大黄素、维生素C及其配伍对团头鲂抗拥挤胁迫的影响

选取1200尾健康的团头鲂(Megalobrama amblycephala Yih),体重为(133.44±2.11)g,随机分成4组,其中1组为对照组,投喂基础日粮(含50.3 mg/kg维生素C,以L-抗坏血酸-2-多聚磷酸酯为Vc源),另外3组为试验组,投喂饲料是在基础日粮中分别添加60 mg/kg大黄素、700 mg/kg Vc、60 mg/kg大黄素+700 mg/kg Vc。饲养60d后,从各池中取25尾规格基本一致的鱼,进行连续48h的拥挤胁迫(100 g/L)实验,分别于0h、12h、24h、48h取样分析团头鲂血液和肝脏的生化指标以及肝脏两种HSP70s mRNA的表达水平,并统计各组鱼的累积死亡率。结果表明,在拥挤胁迫前,与对照组相比,大黄素、Vc组显著提高了团头鲂血清总蛋白(TP)、溶菌酶(LSZ)和碱性磷酸酶(AKP)的水平,肝脏超氧化物歧化酶(SOD)的活性和诱导型HSP70 mRNA的表达水平,降低了血清皮质醇(COR)、甘油三酯(TG)以及肝脏丙二醛(MDA)的含量(P<0.05),且大黄素组还显著提高了肝脏过氧化氢酶(CAT)的活性(P<0.05);配伍组虽然血清TP、LSZ以及肝脏HSP70 mRNA的水平显著升高,肝脏MDA的含量也显著降低(P<0.05),但均未表现出协同增效作用。在拥挤胁迫后,与对照组相比,大黄素、Vc组不同程度地提高了团头鲂血清TP和AKP的水平,肝脏SOD和CAT的活性以及HSC70和HSP70 mRNAs的表达水平,降低了血清COR、葡萄糖(GLU)、谷丙转氨酶(GPT)、谷草转氨酶(GOT)、TG以及肝脏MDA的水平,而LSZ活性表现为先下降后升高;在配伍组中,这些指标虽然有类似以上的变化趋势,但大多差异不显著(P>0.05),且同样未表现出协同增效作用。统计表明,大黄素和Vc组鱼的累积死亡率在拥挤胁迫24h、48h均显著低于对照组(P<0.05),而配伍组与对照组的差异均不显著(P>0.05)。由此可见,在基础日粮中添加大黄素60 mg/kg或Vc 700 mg/kg,可提高团头鲂的非特异性免疫力、抗氧化能力以及两种HSP70s mRNA的表达水平,增强鱼体的抗应激能力。二者配伍则效果不佳,其相互作用的机理还有待于进一步研究。     

高能量膳食对中老年食蟹猴糖尿病相关基因表达的影响

本研究选择空腹血糖值(FPG)在正常范围内(3.20 mmol/L≤FPG<5.50 mmol/L)的中老年食蟹猴(Macaca fascicularis)60只,高能量膳食诱导12个月后,将其分为正常血糖组和诱高血糖组(FPG≥5.50 mmol/L)。采用荧光定量PCR技术对2组中36个糖尿病相关基因在诱导前后外周血白细胞中的mRNA表达量进行分析。结果表明,高能量膳食诱导后,诱高血糖组FPG和甘油三酯(TG)显著高于正常血糖组(P<0.05),而胆固醇(TCHO)、高密度脂蛋白(HDL-C)、低密度脂蛋白(LDL-C)与分组无显著相关性(P>0.05)。基因表达水平上,诱高血糖组和正常血糖组均有血管紧张素转换酶基因(ACE)、肝糖原磷酸化酶基因(PYGL)、水通道蛋白基因(AQP2)等19个基因的mRNA表达量在高能量膳食诱导前后存在显著差异(P<0.05),且基因的表达模式变化一致,但诱高血糖组的mRNA表达量变化大,且三磷酸腺苷柠檬酸裂解酶(ACLY)、选择素L(SELL)、突触相关蛋白23(SNAP23)、突触融合蛋白(STX4)这4个基因的mRNA表达水平仅在诱高血糖组高能量膳食诱导前后呈差异表达(P<0.05)。     

