纳米氧化锌颗粒对大鼠气管上皮细胞动态变化的影响及毒性机理

纳米氧化锌(Zn O NPs)对呼吸道的毒性损伤作用备受关注,但有关其对呼吸道上皮细胞动态变化的影响机理还有待深入研究.因此,本研究通过将大鼠气管上皮细胞(RTE)暴露于不同浓度(1和10 mg·L~(-1))和不同粒径(50和200 nm)的Zn O NPs中,利用细胞电阻抗检测技术(ECIS)检测细胞动态变化,采用CCK8法检测细胞生长抑制效应,并通过胞内ROS和MDA含量变化探讨其影响机制.ECIS检测结果显示,Zn O NPs暴露下,RTE细胞生长和增殖受到明显抑制,且具有浓度依赖效应.当暴露浓度为1 mg·L~(-1)时,与对照组相比,50 nm暴露组细胞电阻抗值的下调幅度为18%,为200 nm暴露组的1.2倍.Zn O NPs诱导的RTE细胞增殖抑制率具有浓度依赖效应,当暴露浓度为10 mg·L~(-1)时,50和200 nm暴露组细胞增殖抑制率分别为暴露浓度为1 mg·L~(-1)时的2.9和1.4倍.Zn O NPs诱导的RTE细胞氧化应激结果显示,胞内ROS和MDA含量随着纳米颗粒暴露浓度的增加而增加,随着纳米颗粒粒径的减小而增加,具有显著的浓度-和剂量-依赖效应.当Zn O NPs浓度分别为1和10 mg·L~(-1)时,ROS含量分别是对照组的2.8和3.7倍;当暴露浓度为10 mg·L~(-1)时,50 nm暴露组细胞内ROS含量是200 nm暴露组的1.7倍;暴露浓度为1和10 mg·L~(-1)时,50 nm氧化锌处理组诱导的细胞内MDA含量分别是对照组的5.4和7.9倍.Zn O NPs能够影响呼吸道上皮细胞动态变化,从而破环RTE细胞屏障并进入细胞,诱导胞内ROS和MDA水平升高,进而抑制细胞的生长与增殖.研究表明,影响Zn O NPs诱导的RTE细胞动态变化和氧化应激的关键因素是颗粒粒径与暴露浓度.     

A mixture of polychlorinated dibenzo-p-dioxins (PCDDs), dibenzofurans (PCDFs), and non-ortho polychlorinated biphenyls (PCBs) changed the lipid content of pregnant Long Evans rats

Pregnant Long Evans rats received 1.0 μg/kg of dioxin toxic equivalents (TEQ) by oral gavage on the 15th gestational day (GD 15), using a dosing mixture that contained two polychlorinated dioxins, four polychlorinated furans and three non-ortho polychlorinated biphenyls (PCBs). Rats were sacrificed on GD 16, GD 21 and postnatal day 4 (PND 4). The lipid content of fetus, pup, placenta and maternal liver, serum and adipose tissue were determined. Treated GD 16 and GD 21 fetuses had identical lipid content to the control group, yet the lipid content of treated pups on PND 4 was 32% higher than that of the control group. On the other hand, the lipid content of placenta, liver, and serum from the treated dams was 44–50%, 24%, and 38% lower than that of the control group, respectively. Thus, a low-dose mixture of dioxin-like compounds can cause changes in lipid content. The lipid content of offspring was not affected until they were exposed via lactation.     

冀东某县农村养殖粪污处理环境风险与对策研究

生猪养殖粪污是农村面源污染来源之一,冀东某县是河北省养猪大县,养殖产生的粪污主要采用能源生态处理方式进行处理与利用,在养殖粪污的现有利用方式中,存在土壤中重金属积累风险、粪便中病菌抗生素风险、生猪散养的环境风险.针对冀东某县生猪养殖粪污处理利用存在的环境风险,对规模化养殖场采用粪便堆肥发酵制有机肥工艺进行处理利用;散养型粪污,依托规模化养殖场进行处理利用;政府依据种养结合的原则,制定养殖规划,确定养殖规模,建立制度化管理,保护农村环境.     

