亚硒酸钠和酵母硒在猪组织中的代谢及其代谢产物对人小细胞肺癌细胞增殖的影响
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摘要
为研究不同水平的酵母硒(Selenium-enriched Yeast, SeY)和亚硒酸钠(Sodium Selenite, SS)对组织硒存留、组织抗氧化系统以及两种硒的代谢产物对人小细胞肺癌细胞株NCI-H446增殖的影响而进行了以下四个试验：试验一：不同日粮水平的SeY和SS对生长猪组织抗氧化系统和硒存留的影响
为研究日粮SeY和SS水平对猪肝脏和肌肉中组织硒存留和抗氧化系统的影响,60头28日龄断奶仔猪随机分配到两期试验中即一期试验和二期试验中。两期试验分别设3个处理,每个处理10个重复。所有仔猪在饲喂用缺硒地区所产玉米-大豆配制的低硒日粮(0.03 mg kg-1 Se) 4周后分成6组,分别饲喂8周的基础日粮(0.04 mg kg-1 Se)添加0,0.3和3.0 mg kg -1的SeY(一期试验)和0,0.3和3.0 mg kg -1的SS(二期试验)。结果表明：与0.3 mg kg-1 SeY和SS相比,日粮0或3.0 mg kg-1 SS和SeY对猪的采食量、日增重和饲料转化率均无显著影响。当SS和SeY的水平从0增加到0.3 mg kg-1,猪肝脏、肌肉和血浆中GPX活性均极显著(P<0.01)提高；但3.0 mg kg-1 SeY和SS均未能(P>0.05)在其相应的0.3 mg kg-1水平上继续提高肝脏、肌肉和血浆中GPX活性。在0-3.0 mg kg-1范围内日粮SeY和SS的水平对猪血浆、肝脏和肌肉中SOD和GST活性无显著影响。随着日粮SeY水平在0-3.0 mg kg-1范围内的升高,猪肝脏、肌肉和血浆中的硒含量均极显著(P<0.01)提高。在0-3.0 mg kg-1范围内日粮SS极显著(P<0.01)提高了猪肝脏硒含量,显著(P<0.05)提高肌肉硒含量,而血浆中硒含量在0.3 mg kg-1 SS水平即达到平台。这表明8周内日粮SeY和SS的缺乏或过量对猪生产性能和组织抗氧化系统无显著影响,但能提高肝脏和肌肉的硒存留。试验二：不同日粮水平的SeY和SS对猪肝脏和肌肉中的硒化物NBS/PBS分布的影响
试验一的结果表明随着日粮SeY和SS的水平增加,肝脏和肌肉中硒存留量显著提高,但两种组织中硒是以蛋白结合硒(Protein-bound Selenium, PBS)或是非蛋白结合硒(Nonprotein-bound Selenium, NBS)的形式存在不清楚。本实验研究日粮SeY和SS水平对猪肝脏和肌肉中PBS& NBS的分布如影响。为此首先建立一套改良的分离猪组织中NBS和PBS的方法,用此方法分离测定了试验一中饲喂0-3.0 mg kg-1 SeY和SS两种硒的猪肝脏和肌肉中NBS和PBS的含量。结果表明：随着日粮SeY的升高,肝脏和肌肉中NBS和PBS的绝对含量均极显著(P<0.01)升高。肝脏中PBS占其总硒的相对含量随日粮SeY的增加而降低(P<0.01),相应地肝脏中NBS占其总硒的相对含量随日粮SeY水平的升高而增加(P<0.05)；与肝脏形成对比的是肌肉中NBS和PBS占其总硒的相对含量随日粮SeY的增加分别保持稳定在6%和94%。与饲喂SeY的猪肝脏NBS和PBS变化规律类似,饲喂SS的猪肝脏中NBS和PBS占其总硒的相对含量随SS的升高分别表现为显著升高(P<0.05)和显著降低(P<0.05),饲喂SS的猪肌肉中NBS和PBS的变化规律与肝脏类似,分别表现为随日粮SS的增加而显著升高(P<0.05)和降低(P<0.05)。这些表明日粮SeY和SS对肝脏中NBS和PBS的分布模式的影响类似,而对肌肉中NBS和PBS的分布模式影响不同。
试验三：饲喂过量SeY和SS的猪肝脏中NBS对人肺癌细胞增殖的影响
上述实验结果表明日粮SeY和SS均能极显著提高肝脏中NBS的绝对含量和相对含量,且对饲喂SeY和SS的猪肝脏中NBS和PBS的分布模式的影响一致。本实验旨在研究饲喂高水平酵母硒和亚硒酸钠16周的猪肝脏中是否含有抑制人肺癌细胞增殖功能的NBS。取饲喂16周基础日粮(0.05mg Se kg-1)及添加3.0mg kg-1 SeY和3.0 mg kg-1 SS的三种猪肝在超纯水中匀浆,匀浆液分别用氯仿和乙醇沉淀去除蛋白后所得清液经冷冻干燥浓缩、超滤和过滤除菌后得到含有NBS的硒含量分别为0.98μM,2.62μM和7.84 pM的肝脏提取物(Liver Extract,LE),分别简写为LEB, LEY和LES。将三种LE分别加入到癌细胞培养体系中,结果表明当添加水平在14%时LEY和LES能显著抑制人肺癌细胞的增殖,且诱导癌细胞的凋亡。亚硒酸钠是硒抗癌研究中的标准物质,而当在LEB中添加亚硒酸钠达到与LEY和LES中同等Se水平再添加到细胞培养体系后发现LEY和LES仍能显著抑制癌细胞的增殖,这说明LEY和LES中的NBS抑制癌细胞的能力高于亚硒酸钠,即饲喂3.0 mg/kg SeY或SS的猪肝脏中含有比亚硒酸钠抗癌活性更强的硒化物。
试验四：饲喂3.0 mg/kg SeY和SS的猪血清对人肺癌细胞增殖的影响
试验三表明饲喂3.0 mg kg-1 SS和3.0 mg kg-1 SeY的猪肝脏中有抑制人肺癌细胞增殖的硒化物。根据硒的代谢规律和抗癌功能,推测饲喂3.0 mg/kg SeY或SS的猪血清中也可能含有抗癌的硒化物。本实验旨在研究饲喂高水平日粮SeY和SS的猪血液中是否含有抑制人肺癌细胞增殖的物质。前腔静脉采集饲喂基础日粮(0.03 mg kg-1 Se)以及基础日粮添加3.0mg kg-1 SeY和3.0mg kg-1 SS 16周的猪清,经0.22μm滤膜过滤除菌后得到总硒含量分别为165 ng ml-1,1316 ng ml-1和834 ng ml-1的三种猪血清,分别简写为Se(0), SeY(3)和SS(3)。为探索猪血清代替FBS培养细胞的最佳添加水平,在培养液中添加7个梯度(8%-20%)的猪血清,根据细胞的MTT OD值随血清添加水平的变化Se(0)、SeY(3)和SS(3)三种猪血清的最适添加比例分别为17.79%,15.39%和14.31%,取三者的均值15.83%作为猪血清的最适添加水平。根据细胞在48h,96h和144h三个时间点的形态变化取144 h作为细胞最适培养时间。试验结果表明Se(0)猪血清培养癌细胞的MTTOD值显著(P<0.05)高于SS(3)猪血清,而SeY(3)与Se(0)和SS(3)猪血清处理细胞的MTT OD值均无显著差异。与SeY(3)和SS(3)相比,Se(缺)比培养细胞的上清液中LDH活性均显著(P<0.05)降低,而SeY(3)和SS(3)两者培养细胞上清中LDH活性无显著差异。细胞经Annexin V和PI双染后经流式细胞仪分析表明,Se(0)比SS(3)培养细胞的凋亡率显著降低(P<0.05)；而SeY(3)培养细胞的凋亡率介于Se(0)和SS(3)之间,与两者差异均不显著。上述表明饲喂3.0 mg kg-1 SS的猪血清中含有抑制人肺癌细胞增殖的硒化物。
本研究通过对日粮SeY和SS在猪肝脏和肌肉中代谢存留的研究机理我们推测饲喂3.0 mg/kg SeY或SS的猪肝脏中可能具有抑制人癌细胞增殖的硒化物。通过将从饲喂过量SeY和SS猪的肝脏中提取的含有NBS猪肝提取物添加到人肺癌细胞培养液中证明了这个推测。最后根据硒的代谢证明饲喂高3.0 mg/kg SS的日粮猪血清中含有抑制人肺癌细胞增殖的硒化物。
To study the metabolism of dietary selenium-enriched yeast (SeY) and sodium selenite (SS) in pig tissues and the effect of selenium metabolites on the proliferation of human small lung cancer cell NCI-H446, four experiments were couducted.
Exp 1. Effect of dietary SeY and SS on the antioxidant system and Se retention in pig liver and muscle.
To study effect of dietary SeY and SS on the antioxidant system and Se retention in pig tissues,60 piglets were allocated into two experiments (eperimentl and experiment 2), with 3 treatments per experiment and 10 replicates per treatment. The 6 treatments were fed basal diet supplemented with 0,0.3,3.0 mg kg-1 SeY (period 1) and 0,0.3 and 3.0 mg kg-1 SS (period 2) for 8 wk, respectively. Results showed there was no influence of dietary SeY or SS level on pig performance. Compared with 0 mg kg-1 dietary 0.3 mg kg-1 SeY and SS remarkably (P< 0.05) increased GPX activity in liver, muscle and plasma, but there was no further increase of GPX activity in these tissues from 0.3 to 3.0 mg kg-1 dietary SS and SeY. There was no remarkable effect of dietary SeY and SS level on tissue SOD and GST activity. With the increase of dietary SeY, Se concentration in pig liver, muscle and plasma was all significantly (P< 0.01) increased. In the range of 0-3.0 mg kg-1 SS, Se concentration in liver and muscle were remarkably increased (P< 0.01), while plasma Se reached plateau at the level of 0.3 mg kg-1. These suggested there was no effect of dietary SeY and SS on pig performance and tissue antioxidant system within 8 wk, but tissue Se level could be increased by the two Se in this period.
Exp 2. Influence of dietary SS and SeY on the distribution of selenocompounds in pig liver and muscle.
There are two forms of Se in mammal tissues, protein-bound selenium (PBS) and nonprotein-bound selenium (NBS). The results of Exp 1 indicated dietary SeY and SS increased Se concentrations in both liver and muscle. To study the effect of dietary SeY and SS on the distribution of NBS and PBS, one procedure for separating NBS and PBS in pig tissues was firstly developed, and then selenocompounds in pig liver and muscle obtained from Exp 1 was separated with this procedure. Results showed the NBS and PBS content in both liver and muscle increased (P< 0.05) with dietary SeY and SS. The relative percentage of PBS amount to total Se in liver decreased (P < 0.05) with the increase of dietary SeY, and that of NBS amount to liver Se increased (P< 0.05), accordingly. In contrasted to liver, the relative percentage of NBS and PBS in pig muscle kept consistent at 6% and 94%, respectively. Similar with SeY, the relative of NBS and PBS in the liver of pig fed SS increased (P< 0.05) and decreased (P< 0.05), respectively, with increase of dietary SS level. The distributing pattern of NBS and PBS in the muscle of pigs fed SS was similar with that in liver. These suggested that the distribution of NBS and PBS were similarly affected in liver and differently affected in muscle by dietary SeY and SS.
Exp 3. Effect of LE containing NBS from the liver of pig fed excess SeY and SS on the proliferation of human lung cancer cell line.
The results of Exp 2 indicated that NBS concentrate and relative percentage in pig liver was increased by ditary SeY and SS, and the distribution of NBS and PBS was similar in liver of pigs fed SS and SeY. To study whether there was potent anticancer potential of NBS, livers from pigs fed basal diet and the diet plus 3.0 mg kg-1 SS and 3.0 mg kg-1 SeY for 16 wk were homogenated, then the liver homogenate was precipitated with chloroform and ethanol. The superant was freez-dried, condensed, then three LE containg NBS was obtained. The LE from pigs fed basal diet, plus SeY and SS Se was expressed as LEB, LEY and LES, which contained 0.98μM,2.62μM and 7.84μM Se. After the addition of LEB, LEY and LES into cell culture medium, results showed that cell viability were decreased by LEY and LES when compared with LEB. The mechanism of LES and LEY inhibiting cancer cell was via the induction of DNA fragmentation. Sodium selenite is the standard material for evaluating the relative anticancer potential. After the balance of Se content between LEB and LEY by addition of SS into LEB, the inhibition effect of LEY was still higher than that of LEB. This is also true for LES. These suggested that there were powerful anticancer NBS in the liver of pigs fed excess SeY and SS.
Exp 4. Effect of pig serum on the proliferation of human lung cancer cell
The results of Exp 3 demonstrated there was potential cancer-preventive NBS in the liver of pigs fed 3.0 mg kg-1 SS. To study whether there was cancer cell suppressing selenocompounds in the serum of pigs fed 3.0 mg kg-1 SS and SeY, blood were collected via vena cava anterior and serum was separated and filter with 0.22 uM membrane. Se concentration in serum of pigs fed basal diet and supplemented with 3.0 mg kg-1 SeY and SS was 165,1316 and 834 ng ml-1, that's Se(deficient), SeY(excess) and SS (excess). To proquest the opitimal addition level of pig serum to cell medium in place of FBS,7 graded levels (8-20%) of pig serum of Se(deficient), SeY(excess) and SS(excess) were added. The optimal addition level of pig serum was 17.79%,15.39% and 14.31%, respectively based on relation between the cell MTT OD value and serum level, and the average 14.31% was chosen as the optimal level. The optimal culture time was 144 h based on cell morphology. After the addition of 15.83% pig serum to cell culture for 144h results showed the MTT OD value of cell cultured with Se (deficient) was significantly higher than that of cell treated with SS(excess), while the MTT OD value of SeY(excess) treatment cell was between Se (deficient) and SS (excess). Compared with SeY (excess) and SS (excess), Se(deficient) treat cell had lower (P< 0.05) LDH activity in medium, while no difference between SeY(excess) and SS(excess) treated cells. After stained with Annexin V and PI and analyzed with cytometry, cell apoptotic rate was significantly lower (P< 0.05) in Se (deficient) than in Se (excess) treated cell, while the treatment of SeY (excess) was still between Se (deficient) and SS (excess) and no difference was observed.
In summary, we found the Se content in the liver and muscle of pigs fed SeY or SS ranging from deficient to excess significantly increased with dietary Se level, but the NBS content and relative perentage increased with dietary SeY and SS level after NBS was separated from PBS. Furthermore, the distribution pattern of NBS/PBS in liver was similar. So, we concluded there would be anticancer NBS in pig liver based on the above finding and the potent anticancer mechanism of Se. After the addition of extracted NBS into lung cancer cell media, we found there were powerful anticancer NBS in the pig liver. Since blood was the carrier from liver to the peripheral tissue, we also suspect there was anticancer selenocompound in the serum of pig fed excess SeY and SS. With serum pharmacology, we found there was anticancer selenocompound in the serum of pig fed 3.0 mg/kg Se as SS.
