检测水中神经类毒素-a,其标准曲线线性范围为0.01～5μg/ml,R2为0.9999,检出限2.9ng/L,加标回收率75.2%～100.6%,相对标准偏差(RSD%)6.38%～8.76%。连续6日加标检测均值与真实值一致,日内和日间RSD%在1.60%～8.90%之间。两水厂原水中均可检出神经类毒素-a,A、B水厂神经类毒素-a含量分别为0.028μg/L和0.137μg/L,出厂水中均未检出神经类毒素-a。结论本方法检出限低,回收率高,精密度高,稳定性好,可靠性强,可用于水中痕量神经类毒素-a检测,并可连续批量检测。为防止神经类毒素-a对人群健康的潜在危害,应加强监测青草沙库区及以此为水源的水厂原水和出厂水神经类毒素-a。     

饮用水碘代消毒副产物的形成及细胞和遗传毒性研究进展

碘代消毒副产物(iodo-disinfection byproducts,Iodo-DBPs)是饮用水消毒过程中新发现的未受控消毒副产物(disinfection byproducts,DBPs)[1],其形成取决于原水碘离子水平和消毒剂,氯胺消毒易形成Iodo-DBPs.     

S型氯代甘油醇对大鼠精子运动和超活化抑制作用研究

目的研究S型氯代甘油醇(SACH)对大鼠精子运动和超活化的影响和可能机制。方法 20只成熟雄性SD大鼠随机分为4组,分别给予0、2.5、5.0和10mg/kg BW SACH,连续灌胃染毒52天。取附睾尾精子,获能条件下培养5h,以计算机辅助精子分析系统检测精子运动和超活化运动,同时检测精子特异的3-磷酸甘油醛脱氢酶(GAPDS)活性、三磷酸腺苷(ATP)和环磷酸腺苷(cAMP)水平,观察己酮可可碱(PTF)对SACH的拮抗作用。结果 SACH染毒组大鼠精子曲线运动速率(VCL)、平均路径速率(VAP)、直线运动速率(VSL)和精子头侧摆幅度(ALH)与对照组相比受到显著抑制(P0.01),运动直线性(LIN)显著增强(P0.01)。SACH使VCL≥400μm/s或LIN≤20%的比例显著减少(P0.01和P0.05),表明精子超活化受抑制。SACH显著抑制GAPDS活性,ATP和cAMP水平也随染毒剂量增加呈下降趋势。PTF可改善精子cAMP水平,部分缓解精子运动和超活化的抑制程度。结论 SACH抑制大鼠精子GAPDS活性导致ATP和cAMP水平下降,可能是影响精子运动和超活化的重要机制。     

双柱定性气相色谱电子捕获检测器测定上海市饮用水中微量碘化物

Objective To modify and optimize gas chromatographic conditions, qualitative and quantitative method on the base of the actual national standard detection method for drinking water on testing iodide with gas chromatography, and detect iodide in drinking water of Shanghai.Methods Iodobutanone derivative was identified by gas chromatography/mass spectrometry, then two capillary columns coupled to electron capture detector were made by qualitative and quantitative analysis.Finally, 100 ml water samples were collected respectively from raw water and finished water of three plants in which water sources were Yangtse river, Huangpu river, and inner river respectively and detected by this developed method.Results The results of mass spectrometry showed that iodide would form iodobutanone which could generate 1-iodo-2-butanone and 3-iodo-2-butanone isomers by derivatization.The data of qualitative analysis by two capillary columns revealed that iodobutanone was separated completely and the total time of chromatogram separation was 19.33 min.3-iodo-2-butanone with the high response value was selected to quantitatively analyse.The linear range was 1 - 100 μg/L,and the coefficient of determination (r2) was 0.9997.The limit of detection was 13 ng/L.Recoveries were between 97.68% and 104.37% ,and relative standard deviations were between 2.14% and 4.41%.The results of iodide detected by this method in raw water and finished water in three plants in Shanghai showed that the ranking of iodide's concentration in raw water was Huangpu river (15.14 μg/L) ＞ inner river (6.97 μg/L) ＞ Yangtse river (3.55 μg/L).The level of iodide in finished water of plant 1 (3.55 μg/L and 5.92 μg/L for raw and finished water respectively) and 3 ( 6.97 μg/L and 9.62 μg/L for raw and finished water respectively) increased slightly except for plant 2( 15.14 μg/L and 1.81 μg/L for raw and finished water respectively).The relative standard deviations of all water samples measured in duplicate were less than 9.73%.Conclusion The improved method possesses higher degree of sensitivity and accuracy of qualitative and quantitative analysis than actual national standard detection method, and it is fit for trace analysis of iodide in water.     

双柱定性气相色谱电子捕获检测器测定上海市饮用水中微量碘化物

Objective To modify and optimize gas chromatographic conditions, qualitative and quantitative method on the base of the actual national standard detection method for drinking water on testing iodide with gas chromatography, and detect iodide in drinking water of Shanghai.Methods Iodobutanone derivative was identified by gas chromatography/mass spectrometry, then two capillary columns coupled to electron capture detector were made by qualitative and quantitative analysis.Finally, 100 ml water samples were collected respectively from raw water and finished water of three plants in which water sources were Yangtse river, Huangpu river, and inner river respectively and detected by this developed method.Results The results of mass spectrometry showed that iodide would form iodobutanone which could generate 1-iodo-2-butanone and 3-iodo-2-butanone isomers by derivatization.The data of qualitative analysis by two capillary columns revealed that iodobutanone was separated completely and the total time of chromatogram separation was 19.33 min.3-iodo-2-butanone with the high response value was selected to quantitatively analyse.The linear range was 1 - 100 μg/L,and the coefficient of determination (r2) was 0.9997.The limit of detection was 13 ng/L.Recoveries were between 97.68% and 104.37% ,and relative standard deviations were between 2.14% and 4.41%.The results of iodide detected by this method in raw water and finished water in three plants in Shanghai showed that the ranking of iodide's concentration in raw water was Huangpu river (15.14 μg/L) ＞ inner river (6.97 μg/L) ＞ Yangtse river (3.55 μg/L).The level of iodide in finished water of plant 1 (3.55 μg/L and 5.92 μg/L for raw and finished water respectively) and 3 ( 6.97 μg/L and 9.62 μg/L for raw and finished water respectively) increased slightly except for plant 2( 15.14 μg/L and 1.81 μg/L for raw and finished water respectively).The relative standard deviations of all water samples measured in duplicate were less than 9.73%.Conclusion The improved method possesses higher degree of sensitivity and accuracy of qualitative and quantitative analysis than actual national standard detection method, and it is fit for trace analysis of iodide in water.