SO2污染激活AT1R改变运动大鼠心肌胶原纤维形态学并导致心功能降低

为了观察SO₂污染环境下运动对大鼠心功能的影响,从心脏局部肾素-血管紧张素系统（renin-angiotensin system,RAS）核心成员——血管紧张素Ⅱ（angiotensin Ⅱ,AngⅡ）Ⅰ型受体（AT₁R）介导的心肌胶原纤维形态结构重塑的角度出发,应用心脏插管技术观察大鼠的心脏功能;采用放射免疫技术（ELISA）、免疫组织化学和胃酶酸解法等方法对心肌局部ρ（AngⅡ）、AT₁R蛋白表达水平、w（HYP）（HYP为羟脯氨酸）及胶原容积分数进行检测.结果表明:①单纯运动组（EG）大鼠主动脉收缩压、左室内压峰值、±dp/dt_max（左室内压最大上升速率/下降速率）显著升高（P < 0.01）,舒张压、AT₁R蛋白表达显著降低（P < 0.01）,w（HYP）、w（CC）（心肌胶原浓度）、PVCA（血管周围胶原面积）、CVF（心肌胶原容积分数）及ρ（AngⅡ）有升高趋势（P>0.01）;②单纯SO₂污染组（SRG）大鼠左室末期舒张压显著升高（P < 0.01）,左室内压±dp/dt_max显著降低（P < 0.01）;w（HYP）、w（CC）、PVCA、CVF、ρ（AngⅡ）及AT₁R蛋白表达水平均显著升高（P < 0.01）;③SO₂污染+运动组（SEG）大鼠左室末期舒张压显著升高（P < 0.01）,主动脉收缩压、左室内压峰值、左室内压±dp/dt_max显著降低（P < 0.01）,w（HYP）、w（CC）、PVCA、CVF、ρ（AngⅡ）及AT₁R蛋白表达水平均显著升高（P < 0.01）,并且较SRG大鼠升高更显著（P < 0.01）.研究显示,SO₂污染导致运动大鼠心肌胶原纤维形态结构发生异常重塑,最终使大鼠的心功能产生显著的负性变力性效应,其机制可能与心脏局部RAS系统的激活有关.      

大气细颗粒物亚慢性染毒对大鼠肺内质网应激相关因子表达的影响

采集冬、夏季太原市大气细颗粒物(PM2.5),并制备PM2.5生理盐水混悬液.将35只雄性SD大鼠随机分为7组:对照组、3个不同剂量夏季PM2.5染毒组(0.2、0.6、1.5 mg·kg~(-1)体重)及3个不同剂量冬季PM2.5染毒组(0.3、1.5、2.7 mg·kg~(-1)体重),每组5只,气道滴注法染毒,每隔2 d染毒1次,共60 d.采用荧光实时定量PCR、Western blot、ELISA方法检测大鼠肺内质网应激指标——葡萄糖调节蛋白78(GRP78)、活化转录因子6(ATF6)、C/EBP同源蛋白(CHOP)、半胱氨酸天冬氨酸蛋白酶12(Caspase12)及血红素氧合酶-1(HO-1)的mRNA和蛋白表达变化.结果表明,与对照组相比,冬季中、高剂量PM2.5染毒组大鼠肺组织GRP78、ATF6、CHOP、Caspase12、HO-1 mRNA和蛋白表达显著增加,夏季高剂量PM2.5染毒组大鼠肺组织这5个基因mRNA和蛋白表达显著增加.冬季和夏季PM2.5组大鼠肺上述5个基因表达有剂量-效应关系.结果表明,太原市PM2.5亚慢性染毒可诱导大鼠肺内质网应激相关基因GRP78/ATF6/CHOP/HO-1表达,说明肺内质网应激反应加强;而CHOP和Caspase-12上调,提示与细胞凋亡关联的内质网相关性死亡途径被激活.冬季和夏季PM2.5引起内质网应激相关因子表达上调的效应没有显著差别.     