为研究日粮SeY和SS水平对猪肝脏和肌肉中组织硒存留和抗氧化系统的影响,60头28日龄断奶仔猪随机分配到两期试验中即一期试验和二期试验中。两期试验分别设3个处理,每个处理10个重复。所有仔猪在饲喂用缺硒地区所产玉米-大豆配制的低硒日粮(0.03 mg kg-1 Se) 4周后分成6组,分别饲喂8周的基础日粮(0.04 mg kg-1 Se)添加0,0.3和3.0 mg kg -1的SeY(一期试验)和0,0.3和3.0 mg kg -1的SS(二期试验)。结果表明：与0.3 mg kg-1 SeY和SS相比,日粮0或3.0 mg kg-1 SS和SeY对猪的采食量、日增重和饲料转化率均无显著影响。当SS和SeY的水平从0增加到0.3 mg kg-1,猪肝脏、肌肉和血浆中GPX活性均极显著(P<0.01)提高；但3.0 mg kg-1 SeY和SS均未能(P>0.05)在其相应的0.3 mg kg-1水平上继续提高肝脏、肌肉和血浆中GPX活性。在0-3.0 mg kg-1范围内日粮SeY和SS的水平对猪血浆、肝脏和肌肉中SOD和GST活性无显著影响。随着日粮SeY水平在0-3.0 mg kg-1范围内的升高,猪肝脏、肌肉和血浆中的硒含量均极显著(P<0.01)提高。在0-3.0 mg kg-1范围内日粮SS极显著(P<0.01)提高了猪肝脏硒含量,显著(P<0.05)提高肌肉硒含量,而血浆中硒含量在0.3 mg kg-1 SS水平即达到平台。这表明8周内日粮SeY和SS的缺乏或过量对猪生产性能和组织抗氧化系统无显著影响,但能提高肝脏和肌肉的硒存留。试验二：不同日粮水平的SeY和SS对猪肝脏和肌肉中的硒化物NBS/PBS分布的影响
试验一的结果表明随着日粮SeY和SS的水平增加,肝脏和肌肉中硒存留量显著提高,但两种组织中硒是以蛋白结合硒(Protein-bound Selenium, PBS)或是非蛋白结合硒(Nonprotein-bound Selenium, NBS)的形式存在不清楚。本实验研究日粮SeY和SS水平对猪肝脏和肌肉中PBS& NBS的分布如影响。为此首先建立一套改良的分离猪组织中NBS和PBS的方法,用此方法分离测定了试验一中饲喂0-3.0 mg kg-1 SeY和SS两种硒的猪肝脏和肌肉中NBS和PBS的含量。结果表明：随着日粮SeY的升高,肝脏和肌肉中NBS和PBS的绝对含量均极显著(P<0.01)升高。肝脏中PBS占其总硒的相对含量随日粮SeY的增加而降低(P<0.01),相应地肝脏中NBS占其总硒的相对含量随日粮SeY水平的升高而增加(P<0.05)；与肝脏形成对比的是肌肉中NBS和PBS占其总硒的相对含量随日粮SeY的增加分别保持稳定在6%和94%。与饲喂SeY的猪肝脏NBS和PBS变化规律类似,饲喂SS的猪肝脏中NBS和PBS占其总硒的相对含量随SS的升高分别表现为显著升高(P<0.05)和显著降低(P<0.05),饲喂SS的猪肌肉中NBS和PBS的变化规律与肝脏类似,分别表现为随日粮SS的增加而显著升高(P<0.05)和降低(P<0.05)。这些表明日粮SeY和SS对肝脏中NBS和PBS的分布模式的影响类似,而对肌肉中NBS和PBS的分布模式影响不同。
试验三：饲喂过量SeY和SS的猪肝脏中NBS对人肺癌细胞增殖的影响
上述实验结果表明日粮SeY和SS均能极显著提高肝脏中NBS的绝对含量和相对含量,且对饲喂SeY和SS的猪肝脏中NBS和PBS的分布模式的影响一致。本实验旨在研究饲喂高水平酵母硒和亚硒酸钠16周的猪肝脏中是否含有抑制人肺癌细胞增殖功能的NBS。取饲喂16周基础日粮(0.05mg Se kg-1)及添加3.0mg kg-1 SeY和3.0 mg kg-1 SS的三种猪肝在超纯水中匀浆,匀浆液分别用氯仿和乙醇沉淀去除蛋白后所得清液经冷冻干燥浓缩、超滤和过滤除菌后得到含有NBS的硒含量分别为0.98μM,2.62μM和7.84 pM的肝脏提取物(Liver Extract,LE),分别简写为LEB, LEY和LES。将三种LE分别加入到癌细胞培养体系中,结果表明当添加水平在14%时LEY和LES能显著抑制人肺癌细胞的增殖,且诱导癌细胞的凋亡。亚硒酸钠是硒抗癌研究中的标准物质,而当在LEB中添加亚硒酸钠达到与LEY和LES中同等Se水平再添加到细胞培养体系后发现LEY和LES仍能显著抑制癌细胞的增殖,这说明LEY和LES中的NBS抑制癌细胞的能力高于亚硒酸钠,即饲喂3.0 mg/kg SeY或SS的猪肝脏中含有比亚硒酸钠抗癌活性更强的硒化物。
试验四：饲喂3.0 mg/kg SeY和SS的猪血清对人肺癌细胞增殖的影响
试验三表明饲喂3.0 mg kg-1 SS和3.0 mg kg-1 SeY的猪肝脏中有抑制人肺癌细胞增殖的硒化物。根据硒的代谢规律和抗癌功能,推测饲喂3.0 mg/kg SeY或SS的猪血清中也可能含有抗癌的硒化物。本实验旨在研究饲喂高水平日粮SeY和SS的猪血液中是否含有抑制人肺癌细胞增殖的物质。前腔静脉采集饲喂基础日粮(0.03 mg kg-1 Se)以及基础日粮添加3.0mg kg-1 SeY和3.0mg kg-1 SS 16周的猪清,经0.22μm滤膜过滤除菌后得到总硒含量分别为165 ng ml-1,1316 ng ml-1和834 ng ml-1的三种猪血清,分别简写为Se(0), SeY(3)和SS(3)。为探索猪血清代替FBS培养细胞的最佳添加水平,在培养液中添加7个梯度(8%-20%)的猪血清,根据细胞的MTT OD值随血清添加水平的变化Se(0)、SeY(3)和SS(3)三种猪血清的最适添加比例分别为17.79%,15.39%和14.31%,取三者的均值15.83%作为猪血清的最适添加水平。根据细胞在48h,96h和144h三个时间点的形态变化取144 h作为细胞最适培养时间。试验结果表明Se(0)猪血清培养癌细胞的MTTOD值显著(P<0.05)高于SS(3)猪血清,而SeY(3)与Se(0)和SS(3)猪血清处理细胞的MTT OD值均无显著差异。与SeY(3)和SS(3)相比,Se(缺)比培养细胞的上清液中LDH活性均显著(P<0.05)降低,而SeY(3)和SS(3)两者培养细胞上清中LDH活性无显著差异。细胞经Annexin V和PI双染后经流式细胞仪分析表明,Se(0)比SS(3)培养细胞的凋亡率显著降低(P<0.05)；而SeY(3)培养细胞的凋亡率介于Se(0)和SS(3)之间,与两者差异均不显著。上述表明饲喂3.0 mg kg-1 SS的猪血清中含有抑制人肺癌细胞增殖的硒化物。
本研究通过对日粮SeY和SS在猪肝脏和肌肉中代谢存留的研究机理我们推测饲喂3.0 mg/kg SeY或SS的猪肝脏中可能具有抑制人癌细胞增殖的硒化物。通过将从饲喂过量SeY和SS猪的肝脏中提取的含有NBS猪肝提取物添加到人肺癌细胞培养液中证明了这个推测。最后根据硒的代谢证明饲喂高3.0 mg/kg SS的日粮猪血清中含有抑制人肺癌细胞增殖的硒化物。
To study the metabolism of dietary selenium-enriched yeast (SeY) and sodium selenite (SS) in pig tissues and the effect of selenium metabolites on the proliferation of human small lung cancer cell NCI-H446, four experiments were couducted.
Exp 1. Effect of dietary SeY and SS on the antioxidant system and Se retention in pig liver and muscle.
To study effect of dietary SeY and SS on the antioxidant system and Se retention in pig tissues,60 piglets were allocated into two experiments (eperimentl and experiment 2), with 3 treatments per experiment and 10 replicates per treatment. The 6 treatments were fed basal diet supplemented with 0,0.3,3.0 mg kg-1 SeY (period 1) and 0,0.3 and 3.0 mg kg-1 SS (period 2) for 8 wk, respectively. Results showed there was no influence of dietary SeY or SS level on pig performance. Compared with 0 mg kg-1 dietary 0.3 mg kg-1 SeY and SS remarkably (P< 0.05) increased GPX activity in liver, muscle and plasma, but there was no further increase of GPX activity in these tissues from 0.3 to 3.0 mg kg-1 dietary SS and SeY. There was no remarkable effect of dietary SeY and SS level on tissue SOD and GST activity. With the increase of dietary SeY, Se concentration in pig liver, muscle and plasma was all significantly (P< 0.01) increased. In the range of 0-3.0 mg kg-1 SS, Se concentration in liver and muscle were remarkably increased (P< 0.01), while plasma Se reached plateau at the level of 0.3 mg kg-1. These suggested there was no effect of dietary SeY and SS on pig performance and tissue antioxidant system within 8 wk, but tissue Se level could be increased by the two Se in this period.
Exp 2. Influence of dietary SS and SeY on the distribution of selenocompounds in pig liver and muscle.
There are two forms of Se in mammal tissues, protein-bound selenium (PBS) and nonprotein-bound selenium (NBS). The results of Exp 1 indicated dietary SeY and SS increased Se concentrations in both liver and muscle. To study the effect of dietary SeY and SS on the distribution of NBS and PBS, one procedure for separating NBS and PBS in pig tissues was firstly developed, and then selenocompounds in pig liver and muscle obtained from Exp 1 was separated with this procedure. Results showed the NBS and PBS content in both liver and muscle increased (P< 0.05) with dietary SeY and SS. The relative percentage of PBS amount to total Se in liver decreased (P < 0.05) with the increase of dietary SeY, and that of NBS amount to liver Se increased (P< 0.05), accordingly. In contrasted to liver, the relative percentage of NBS and PBS in pig muscle kept consistent at 6% and 94%, respectively. Similar with SeY, the relative of NBS and PBS in the liver of pig fed SS increased (P< 0.05) and decreased (P< 0.05), respectively, with increase of dietary SS level. The distributing pattern of NBS and PBS in the muscle of pigs fed SS was similar with that in liver. These suggested that the distribution of NBS and PBS were similarly affected in liver and differently affected in muscle by dietary SeY and SS.
Exp 3. Effect of LE containing NBS from the liver of pig fed excess SeY and SS on the proliferation of human lung cancer cell line.
The results of Exp 2 indicated that NBS concentrate and relative percentage in pig liver was increased by ditary SeY and SS, and the distribution of NBS and PBS was similar in liver of pigs fed SS and SeY. To study whether there was potent anticancer potential of NBS, livers from pigs fed basal diet and the diet plus 3.0 mg kg-1 SS and 3.0 mg kg-1 SeY for 16 wk were homogenated, then the liver homogenate was precipitated with chloroform and ethanol. The superant was freez-dried, condensed, then three LE containg NBS was obtained. The LE from pigs fed basal diet, plus SeY and SS Se was expressed as LEB, LEY and LES, which contained 0.98μM,2.62μM and 7.84μM Se. After the addition of LEB, LEY and LES into cell culture medium, results showed that cell viability were decreased by LEY and LES when compared with LEB. The mechanism of LES and LEY inhibiting cancer cell was via the induction of DNA fragmentation. Sodium selenite is the standard material for evaluating the relative anticancer potential. After the balance of Se content between LEB and LEY by addition of SS into LEB, the inhibition effect of LEY was still higher than that of LEB. This is also true for LES. These suggested that there were powerful anticancer NBS in the liver of pigs fed excess SeY and SS.
Exp 4. Effect of pig serum on the proliferation of human lung cancer cell
The results of Exp 3 demonstrated there was potential cancer-preventive NBS in the liver of pigs fed 3.0 mg kg-1 SS. To study whether there was cancer cell suppressing selenocompounds in the serum of pigs fed 3.0 mg kg-1 SS and SeY, blood were collected via vena cava anterior and serum was separated and filter with 0.22 uM membrane. Se concentration in serum of pigs fed basal diet and supplemented with 3.0 mg kg-1 SeY and SS was 165,1316 and 834 ng ml-1, that's Se(deficient), SeY(excess) and SS (excess). To proquest the opitimal addition level of pig serum to cell medium in place of FBS,7 graded levels (8-20%) of pig serum of Se(deficient), SeY(excess) and SS(excess) were added. The optimal addition level of pig serum was 17.79%,15.39% and 14.31%, respectively based on relation between the cell MTT OD value and serum level, and the average 14.31% was chosen as the optimal level. The optimal culture time was 144 h based on cell morphology. After the addition of 15.83% pig serum to cell culture for 144h results showed the MTT OD value of cell cultured with Se (deficient) was significantly higher than that of cell treated with SS(excess), while the MTT OD value of SeY(excess) treatment cell was between Se (deficient) and SS (excess). Compared with SeY (excess) and SS (excess), Se(deficient) treat cell had lower (P< 0.05) LDH activity in medium, while no difference between SeY(excess) and SS(excess) treated cells. After stained with Annexin V and PI and analyzed with cytometry, cell apoptotic rate was significantly lower (P< 0.05) in Se (deficient) than in Se (excess) treated cell, while the treatment of SeY (excess) was still between Se (deficient) and SS (excess) and no difference was observed.
In summary, we found the Se content in the liver and muscle of pigs fed SeY or SS ranging from deficient to excess significantly increased with dietary Se level, but the NBS content and relative perentage increased with dietary SeY and SS level after NBS was separated from PBS. Furthermore, the distribution pattern of NBS/PBS in liver was similar. So, we concluded there would be anticancer NBS in pig liver based on the above finding and the potent anticancer mechanism of Se. After the addition of extracted NBS into lung cancer cell media, we found there were powerful anticancer NBS in the pig liver. Since blood was the carrier from liver to the peripheral tissue, we also suspect there was anticancer selenocompound in the serum of pig fed excess SeY and SS. With serum pharmacology, we found there was anticancer selenocompound in the serum of pig fed 3.0 mg/kg Se as SS.
引文
[1]Schweizer U, Schomburg L, Savaskan NE. The neurobiology of selenium:lessons from transgenic mice[J]. Journal of nutrition,2004;134(4):707-710.
[2]Derumeaux H, Valeix P, Castetbon K, Bensimon M, Boutron-Ruault MC, Arnaud J, et al. Association of selenium with thyroid volume and echostructure in 35-to 60-year-old French adults[J]. European Journal of Endocrinology,2003; 148(3):309-315.
[3]Camins A, Jimenez A, Sureda FX, Pallas M, Escubedo E, Camarasa J. Characterization of nitrobenzylthioinosine binding sites in the mitochondrial fraction of rat testis[J]. Life Sciences,1996;58(9):753-759.
[4]Schwarz K, Foltz CM. Selenium as an integral part of factor 3 against dietary necrotic liver degeneration[J]. J Am Chem Soc,1957;79(12):3292-3293.
[5]Brown MR, Cohen HJ, Lyons JM, Curtis TW, Thunberg B, Cochran WJ, et al. Proximal muscle weakness and selenium deficiency associated with long term parenteral nutrition[J]. The American journal of clinical nutrition,1986;43(4):549.
[6]Rayman MP. The importance of selenium to human health[J]. The Lancet,2000;356(9225):233-241.
[7]Sheridan PA, Zhong N, Carlson BA, Perella CM, Hatfield DL, Beck MA. Decreased selenoprotein expression alters the immune response during influenza virus infection in mice[J]. Journal of Nutrition,2007,131(6):1466.
[8]Shamberger RJ, Tytko SA, Willis CE. Antioxidants and cancer. Part VI. Selenium and age-adjusted human cancer mortality[J]. Archives of environmental health,1977;31 (5):231.
[9]Clark LC, Cantor KP, Allaway WH. Selenium in forage crops and cancer mortality in US counties[J]. Archives of environmental health (USA),1991.
[10]Ip C, Dong Y, Ganther HE. New concepts in selenium chemoprevention[J]. Cancer and Metastasis Reviews,2002;21 (3):281-289.
[11]Rubins HB, Robins SJ, Collins D, Fye CL, Anderson JW, Elam MB, et al. Gemfibrozil for the secondary prevention of coronary heart disease in men with low levels of high-density lipoprotein cholesterol.1999:410-418.
[12]Finley JW. Bioavailability of selenium from foods[J]. Nutrition reviews,2006;64(3):146-151.
[13]Clark LC, F.Combs G, B. W.Urnbull, H.Slate E, Chalker DK, Chow J, et al. Effect of selenium supplementation for cancer prevention in patients with carcinoma of the skin:a randomized controlled trial.[J]. J Am Med Assoc,1996;276:1957-1963.
[14]Combs GF. Current Evidence and Research Needs to Support a Health Claim for Selenium and Cancer Prevention 1. Am Soc Nutrition,2005:343-347.
