川金丝猴尿液类固醇激素的保存时效性

尿液类固醇激素检测已经广泛运用于野生动物的生理机能、健康状况和繁殖状态的研究,由于分析条件以及样品采集时间限制,一般尿液样品需保存一段时间后才能在实验室分析,不同物种尿液类固醇在不同保存条件的保存时效有差异。本研究利用2只成年雄性和4只成年雌性的尿液样品共21份,确定川金丝猴尿液类固醇的保存时效和保存方法。结果表明:4℃时,雄性川金丝猴尿液睾酮、雌性尿液睾酮、孕酮的保存时效为7 d左右;雌性尿液雌二醇的保存时效在3 d以内。-20℃时,雄性川金丝猴尿液中睾酮的含量在60 d内保持稳定;雌性尿液中睾酮、孕酮和雌二醇的含量在60 d、45 d、60 d内保持稳定。-20℃保存川金丝猴尿液样品是一种有效、可靠的方法。     

秦岭野生雌性川金丝猴粪便睾酮水平与邀配频次的季节性变化

为研究野生雌性川金丝猴(Rhinopithecus roxellana)不同季节体内睾酮水平与性行为频次的变化及二者的关系,我们以秦岭一野生川金丝猴种群为研究对象,通过长时间连续跟踪观察,完成了该种群中所有87只金丝猴的个体识别。用无损伤取样法采集了三只成年雌性川金丝猴不同季节的粪样,对粪样中的睾酮进行抽提测定,同时对采样个体的性行为进行记录。结果显示:1)成年雌猴在交配盛期与非交配盛期,粪样中睾酮水平均呈明显的周期性波动,交配盛期时睾酮水平的变化周期为24.56 d±4.07 d,非交配盛期时为43.67 d±2.89 d,显著长于交配盛期;2)成年雌猴在交配盛期的粪样睾酮水平为2.85 ng±1.12 ng/g,邀配频次为0.98±1.04次/d,非交配盛期时睾酮水平为1.71 ng±0.77 ng/g,邀配频次为0.28±0.53次/d,与交配盛期相比,非交配盛期的睾酮水平和邀配频次均明显降低;3)交配盛期各成年雌猴的粪样睾酮水平与邀配频次均有较显著的正相关关系(相关系数分别为0.631、0.683和0.659),而在非交配盛期这种相关关系并不存在(相关系数分别为0.091、0.493和0.205)。本研究表明,成年雌性川金丝猴体内睾酮水平、睾酮水平变化周期及邀配行为频次均具有明显的季节性差异,这种季节性差异可能是川金丝猴行季节性繁殖的内在原因和外在表现之一。本研究亦表明交配盛期雌猴的性行为频次在一定程度上可能受体内睾酮水平的控制,而在非交配盛期这种控制作用较弱。     

川金丝猴粪尿中类固醇性激素抽提方法比较

本文旨在探讨川金丝猴粪尿中类固醇性激素的抽提方法, 实验比较了3 种尿液抽提溶剂和5 种粪便抽提方法的抽提回收率。结果显示, 以二氯甲烷为溶剂抽提雄猴尿液中的睾酮、雌猴尿液中的雌二醇和孕酮, 回收率分别为79.27%、73.39%和80.28%, 抽提效果较其它两种溶剂为好; 以乙醇加热法抽提雄猴粪便中的睾酮和雌二醇、雌猴粪便中的睾酮、雌二醇和孕酮, 回收率分别为1.33%、85.12%与67.41%、79.58%和77.71%, 抽提效果优于其它4 种方法,且该方法简便易于操作。实验表明, 以二氯甲烷和乙醇加热法分别抽提川金丝猴尿液和粪便中的类固醇性激素是较理想的抽提方法。     

圈养林麝粪便类固醇激素保存时效性研究

用非损伤性采样方法对圈养林麝在三个条件:1)未处理,2)90%甲醇,3)0.9%抗生素,15个时间梯度下(20℃)的粪便中类固醇激素及其代谢物的保存时间及方法进行了比较,结果表明:林麝粪便在室温下(20℃),孕酮及其代谢物48 h有明显的上升趋势;雌二醇、睾酮和皮质醇及其代谢物在18 h均出现显著变化;对比抗生素保存,用甲醇保存是一种较好的保存林麝粪便类固醇激素的方法;然而不同激素用甲醇保存的时间不同,孕酮不宜超过1个月,皮质醇不宜超过半个月,睾酮则不宜超过10天,而雌二醇可以保存2个月之久.     