Responses of Tuatara (Sphenodon punctatus) to Removal of Introduced Pacific Rats from Islands

Abstract:  Invasive mammalian predators such as rats are now widespread on islands, but hypotheses about their effects have rarely been tested. Circumstantial evidence from New Zealand indicates that, when introduced to islands, Pacific rats ( Rattus exulans ) have negative effects on endemic plants, invertebrates, birds, and reptiles, including the tuatara ( Sphenodon punctatus ). We tested the effects of Pacific rats on tuatara by comparing the demographic structure and body condition of tuatara populations on three islands before and after removal of rats and on a fourth island where rats remained. In the presence of rats, juvenile tuatara constituted on average 0–5% of the sample tuatara populations. When Pacific rats were removed after at least 200 years' occupancy, the proportion of juvenile tuatara increased 3.5- to 17-fold and body condition of adult males and females also improved (sometimes dramatically). We predict that, unless Pacific rats are removed from Taranga Island, the tuatara population will collapse because of low population density and the lack of juvenile recruitment. Our results demonstrate that when invasive species exert subtle effects on recruitment and body condition, the effects on populations of long-lived endemic species may only become apparent long after the invasion.     

Effect of nitroethane and nitroethanol on the production of indole and 3-methylindole (skatole) from bacteria in swine feces by gas chromatography

Indole and 3-methylindole (skatole) are odor pollutants in livestock waste, and skatole is a major component of boar taint. Skatole causes pulmonary edema and emphysema in ruminants and causes damage to lung Clara cells in animals and humans. A gas chromatographic method that originally used a nitrogen–phosphorus detector to increase sensitivity was modified resulting in an improved flame ionization detection response for indole and skatole of 236% and 207%, respectively. The improved method eliminates the large amount of indole decomposition in the injector. A 10 μ g mL^–1 spike of indole and skatole in water and swine fecal slurries resulted in recovery of 78.5% and 96% in water and 76.1% and 85.8% in fecal slurries, respectively. The effect of the addition of nitroethane and nitroethanol at 21.8 mM in swine fecal slurries was studied on the microbial production of indole and skatole. Nitroethane and nitroethanol decreased the production of skatole in swine fecal slurries at 24 h. The nitroethane effect on l-tryptophan-supplemented fecal slurries after 6 and 24 h incubation resulted in a decrease of 69.0% (P = 0.02) and 23.5% skatole production, respectively, and a decrease of 14.9% indole at 6 h, but an increase in indole production of 81.1% at 24 h.     

PFOS致大鼠肝毒性及其作用机制研究

通过全氟辛烷磺酸(perfluomoctane sultanate,PFOS)大鼠灌胃染毒实验评价PFOS对肝功能的影响,探讨PFOS肝毒性反应的潜在机制与可能途径。将Sprague Dawley (SD)雄性大鼠随机分为3组,分别以0 mg·kg~(-1)、5 mg·kg~(-1)和10 mg·kg~(-1)PFOS灌胃染毒28 d。以HE和油红染色法观察大鼠肝脏形态改变。ELISA法测定各组谷丙转氨酶(alanine aminotransferase,ALT)、谷草转氨酶(aspartate transaminase,AST)、碱性磷酸酶(alkaline phosphatase,ALP)含量变化。化学比色法测定肝匀浆脂代谢水平和氧化产物含量。RT-PCR法检测肝脏内氧化应激以及脂代谢相关基因表达水平。结果表明,PFOS暴露大鼠体重显著降低而肝脏系数显著增加(P0.05),与对照组相比PFOS组血清肝功能酶均出现随PFOS浓度增加而升高(P0.05)。同时大鼠肝脏谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-px)和丙二醛(malondialdehyde,MDA)水平在高剂量组显著升高(P0.05),超氧化物歧化酶(superoxide dismutase,SOD)含量先显著升高(P0.05)后显著降低(P0.05)。且肝脏中脂代谢水平也随PFOS浓度的增加而出现显著改变(P0.05)。PFOS组基因表达均较对照组显著上升(P0.05)。以上结果说明PFOS具有明显的肝毒性作用,可影响肝脂代谢水平,这可能与PFOS引起的氧化应激所导致的损伤有关。     