[15]Yu Yu S, Zhu YJ, Li WG. Protective role of selenium against hepatitis B virus and primary liver cancer in Qidong[J]. Biological trace element research,1997;56(1):117-124.
[16]Taylor PR, Li B, Dawsey SM, Li JY, Yang CS, Guo W, et al. Prevention of esophageal cancer:the nutrition intervention trials in Linxian, China[J]. Cancer Research,1994;54(7 Supplement):2029s.
[17]Blot WJ, Li JY, Taylor PR, Guo W, Dawsey SM, Li B. The Linxian trials:mortality rates by vitamin-mineral intervention group. Am Soc Nutrition,1995:1424-1426.
[18]Thompson HJ. Effect of an inorganic and organic form of dietary selenium on the promotional stage of mammary carcinogenesis in the rat[J]. Cancer Research,1984;44(7):2803-2806.
[19]Birt DF. Inhibition by dietary selenium of colon cancer induced in the rat by bis (2-oxopropyl) nitrosamine[J]. Cancer Research,1982;42(11):4455-4459.
[20]Li DH, Graef GL, Yee JA, Yan L. Dietary supplementation with high-selenium soy protein reduces pulmonary metastasis of melanoma cells in mice[J]. Journal of Nutrition,2004; 134(6):1536-1540.
[21]Jiang C, Wang Z, Ganther H, Lu J. Caspases as Key Executors of Methyl Selenium-induced Apoptosis (Anoikis) of DU-145 Prostate Cancer Cells 1. AACR,2001:3062-3070.
[22]Jiang C, Wang Z, Ganther H, Lu J. Distinct effects of methylseleninic acid versus selenite on apoptosis, cell cycle, and protein kinase pathways in DU145 human prostate cancer cells[J]. Molecular cancer therapeutics,2002;1(12):1059.
[23]Ip C. Lessons from basic research in selenium and cancer prevention[J]. Journal of Nutrition,1998;128(11):1845-1854.
[24]Beilstein MA, Whanger PD. Effect of Vitamin B-6 Deficiency on Selenium Metabolism in the rat.[J]. J Nutr,1989; 119:1962-1972.
[25]Behne D, Kyriakopoulos A, Scheid S, Gessner H. Effects of chemical form and dosage on the incorporation of selenium into tissue proteins in rats[J]. Journal of Nutrition,1991;121(6):806.
[26]Kim YY, Mahan DC. Comparative effects of high dietary levels of organic and inorganic selenium on selenium toxicity of growing-finishing pigs[J]. Journal of animal science,2001;79(4):942.
[27]Mahan DC, Cline TR, Richert B. Effects of dietary levels of selenium-enriched yeast and sodium selenite as selenium sources fed to growing-finishing pigs on performance, tissue selenium, serum glutathione peroxidase activity, carcass characteristics, and loin quality[J]. Journal of Animal Science,1999;77(8):2172-2179.
[28]Mahan DC, Parrett NA. Evaluating the efficacy of selenium-enriched yeast and sodium selenite on tissue selenium retention and serum glutathione peroxidase activity in grower and finisher swine[J]. Journal of Animal Science,1996;74(12):2967-2974.
[29]Thompson HJ. Effect of short-term feeding of sodium selenite on 7,12-dimethylbenz (a) anthracene-induced mammary carcinogenesis in the rat[J]. Cancer Research,1982;42(12):4954-4958.
[30]Arthur JR, McKenzie RC, Beckett GJ. Selenium in the immune system[J]. The Journal of nutrition,2003;133(5):1457-1459.
[31]Lu J, Jiang C. Antiangiogenic activity of selenium in cancer chemoprevention:metabolite-specific effects[J]. Nutrition and cancer,2001;40(1):64-73.
[32]Hu YJ, Diamond AM. Role of Glutathione Peroxidase 1 in Breast Cancer Loss of Heterozygosity and Allelic Differences in the Response to Selenium 1. AACR,2003:3347-3351.
[33]Hardie LJ, Briggs JA, Davidson LA, Allan JM, King R, Williams GI, et al. The effect of hOGGl and glutathione peroxidase I genotypes and 3p chromosomal loss on 8-hydroxydeoxyguanosine levels in lung cancer[J]. Carcinogenesis,2000;21(2):167.
[34]Sinha R, El-Bayoumy K. Apoptosis is a critical cellular event in cancer chemoprevention and chemotherapy by selenium compounds[J]. Current Cancer Drug Targets,2004;4:13-28.
[35]Spallholz JE, Hoffman DJ. Selenium toxicity:cause and effects in aquatic birds[J]. Aquatic Toxicology,2002;57(1-2):27-37.
[36]Zhong W, Oberley TD. Redox-mediated Effects of Selenium on Apoptosis and Cell Cycle in the LNCaP Human Prostate Cancer Cell Line 1. AACR,2001:7071-7078.
[37]Whanger PD. Selenium and its relationship to cancer:an update[J]. British journal of nutrition,2004;91(01):11-28.
[38]Ganther HE. Selenium metabolism, selenoproteins and mechanisms of cancer prevention: complexities with thioredoxin reductase[J]. Carcinogenesis,1999;20(9):1657.
[39]Miller ER, Ullrey DE. The pig as a model for human nutrition[J]. Annual review of nutrition,1987;7(1):361-382.
[40]Baquerizo A, Mhoyan A, Kearns-Jonker M, Arnaout WS, Shackleton C, Busuttil RW, et al. Characterization of human xenoreactive antibodies in liver failure patients exposed to pig hepatocytes after bioartificial liver treatment:an ex vivo model of pig to human xenotransplantation[J]. Transplantation,1999;67(1):5.
[41]Van Rij AM, Thomson CD, McKenzie JM, Robinson MF. Selenium deficiency in total parenteral nutrition[J]. American Journal of Clinical Nutrition,1979;32(10):2076.
[42]Yang F, Lin Z, Xing JRU, Li SG, Yang J, San SUN, et al. Se deficiency is a necessary but not sufficient factor required for the pathogenesis of Keshan disease[J]. Journal of clinical biochemistry and nutrition,1994;16(2):101-110.
[43]Moreno-Reyes R, Suetens C, Mathieu F, Begaux F, Zhu D, Rivera MT, et al. Kashin-Beck osteoarthropathy in rural Tibet in relation to selenium and iodine status.1998:1112-1120.
[44]Yang C, Niu C, Bodo M, Gabriel E, Notbohm H, Wolf E, et al. Fulvic acid supplementation and selenium deficiency disturb the structural integrity of mouse skeletal tissue. An animal model to study the molecular defects of Kashin-Beck disease[J]. Biochemical Journal,1993;289(Pt 3):829.
[45]Medina D, Oborn CJ. Selenium Inhibition of DNA Synthesis in Mouse Mammary Epithelial Cell Line YN-4[J]. Cancer Research,1984;44:4361-4365.
[46]Zeng H. Selenite and Selenomethionine Promote HL-60 Cell Cycle Progression 1 2[J]. Journal of Nutrition,2002;132(4):674-679.
[47]Thomson CD. Assessment of requirements for selenium and adequacy of selenium status:a review[J]. European journal of clinical nutrition,2004;58(3):391-402.
[48]Behne D, Pfeifer H, Rothlein D, Kyriakopoulos A. Cellular and subcellular distribution of selenium and selenium-containing proteins in the rat[J]. Trace elements in man and animals,2000;10:29-34.
[49]Behne D, Hofer-Bosse T. Effects of a low selenium status on the distribution and retention of selenium in the rat[J]. Journal of Nutrition,1984;114(7):1289.
[50]Whanger PD. Selenocompounds in plants and animals and their biological significance[J]. Journal of the American College ofNutrition,2002;21(3):223-232.
[51]Terry N, Zayed AM, De Souza MP, Tarun AS. S ELENIUM IN H IGHER P LANTS[J]. Annual Review of Plant Biology,2000;51(1):401-432.
[52]Finley JW. Selenium accumulation in plant foods[J]. Nutrition reviews,2005;63(6):196-202.
[53]Cases J, Vacchina V, Napolitano A, Caporiccio B, Besancon P, Lobinski R, et al. Selenium from Selenium-Rich Spirulina Is Less Bioavailable than Selenium from Sodium Selenite and Selenomethionine in Selenium-Deficient Rats 1[J]. Journal of nutrition,2001;131(9):2343-2350.
[54]Cai XJ, Block E, Uden PC, Zhang X, Quimby BD, Sullivan JJ. Allium chemistry:identification of selenoamino acids in ordinary and selenium-enriched garlic, onion, and broccoli using gas chromatography with atomic emission detection[J]. Journal of Agricultural and Food Chemistry,1995;43(7):1754-1757.
[55]Bell JG, Cowey CB. Digestibility and bioavailability of dietary selenium from fishmeal, selenite, selenomethionine and selenocystine in Atlantic salmon (Salmo salar)[J]. Aquaculture (Netherlands),1989.
[56]Ip C, Birringer M, Block E, Kotrebai M, Tyson JF, Uden PC, et al. Chemical speciation influences comparative activity of selenium-enriched garlic and yeast in mammary cancer prevention[J]. J Agric Food Chem,2000;48(6):2062-2070.
[57]Bird SM, Uden PC, Tyson JF, Block E, Denoyer E. Speciation of selenoamino acids and organoselenium compounds in selenium-enriched yeast using high-performance liquid chromatography-inductively coupled plasma mass spectrometry[J]. Journal of Analytical Atomic Spectrometry,1997;12(7):785-788.
[58]Chassaigne H, Chery CC, Bordin G, Rodriguez AR. Development of new analytical methods for selenium speciation in selenium-enriched yeast material[J]. Journal of Chromatography A,2002;976(1-2):409-422.
[59]Rayman MP. Selenium in cancer prevention:a review of the evidence and mechanism of action[J]. Proceedings of the Nutrition Society,2005;64(04):527-542.
[60]Hawkes WC, Alkan FZ, Oehler L, Usda ARS. Absorption, distribution and excretion of selenium from beef and rice in healthy North American men[J].2003.
[61]Xu XM, Carlson BA, Irons R, Mix H, Zhong N, Gladyshev VN, et al. Selenophosphate synthetase 2 is essential for selenoprotein biosynthesis[J]. Biochemical Journal,2007;404(Pt 1):115.
[62]Kobayashi Y, Ogra Y, Ishiwata K, Takayama H, Aimi N, Suzuki KT. Selenosugars are key and urinary metabolites for selenium excretion within the required to low-toxic range[J]. Proceedings of the National Academy of Sciences,2002;99(25):15932-15936.
[63]Schrauzer GN. Selenomethionine:A Review of Its Nutritional Significance, Metabolism and Toxicity 1[J]. Journal of nutrition,2000;130(7):1653-1656.
[64]Olson OE, Palmer IS. Selenoamino acids in tissues of rats administered inorganic selenium[J]. Metabolism:clinical and experimental,1976;25(3):299.
[65]Beilstein MA, Whanger PD. Glutathione peroxidase activity and chemical forms of selenium in tissues of rats given selenite or selenomethionine[J]. J Inorg Biochem,1988;33(1).
[66]Sunde RA, Gutzke GE, Hoekstra WG Effect of dietary methionine on the biopotency of selenite and selenomethionine in the rat[J]. Journal of Nutrition,1981;111(1):76.
[67]Luo XM, Wei HJ, Yang CL, Xing J, Liu X, Qiao CH, et al. Bioavailability of selenium to residents in a low-selenium area of China[J]. American Journal of Clinical Nutrition,1985;42(3):439.
[68]Vendeland SC, Deagen JT, Butler JA, Whanger PD. Uptake of selenite, selenomethionine and selenate by brush border membrane vesicles isolated from rat small intestine[J]. Biometals,1994;7(4):305-312.
[69]Deagen JT, Butler JA, Beilstein MA, Whanger PD. Effects of dietary selenite, selenocystine and selenomethionine on selenocysteine lyase and glutathione peroxidase activities and on selenium levels in rat tissues[J]. Journal of Nutrition,1987;117(1):91.
[70]Waschulewski IH, Sunde RA. Effect of dietary methionine on utilization of tissue selenium from dietary selenomethionine for glutathione peroxidase in the rat[J]. Journal ofNutrition,1988;118(3):367.
[71]Whanger PD, Butler JA, Xia Y. Can physiological responses be used to assess the chemical forms of selenium consumed by humans.1994; 1994. p.8-10.
[72]McGuire MK, Burgert SL, Milner JA, Glass L, Kummer R, Deering R, et al. Selenium status of lactating women is affected by the form of selenium consumed[J]. American Journal of Clinical Nutrition,1993;58:649-649.
[73]Alaejos MS, Romero CD. Selenium in human lactation[J]. Nutrition Reviews,1995;53(6):159-166.
[74]Boles JO, Cisneros RJ, Weir MS, Odom JD, Villafranca JE, Dunlap RB. Purification and characterization of selenomethionyl thymidylate synthase from Escherichia coli:comparison with the wild-type enzyme[J]. Biochemistry,1991;30(46):11073-11080.
[75]Huber RE, Criddle RS. The isolation and properties of beta-galactosidase from Escherichia coli grown on sodium selenate[J]. Biochim Biophys Acta,1967;141(3):587-599.
[76]Soda K, Oikawa T, Esaki N. Vitamin B6 enzymes participating in selenium amino acid metabolism[J]. Biofactors,1999; 10.
[77]Kajander EO, Harvima RJ, Eloranta TO, Martikainen H, Kantola M, K renlampi SO, et al. Metabolism, cellular actions, and cytotoxicity of selenomethionine in cultured cells[J]. Biological trace element research,1991;28(1):57-68.
[78]Rotruck JT, Pope AL, Ganther HE, Swanson AB, Hafeman DG, Hoekstra WG. Selenium: biochemical role as a component of glutathione peroxidase[J]. Science,1973;179(4073):588.
[79]Letavayovi L, Vl?kova V, Brozmanova J. Selenium:from cancer prevention to DNA damage[J]. Toxicology,2006;227(1-2):1-14.
[80]Spallholz JE, Palace VP, Reid TW. Methioninase and selenomethionine but not Se-methylselenocysteine generate methylselenol and superoxide in an in vitro chemiluminescent assay: implications for the nutritional carcinostatic activity of selenoamino acids [J]. Biochemical pharmacology,2004;67(3):547-554.
[81]Shen HM, Yang CF, ONG CN. Sodium selenite-induced oxidative stress and apoptosis in human hepatoma HepG2 cells[J]. Int J Cancer,1999;81:820-828.
[82]Shen HM, Yang CF, Liu J, Ong CN. Dual role of glutathione in selenite-induced oxidative stress and apoptosis in human hepatoma cells[J]. Free Radical Biology and Medicine,2000;28(7):1115-1124.
[83]Kim Y, Jhon D, Lee KY. Involvement of ROS and JNK1 in selenite-induced apoptosis in Chang liver cells[J]. Experimental and molecular medicine,2004;36(2):151-164.
[84]Spallholz JE. Free radical generation by selenium compounds and their prooxidant toxicity[J]. Biomedical and environmental sciences:BES,1997;10(2-3):260.
[85]Drake EN. Cancer chemoprevention:selenium as a prooxidant, not an antioxidant[J]. Medical hypotheses,2006;61(2):318-322.
[86]Miki K, Xu M, Gupta A, Ba Y, Tan Y, Al-Refaie W, et al. Methioninase Cancer Gene Therapy with Selenomethionine as Suicide Prodrug Substrate 1[J]. Cancer Research,2001;61(18):6805-6810.
[87]Zeng H, Briske-Anderson M, Idso JP, Hunt CD. The selenium metabolite methylselenol inhibits the migration and invasion potential of HT1080 tumor cells[J]. Journal of Nutrition,2006; 136(6):1528-1532.
[88]Inoue H, Inagaki K, Sugimoto M, Esaki N, Soda K, Tanaka H. Structural analysis of the L-methionine{gamma}-lyase gene from Pseudomonas putida[J]. Journal of Biochemistry,1995; 117(5):1120.
[89]Smith LL. Mechanism of paraquat toxicity in lung and its relevance to treatment[J]. Human & Experimental Toxicology,1987,6(1):31.