四个罗非鱼选育品种抗链球菌病能力差异研究

为筛选出抗病力优良的罗非鱼品种, 以奥利亚罗非鱼“夏奥1号”、尼罗罗非鱼“99”埃及品系、吉富罗非鱼“中威1号”和奥尼罗非鱼为研究对象, 33℃水温暂养7d后分别进行无乳链球菌人工感染实验, 连续7d统计累计死亡率, 并于人工感染后0、24h、48h和72h采集血液和组织样本, 研究这4个罗非鱼选育品种抗链球菌病能力的差异。结果显示: 感染7d后奥尼罗非鱼的累计死亡率最低; 奥尼罗非鱼的谷草转氨酶(AST)感染前后始终都低于其余3个品种罗非鱼(P<0.05); 埃及尼罗和奥尼在感染72h后球蛋白(GLO)分别显著升高1.13倍和1.41倍; 奥尼罗非鱼白蛋白/球蛋白(A/G)在感染前后没有显著性变化(P>0.05), 而其余3个品种罗非鱼A/G比值在感染后都显著性降低(P<0.05); 埃及尼罗的碱性磷酸酶(AKP)在感染72h后显著降低(P<0.05), 奥利亚和吉富的AKP表现为先上升后下降, 奥尼的AKP感染前后没有显著性变化(P>0.05); 各品种罗非鱼血清中的乳酸脱氢酶(LDH)感染后都显著升高(P<0.05); 奥利亚、吉富和奥尼罗非鱼的超氧化歧化酶(SOD)感染48h时都显著升高(P<0.05); 奥尼罗非鱼在感染前后溶菌酶(LZM)活性都显著高于其余3个品种罗非鱼(P<0.05)。组织病理学结果显示:吉富和奥尼肝细胞水肿变性, 而奥利亚和埃及尼罗出现大面积肝细胞脂肪变性; 每个品种罗非鱼均呈现严重的脾炎, 奥利亚、埃及尼罗和吉富的脾脏中大量铁血黄素沉积; 每种罗非鱼呈现不同程度的肾小球萎缩, 肾小管上皮细胞变性、坏死。研究表明奥尼罗非鱼抗链球菌病能力最强, 感染后血清中AST水平与肝受损程度呈一定的正相关, LZM水平和罗非鱼抗链球菌病能力呈一定的正相关。     

三峡库区172m蓄水期间泥、沙沉降对低位狗牙根种群的影响

三峡库区消落带植物恢复不仅面临长期淹水逆境,还面临泥沙、干旱等环境因素的胁迫。2009年实验、调查研究了三峡库区长寿段低位消落带的狗牙根(Cynodon dactylon L.)种群,探讨了泥、沙沉降对狗牙根种群的影响。结果表明,泥沉降和沙沉降均显著促进了狗牙根芽的萌发,而抑制了狗牙根的芽形成和萌发苗的生长,且前者的抑制作用显著大于后者。泥沉降导致总芽数、萌发苗的茎长和茎宽分别比对照低65.4%(P<0.05)、97.0%(P <0.05)、31.2%(P <0.05),而沙沉降导致前述参数分别比对照低17.1%(P<0.05)、21.2%(P<0.05)、1.0%(P>0.05)。分别将被沉降泥掩埋的狗牙根移植、掩埋于12cm厚的泥和沙中,40d后对照和S'_d组(覆盖沙)狗牙根芽萌发后均能在地面上形成正常的分株,而S'_l组(覆盖泥)的萌发芽无法穿透覆盖层,形成分株。S'_l组狗牙根芽的萌发率显著高于对照和S'_d组,但其萌发苗的茎长和茎节数均显著低于对照,分别为对照的30.3%(P<0.05)、80.4%(P<0.05);而S'_d组分株的茎长、茎节数、叶片数、叶长和叶宽均显著高于对照组,分别比后者高87.9%(P<0.05)、53.0%(P<0.05)、24.2%(P<0.05)、23.4%(P<0.05)和24.1%(P<0.05)。S'_l组萌发苗的鲜质量、干质量和干鲜质量比分别比对照组低83.3%(P<0.05)、86.2%(P<0.05)、15.8%(P<0.05);而S'_d组萌发苗的鲜质量、干质量和干鲜质量比分别比对照组高76.8%(P<0.05)、110.1%(P<0.05)、20.0%(P<0.05)。表明,低位消落带的狗牙根对沙沉降具有较强的适应能力,而对泥沉降的适应能力较低,泥沉降是库区低位消落带狗牙根种群恢复的主要影响因素之一。     