三(1,3-二氯-2-丙基)磷酸酯诱发肝脏损害及病理改变研究

本研究观测有机磷酯阻燃剂(OPFRs)污染是否可以诱发肝脏损害,考察其发生及发展程度,并探讨其发生机理,为有机磷阻燃剂污染的防治和相关疾病的有效治疗提供基础数据和科学依据。实验以大鼠为动物模型,将60只SPF级SD雄性大鼠分为5组,每组12只,选取典型的氯代有机磷阻燃剂三(1,3-二氯-2-丙基)磷酸酯(TDCPP)对大鼠进行染毒,空白对照组不做任何处理,溶剂对照组以相同体积的橄榄油灌胃,染毒组以不同剂量的TDCPP进行灌胃(125 mg·kg~(-1)·d~(-1)、250 mg·kg~(-1)·d~(-1)和500 mg·kg~(-1)·d~(-1)),每周测量体重,于第4周和第8周取血检测肝功及其他生化指标,在第8周每组抽取3只大鼠取肝脏组织做HE染色,并用透射电镜观察分析肝组织病理学改变。TDCPP对大鼠染毒8周后,结果表明:(1)体重指标在灌胃1周后开始发生差异,TDCPP处理组大鼠的体重有下降的趋势,染毒组与空白对照组和溶剂对照组相比较,差异显著(*P0.05,**P0.01),其中高剂量灌胃组的体重下降最为明显(**P0.01);(2)血清肝功指标表现出显著变化,血清谷丙转氨酶、谷草转氨酶、胆固醇和甘油三脂水平在第8周呈现明显下降趋势,染毒组与空白对照组和溶剂对照组比较,差异明显(*P0.05,**P0.01);(3) TDCPP暴露组生理生化指标变化明显,血清乙酰胆碱酯酶活性显著降低,MDA含量显著升高,SOD活力显著性降低,造成氧化损伤,与空白对照组和溶剂对照组比较,差异显著(*P0.05,**P0.01);(4)病理切片结果显示染毒组与对照组比较,细胞坏死现象明显,且高剂量组坏死更为严重。研究结果显示:TDCPP可引起大鼠体重明显下降,大鼠肝脏细胞损伤、合成功能下降,造成肝脏代谢功能紊乱,造成较为严重的肝损伤。     

Estimating Population Size of Elusive Animals with DNA from Hunter-Collected Feces: Four Methods for Brown Bears

Abstract:  Noninvasive genetic methods can be used to estimate animal abundances and offer several advantages over conventional methods. Few attempts have been made, however, to evaluate the accuracy and precision of the estimates. We compared four methods of estimating population size based on fecal sampling. Two methods used rarefaction indices and two were based on capture-mark-recapture (CMR) estimators, one combining genetic and field data. Volunteer hunters and others collected 1904 fecal samples over 2 consecutive years in a large area containing a well-studied population of brown bears ( Ursus arctos ). On our 49,000-km² study area in south-central Sweden, population size estimates ranged from 378 to 572 bears in 2001 and 273 to 433 bears in 2002, depending on the method of estimation used. The estimates from the best model in the program MARK appeared to be the most accurate, based on the minimum population size estimate from radio-marked bears in a subsection of our sampling area. In addition, MARK models included heterogeneity and temporal variation in detection probabilities, which appeared to be present in our samples. All methods, though, incorrectly suggested a biased sex ratio, probably because of sex differences in detection probabilities and low overall detection probabilities. The population size of elusive animals can be estimated reliably over large areas with noninvasive genetic methods, but we stress the importance of an adequate and well-distributed sampling effort. In cases of biased sampling, calibration with independent estimates may be necessary. We recommend that this noninvasive genetic approach, using the MARK models, be used in the future in areas where sufficient numbers of volunteers can be mobilized.