[90]Cheng WH, Fu YX, Porres JM, Ross DA, Lei XG. Selenium-dependent cellular glutathione peroxidase protects mice against a pro-oxidant-induced oxidation of NADPH, NADH, lipids, and protein. FASEB,1999:1467-1475.
[91]Bray RC, Cockle SA, Fielden EM, Roberts PB, Rotilio G, Calabrese L. Reduction and inactivation of superoxide dismutase by hydrogen peroxide[J]. Biochemical Journal,1974;139(1):43.
[92]de Haan JB, Bladier C, Griffiths P, Kelner M, O'Shea RD, Cheung NS, et al. Mice with a homozygous null mutation for the most abundant glutathione peroxidase, Gpxl, show increased susceptibility to the oxidative stress-inducing agents paraquat and hydrogen peroxide[J]. Journal of Biological Chemistry,1998;273(35):22528-22536.
[93]Sugiyama M, Ando A, Ogura R. Effect of vitamin E on survival, glutathione reductase and formation of chromium (V) in Chinese hamster V-79 cells treated with sodium chromate (VI)[J]. Carcinogenesis,1989; 10(4):737.
[94]Sies H. Glutathione and its role in cellular functions[J]. Free Radical Biology and Medicine,1999;27(9-10):916-921.
[95]Burk RF, Nishiki K, Lawrence RA, Chance B. Peroxide removal by selenium-dependent and selenium-independent glutathione peroxidases in hemoglobin-free perfused rat liver[J]. Journal of Biological Chemistry,1978;253(1):43-46.
[96]Reiter R, Wendel A. Selenium and drug metabolism--II. Independence of glutathione peroxidase and reversibility of hepatic enzyme modulations in deficient mice[J]. Biochemical pharmacology,1984;33(12):1923.
[97]Cheng WH, Ho YS, Ross DA, Valentine BA, Combs GF, Lei XG. Cellular glutathione peroxidase knockout mice express normal levels of selenium-dependent plasma and phospholipid hydroperoxide glutathione peroxidases in various tissues[J]. Journal of nutrition,1997;127(8):1445-1450.
[98]Lei XG, Dann HM, Ross DA, Cheng WH, Combs GF, Roneker KR. Dietary selenium supplementation is required to support full expression of three selenium-dependent glutathione peroxidases in various tissues of weanling pigs[J]. Journal of Nutrition,1998;128(1):130-135.
[99]Butler JA, Whanger PD, Kaneps AJ, Patton NM. Metabolism of selenite and selenomethionine in the rhesus monkey[J]. Journal ofNutrition,1990;120(7):751.
[100]Payne RL, Southern LL. Comparison of inorganic and organic selenium sources for broilers[J]. Poultry Science,2005;84(6):898.
[101]Gladyshev VN, Stadtman TC, Hatfield DL, Jeang KT. Levels of major selenoproteins in T cells decrease during HIV infection and low molecular mass selenium compounds increase. National Acad Sciences,1999:835-839.
[102]Irons R, Carlson BA, Hatfield DL, Davis CD. Both Selenoproteins and Low Molecular Weight Selenocompounds Reduce Colon Cancer Risk in Mice with Genetically Impaired Selenoprotein Expression 1 [J]. Journal ofNutrition,2006,136(5):1311-1317.
[103]Combs Jr GF, Clark LC, Turnbull BW. An analysis of cancer prevention by selenium[J]. Biofactors,2001;14(1):153-159.
[104]Behne D, Kyriakopoulos A. Mammalian Selenium-containing proteins[J]. Annu Rev Nutr,2001;21:453-473.
[105]Kryukov GV, Castellano S, Novoselov SV, Lobanov AV, Zehtab O, Guigo R, et al. Characterization of mammalian selenoproteomes[J]. Science,2003;300(5624):1439-1443.
[106]Bansal MP, Cook RG, Danielson KG, Medina D. A 14-kilodalton selenium-binding protein in mouse liver is fatty acid-binding protein[J]. Journal of Biological Chemistry,1989;264(23):13780-13784.
[107]Pumford NR, Martin BM, Hinson JA. A metabolite of acetaminophen covalently binds to the 56 kDa selenium binding protein[J]. Biochemical and biophysical research communications,1992; 182(3):1348.
[108]Li ZY, Guo SY, Li L. Bioeffects of selenite on the growth of Spirulina platensis and its biotransformation[J]. Bioresource technology,2003;89(2):171-176.
[109]Lunde G. Location of lipid-soluble selenium in marine fish to the lipoproteins[J]. Journal of the Science of Food and Agriculture,1972;23(8).
[110]Shamberger RJ, Frost DV. Possible protective effect of selenium against human cancer[J]. Canadian Medical Association Journal,1969;100(14):682.
[111]Shamberger RJ, Willis CE. Selenium distribution and human cancer mortality[J]. CRC Crit Rev Clin Lab Sci,1971;2(2):211-221.
[112]Li JY, Taylor PR, Li B, Dawsey S, Wang GQ, Ershow AG, et al. Nutrition intervention trials in Linxian, China:multiple vitamin/mineral supplementation, cancer incidence, and disease-specific mortality among adults with esophageal dysplasia[J].jnci,1993;85(18):1492-1498.
[113]Yu SY, Zhu YJ, Li WG, Huang QS, Huang CZ, Zhang QN, et al. A preliminary report on the intervention trials of primary liver cancer in high-risk populations with nutritional supplementation of selenium in China[J]. Biol Trace Elem Res,1991;29(3):289-294.
[114]Van den Brandt PA. A prospective cohort study on selenium status and the risk of lung cancer[J]. Cancer Research,1993;53(20):4860-4865.
[115]Coates RJ, Weiss NS, Daling JR, Morris JS, Labbe RF. Serum levels of selenium and retinol and the subsequent risk of cancer[J]. American journal of epidemiology,1988;128(3):515-523.
[116]Knekt P, Aromaa A, Maatela J, Alfthan G, Aaran RK, Teppo L, et al. Serum vitamin E, serum selenium and the risk of gastrointestinal cancer [J]. International journal of cancer Journal international du cancer,1988;42(6):846.
[117]Taylor PR, Li B, Dawsey S,. Prevention of esophageal cancer:the nutrition intervention trials in Linxian, China.[J]. Cancer Reserch,1994;54:2029s-2031s.
[118]Comstock GW, Bush TL, Helzlsouer K. Serum retinol, beta-carotene, vitamin E, and selenium as related to subsequent cancer of specific sites[J]. American journal of epidemiology,1992;135(2):115-121.
[119]Zhuo H, Smith AH, Steinmaus C. Selenium and lung cancer:a quantitative analysis of heterogeneity in the current epidemiological literature[J]. Cancer Epidemiology Biomarkers & Prevention,2004;13(5):771-778.
[120]Finley JW. The retention and distribution by healthy young men of stable isotopes of selenium consumed as selenite, selenate or hydroponically-grown broccoli are dependent on the isotopic form[J]. The Journal of nutrition,1999;129(4):865-871.
[121]Yang G, Ge K, Chen JCX. Selenium-related endemic diseases and the daily selenium requirement of humans[J]. World review of nutrition and dietetics,1988;55:91-152.
[122]Raines AM, Sunde RA. Selenium-regulated expression of non-selenoprotein genes [J]. FASEB J,2009;23:346(344).
[123]Ip C. Factors influencing the anticarcinogenic efficacy of selenium in dimethylbenz [a] anthracene-induced mammary tumorigenesis in rats[J]. Cancer Research,1981;41(7):2683-2686.
[124]Jacobs MM. Selenium inhibition of 1,2-dimethylhydrazine-induced colon carcinogenesis[J]. Cancer Research,1983;43(4):1646-1649.
[125]El-Bayoumy K. Inhibition of 7,12-dimethylbenz (a) anthracene-induced tumors and DNA adduct formation in the mammary glands of female Sprague-Dawley rats by the synthetic organoselenium compound,1,4-phenylenebis (methylene) selenocyanate[J]. Cancer Research,1992;52(9):2402-2407.
[126]Ip C, Lisk DJ, Thompson HJ. Selenium-enriched garlic inhibits the early stage but not the late stage of mammary carcinogenesis[J]. Carcinogenesis,1996;17(9):1979.
[127]Davis CD, Zeng H, Finley JW. Selenium-enriched broccoli decreases intestinal tumorigenesis in multiple intestinal neoplasia mice[J].J Nutr,2002;132(2):307-309.
[128]Ganther HE. Selenium metabolism and mechanisms of cancer prevention[J]. Nutrition and Cancer Prevention:New Insights Into the Role of Phytochemicals,2001: 119.
[129]Feng Y, Finley JW, Davis CD, Becker WK, Fretland AJ, Hein DW. Dietary selenium reduces the formation of aberrant crypts in rats administered 3,2'-dimethyl-4-aminobiphenyl[J]. Toxicology and Applied Pharmacology,1999;151(1):36-42.
[130]Ip C. Chemical form of selenium, critical metabolites, and cancer prevention[J]. Cancer Research,1991;51 (2):595-600.
[131]Ip C, Thompson HJ, Zhu Z, Ganther HE. In Vitro and in Vivo Studies of Methylseleninic Acid:Evidence That a Monomethylated Selenium Metabolite Is Critical for Cancer Chemoprevention [J]. Cancer Research,2000;60(11):2882-2886.
[132]Legrand C, Bour JM, Jacob C, Capiaumont J, Martial A, Marc A, et al. Lactate dehydrogenase (LDH) activity of the cultured eukaryotic cells as marker of the number of dead cells in the medium [corrected] [J]. Journal of biotechnology,1992;25(3):231.
[133]Wang Z, Jiang C, Ganther H, Lu J. Antimitogenic and proapoptotic activities of methylseleninic acid in vascular endothelial cells and associated effects on PI3K-AKT, ERK, JNK and p38 MAPK signaling[J]. Cancer research,2001;61(19):7171-7178.
[134]Jiang C, Wang ZS, Ganther HE, Lv JX. Distinct effects of methylseleninic acid vers selenite on apoptosis, cell cycle, and protein kinase pathways in DU145 human prostate cancer cells.[J]. Molecular Cancer Therapeutics,2002;1:1059-1066.
[135]Hu H, Jiang C, Li G, Lu J. PKB/AKT and ERK regulation of caspase-mediated apoptosis by methylseleninic acid in LNCaP prostate cancer cells[J]. Carcinogenesis,2005;26(8):1374.
[136]Batist G. Selenium-induced cytotoxicity of human leukemia cells:interaction with reduced glutathione[J]. Cancer Research,1986;46(11):5482-5485.
[137]Spyrou G. Selenite and selenate inhibit human lymphocyte growth via different mechanisms[J]. Cancer Research,1996;56(19):4407-4412.
[138]Chen J, Goetchius MP, Combs Jr GF, Campbell TC. Effects of dietary selenium and vitamin E on covalent binding of aflatoxin to chick liver cell macromolecules[J]. Journal of Nutrition,1982;112(2):350.
[139]Fiala ES, Joseph C, Sohn OS, El-Bayoumy K, Reddy BS. Mechanism of benzylselenocyanate inhibition of azoxymethane-induced colon carcinogenesis in F344 rats[J]. Cancer Research,1991;51(11):2826-2830.
[140]Jacobs MM, Forst CF, Beams FA. Biochemical and clinical effects of selenium on dimethylhydrazine-induced colon cancer in rats[J]. Cancer Res,1981;41(11 Pt 1):4458-4465.
[141]Weindruch R, Walford RL, Fligiel S, Guthrie D. The retardation of aging in mice by dietary restriction:longevity, cancer, immunity and lifetime energy intake[J]. Journal of Nutrition,1986; 116(4):641.
[142]Darnell RB. Onconeural antigens and the paraneoplastic neurologic disorders:at the intersection of cancer, immunity, and the brain[J]. Proceedings of the National Academy of Sciences,1996;93(10):4529-4536.
[143]Koller LD, Exon JH, Talcott PA, Osborne CA, Henningsen GM. Immune responses in rats supplemented with selenium[J]. Clinical and Experimental Immunology,1986;63(3):570.
[144]Roy M, Kiremidjian-Schumacher L, Wishe HI, Cohen MW, Stotzky G. Supplementation with selenium and human immune cell functions[J]. Biological trace element research,1994;41(1):103-114.
[145]Kiremidjian-Schumacher L, Roy M, Wishe HI, Cohen MW, Stotzky G. Supplementation with selenium augments the functions of natural killer and lymphokine-activated killer cells[J]. Biological trace element research,1996;52(3):221-239.
[146]Taylor EW. Selenium and cellular immunity [J]. Biological trace element research,1995;49(2):85-95.
[147]Diamond AM, Hu YJ, Mansur DB. Glutathione peroxidase and viral replication:implications for viral evolution and chemoprevention[J].Biofactors,2001;14(1):205-210.
[148]Buettner C, Harney JW, Berry MJ. The Caenorhabditis elegans homologue of thioredoxin reductase contains a selenocysteine insertion sequence (SECIS) element that differs from mammalian SECIS elements but directs selenocysteine incorporation[J]. Journal of Biological Chemistry,1999;274(31):21598-21602.
[149]Nordberg J, Zhong L, Holmgren A, Arner ESJ. Mammalian thioredoxin reductase is irreversibly inhibited by dinitrohalobenzenes by alkylation of both the redox active selenocysteine and its neighboring cysteine residue[J]. Journal of Biological Chemistry,1998;273(18):10835-10842.
[150]Rederstorff M, Krol A, Lescure A. Understanding the importance of selenium and selenoproteins in muscle function[J]. Cellular and Molecular Life Sciences,2005;63:52-59.
[151]Zhang D, Okada S, Yu Y, Zheng P, Yamaguchi R, Kasai H. Vitamin E inhibits apoptosis, DNA modification, and cancer incidence induced by iron-mediated peroxidation in Wistar rat kidney[J]. Cancer Research,1997;57(12):2410-2414.
[152]Ray Q Batra S, Shukla NK, Deo S, Raina V, Ashok S, et al. Lipid peroxidation, free radical production and antioxidant status in breast cancer [J]. Breast cancer research and treatment,2000;59(2):163-170.
[153]Sangeetha P, Das UN, Koratkar R, Suryaprabha P. Increase in free radical generation and lipid peroxidation following chemotherapy in patients with cancer[J]. Free radical biology & medicine,1990;8(1):15.
[154]Cheng WH, Ho YS, Valentine BA, Ross DA, Combs GF, Lei XG. Cellular glutathione peroxidase is the mediator of body selenium to protect against paraquat lethality in transgenic mice[J]. Journal of Nutrition,1998; 128(7):1070-1076.
[155]Cheng WH, Ho YS, Ross DA, Han Y, Combs GF, Lei XG. Overexpression of cellular glutathione peroxidase does not affect expression of plasma glutathione peroxidase or phospholipid hydroperoxide glutathione peroxidase in mice offered diets adequate or deficient in selenium[J]. Journal of Nutrition,1997; 127(5):675-680.
[156]Fu Y, Cheng WH, Porres JM, Ross DA, Lei XG. Knockout of cellular glutathione peroxidase gene renders mice susceptible to diquat-induced oxidative stress[J]. Free Radical Biology and Medicine,1999;27(5-6):605-611.
[157]Ratnasinghe D, Tangrea JA, Andersen MR, Barrett MJ, Virtamo J, Taylor PR, et al. Glutathione Peroxidase Codon 198 Polymorphism Variant Increases Lung Cancer Risk 1. AACR,2000: 6381-6383.
[158]Zhou JC, Zhao H, Li JG, Xia XJ, Wang KN, Zhang YJ, et al. Selenoprotein Gene Expression in Thyroid and Pituitary of Young Pigs Is Not Affected by Dietary Selenium Deficiency or Excess[J]. Journal ofNutrition,2009;139(6):1061.
[159]Kumaraswamy E, Malykh A, Korotkov KV, Kozyavkin S, Hu Y, Kwon SY, et al. Structure-expression relationships of the 15-kDa selenoprotein gene Possible role of the protein in cancer etiology[J]. Journal of Biological Chemistry,2000;275(45):35540-35547.
[160]Chen C, Zhao J, Zhang P, Chai Z. Speciation and subcellular location of Se-containing proteins in human liver studied by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and hydride generation-atomic fluorescence spectrometric detection[J]. Analytical and bioanalytical chemistry,2002;372(3):426-430.