饲料蛋白水平对低温应激下吉富罗非鱼血清生化指标和HSP70 mRNA表达的影响

在水温27℃条件下, 分别对吉富罗非鱼(27.642.79) g投喂3组不同蛋白水平的饲料, 养殖周期56d。饲养结束后, 进行14℃低温应激实验, 研究低温应激后0-24h内, 鱼体血清生化指标与肝脏HSP70　mRNA表达量的变化。结果表明, 短期投喂50%蛋白水平的饲料可以提高罗非鱼血清中葡萄糖、总蛋白、甘油三酯与胆固醇水平以及血清谷丙转氨酶与谷草转氨酶活力。在低温应激后, 50%蛋白组血清葡萄糖、总蛋白、甘油三酯与胆固醇水平呈下降趋势, 而血清皮质醇水平与谷草转氨酶活力表现为先上升后下降变化。25%和38%蛋白组血清葡萄糖水平与谷丙转氨酶活力在应激期间呈上升趋势, 而谷草转氨酶与碱性磷酸酶活力和总蛋白与胆固醇水平呈先上升后下降变化。各实验组溶菌酶活力与肝脏HSP70 mRNA的表达量呈先上升后下降的变化。由此可知, 短期投喂高蛋白饲料可以提高罗非鱼血液中蛋白、血糖与脂肪含量, 增强鱼体抗应激能力, 同时, 高蛋白水平的饲料也会引起肝脏产生分解压力以及增加饲料成本。在生产中, 需要根据实际需要合理确定饲料的蛋白含量。       

LHRH-A对尼罗罗非鱼生长及生长轴相关基因表达的影响

促性腺激素释放激素(GnRH)的主要作用是刺激脑垂体促性激素(GtH)的释放, 亦可促进鱼类生长激素(GH)的释放。促黄体素释放激素类似物(LHRH-A)是哺乳类GnRH的类似物, 为了分析LHRH-A对尼罗罗非鱼生长调节的作用, 设计了长期和短期2个实验, 采用腹腔注射(剂量0.1 μg/g体重)方法, 分析LHRH-A对尼罗罗非鱼绝对生长率、特定生长率、肝体系数和肥满度的影响, 并应用荧光实时定量PCR方法检测在注射LHRH-A后不同时段(6h、12h、24h、2周)尼罗罗非鱼垂体GH、肝脏GHR和肝脏IGF-I基因的表达变化。结果表明, LHRH-A组尼罗罗非鱼的绝对生长率、特定生长率、肝体系数、肥满度均显著高于对照组(P<0.05); 注射LHRH-A后12h、24h垂体GH mRNA表达水平均显著升高(P<0.05), 2周后恢复到对照组水平; 注射LHRH-A后24h和2周肝脏GHR mRNA表达水平显著上升(P<0.05); 注射LHRH-A后6h肝脏IGF-I mRNA表达水平显著升高(P<0.05), 12h、24h和2周恢复到对照组水平。以上结果提示, LHRH-A可显著上调尼罗罗非鱼生长轴相关基因的表达, 从而促进鱼类的生长。     