新疆塔里木马鹿粪便激素含量季节性变化的初步研究

目的:检测雄性塔里木马鹿的睾酮,雌鹿的雌二醇、雌三醇、孕酮等粪便激素含量,并比较不同月份之间的变化。方法:利用放射免疫分析法(RIA)在且末和尉犁县马鹿场采集塔里木马鹿粪便,检测了2004年2～12月间塔里木马鹿粪便中的激素水平。结果:粪便性激素水平有明显的季节变化(P<0.05)。雄鹿粪便睾酮水平月间差异显著(P<0.05),浓度在8月有2个高峰。雌鹿粪便雌二醇在3月和6月有2个高峰;粪便孕酮水平在3月有2个高峰,在8月有1个高峰;雌三醇差异不显著,但4月的最高值也显著高于其他各时间点(P<0.05)。结论:利用动物粪便研究激素具有对保护动物完全无伤害、材料收集容易等优点,并且有助于评估动物青春期的健康状况、判断动物排卵方式、早孕检查、产前预测等,因而是目前研究野生珍稀动物性激素的一种较为理想的方法。     

繁殖期雌性蓝狐性激素检测与 性器官组织学观察

为了研究繁殖期雌性蓝狐(Alopexlagopus)类固醇激素含量、发情表现与性器官组织学结构之间的关系,于2019年选取繁殖期不同发情状况的雌性蓝狐,共18只,分为典型发情、非典型发情和不发情3组。通过无损伤取样法采集粪便与尿液并通过放射免疫法(RIA)测定孕酮(P)和雌二醇(E2)的含量;每组选取1只在发情并输精的当天取卵巢和子宫,进行组织学观察。结果显示,所测定典型发情、非典型发情、不发情蓝狐类固醇激素水平,雌二醇在尿液中的含量显著高于粪便,且在不同组间差异明显,尤其在典型发情组更高,达到(11 065.17±546.76)ng/L;孕酮含量尿液与粪样差异不大,孕酮含量在典型发情雌性蓝狐粪便与尿液含量差异不显著,但在不发情雌性蓝狐中差异显著,为(16.61±0.63)μg/L。粪尿测定孕酮和雌二醇激素含量与三组不同情况的发情表现相比相关性显著。粪、尿作为类固醇激素检测样本,虽然含量有差异,但变化趋势一致,都可使用。卵巢与子宫在典型发情雌性蓝狐中体积较大,卵巢可见各级卵泡和多个黄体,子宫黏膜上皮为柱状上皮,排列紧密,固有层内可见大量腺体;非典型发情雌性蓝狐卵巢、子宫发育状况与典型发情雌性蓝狐相类似;不发情雌性蓝狐卵巢、子宫呈静息状态,卵巢中卵泡多处于闭锁状态,无卵母细胞,也无黄体,固有层间质细胞及肌层肌细胞排列更为紧密。说明检测孕酮和雌二醇激素的含量,可以准确判定蓝狐发情的状态。     

短尾猴粪便雌二醇和孕酮处理与 提取方法比较

非损伤性的粪便样本已在监测动物性激素水平中得到广泛应用,但粪便处理和激素的提取方法在不同物种中因其食性与代谢不同具有明显的物种特异性。本研究以野生和笼养两种环境下的短尾猴(Macacathibetana)为研究对象,采集粪便样品,利用放射免疫法(RIA)检测激素含量,比较不同粪便处理方式(湿便法、冷冻干燥粉碎法、冷冻干燥研筛法)和激素提取方法(乙醇加热法、甲醇乙醚法、乙醇丙酮法、甲醇乙醇法)的效果,探讨不同取食条件下短尾猴雌二醇(E2)和孕酮(P4)水平的最佳测定方法。粪便不同处理方法的比较中,冷冻干燥研筛法测得雌二醇和孕酮含量最高,冷冻干燥研筛法与冷冻干燥粉碎法在野生短尾猴中具有显著差异(P0.01),但在笼养短尾猴中无显著差异(P 0.05)。湿便法测得含量最低,与另两种处理方法均具有显著差异性(P 0.01)。激素提取方法的比较中,乙醇加热法提取雌二醇和孕酮含量最高,与甲醇乙醚法(P 0.01)、乙醇丙酮法(P 0.01)、甲醇乙醇法(P 0.01)之间均具有显著性差异。综合考虑激素平均含量、处理过程的简便性、抽提步骤的可行性,野生短尾猴粪便处理建议选用冷冻干燥研筛法,笼养短尾猴粪便由于冷冻干燥粉碎法操作简便且与冷冻干燥研筛法无显著差异,因此笼养短尾猴建议选用冷冻干燥粉碎法。野生与笼养短尾猴粪便激素的提取均建议采用乙醇加热法。     