[161]Apostolou S, Klein JO, Mitsuuchi Y, Shetler JN, Poulikakos PI, Jhanwar SC, et al. Growth inhibition and induction of apoptosis in mesothelioma cells by selenium and dependence on selenoprotein SEP15 genotype[J]. Oncogene,2004;23(29):5032-5040.
[162]Diwadkar-Navsariwala V, Diamond AM. The Link between Selenium and Chemoprevention: A Case for Selenoproteins 1[J]. Journal of Nutrition,2004;134(11):2899-2902.
[163]Tsuji PA, Irons R, Ouyang P, Carlson B, Yoo MH, XMX, et al. Sep15 knockdown decreases tumorigenicity and metastasis of a colon cancer cell line[J]. FASEB J,2009;23:338.337.
[164]Mustacich D, Powis G. Thioredoxin reductase[J]. Biochemical Journal,2000;346(Pt 1):1.
[165]Gallegos A, Berggren M, Gasdaska JR, Powis G. Mechanisms of the regulation of thioredoxin reductase activity in cancer cells by the chemopreventive agent selenium[J]. Cancer Research,1997;57(21):4965-4970.
[166]Berggren MM, Mangin JF, Gasdaska JR, Powis G Effect of selenium on rat thioredoxin reductase activity Increase by supranutritional selenium and decrease by selenium deficiency[J]. Biochemical pharmacology,1999;57(2):187-193.
[167]Allred DC, Clark GM, Elledge R, Fuqua SAW, Brown RW, Chamness GC, et al. Association of p53 protein expression with tumor cell proliferation rate and clinical outcome in node-negative breast cancer[J]. JNCI Cancer Spectrum,1993,85(3):200.
[168]Levine AJ, Finlay CA, Hinds PW. P53 is a tumor suppressor gene[J]. Cell,2004; 116:67-70.
[169]Cuddihy AR, Wong AH, Tam NW, Li S, Koromilas AE. The double-stranded RNA activated protein kinase PKR physically associates with the tumor suppressor p53 protein and phosphorylates human p53 on serine 392 in vitro[J]. Oncogene,1999;18(17):2690.
[170]Harris MP, Sutjipto S, Wills KN, Hancock W, Cornell D, Johnson DE, et al. Adenovirus-mediated p53 gene transfer inhibits growth of human tumor cells expressing mutant p53 protein[J]. Cancer gene therapy;3(2):121.
[171]Louis DN, Von Deimling A, Chung RY, Rubio MP, Whaley JM, Eibl RH, et al. Comparative study of p53 gene and protein alterations in human astrocytic tumors[J]. Journal of neuropathology and experimental neurology,1993;52(1):31.
[172]Pearson GD, Merrill GF. Deletion of the Saccharomyces cerevisiae TRR1 gene encoding thioredoxin reductase inhibits p53-dependent reporter gene expression. ASBMB,1998:5431-5434.
[173]Ueno M, Masutani H, Arai RJ, Yamauchi A, Hirota K, Sakai T, et al. Thioredoxin-dependent redox regulation of p53-mediated p21 activation[J]. Journal of Biological Chemistry,1999;274(50):35809-35815.
[174]Shibata T, Yamada T, Ishii T, Kumazawa S, Nakamura H, Masutani H, et al. Thioredoxin as a Molecular Target of Cyclopentenone Prostaglandins*[J]. Journal of Biological Chemistry.2003;278(28):26046-26054.
[175]Patterson BH, Levander OA, Helzlsouer K, McAdam PA, Lewis SA, Taylor PR, et al. Human selenite metabolism:a kinetic model[J]. American Journal of Physiology-Regulatory, Integrative and Comparative Physiology,1989;257(3):556-561.
[176]Behne D, Kyriakopoulos A. M AMMALIAN S ELENIUM-C ONTAINING P ROTEINS[J]. Annual review ofnutrition,2001;21 (1):453-473.
[177]Weiss SL, Sunde RA. Selenium regulation of classical glutathione peroxidase expression requires the 3'untranslated region in Chinese hamster ovary cells[J]. Journal of Nutrition,1997;127(7):1304-1310.
[178]Simon HU, Haj-Yehia A, Levi-Schaffer F. Role of reactive oxygen species (ROS) in apoptosis induction[J]. Apoptosis,2000;5(5):415-418.
[179]Zhao R, Xiang N, Domann FE, Zhong W. Expression of p53 enhances selenite-induced superoxide production and apoptosis in human prostate cancer cells[J]. Cancer Research,2006;66(4):2296-2304.
[180]Ip C. Activity of methylated forms of selenium in cancer prevention[J]. Cancer Research,1990;50(4):1206-1211.
[181]Cai XJ, Block E, Uden PC, Zhang X, Quimby BD, Sullivan JJ. Allium chemistry: identification of selenoamino acids in ordinary and selenium-enriched garlic, onion, and broccoli using gas chromatography with atomic emission detection[J]. Journal of agricultural and food chemistry(Print);,1995;43(7):1754-1757.
[182]Lu J, Pei H, Ip C, Lisk DJ, Ganther H, Thompson HJ. Effect of an aqueous extract of selenium-enriched garlic on in vitro markers and in vivo efficacy in cancer prevention[J]. Carcinogenesis,1996; 17(9):1903.
[183]Nagata S. Apoptosis by death factor[J]. Cell,1997;88(3):355-366.
[184]Chawla-Sarkar M, Lindner DJ, Liu YF, Williams BR, Sen GC, Silverman RH, et al. Apoptosis and interferons:role of interferon-stimulated genes as mediators of apoptosis[J]. Apoptosis,2003;8(3):237-249.
[185]Shapiro AB, Ling V. Extraction of Hoechst 33342 from the cytoplasmic leaflet of the plasma membrane by P-glycoprotein[J].European Journal of Biochemistry,1997;250(1):122-129.
[186]Vermes I, Haanen C, Steffens-Nakken H, Reutellingsperger C. A novel assay for apoptosis flow cytometric detection of phosphatidylserine expression on early apoptotic cells using fluorescein labelled annexin V[J]. Journal of immunological methods,1995;184(1):39-51.
[187]周明眉,杨奎.中药血清药理学的方法学研究—反应体系中含药血清加入量的研究[J].中药药理与临床,1998；14(006)：43-44.
[188]李仪奎,吴健宇.小青龙汤药物血清对致敏大鼠腹腔肥大细胞脱颗粒以及组…[J].中国中西医结合杂志，2000；20(001)：40-42.
[189]Bochu W, Liancai Z, Qi C. Primary study on the application of Serum Pharmacology in Chinese traditional medicine[J]. Colloids and Surfaces B:Biointerfaces,2005;43(3-4):194-197.
[190]张丽英.饲料分析及饲料质量检测技术.北京：中国农业大学出版社,2003.
[191]Li Z, Guo Y. Simultaneous determination of trace arsenic, antimony, bismuth and selenium in biological samples by hydride generation-four-channel atomic fluorescence spectrometry[J]. Talanta,2005;65(5):1318-1325.
[192]Lawrence RA, Sunde RA, Schwartz GL, Hoekstra WG Glutathione peroxidase activity in rat lens and other tissues in relation to dietary selenium intake[J]. Experimental eye research,1974;18(6):563.
[193]Massey V, Williams CH. On the reaction mechanism of yeast glutathione reductase[J]. Journal of Biological Chemistry,1965;240(11):4470-4480.
[194]Storch J, Ferber E. Detergent-amplified chemiluminescence of lucigenin for determination of superoxide anion production by NADPH oxidase and xanthine oxidase[J]. Analytical biochemistry,1988; 169(2):262-267.
[195]Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding[J]. Analytical biochemistry,1976;72(1-2):248-254.
[196]Wilkinson JE, Bell MC, Bacon JA, Masincupp FB. Effects of Supplemental Selenium on Swine I. Gestation and Lactation[J]. Journal ofAnimal Science,1977;44(2):224.
[197]Spears JW. Trace Mineral Bioavailability in Ruminants 1. Am Soc Nutrition,2003: 1506-1509.
[198]Swanson CA, Patterson BH, Levander OA, Veillon C, Taylor PR, Helzlsouer K, et al. Human [74Se] selenomethionine metabolism:a kinetic model. Am Soc Nutrition,1991:917-926.
[199]Bunk MJ, Combs Jr GF. Effect of selenium on appetite in the selenium-deficient chick[J]. Journal of nutrition,1980; 110(4):743.
[200]Lawler TL, Taylor JB, Finley JW, Caton JS. Effect of supranutritional and organically bound selenium on performance, carcass characteristics, and selenium distribution in finishing beef steers 1. Am Soc Animal Sci,2004:1488-1493.
[201]Soto-Navarro SA, Lawler TL, Taylor JB, Reynolds LP, Reed JJ, Finley JW, et al. Effect of high-selenium wheat on visceral organ mass, and intestinal cellularity and vascularity in finishing beef steers 1. Am Soc Animal Sci,2004:1788-1793.
[202]Zhu JH, Zhang X, McClung JP, Lei XG Impact of Cu, Zn-superoxide dismutase and Se-dependent glutathione peroxidase-1 knockouts on acetaminophen-induced cell death and related signaling in murine liver[J]. Experimental Biology and Medicine,2006;231(11):1726.
[203]Hussain SP, Amstad P, He P, Robles A, Lupold S, Kaneko I, et al. p53-induced up-regulation of MnSOD and GPx but not catalase increases oxidative stress and apoptosis. AACR,2004: 2350-2356.
[204]Spallholz JE. On the nature of selenium toxicity and carcinostatic activity[J]. Free Radical Biology and Medicine,1994;17(1):45-64.
[205]Christensen MJ, Nelson BL, Wray CD. Regulation of glutathione S-transferase gene expression and activity by dietary selenium[J]. Biochemical and biophysical research communications,1994;202(1):271-277.
[206]Lei XG, Ross DA, Roneker KR. Comparison of age-related differences in expression of phospholipid hydroperoxide gltutathione peroxidase mRNA and activity in various tissues of pigs[J]. Comp Biochem Physiol,1997;117:109-114.
[207]Dickinson DA, Forman HJ. Cellular glutathione and thiols metabolism[J]. Biochemical pharmacology,2002;64(5-6):1019-1026.
[208]Jenkinson SG, Lawrence RA, Burk RF, Gregory PE. Non-selenium-dependent glutathione peroxidase activity in rat lung:Association with lung glutathione S-transferase activity and the effects of hyperoxia[J]. Toxicology and Applied Pharmacology,1983;68(3):399-404.
[209]Liu Q, Lauridsem E, Clausen J. The major selenium-containing protein in human peripheral granulocytes[J]. Biological trace element research,1999;68(3):193-207.
[210]Goodman GE, Schaffer S, Bankson DD, Hughes MP, Omenn GS. Predictors of Serum Selenium in Cigarette Smokers and the Lack of Association with Lung and Prostate Cancer Risk 1[J]. Cancer Epidemiology Biomarkers & Prevention,2001; 10(10):1069-1076.
[211]Ip C. Differential Effect of Dietary Methionine on the Biopotency of Selenomethionine and Selenite in Cancer Chemopreventionl[J]. JNCI Cancer Spectrum,1988;80(4):258.
[212]Schmidt HHH, Pollock JS, Nakane M, Gorsky LD, Forstermann U, Murad F. Purification of a soluble isoform of guanylyl cyclase-activating factor synthase[J]. Proceedings of the National Academy of Sciences,1991;88(2):365-369.
[213]Casiot C, Szpunar J, obinski R, Potin-Gautier M. Sample preparation and HPLC separation approaches to speciation analysis of selenium in yeast by ICP-MS[J]. Journal of Analytical Atomic Spectrometry,1999;14(4):645-650.
[214]Infante HG, O'Connor G, Rayman M, Wahlen R, Entwisle J, Norris P, et al. Selenium speciation analysis of selenium-enriched supplements by HPLC with ultrasonic nebulisation ICP-MS and electrospray MS/MS detection[J]. Journal of Analytical Atomic Spectrometry,2004;19(12):1529-1538.
[215]Burk RF, Hill KE. Regulation of selenoproteins[J]. Annual review of nutrition,1993;13(1):65-81.
[216]Patterson JK, Lei XG, Miller DD. The Pig as an Experimental Model for Elucidating the Mechanisms Governing Dietary Influence on Mineral Absorption[J]. Experimental Biology and Medicine,2008;233(6):651.
[217]Clark LC, Combs GF, Turnbull BW, Slate EH, Chalker DK, Chow J, et al. Effects of selenium supplementation for cancer prevention in patients with carcinoma of the skin. A randomized controlled trial. Nutritional Prevention of Cancer Study Group[J].Jama,1996;276(24):1957-1963.
[218]薛庆善.体外培养的原理与技术.北京：科学出版社,2001.
[219]Yangxin FU, Porres JM, Lei XG Comparative impacts of glutathione peroxidase-1 gene knockout on oxidative stress induced by reactive oxygen and nitrogen species in mouse hepatocytes[J]. Biochem J,2001;359:687-695.
[220]Babson AL, Babson SR. Kinetic colorimetric measurement of serum lactate dehydrogenase activity[J]. Clinical Chemistry,1973; 19(7):766-769.
[221]Olie R, Simoes-Wust A, Baumann B, Leech S, Fabbro D, Stahel R, et al. A novel antisense oligonucleotide targeting survivin expression induces apoptosis and sensitizes lung cancer cells to chemotherapy[J]. Cancer Research,2000;60(11):2805.
[222]Gavrieli Y, Sherman Y, Ben-Sasson SA. Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation[J]. Journal of cell Biology,1992;119(3):493-501.
[223]Jiang C, Hu H, Malewicz B, Wang Z, Lu J. Selenite-induced p53 Ser-15 phosphorylation and caspase-mediated apoptosis in LNCaP human prostate cancer cells[J]. Molecular Cancer Therapeutics,2004;3(7):877-884.
[224]Wang. BC, Zhu. LC, Chen. Q. Primary study on the application of Serum Pharmacology in Chinese traditional medicine[J]. Colloids and Surfaces B:Biointerfaces,2005;43(3-4):194-197.
[225]江岩,黄承钰,陶德明,于静.盐亭食管癌高发区饮食对人食管癌细胞增殖的影响[J].卫生研究,2007；36(002)：226-230.
[226]刘建文.抗肿瘤作用的实验方法[J].药理实.验方法学一新技术与新方法，2003：21—75.
[227]Freshney RI. Culture of epithelial cells[M]:Wiley-Liss,2002.
[228]Whanger PD, Muth OH, Oldfield JE, Weswig PH. Influence of sulfur on incidence of white muscle disease in lambs[J]. Journal ofNutrition,1969;97(4):553.
[229]Ip C, White G Mammary cancer chemoprevention by inorganic and organic selenium:single agent treatment or in combination with vitamin E and their effects on in vitro immune functions[J]. Carcinogenesis,1987;8(12):1763.
[2]Derumeaux H, Valeix P, Castetbon K, Bensimon M, Boutron-Ruault MC, Arnaud J, et al. Association of selenium with thyroid volume and echostructure in 35-to 60-year-old French adults[J]. European Journal of Endocrinology,2003; 148(3):309-315.
[3]Camins A, Jimenez A, Sureda FX, Pallas M, Escubedo E, Camarasa J. Characterization of nitrobenzylthioinosine binding sites in the mitochondrial fraction of rat testis[J]. Life Sciences,1996;58(9):753-759.
[4]Schwarz K, Foltz CM. Selenium as an integral part of factor 3 against dietary necrotic liver degeneration[J]. J Am Chem Soc,1957;79(12):3292-3293.
[5]Brown MR, Cohen HJ, Lyons JM, Curtis TW, Thunberg B, Cochran WJ, et al. Proximal muscle weakness and selenium deficiency associated with long term parenteral nutrition[J]. The American journal of clinical nutrition,1986;43(4):549.
[6]Rayman MP. The importance of selenium to human health[J]. The Lancet,2000;356(9225):233-241.
[7]Sheridan PA, Zhong N, Carlson BA, Perella CM, Hatfield DL, Beck MA. Decreased selenoprotein expression alters the immune response during influenza virus infection in mice[J]. Journal of Nutrition,2007,131(6):1466.