副猪嗜血杆菌外膜囊泡激活caspase-11和NLRP3炎性体诱导炎性细胞因子IL-1β和IL-18分泌

【背景】副猪嗜血杆菌可引起多种炎性反应和较高死亡率,其致炎机制目前尚不清楚。【目的】探究副猪嗜血杆菌外膜囊泡(outer membrane vesicles,OMVs)诱导RAW264.7细胞caspase-11及NLRP3炎性体活化的功能,以及caspase-11在OMVs活化炎性体诱导炎性因子表达过程中的关键作用。【方法】副猪嗜血杆菌OMVs感染RAW264.7细胞,收集6、12和24h细胞,RT-PCR检测caspase-11、NLRP3、ASC和caspase-1 mRNA的表达;收集感染后48 h细胞,Western blotting检测caspase-11、NLRP3、ASC和caspase-1蛋白表达。收集感染后6、12、24、48和72h细胞上清,ELISA检测interleukin(IL)-1β和IL-18表达水平;不同浓度OMVs(0、2.5、10、25、50和100μg/mL)感染RAW264.7细胞24h,ELISA检测上清中IL-1β和IL-18水平。构建caspase-11沉默RAW264.7细胞株,收集OMVs感染后12、24和48h细胞培养上清检测IL-1β和IL-18水平。【结果】Caspase-11 mRNA转录水平在OMVs感染6、12和24h均极显著高于对照组(P<0.01);NLRP3 mRNA转录水平在感染后6、24h极显著高于对照组(P<0.01);ASC mRNA转录水平在感染后12、24h显著低于对照组(P<0.05);caspase-1 mRNA转录水平在感染后6、12和24h显著高于对照组(P<0.05)。Western blotting结果显示,与空白对照组相比,OMVs组caspase-11、NLRP3、ASC和caspase-1蛋白表达均明显升高。OMVs感染RAW264.7细胞后12、24、48和72h,IL-1β表达量极显著高于对照组(P<0.01),而且呈现时间依赖性升高,IL-18表达水平在感染后6、12、24、48和72h显著高于对照组(P<0.05);不同浓度OMVs感染细胞时,OMVs对细胞的致炎作用呈现剂量依赖性增强。Caspase-11沉默后,IL-1β水平在感染后12、24和48h显著低于未沉默组;Caspase-11沉默组IL-18表达量在感染后24、48h均显著低于未沉默组(P<0.05)。【结论】副猪嗜血杆菌OMVs在副猪嗜血杆菌诱导的炎症反应中发挥重要作用,OMVs可诱导RAW264.7细胞caspase-11介导的非经典NLRP3炎性体信号通路活化。     

Effects of active and passive hyperthermia on heat shock protein 70 (HSP70)

Summary. The purpose of this study was to delineate the effects of hyperthermia and physical exercise on the heat shock protein 70 (HSP70) response in circulating peripheral blood mononuclear cells (PBMCs). Six healthy, young (age: 24 ± 3 yrs), moderately trained males (VO_2max: 48.9 ± 2.7 ml · kg · min⁻¹) undertook two experimental trials in a randomised fashion in which the core temperature (T _c) was increased and then maintained at 39 °C during a 90 min bout by either active (AH) or passive (PH) means. AH involved subjects cycling at 90% of their lactate threshold in attire designed to impede heat loss mechanisms. In the PH trial, subjects were immersed up to the neck in a hot bath (40.2 ± 0.4 °C), once the critical T _c was achieved, intermittent cycling and water immersions were prescribed for the AH and PH conditions, respectively, to maintain the T _c at 39 °C. HSP70 was measured intracellularly pre, post and 4 h after trials, from circulating PBMCs using an ELISA technique. T _c reached 39 °C quicker in PH than during AH trials (PH: 21 ± 4 min vs. AH: 39 ± 6 min; P < 0.01), thereafter T _c was maintained around 39 °C (PH: 39.1 ± 0.2 °C; AH: 38.8 ± 0.3 °C; P > 0.05). AH induced a marked leukocytosis in all sub-sets (P < 0.05). PH generated significant monocytosis and granulocytosis (P < 0.05), without changes in lymphocyte counts (P > 0.05). There were no significant increases in intracellular HSP70 at 0 h (AH: Δ − 21.1 ± 44.8; PH: Δ + 12.5 ± 32.4 ng/mg TP/10³/μl PBMCs; P > 0.05) and 4 h (AH: Δ − 30.0 ± 40.1; PH: Δ + 36.3 ± 70.4 ng/mg TP/10³/μl PBMCs; P > 0.05) post active and passive heating. Peak HSP70 expressed as a fold-change from rest was also not increased by AH (1.1 ± 0.9; P > 0.05) or PH (3.2 ± 4.8; P > 0.05). There were no significant differences between the AH and PH trials at any time-point, and the HSP70 response appeared to be individual specific. These results did not allow us to delineate the effects of hyperthermia and other exercise associated stressors on the heat shock response and therefore further work is warranted. Authors’ address: Ric Lovell, Department of Sport, Health and Exercise Science, University of Hull, Hull HU6 7RX, U.K.     