[8]Shamberger RJ, Tytko SA, Willis CE. Antioxidants and cancer. Part VI. Selenium and age-adjusted human cancer mortality[J]. Archives of environmental health,1977;31 (5):231.
[9]Clark LC, Cantor KP, Allaway WH. Selenium in forage crops and cancer mortality in US counties[J]. Archives of environmental health (USA),1991.
[10]Ip C, Dong Y, Ganther HE. New concepts in selenium chemoprevention[J]. Cancer and Metastasis Reviews,2002;21 (3):281-289.
[11]Rubins HB, Robins SJ, Collins D, Fye CL, Anderson JW, Elam MB, et al. Gemfibrozil for the secondary prevention of coronary heart disease in men with low levels of high-density lipoprotein cholesterol.1999:410-418.
[12]Finley JW. Bioavailability of selenium from foods[J]. Nutrition reviews,2006;64(3):146-151.
[13]Clark LC, F.Combs G, B. W.Urnbull, H.Slate E, Chalker DK, Chow J, et al. Effect of selenium supplementation for cancer prevention in patients with carcinoma of the skin:a randomized controlled trial.[J]. J Am Med Assoc,1996;276:1957-1963.
[14]Combs GF. Current Evidence and Research Needs to Support a Health Claim for Selenium and Cancer Prevention 1. Am Soc Nutrition,2005:343-347.
[15]Yu Yu S, Zhu YJ, Li WG. Protective role of selenium against hepatitis B virus and primary liver cancer in Qidong[J]. Biological trace element research,1997;56(1):117-124.
[16]Taylor PR, Li B, Dawsey SM, Li JY, Yang CS, Guo W, et al. Prevention of esophageal cancer:the nutrition intervention trials in Linxian, China[J]. Cancer Research,1994;54(7 Supplement):2029s.
[17]Blot WJ, Li JY, Taylor PR, Guo W, Dawsey SM, Li B. The Linxian trials:mortality rates by vitamin-mineral intervention group. Am Soc Nutrition,1995:1424-1426.
[18]Thompson HJ. Effect of an inorganic and organic form of dietary selenium on the promotional stage of mammary carcinogenesis in the rat[J]. Cancer Research,1984;44(7):2803-2806.
[19]Birt DF. Inhibition by dietary selenium of colon cancer induced in the rat by bis (2-oxopropyl) nitrosamine[J]. Cancer Research,1982;42(11):4455-4459.
[20]Li DH, Graef GL, Yee JA, Yan L. Dietary supplementation with high-selenium soy protein reduces pulmonary metastasis of melanoma cells in mice[J]. Journal of Nutrition,2004; 134(6):1536-1540.
[21]Jiang C, Wang Z, Ganther H, Lu J. Caspases as Key Executors of Methyl Selenium-induced Apoptosis (Anoikis) of DU-145 Prostate Cancer Cells 1. AACR,2001:3062-3070.
[22]Jiang C, Wang Z, Ganther H, Lu J. Distinct effects of methylseleninic acid versus selenite on apoptosis, cell cycle, and protein kinase pathways in DU145 human prostate cancer cells[J]. Molecular cancer therapeutics,2002;1(12):1059.
[23]Ip C. Lessons from basic research in selenium and cancer prevention[J]. Journal of Nutrition,1998;128(11):1845-1854.
[24]Beilstein MA, Whanger PD. Effect of Vitamin B-6 Deficiency on Selenium Metabolism in the rat.[J]. J Nutr,1989; 119:1962-1972.
[25]Behne D, Kyriakopoulos A, Scheid S, Gessner H. Effects of chemical form and dosage on the incorporation of selenium into tissue proteins in rats[J]. Journal of Nutrition,1991;121(6):806.
[26]Kim YY, Mahan DC. Comparative effects of high dietary levels of organic and inorganic selenium on selenium toxicity of growing-finishing pigs[J]. Journal of animal science,2001;79(4):942.
[27]Mahan DC, Cline TR, Richert B. Effects of dietary levels of selenium-enriched yeast and sodium selenite as selenium sources fed to growing-finishing pigs on performance, tissue selenium, serum glutathione peroxidase activity, carcass characteristics, and loin quality[J]. Journal of Animal Science,1999;77(8):2172-2179.
[28]Mahan DC, Parrett NA. Evaluating the efficacy of selenium-enriched yeast and sodium selenite on tissue selenium retention and serum glutathione peroxidase activity in grower and finisher swine[J]. Journal of Animal Science,1996;74(12):2967-2974.
[29]Thompson HJ. Effect of short-term feeding of sodium selenite on 7,12-dimethylbenz (a) anthracene-induced mammary carcinogenesis in the rat[J]. Cancer Research,1982;42(12):4954-4958.
[30]Arthur JR, McKenzie RC, Beckett GJ. Selenium in the immune system[J]. The Journal of nutrition,2003;133(5):1457-1459.
[31]Lu J, Jiang C. Antiangiogenic activity of selenium in cancer chemoprevention:metabolite-specific effects[J]. Nutrition and cancer,2001;40(1):64-73.
[32]Hu YJ, Diamond AM. Role of Glutathione Peroxidase 1 in Breast Cancer Loss of Heterozygosity and Allelic Differences in the Response to Selenium 1. AACR,2003:3347-3351.
[33]Hardie LJ, Briggs JA, Davidson LA, Allan JM, King R, Williams GI, et al. The effect of hOGGl and glutathione peroxidase I genotypes and 3p chromosomal loss on 8-hydroxydeoxyguanosine levels in lung cancer[J]. Carcinogenesis,2000;21(2):167.
[34]Sinha R, El-Bayoumy K. Apoptosis is a critical cellular event in cancer chemoprevention and chemotherapy by selenium compounds[J]. Current Cancer Drug Targets,2004;4:13-28.
[35]Spallholz JE, Hoffman DJ. Selenium toxicity:cause and effects in aquatic birds[J]. Aquatic Toxicology,2002;57(1-2):27-37.
[36]Zhong W, Oberley TD. Redox-mediated Effects of Selenium on Apoptosis and Cell Cycle in the LNCaP Human Prostate Cancer Cell Line 1. AACR,2001:7071-7078.
[37]Whanger PD. Selenium and its relationship to cancer:an update[J]. British journal of nutrition,2004;91(01):11-28.
[38]Ganther HE. Selenium metabolism, selenoproteins and mechanisms of cancer prevention: complexities with thioredoxin reductase[J]. Carcinogenesis,1999;20(9):1657.
[39]Miller ER, Ullrey DE. The pig as a model for human nutrition[J]. Annual review of nutrition,1987;7(1):361-382.
[40]Baquerizo A, Mhoyan A, Kearns-Jonker M, Arnaout WS, Shackleton C, Busuttil RW, et al. Characterization of human xenoreactive antibodies in liver failure patients exposed to pig hepatocytes after bioartificial liver treatment:an ex vivo model of pig to human xenotransplantation[J]. Transplantation,1999;67(1):5.
[41]Van Rij AM, Thomson CD, McKenzie JM, Robinson MF. Selenium deficiency in total parenteral nutrition[J]. American Journal of Clinical Nutrition,1979;32(10):2076.
[42]Yang F, Lin Z, Xing JRU, Li SG, Yang J, San SUN, et al. Se deficiency is a necessary but not sufficient factor required for the pathogenesis of Keshan disease[J]. Journal of clinical biochemistry and nutrition,1994;16(2):101-110.
[43]Moreno-Reyes R, Suetens C, Mathieu F, Begaux F, Zhu D, Rivera MT, et al. Kashin-Beck osteoarthropathy in rural Tibet in relation to selenium and iodine status.1998:1112-1120.
[44]Yang C, Niu C, Bodo M, Gabriel E, Notbohm H, Wolf E, et al. Fulvic acid supplementation and selenium deficiency disturb the structural integrity of mouse skeletal tissue. An animal model to study the molecular defects of Kashin-Beck disease[J]. Biochemical Journal,1993;289(Pt 3):829.
[45]Medina D, Oborn CJ. Selenium Inhibition of DNA Synthesis in Mouse Mammary Epithelial Cell Line YN-4[J]. Cancer Research,1984;44:4361-4365.
[46]Zeng H. Selenite and Selenomethionine Promote HL-60 Cell Cycle Progression 1 2[J]. Journal of Nutrition,2002;132(4):674-679.
[47]Thomson CD. Assessment of requirements for selenium and adequacy of selenium status:a review[J]. European journal of clinical nutrition,2004;58(3):391-402.
[48]Behne D, Pfeifer H, Rothlein D, Kyriakopoulos A. Cellular and subcellular distribution of selenium and selenium-containing proteins in the rat[J]. Trace elements in man and animals,2000;10:29-34.
[49]Behne D, Hofer-Bosse T. Effects of a low selenium status on the distribution and retention of selenium in the rat[J]. Journal of Nutrition,1984;114(7):1289.
[50]Whanger PD. Selenocompounds in plants and animals and their biological significance[J]. Journal of the American College ofNutrition,2002;21(3):223-232.
[51]Terry N, Zayed AM, De Souza MP, Tarun AS. S ELENIUM IN H IGHER P LANTS[J]. Annual Review of Plant Biology,2000;51(1):401-432.
[52]Finley JW. Selenium accumulation in plant foods[J]. Nutrition reviews,2005;63(6):196-202.
[53]Cases J, Vacchina V, Napolitano A, Caporiccio B, Besancon P, Lobinski R, et al. Selenium from Selenium-Rich Spirulina Is Less Bioavailable than Selenium from Sodium Selenite and Selenomethionine in Selenium-Deficient Rats 1[J]. Journal of nutrition,2001;131(9):2343-2350.
[54]Cai XJ, Block E, Uden PC, Zhang X, Quimby BD, Sullivan JJ. Allium chemistry:identification of selenoamino acids in ordinary and selenium-enriched garlic, onion, and broccoli using gas chromatography with atomic emission detection[J]. Journal of Agricultural and Food Chemistry,1995;43(7):1754-1757.
[55]Bell JG, Cowey CB. Digestibility and bioavailability of dietary selenium from fishmeal, selenite, selenomethionine and selenocystine in Atlantic salmon (Salmo salar)[J]. Aquaculture (Netherlands),1989.
[56]Ip C, Birringer M, Block E, Kotrebai M, Tyson JF, Uden PC, et al. Chemical speciation influences comparative activity of selenium-enriched garlic and yeast in mammary cancer prevention[J]. J Agric Food Chem,2000;48(6):2062-2070.
[57]Bird SM, Uden PC, Tyson JF, Block E, Denoyer E. Speciation of selenoamino acids and organoselenium compounds in selenium-enriched yeast using high-performance liquid chromatography-inductively coupled plasma mass spectrometry[J]. Journal of Analytical Atomic Spectrometry,1997;12(7):785-788.
[58]Chassaigne H, Chery CC, Bordin G, Rodriguez AR. Development of new analytical methods for selenium speciation in selenium-enriched yeast material[J]. Journal of Chromatography A,2002;976(1-2):409-422.
[59]Rayman MP. Selenium in cancer prevention:a review of the evidence and mechanism of action[J]. Proceedings of the Nutrition Society,2005;64(04):527-542.
[60]Hawkes WC, Alkan FZ, Oehler L, Usda ARS. Absorption, distribution and excretion of selenium from beef and rice in healthy North American men[J].2003.
[61]Xu XM, Carlson BA, Irons R, Mix H, Zhong N, Gladyshev VN, et al. Selenophosphate synthetase 2 is essential for selenoprotein biosynthesis[J]. Biochemical Journal,2007;404(Pt 1):115.
[62]Kobayashi Y, Ogra Y, Ishiwata K, Takayama H, Aimi N, Suzuki KT. Selenosugars are key and urinary metabolites for selenium excretion within the required to low-toxic range[J]. Proceedings of the National Academy of Sciences,2002;99(25):15932-15936.
[63]Schrauzer GN. Selenomethionine:A Review of Its Nutritional Significance, Metabolism and Toxicity 1[J]. Journal of nutrition,2000;130(7):1653-1656.
[64]Olson OE, Palmer IS. Selenoamino acids in tissues of rats administered inorganic selenium[J]. Metabolism:clinical and experimental,1976;25(3):299.
[65]Beilstein MA, Whanger PD. Glutathione peroxidase activity and chemical forms of selenium in tissues of rats given selenite or selenomethionine[J]. J Inorg Biochem,1988;33(1).
[66]Sunde RA, Gutzke GE, Hoekstra WG Effect of dietary methionine on the biopotency of selenite and selenomethionine in the rat[J]. Journal of Nutrition,1981;111(1):76.
[67]Luo XM, Wei HJ, Yang CL, Xing J, Liu X, Qiao CH, et al. Bioavailability of selenium to residents in a low-selenium area of China[J]. American Journal of Clinical Nutrition,1985;42(3):439.
[68]Vendeland SC, Deagen JT, Butler JA, Whanger PD. Uptake of selenite, selenomethionine and selenate by brush border membrane vesicles isolated from rat small intestine[J]. Biometals,1994;7(4):305-312.
[69]Deagen JT, Butler JA, Beilstein MA, Whanger PD. Effects of dietary selenite, selenocystine and selenomethionine on selenocysteine lyase and glutathione peroxidase activities and on selenium levels in rat tissues[J]. Journal of Nutrition,1987;117(1):91.
[70]Waschulewski IH, Sunde RA. Effect of dietary methionine on utilization of tissue selenium from dietary selenomethionine for glutathione peroxidase in the rat[J]. Journal ofNutrition,1988;118(3):367.
[71]Whanger PD, Butler JA, Xia Y. Can physiological responses be used to assess the chemical forms of selenium consumed by humans.1994; 1994. p.8-10.
[72]McGuire MK, Burgert SL, Milner JA, Glass L, Kummer R, Deering R, et al. Selenium status of lactating women is affected by the form of selenium consumed[J]. American Journal of Clinical Nutrition,1993;58:649-649.
[73]Alaejos MS, Romero CD. Selenium in human lactation[J]. Nutrition Reviews,1995;53(6):159-166.
[74]Boles JO, Cisneros RJ, Weir MS, Odom JD, Villafranca JE, Dunlap RB. Purification and characterization of selenomethionyl thymidylate synthase from Escherichia coli:comparison with the wild-type enzyme[J]. Biochemistry,1991;30(46):11073-11080.
[75]Huber RE, Criddle RS. The isolation and properties of beta-galactosidase from Escherichia coli grown on sodium selenate[J]. Biochim Biophys Acta,1967;141(3):587-599.
[76]Soda K, Oikawa T, Esaki N. Vitamin B6 enzymes participating in selenium amino acid metabolism[J]. Biofactors,1999; 10.
[77]Kajander EO, Harvima RJ, Eloranta TO, Martikainen H, Kantola M, K renlampi SO, et al. Metabolism, cellular actions, and cytotoxicity of selenomethionine in cultured cells[J]. Biological trace element research,1991;28(1):57-68.
[78]Rotruck JT, Pope AL, Ganther HE, Swanson AB, Hafeman DG, Hoekstra WG. Selenium: biochemical role as a component of glutathione peroxidase[J]. Science,1973;179(4073):588.
[79]Letavayovi L, Vl?kova V, Brozmanova J. Selenium:from cancer prevention to DNA damage[J]. Toxicology,2006;227(1-2):1-14.
[80]Spallholz JE, Palace VP, Reid TW. Methioninase and selenomethionine but not Se-methylselenocysteine generate methylselenol and superoxide in an in vitro chemiluminescent assay: implications for the nutritional carcinostatic activity of selenoamino acids [J]. Biochemical pharmacology,2004;67(3):547-554.
[81]Shen HM, Yang CF, ONG CN. Sodium selenite-induced oxidative stress and apoptosis in human hepatoma HepG2 cells[J]. Int J Cancer,1999;81:820-828.
[82]Shen HM, Yang CF, Liu J, Ong CN. Dual role of glutathione in selenite-induced oxidative stress and apoptosis in human hepatoma cells[J]. Free Radical Biology and Medicine,2000;28(7):1115-1124.
[83]Kim Y, Jhon D, Lee KY. Involvement of ROS and JNK1 in selenite-induced apoptosis in Chang liver cells[J]. Experimental and molecular medicine,2004;36(2):151-164.
[84]Spallholz JE. Free radical generation by selenium compounds and their prooxidant toxicity[J]. Biomedical and environmental sciences:BES,1997;10(2-3):260.
[85]Drake EN. Cancer chemoprevention:selenium as a prooxidant, not an antioxidant[J]. Medical hypotheses,2006;61(2):318-322.
[86]Miki K, Xu M, Gupta A, Ba Y, Tan Y, Al-Refaie W, et al. Methioninase Cancer Gene Therapy with Selenomethionine as Suicide Prodrug Substrate 1[J]. Cancer Research,2001;61(18):6805-6810.
[87]Zeng H, Briske-Anderson M, Idso JP, Hunt CD. The selenium metabolite methylselenol inhibits the migration and invasion potential of HT1080 tumor cells[J]. Journal of Nutrition,2006; 136(6):1528-1532.
[88]Inoue H, Inagaki K, Sugimoto M, Esaki N, Soda K, Tanaka H. Structural analysis of the L-methionine{gamma}-lyase gene from Pseudomonas putida[J]. Journal of Biochemistry,1995; 117(5):1120.
[89]Smith LL. Mechanism of paraquat toxicity in lung and its relevance to treatment[J]. Human & Experimental Toxicology,1987,6(1):31.
[90]Cheng WH, Fu YX, Porres JM, Ross DA, Lei XG. Selenium-dependent cellular glutathione peroxidase protects mice against a pro-oxidant-induced oxidation of NADPH, NADH, lipids, and protein. FASEB,1999:1467-1475.
[91]Bray RC, Cockle SA, Fielden EM, Roberts PB, Rotilio G, Calabrese L. Reduction and inactivation of superoxide dismutase by hydrogen peroxide[J]. Biochemical Journal,1974;139(1):43.
[92]de Haan JB, Bladier C, Griffiths P, Kelner M, O'Shea RD, Cheung NS, et al. Mice with a homozygous null mutation for the most abundant glutathione peroxidase, Gpxl, show increased susceptibility to the oxidative stress-inducing agents paraquat and hydrogen peroxide[J]. Journal of Biological Chemistry,1998;273(35):22528-22536.
[93]Sugiyama M, Ando A, Ogura R. Effect of vitamin E on survival, glutathione reductase and formation of chromium (V) in Chinese hamster V-79 cells treated with sodium chromate (VI)[J]. Carcinogenesis,1989; 10(4):737.
[94]Sies H. Glutathione and its role in cellular functions[J]. Free Radical Biology and Medicine,1999;27(9-10):916-921.
[95]Burk RF, Nishiki K, Lawrence RA, Chance B. Peroxide removal by selenium-dependent and selenium-independent glutathione peroxidases in hemoglobin-free perfused rat liver[J]. Journal of Biological Chemistry,1978;253(1):43-46.
[96]Reiter R, Wendel A. Selenium and drug metabolism--II. Independence of glutathione peroxidase and reversibility of hepatic enzyme modulations in deficient mice[J]. Biochemical pharmacology,1984;33(12):1923.
[97]Cheng WH, Ho YS, Ross DA, Valentine BA, Combs GF, Lei XG. Cellular glutathione peroxidase knockout mice express normal levels of selenium-dependent plasma and phospholipid hydroperoxide glutathione peroxidases in various tissues[J]. Journal of nutrition,1997;127(8):1445-1450.
[98]Lei XG, Dann HM, Ross DA, Cheng WH, Combs GF, Roneker KR. Dietary selenium supplementation is required to support full expression of three selenium-dependent glutathione peroxidases in various tissues of weanling pigs[J]. Journal of Nutrition,1998;128(1):130-135.
[99]Butler JA, Whanger PD, Kaneps AJ, Patton NM. Metabolism of selenite and selenomethionine in the rhesus monkey[J]. Journal ofNutrition,1990;120(7):751.
[100]Payne RL, Southern LL. Comparison of inorganic and organic selenium sources for broilers[J]. Poultry Science,2005;84(6):898.
[101]Gladyshev VN, Stadtman TC, Hatfield DL, Jeang KT. Levels of major selenoproteins in T cells decrease during HIV infection and low molecular mass selenium compounds increase. National Acad Sciences,1999:835-839.
[102]Irons R, Carlson BA, Hatfield DL, Davis CD. Both Selenoproteins and Low Molecular Weight Selenocompounds Reduce Colon Cancer Risk in Mice with Genetically Impaired Selenoprotein Expression 1 [J]. Journal ofNutrition,2006,136(5):1311-1317.
[103]Combs Jr GF, Clark LC, Turnbull BW. An analysis of cancer prevention by selenium[J]. Biofactors,2001;14(1):153-159.
[104]Behne D, Kyriakopoulos A. Mammalian Selenium-containing proteins[J]. Annu Rev Nutr,2001;21:453-473.
[105]Kryukov GV, Castellano S, Novoselov SV, Lobanov AV, Zehtab O, Guigo R, et al. Characterization of mammalian selenoproteomes[J]. Science,2003;300(5624):1439-1443.
[106]Bansal MP, Cook RG, Danielson KG, Medina D. A 14-kilodalton selenium-binding protein in mouse liver is fatty acid-binding protein[J]. Journal of Biological Chemistry,1989;264(23):13780-13784.
[107]Pumford NR, Martin BM, Hinson JA. A metabolite of acetaminophen covalently binds to the 56 kDa selenium binding protein[J]. Biochemical and biophysical research communications,1992; 182(3):1348.
[108]Li ZY, Guo SY, Li L. Bioeffects of selenite on the growth of Spirulina platensis and its biotransformation[J]. Bioresource technology,2003;89(2):171-176.
[109]Lunde G. Location of lipid-soluble selenium in marine fish to the lipoproteins[J]. Journal of the Science of Food and Agriculture,1972;23(8).
[110]Shamberger RJ, Frost DV. Possible protective effect of selenium against human cancer[J]. Canadian Medical Association Journal,1969;100(14):682.
[111]Shamberger RJ, Willis CE. Selenium distribution and human cancer mortality[J]. CRC Crit Rev Clin Lab Sci,1971;2(2):211-221.
[112]Li JY, Taylor PR, Li B, Dawsey S, Wang GQ, Ershow AG, et al. Nutrition intervention trials in Linxian, China:multiple vitamin/mineral supplementation, cancer incidence, and disease-specific mortality among adults with esophageal dysplasia[J].jnci,1993;85(18):1492-1498.
[113]Yu SY, Zhu YJ, Li WG, Huang QS, Huang CZ, Zhang QN, et al. A preliminary report on the intervention trials of primary liver cancer in high-risk populations with nutritional supplementation of selenium in China[J]. Biol Trace Elem Res,1991;29(3):289-294.
[114]Van den Brandt PA. A prospective cohort study on selenium status and the risk of lung cancer[J]. Cancer Research,1993;53(20):4860-4865.
[115]Coates RJ, Weiss NS, Daling JR, Morris JS, Labbe RF. Serum levels of selenium and retinol and the subsequent risk of cancer[J]. American journal of epidemiology,1988;128(3):515-523.
[116]Knekt P, Aromaa A, Maatela J, Alfthan G, Aaran RK, Teppo L, et al. Serum vitamin E, serum selenium and the risk of gastrointestinal cancer [J]. International journal of cancer Journal international du cancer,1988;42(6):846.
[117]Taylor PR, Li B, Dawsey S,. Prevention of esophageal cancer:the nutrition intervention trials in Linxian, China.[J]. Cancer Reserch,1994;54:2029s-2031s.
[118]Comstock GW, Bush TL, Helzlsouer K. Serum retinol, beta-carotene, vitamin E, and selenium as related to subsequent cancer of specific sites[J]. American journal of epidemiology,1992;135(2):115-121.
[119]Zhuo H, Smith AH, Steinmaus C. Selenium and lung cancer:a quantitative analysis of heterogeneity in the current epidemiological literature[J]. Cancer Epidemiology Biomarkers & Prevention,2004;13(5):771-778.
[120]Finley JW. The retention and distribution by healthy young men of stable isotopes of selenium consumed as selenite, selenate or hydroponically-grown broccoli are dependent on the isotopic form[J]. The Journal of nutrition,1999;129(4):865-871.
[121]Yang G, Ge K, Chen JCX. Selenium-related endemic diseases and the daily selenium requirement of humans[J]. World review of nutrition and dietetics,1988;55:91-152.
[122]Raines AM, Sunde RA. Selenium-regulated expression of non-selenoprotein genes [J]. FASEB J,2009;23:346(344).
[123]Ip C. Factors influencing the anticarcinogenic efficacy of selenium in dimethylbenz [a] anthracene-induced mammary tumorigenesis in rats[J]. Cancer Research,1981;41(7):2683-2686.
[124]Jacobs MM. Selenium inhibition of 1,2-dimethylhydrazine-induced colon carcinogenesis[J]. Cancer Research,1983;43(4):1646-1649.
[125]El-Bayoumy K. Inhibition of 7,12-dimethylbenz (a) anthracene-induced tumors and DNA adduct formation in the mammary glands of female Sprague-Dawley rats by the synthetic organoselenium compound,1,4-phenylenebis (methylene) selenocyanate[J]. Cancer Research,1992;52(9):2402-2407.
[126]Ip C, Lisk DJ, Thompson HJ. Selenium-enriched garlic inhibits the early stage but not the late stage of mammary carcinogenesis[J]. Carcinogenesis,1996;17(9):1979.
[127]Davis CD, Zeng H, Finley JW. Selenium-enriched broccoli decreases intestinal tumorigenesis in multiple intestinal neoplasia mice[J].J Nutr,2002;132(2):307-309.
[128]Ganther HE. Selenium metabolism and mechanisms of cancer prevention[J]. Nutrition and Cancer Prevention:New Insights Into the Role of Phytochemicals,2001: 119.
[129]Feng Y, Finley JW, Davis CD, Becker WK, Fretland AJ, Hein DW. Dietary selenium reduces the formation of aberrant crypts in rats administered 3,2'-dimethyl-4-aminobiphenyl[J]. Toxicology and Applied Pharmacology,1999;151(1):36-42.
[130]Ip C. Chemical form of selenium, critical metabolites, and cancer prevention[J]. Cancer Research,1991;51 (2):595-600.
[131]Ip C, Thompson HJ, Zhu Z, Ganther HE. In Vitro and in Vivo Studies of Methylseleninic Acid:Evidence That a Monomethylated Selenium Metabolite Is Critical for Cancer Chemoprevention [J]. Cancer Research,2000;60(11):2882-2886.
[132]Legrand C, Bour JM, Jacob C, Capiaumont J, Martial A, Marc A, et al. Lactate dehydrogenase (LDH) activity of the cultured eukaryotic cells as marker of the number of dead cells in the medium [corrected] [J]. Journal of biotechnology,1992;25(3):231.
[133]Wang Z, Jiang C, Ganther H, Lu J. Antimitogenic and proapoptotic activities of methylseleninic acid in vascular endothelial cells and associated effects on PI3K-AKT, ERK, JNK and p38 MAPK signaling[J]. Cancer research,2001;61(19):7171-7178.
[134]Jiang C, Wang ZS, Ganther HE, Lv JX. Distinct effects of methylseleninic acid vers selenite on apoptosis, cell cycle, and protein kinase pathways in DU145 human prostate cancer cells.[J]. Molecular Cancer Therapeutics,2002;1:1059-1066.
[135]Hu H, Jiang C, Li G, Lu J. PKB/AKT and ERK regulation of caspase-mediated apoptosis by methylseleninic acid in LNCaP prostate cancer cells[J]. Carcinogenesis,2005;26(8):1374.
[136]Batist G. Selenium-induced cytotoxicity of human leukemia cells:interaction with reduced glutathione[J]. Cancer Research,1986;46(11):5482-5485.
[137]Spyrou G. Selenite and selenate inhibit human lymphocyte growth via different mechanisms[J]. Cancer Research,1996;56(19):4407-4412.
[138]Chen J, Goetchius MP, Combs Jr GF, Campbell TC. Effects of dietary selenium and vitamin E on covalent binding of aflatoxin to chick liver cell macromolecules[J]. Journal of Nutrition,1982;112(2):350.
[139]Fiala ES, Joseph C, Sohn OS, El-Bayoumy K, Reddy BS. Mechanism of benzylselenocyanate inhibition of azoxymethane-induced colon carcinogenesis in F344 rats[J]. Cancer Research,1991;51(11):2826-2830.
[140]Jacobs MM, Forst CF, Beams FA. Biochemical and clinical effects of selenium on dimethylhydrazine-induced colon cancer in rats[J]. Cancer Res,1981;41(11 Pt 1):4458-4465.
[141]Weindruch R, Walford RL, Fligiel S, Guthrie D. The retardation of aging in mice by dietary restriction:longevity, cancer, immunity and lifetime energy intake[J]. Journal of Nutrition,1986; 116(4):641.
[142]Darnell RB. Onconeural antigens and the paraneoplastic neurologic disorders:at the intersection of cancer, immunity, and the brain[J]. Proceedings of the National Academy of Sciences,1996;93(10):4529-4536.
[143]Koller LD, Exon JH, Talcott PA, Osborne CA, Henningsen GM. Immune responses in rats supplemented with selenium[J]. Clinical and Experimental Immunology,1986;63(3):570.
[144]Roy M, Kiremidjian-Schumacher L, Wishe HI, Cohen MW, Stotzky G. Supplementation with selenium and human immune cell functions[J]. Biological trace element research,1994;41(1):103-114.
[145]Kiremidjian-Schumacher L, Roy M, Wishe HI, Cohen MW, Stotzky G. Supplementation with selenium augments the functions of natural killer and lymphokine-activated killer cells[J]. Biological trace element research,1996;52(3):221-239.
[146]Taylor EW. Selenium and cellular immunity [J]. Biological trace element research,1995;49(2):85-95.
[147]Diamond AM, Hu YJ, Mansur DB. Glutathione peroxidase and viral replication:implications for viral evolution and chemoprevention[J].Biofactors,2001;14(1):205-210.
[148]Buettner C, Harney JW, Berry MJ. The Caenorhabditis elegans homologue of thioredoxin reductase contains a selenocysteine insertion sequence (SECIS) element that differs from mammalian SECIS elements but directs selenocysteine incorporation[J]. Journal of Biological Chemistry,1999;274(31):21598-21602.
[149]Nordberg J, Zhong L, Holmgren A, Arner ESJ. Mammalian thioredoxin reductase is irreversibly inhibited by dinitrohalobenzenes by alkylation of both the redox active selenocysteine and its neighboring cysteine residue[J]. Journal of Biological Chemistry,1998;273(18):10835-10842.
[150]Rederstorff M, Krol A, Lescure A. Understanding the importance of selenium and selenoproteins in muscle function[J]. Cellular and Molecular Life Sciences,2005;63:52-59.
[151]Zhang D, Okada S, Yu Y, Zheng P, Yamaguchi R, Kasai H. Vitamin E inhibits apoptosis, DNA modification, and cancer incidence induced by iron-mediated peroxidation in Wistar rat kidney[J]. Cancer Research,1997;57(12):2410-2414.
[152]Ray Q Batra S, Shukla NK, Deo S, Raina V, Ashok S, et al. Lipid peroxidation, free radical production and antioxidant status in breast cancer [J]. Breast cancer research and treatment,2000;59(2):163-170.
[153]Sangeetha P, Das UN, Koratkar R, Suryaprabha P. Increase in free radical generation and lipid peroxidation following chemotherapy in patients with cancer[J]. Free radical biology & medicine,1990;8(1):15.
[154]Cheng WH, Ho YS, Valentine BA, Ross DA, Combs GF, Lei XG. Cellular glutathione peroxidase is the mediator of body selenium to protect against paraquat lethality in transgenic mice[J]. Journal of Nutrition,1998; 128(7):1070-1076.
[155]Cheng WH, Ho YS, Ross DA, Han Y, Combs GF, Lei XG. Overexpression of cellular glutathione peroxidase does not affect expression of plasma glutathione peroxidase or phospholipid hydroperoxide glutathione peroxidase in mice offered diets adequate or deficient in selenium[J]. Journal of Nutrition,1997; 127(5):675-680.