体重和盐度对中国蛤蜊耗氧率和排氨率的影响

采用室内实验生态方法研究了不同软体部干重((1.022 ±0.821)、(0.557±0.266)、(0.303±0.277) g)和盐度(13、18、23、28、33)对中国蛤蜊(Mactra chinensis Philippi)耗氧率和排氨率的影响。结果表明:盐度、个体大小对中国蛤蜊耗氧率的影响极显著(P＜0.01),二者的交互作用对中国蛤蜊耗氧率影响显著(P＜0.05);中国蛤蜊单位体重耗氧率(R₀)与软体部干重(W)负相关,符合幂函数方程R₀=aW^-b, 其中a值的取值范围是0.695-1.762,平均值为1.449,b值的取值范围是0.446-0.587,平均值为0.542。盐度、个体大小对中国蛤蜊排氨率影响也极显著(P＜0.01);随着中国蛤蜊个体的增大,其单位体重排氨率逐渐降低;排氨率与其软体部干重呈负相关,它们之间可以用幂函数R_N=a₀W^-b₀表示。单位体重耗氧率和排氨率与盐度(S)、软体部干重(W)的二元线性回归方程分别为: R_O=2.111-1.817W+0.49S (R²=0.546, F=34.294, P＜0.001);R_N=168.186-120.589W+1.734S (R²=0.561, F=36.418, P＜0.001)。     

绿原酸对凡纳滨对虾抗氧化系统及抗低盐度胁迫的影响

对绿原酸调节凡纳滨对虾(Litopenaeus vannamei)血淋巴抗氧化系统功能及抗低盐度胁迫的效果进行了评价。360尾凡纳滨对虾随机分为4组,分别投喂含有0、100、200和400 mg/kg绿原酸的饲料28 d,随后将对虾从盐度为32的天然海水直接转入至盐度为10的水中胁迫72 h。结果表明,在正常养殖条件下,绿原酸对凡纳滨对虾的成活率、血淋巴总抗氧化能力(Total antioxidative capacity, T-AOC)、超氧化物歧化酶(Superoxide dismutase, SOD)及过氧化氢酶(Catalase, CAT)活力均无明显影响,然而投喂含有绿原酸的饲料14 d,对虾血淋巴谷胱甘肽过氧化物酶(Glutathione peroxidase, GPx)活性和血淋巴细胞GPx和CAT基因表达水平均显著高于对照组(P<0.05);低盐度胁迫24 h,绿原酸组凡纳滨对虾的存活率较对照组提高10%,但各组之间无显著性差异(P>0.05);低盐度胁迫24 h,各组凡纳滨对虾血淋巴T-AOC、SOD和GPx活性与胁迫前相比均显著增加,说明低盐度胁迫条件下机体产生了抗氧化胁迫反应,同时绿原酸组对虾血淋巴GPx、CAT活性均显著高于对照组(P<0.05);低盐度胁迫72 h,绿原酸组对虾血淋巴T-AOC、GPx和CAT活性和血淋巴细胞GPx和CAT基因表达水平均明显高于对照组。上述结果表明绿原酸可有效调节凡纳滨对虾的抗氧化系统功能,增强对虾对于低盐度胁迫下的适应能力。     

壳聚糖酶的克隆表达与壳寡糖的制备分析

目的:克隆壳聚糖酶基因于大肠杆菌中实现高表达,制备壳寡糖。方法:以枯草芽孢杆菌总DNA为模板扩增壳聚糖酶基因(CSN),克隆至载体pET23a(+)上,转化菌株BL21(DE3)。重组子经0.5 mmol/L IPTG诱导后,SDS-PAGE和质谱检测与鉴定重组酶。酶纯化后水解壳聚糖,薄层色谱分析其水解产物。结果:质谱证明壳聚糖酶(31.5kDa)成功表达,表达量占菌体总蛋白的45%左右。纯化后重组酶浓度为900 mg/L,纯度95%、回收率85%,酶活力为10 000 U/mg。壳聚糖降解产物为壳二糖至壳四糖。结论:原核表达载体pET23a(+)-CSN构建正确,壳聚糖酶表达量与活性高,适用于水解壳聚糖制备壳寡糖。     

国内外蝗害治理技术现状与展望

本文首先概述了国内外蝗虫发生与为害的态势,总结了现阶段我国蝗虫发生与为害的主要特点:即农田飞蝗暴发频繁而且严重,草原土蝗的发生时常造成严重的经济损失,而且侵入城市干扰市民生活,我国与周边国家之间蝗虫过境迁移频繁,使用化学农药污染环境和农产品;分析了国内外蝗虫防治对策与技术的发展现状,重点介绍了应急防治和可持续治理对策、...     