[156]Fu Y, Cheng WH, Porres JM, Ross DA, Lei XG. Knockout of cellular glutathione peroxidase gene renders mice susceptible to diquat-induced oxidative stress[J]. Free Radical Biology and Medicine,1999;27(5-6):605-611.
[157]Ratnasinghe D, Tangrea JA, Andersen MR, Barrett MJ, Virtamo J, Taylor PR, et al. Glutathione Peroxidase Codon 198 Polymorphism Variant Increases Lung Cancer Risk 1. AACR,2000: 6381-6383.
[158]Zhou JC, Zhao H, Li JG, Xia XJ, Wang KN, Zhang YJ, et al. Selenoprotein Gene Expression in Thyroid and Pituitary of Young Pigs Is Not Affected by Dietary Selenium Deficiency or Excess[J]. Journal ofNutrition,2009;139(6):1061.
[159]Kumaraswamy E, Malykh A, Korotkov KV, Kozyavkin S, Hu Y, Kwon SY, et al. Structure-expression relationships of the 15-kDa selenoprotein gene Possible role of the protein in cancer etiology[J]. Journal of Biological Chemistry,2000;275(45):35540-35547.
[160]Chen C, Zhao J, Zhang P, Chai Z. Speciation and subcellular location of Se-containing proteins in human liver studied by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and hydride generation-atomic fluorescence spectrometric detection[J]. Analytical and bioanalytical chemistry,2002;372(3):426-430.
[161]Apostolou S, Klein JO, Mitsuuchi Y, Shetler JN, Poulikakos PI, Jhanwar SC, et al. Growth inhibition and induction of apoptosis in mesothelioma cells by selenium and dependence on selenoprotein SEP15 genotype[J]. Oncogene,2004;23(29):5032-5040.
[162]Diwadkar-Navsariwala V, Diamond AM. The Link between Selenium and Chemoprevention: A Case for Selenoproteins 1[J]. Journal of Nutrition,2004;134(11):2899-2902.
[163]Tsuji PA, Irons R, Ouyang P, Carlson B, Yoo MH, XMX, et al. Sep15 knockdown decreases tumorigenicity and metastasis of a colon cancer cell line[J]. FASEB J,2009;23:338.337.
[164]Mustacich D, Powis G. Thioredoxin reductase[J]. Biochemical Journal,2000;346(Pt 1):1.
[165]Gallegos A, Berggren M, Gasdaska JR, Powis G. Mechanisms of the regulation of thioredoxin reductase activity in cancer cells by the chemopreventive agent selenium[J]. Cancer Research,1997;57(21):4965-4970.
[166]Berggren MM, Mangin JF, Gasdaska JR, Powis G Effect of selenium on rat thioredoxin reductase activity Increase by supranutritional selenium and decrease by selenium deficiency[J]. Biochemical pharmacology,1999;57(2):187-193.
[167]Allred DC, Clark GM, Elledge R, Fuqua SAW, Brown RW, Chamness GC, et al. Association of p53 protein expression with tumor cell proliferation rate and clinical outcome in node-negative breast cancer[J]. JNCI Cancer Spectrum,1993,85(3):200.
[168]Levine AJ, Finlay CA, Hinds PW. P53 is a tumor suppressor gene[J]. Cell,2004; 116:67-70.
[169]Cuddihy AR, Wong AH, Tam NW, Li S, Koromilas AE. The double-stranded RNA activated protein kinase PKR physically associates with the tumor suppressor p53 protein and phosphorylates human p53 on serine 392 in vitro[J]. Oncogene,1999;18(17):2690.
[170]Harris MP, Sutjipto S, Wills KN, Hancock W, Cornell D, Johnson DE, et al. Adenovirus-mediated p53 gene transfer inhibits growth of human tumor cells expressing mutant p53 protein[J]. Cancer gene therapy;3(2):121.
[171]Louis DN, Von Deimling A, Chung RY, Rubio MP, Whaley JM, Eibl RH, et al. Comparative study of p53 gene and protein alterations in human astrocytic tumors[J]. Journal of neuropathology and experimental neurology,1993;52(1):31.
[172]Pearson GD, Merrill GF. Deletion of the Saccharomyces cerevisiae TRR1 gene encoding thioredoxin reductase inhibits p53-dependent reporter gene expression. ASBMB,1998:5431-5434.
[173]Ueno M, Masutani H, Arai RJ, Yamauchi A, Hirota K, Sakai T, et al. Thioredoxin-dependent redox regulation of p53-mediated p21 activation[J]. Journal of Biological Chemistry,1999;274(50):35809-35815.
[174]Shibata T, Yamada T, Ishii T, Kumazawa S, Nakamura H, Masutani H, et al. Thioredoxin as a Molecular Target of Cyclopentenone Prostaglandins*[J]. Journal of Biological Chemistry.2003;278(28):26046-26054.
[175]Patterson BH, Levander OA, Helzlsouer K, McAdam PA, Lewis SA, Taylor PR, et al. Human selenite metabolism:a kinetic model[J]. American Journal of Physiology-Regulatory, Integrative and Comparative Physiology,1989;257(3):556-561.
[176]Behne D, Kyriakopoulos A. M AMMALIAN S ELENIUM-C ONTAINING P ROTEINS[J]. Annual review ofnutrition,2001;21 (1):453-473.
[177]Weiss SL, Sunde RA. Selenium regulation of classical glutathione peroxidase expression requires the 3'untranslated region in Chinese hamster ovary cells[J]. Journal of Nutrition,1997;127(7):1304-1310.
[178]Simon HU, Haj-Yehia A, Levi-Schaffer F. Role of reactive oxygen species (ROS) in apoptosis induction[J]. Apoptosis,2000;5(5):415-418.
[179]Zhao R, Xiang N, Domann FE, Zhong W. Expression of p53 enhances selenite-induced superoxide production and apoptosis in human prostate cancer cells[J]. Cancer Research,2006;66(4):2296-2304.
[180]Ip C. Activity of methylated forms of selenium in cancer prevention[J]. Cancer Research,1990;50(4):1206-1211.
[181]Cai XJ, Block E, Uden PC, Zhang X, Quimby BD, Sullivan JJ. Allium chemistry: identification of selenoamino acids in ordinary and selenium-enriched garlic, onion, and broccoli using gas chromatography with atomic emission detection[J]. Journal of agricultural and food chemistry(Print);,1995;43(7):1754-1757.
[182]Lu J, Pei H, Ip C, Lisk DJ, Ganther H, Thompson HJ. Effect of an aqueous extract of selenium-enriched garlic on in vitro markers and in vivo efficacy in cancer prevention[J]. Carcinogenesis,1996; 17(9):1903.
[183]Nagata S. Apoptosis by death factor[J]. Cell,1997;88(3):355-366.
[184]Chawla-Sarkar M, Lindner DJ, Liu YF, Williams BR, Sen GC, Silverman RH, et al. Apoptosis and interferons:role of interferon-stimulated genes as mediators of apoptosis[J]. Apoptosis,2003;8(3):237-249.
[185]Shapiro AB, Ling V. Extraction of Hoechst 33342 from the cytoplasmic leaflet of the plasma membrane by P-glycoprotein[J].European Journal of Biochemistry,1997;250(1):122-129.
[186]Vermes I, Haanen C, Steffens-Nakken H, Reutellingsperger C. A novel assay for apoptosis flow cytometric detection of phosphatidylserine expression on early apoptotic cells using fluorescein labelled annexin V[J]. Journal of immunological methods,1995;184(1):39-51.
[187]周明眉,杨奎.中药血清药理学的方法学研究—反应体系中含药血清加入量的研究[J].中药药理与临床,1998；14(006)：43-44.
[188]李仪奎,吴健宇.小青龙汤药物血清对致敏大鼠腹腔肥大细胞脱颗粒以及组…[J].中国中西医结合杂志，2000；20(001)：40-42.
[189]Bochu W, Liancai Z, Qi C. Primary study on the application of Serum Pharmacology in Chinese traditional medicine[J]. Colloids and Surfaces B:Biointerfaces,2005;43(3-4):194-197.
[190]张丽英.饲料分析及饲料质量检测技术.北京：中国农业大学出版社,2003.
[191]Li Z, Guo Y. Simultaneous determination of trace arsenic, antimony, bismuth and selenium in biological samples by hydride generation-four-channel atomic fluorescence spectrometry[J]. Talanta,2005;65(5):1318-1325.
[192]Lawrence RA, Sunde RA, Schwartz GL, Hoekstra WG Glutathione peroxidase activity in rat lens and other tissues in relation to dietary selenium intake[J]. Experimental eye research,1974;18(6):563.
[193]Massey V, Williams CH. On the reaction mechanism of yeast glutathione reductase[J]. Journal of Biological Chemistry,1965;240(11):4470-4480.
[194]Storch J, Ferber E. Detergent-amplified chemiluminescence of lucigenin for determination of superoxide anion production by NADPH oxidase and xanthine oxidase[J]. Analytical biochemistry,1988; 169(2):262-267.
[195]Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding[J]. Analytical biochemistry,1976;72(1-2):248-254.
[196]Wilkinson JE, Bell MC, Bacon JA, Masincupp FB. Effects of Supplemental Selenium on Swine I. Gestation and Lactation[J]. Journal ofAnimal Science,1977;44(2):224.
[197]Spears JW. Trace Mineral Bioavailability in Ruminants 1. Am Soc Nutrition,2003: 1506-1509.
[198]Swanson CA, Patterson BH, Levander OA, Veillon C, Taylor PR, Helzlsouer K, et al. Human [74Se] selenomethionine metabolism:a kinetic model. Am Soc Nutrition,1991:917-926.
[199]Bunk MJ, Combs Jr GF. Effect of selenium on appetite in the selenium-deficient chick[J]. Journal of nutrition,1980; 110(4):743.
[200]Lawler TL, Taylor JB, Finley JW, Caton JS. Effect of supranutritional and organically bound selenium on performance, carcass characteristics, and selenium distribution in finishing beef steers 1. Am Soc Animal Sci,2004:1488-1493.
[201]Soto-Navarro SA, Lawler TL, Taylor JB, Reynolds LP, Reed JJ, Finley JW, et al. Effect of high-selenium wheat on visceral organ mass, and intestinal cellularity and vascularity in finishing beef steers 1. Am Soc Animal Sci,2004:1788-1793.
[202]Zhu JH, Zhang X, McClung JP, Lei XG Impact of Cu, Zn-superoxide dismutase and Se-dependent glutathione peroxidase-1 knockouts on acetaminophen-induced cell death and related signaling in murine liver[J]. Experimental Biology and Medicine,2006;231(11):1726.
[203]Hussain SP, Amstad P, He P, Robles A, Lupold S, Kaneko I, et al. p53-induced up-regulation of MnSOD and GPx but not catalase increases oxidative stress and apoptosis. AACR,2004: 2350-2356.
[204]Spallholz JE. On the nature of selenium toxicity and carcinostatic activity[J]. Free Radical Biology and Medicine,1994;17(1):45-64.
[205]Christensen MJ, Nelson BL, Wray CD. Regulation of glutathione S-transferase gene expression and activity by dietary selenium[J]. Biochemical and biophysical research communications,1994;202(1):271-277.
[206]Lei XG, Ross DA, Roneker KR. Comparison of age-related differences in expression of phospholipid hydroperoxide gltutathione peroxidase mRNA and activity in various tissues of pigs[J]. Comp Biochem Physiol,1997;117:109-114.
[207]Dickinson DA, Forman HJ. Cellular glutathione and thiols metabolism[J]. Biochemical pharmacology,2002;64(5-6):1019-1026.
[208]Jenkinson SG, Lawrence RA, Burk RF, Gregory PE. Non-selenium-dependent glutathione peroxidase activity in rat lung:Association with lung glutathione S-transferase activity and the effects of hyperoxia[J]. Toxicology and Applied Pharmacology,1983;68(3):399-404.
[209]Liu Q, Lauridsem E, Clausen J. The major selenium-containing protein in human peripheral granulocytes[J]. Biological trace element research,1999;68(3):193-207.
[210]Goodman GE, Schaffer S, Bankson DD, Hughes MP, Omenn GS. Predictors of Serum Selenium in Cigarette Smokers and the Lack of Association with Lung and Prostate Cancer Risk 1[J]. Cancer Epidemiology Biomarkers & Prevention,2001; 10(10):1069-1076.
[211]Ip C. Differential Effect of Dietary Methionine on the Biopotency of Selenomethionine and Selenite in Cancer Chemopreventionl[J]. JNCI Cancer Spectrum,1988;80(4):258.
[212]Schmidt HHH, Pollock JS, Nakane M, Gorsky LD, Forstermann U, Murad F. Purification of a soluble isoform of guanylyl cyclase-activating factor synthase[J]. Proceedings of the National Academy of Sciences,1991;88(2):365-369.
[213]Casiot C, Szpunar J, obinski R, Potin-Gautier M. Sample preparation and HPLC separation approaches to speciation analysis of selenium in yeast by ICP-MS[J]. Journal of Analytical Atomic Spectrometry,1999;14(4):645-650.
[214]Infante HG, O'Connor G, Rayman M, Wahlen R, Entwisle J, Norris P, et al. Selenium speciation analysis of selenium-enriched supplements by HPLC with ultrasonic nebulisation ICP-MS and electrospray MS/MS detection[J]. Journal of Analytical Atomic Spectrometry,2004;19(12):1529-1538.
[215]Burk RF, Hill KE. Regulation of selenoproteins[J]. Annual review of nutrition,1993;13(1):65-81.
[216]Patterson JK, Lei XG, Miller DD. The Pig as an Experimental Model for Elucidating the Mechanisms Governing Dietary Influence on Mineral Absorption[J]. Experimental Biology and Medicine,2008;233(6):651.
[217]Clark LC, Combs GF, Turnbull BW, Slate EH, Chalker DK, Chow J, et al. Effects of selenium supplementation for cancer prevention in patients with carcinoma of the skin. A randomized controlled trial. Nutritional Prevention of Cancer Study Group[J].Jama,1996;276(24):1957-1963.
[218]薛庆善.体外培养的原理与技术.北京：科学出版社,2001.
[219]Yangxin FU, Porres JM, Lei XG Comparative impacts of glutathione peroxidase-1 gene knockout on oxidative stress induced by reactive oxygen and nitrogen species in mouse hepatocytes[J]. Biochem J,2001;359:687-695.
[220]Babson AL, Babson SR. Kinetic colorimetric measurement of serum lactate dehydrogenase activity[J]. Clinical Chemistry,1973; 19(7):766-769.
[221]Olie R, Simoes-Wust A, Baumann B, Leech S, Fabbro D, Stahel R, et al. A novel antisense oligonucleotide targeting survivin expression induces apoptosis and sensitizes lung cancer cells to chemotherapy[J]. Cancer Research,2000;60(11):2805.
[222]Gavrieli Y, Sherman Y, Ben-Sasson SA. Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation[J]. Journal of cell Biology,1992;119(3):493-501.
[223]Jiang C, Hu H, Malewicz B, Wang Z, Lu J. Selenite-induced p53 Ser-15 phosphorylation and caspase-mediated apoptosis in LNCaP human prostate cancer cells[J]. Molecular Cancer Therapeutics,2004;3(7):877-884.
[224]Wang. BC, Zhu. LC, Chen. Q. Primary study on the application of Serum Pharmacology in Chinese traditional medicine[J]. Colloids and Surfaces B:Biointerfaces,2005;43(3-4):194-197.
[225]江岩,黄承钰,陶德明,于静.盐亭食管癌高发区饮食对人食管癌细胞增殖的影响[J].卫生研究,2007；36(002)：226-230.
[226]刘建文.抗肿瘤作用的实验方法[J].药理实.验方法学一新技术与新方法，2003：21—75.
[227]Freshney RI. Culture of epithelial cells[M]:Wiley-Liss,2002.
[228]Whanger PD, Muth OH, Oldfield JE, Weswig PH. Influence of sulfur on incidence of white muscle disease in lambs[J]. Journal ofNutrition,1969;97(4):553.
[229]Ip C, White G Mammary cancer chemoprevention by inorganic and organic selenium:single agent treatment or in combination with vitamin E and their effects on in vitro immune functions[J]. Carcinogenesis,1987;8(12):1763.