Coiled-coil nanomechanics and uncoiling and unfolding of the superhelix and alpha-helices of myosin

The nanomechanical properties of the coiled-coils of myosin are fundamentally important in understanding muscle assembly and contraction. Force spectra of single molecules of double-headed myosin, single-headed myosin, and coiled-coil tail fragments were acquired with an atomic force microscope and displayed characteristic triphasic force-distance responses to stretch: a rise phase (R) and a plateau phase (P) and an exponential phase (E). The R and P phases arise mainly from the stretching of the coiled-coils, with the hinge region being the main contributor to the rise phase at low force. Only the E phase was analyzable by the worm-like chain model of polymer elasticity. Restrained molecular mechanics simulations on an existing x-ray structure of scallop S2 yielded force spectra with either two or three phases, depending on the mode of stretch. It revealed that coiled-coil chains separate completely near the end of the P phase and the stretching of the unfolded chains gives rise to the E phase. Extensive conformational searching yielded a P phase force near 40 pN that agreed well with the experimental value. We suggest that the flexible and elastic S2 region, particularly the hinge region, may undergo force-induced unfolding and extend reversibly during actomyosin powerstroke.     

放牧对牧草光合作用、呼吸作用和氮、碳吸收与转运的影响

研究放牧对草地植物生理活动的影响,对于揭示草地放牧演替的生理机制有重要意义.大量研究表明,家畜放牧对牧草光合作用、呼吸作用以及C和N吸收与转运的影响,可以分为生理伤害和生理恢复2个阶段.放牧通过改变草地冠层结构影响牧草光合作用,净光合作用速率短期内迅速下降,随着叶面积指数增加又逐渐上升,呼吸作用有相似的变化趋势.牧草放牧后再生长所需的C和N最初主要来自根系和留茬中的贮藏物质,此后随着牧草生长恢复逐渐由同化作用供给,C代谢与土壤N水平负相关.放牧后牧草生理活动变化与牧草遗传特性、种间竞争、家畜放牧特征、非生物环境等因素密切相关.     

Kinetics and thermodynamics of peroxidase- and laccase-catalyzed oxidation of N-substituted phenothiazines and phenoxazines

N -substituted phenothiazines (PTs) and phenoxazines (POs) catalyzed by fungal Coprinus cinereus peroxidase and Polyporus pinsitus laccase were investigated at pH 4–10. In the case of peroxidase, an apparent bimolecular rate constant (expressed as k _cat/K _m) varied from 1 ×10⁷M⁻¹s⁻¹to 2.6×10⁸M⁻¹s⁻¹ at pH 7.0. The constants for PO oxidation were higher in comparison to PT. pH dependence revealed two or three ionizable groups with pK _a values of 4.9–5.7 and 7.7–9.7 that significantly affected the activity of peroxidase. Single-turnover experiments showed that the limiting step of PT oxidation was reduction of compound II and second-order rate constants were obtained which were consistent with the constants at steady-state conditions. Laccase-catalyzed PT and PO oxidation rates were lower; apparent bimolecular rate constants varied from 1.8×10⁵M⁻¹s⁻¹ to 2.0×10⁷M⁻¹s⁻¹ at pH 5.3. PO constants were higher in comparison to PT, as was the case with peroxidase. The dependence of the apparent bimolecular constants of compound II or copper type 1 reduction, in the case of peroxidase or laccase, respectively, was analyzed in the framework of the Marcus outer-sphere electron-transfer theory. Peroxidase-catalyzed reactions with PT, as well as PO, fitted the same hyperbolic dependence with a maximal oxidation rate of 1.6×10⁸M⁻¹s⁻¹ and a reorganization energy of 0.30 eV. The respective parameters for laccase were 5.0×10⁷M⁻¹s⁻¹ and 0.29 eV. Received: 20 September 1999 / Accepted: 24 February 2000     

Synthesis of hapten and conjugates of coumestrol and development of immunoassay

3-O-Carboxymethylcoumestrol was prepared as the hapten for immunoassay by a partial alkylation of coumestrol with ethyl chloroacetate in acetone alkalized with potassium carbonate. 3-O-Ethoxycarbonylmethylcoumestrol was separated by column chromatography and finally was hydrolyzed with formic acid. 1H and 13C NMR data (APT, COSY, HMQC, and HMBC) revealed that the reaction was regioselective, as 3-O-ethoxycarboxymethylcoumestrol was the only monosubstituted derivative. The hapten was then conjugated to bovine serum albumin and used for immunization of rabbits. A radioimmunoassay (RIA) system was established based on the polyclonal antiserum and a 125I-labeled hapten-tyrosine methyl ester conjugate as the radioligand. Parameters of the RIA: sensitivity: 12 pg per tube, 50% intercept: 140 pg per tube, working range: 20-4000 pg per tube. The cross-reactivity of a panel isoflavonoid and lignan phytoestrogens was either negligible (e.g. formononetin 0.07%; biochanin A 0.06%) or not detectable at all. The major immunoreactive peak in HPLC fractions from an alfalfa extract had the same retention time as coumestrol standard and represented 94.8% of the signal. The remaining 5.2% of immunoreactivity was distributed between five minor peaks. We conclude that after the validation for particular matrices, the method will be a useful tool for analysis of coumestrol, especially in low volume and low concentration samples.     

白术同源四倍体的诱导和鉴定及其与二倍体过氧化物酶的比较

以白术（Atractylodes macrooephala Koidz.）二倍体组培苗为材料,对其四倍体诱导方法进行研究,共获得45个白术同源四倍体株系,为优良株系的选育提供了材料。此外,还分析比较了其中8个白术四倍体株系与二倍体的过氧化物酶同工酶（POD）的酶谱差异,发现四倍体各株系过氧化物酶同工酶谱比二倍体的均多了Rf0.310的谱带,且总过氧化物酶比活力也发生了很大改变,对探讨白术四倍体优良株系的生理生化机理具有一定的参考价值。     

Morphology and putative function of the colon and cloaca of marine and freshwater snakes

Among tetrapods, evidence for postrenal modification of the urine by the distal digestive tract (including the colon and cloaca) is highly variable. Birds and bladderless reptiles are of interest because the colon and cloaca represent the only sites from which water and ions can be reclaimed from the urine secreted by the kidney. For animals occupying desiccating environments (e.g., deserts and marine environments), postrenal modification of the urine may directly contribute to the maintenance of hypo‐osmotic body fluids. We compared the morphology and distribution of key proteins in the colon, cloaca, and urogenital ducts of watersnakes from marine (Nerodia clarkii clarkii) and freshwater (Nerodia fasciata) habitats. Specifically, we examined the epithelia of each tissue for evidence of mucus production by examining the distribution of mucopolysaccharides, and for evidence of water/ion regulation by examining the distribution of Na⁺/K⁺‐ATPase (NKA), Na⁺/K⁺/Cl^? cotransporter (NKCC), and aquaporin 3 (AQP3). NKCC localized to the basolateral epithelium of the colon, urodeal sphincter, and proctodeum, consistent with a role in secretion of Na⁺, Cl^?, and K⁺ from the tissue, but NKA was not detected in the colon or any compartment of the cloaca. Interestingly, NKA was detected in the basolateral epithelium of the ureters, suggesting the urothelium may play a role in active ion transport. AQP3 was detected in the ureters and coprodeal complex, consistent with a role in urinary and fecal dehydration or, potentially, in the production of the watery component of the mucus secreted by the coprodeal complex. Since no differences in general cloacal morphology, production of mucus, or the distribution of ion transporters/water channels were detected between the two species, cloacal osmoregulation may either be regulated by proteins not examined in this study or may not be responsible for the differential success of N. c. clarkii and N. fasciata in marine habitats. J. Morphol. 2011. © 2011 Wiley Periodicals, Inc.