大熊猫粪便样本处理与激素萃取方法比较

通过测定圈养动物粪便中的激素水平,监测其发情与怀孕状态和评估其福利状况是目前普遍采用的一种实验手段。然而,现有研究对于圈养野生动物粪便样本的处理及其激素萃取方法存在很大分歧,研究结果也不尽一致。本研究采集了10只(雌∶雄=1∶1)圈养大熊猫(Ailuropoda melanoleuca)的粪便样本,分别采用冷冻干燥-研筛法和冷冻干燥-粉碎法处理粪便样本,然后分别使用80%甲醇、90%乙醇和95%乙醇3种萃取剂进行激素萃取,最后比较2种粪便处理方法和3种溶剂萃取方法所获得的粪便样本中孕酮、雌二醇、睾酮和皮质醇的浓度。结果表明,冷冻干燥-研筛法结合80%甲醇溶液萃取处理的样本中雌二醇和孕酮检测值最高,冷冻干燥-粉碎法结合80%甲醇溶液萃取处理的样本中睾酮检测值最高。冷冻干燥-粉碎法结合90%乙醇溶液萃取处理的样本与冷冻干燥-研筛法结合80%甲醇溶液萃取处理的样本中皮质醇检测值更高,且2种方法的检测结果差异不显著。综合粪便处理难易程度和萃取溶剂性质,建议雌二醇、孕酮和睾酮激素检测根据最高检测浓度的方法进行粪便前处理,皮质醇激素检测使用冷冻干燥-粉碎法结合90%乙醇溶液萃取方法进行粪便前处理。...     

注射LHRHa对泥鳅血清性类固醇激素变化研究

研究了注射促黄体激素类似物(LHRHa)后,泥鳅(Misgurnus anguillicaudatus)血清性类同醇激素的变化规律,并探讨在泥鳅繁殖季节时.孕酮(P)、睾酮(T)和雌二醇(E2)对性腺发育的作用及调节机制.实验共分两组,对照组和实验组;对照组只注射生理盐水;实验组注射LHRHa,雌鱼0.2 μg/g,雄鱼减半.注射前尾静脉采血,作为血液样本分析基础水平(Oh),注射药物后分别在7、24、48、72和96h尾静脉采血.测定雌鱼睾酮、雌二醇和孕酮,雄鱼孕酮和睾酬血清浓度.实验结果表明:注射LHRHa,雌鱼血清睾酮和雌二醇浓度显著高于对照组,雄鱼血清睾酮和孕酮显著高于对照组(P<0.05);24h浓度较高.雌鱼孕酬、睾酮和雌二醇分别为(0.710±0.082)ng/mL、(9.00±0.57)ng/mL和(696.4±26.2)pg/mL,雄鱼孕酮和睾酮分别为(0.527±0.121)ng/mL和(9.62±0.62)ng/mL.实验组雌鱼孕酮变化基本规律为,基础水平(0-7h)-逐渐升至最高(7-24h)-逐渐降至基础水平(24-48h)-维持基础水平(48-96h).实验组雌鱼睾酮和雌二醇与雄鱼孕酮和睾酮变化规律基本相似,其规律为,逐渐上升至最高(0-24h)-逐渐降至基础水平(24-72h)-维持基础水(72-96h).24h对照组雌鱼睾酮和雌二醇显著升高,浓度分别为:睾酮(2.20±0.18)ng/mL,雌二醇(269.1±36.6)pg/mL.对照组雄鱼血清孕酮和睾酮浓度实验期间均无显著变化.研究认为:LHRHa能够刺激泥鳅性类同醇激素分泌,特别是睾酮的分泌,显著提高雌鱼性腺指数(GSI).但刺激P的分泌调控能力有限,实验期间处于较低水平,诱导排卵效果差,泥鳅的性类固醇激素可能有特殊的调节机制.雌二醇和睾酮对性腺成熟有重要作用,孕酮对介导卵细胞最终成熟和排卵可能起重要作用,而雌二醇和睾酮无明显效果.     

秦岭山中的川金丝猴

川金丝猴(Rhinopithecus roxellana)是我国特有物种。秦岭地区是川金丝猴的主要分布区之一．也是金丝猴分布的最北限依据近年来对笼养川金丝猴繁殖生理学和野生川金丝猴野外实地观察研究的成果,并结合已发表的资料,对秦岭地区川金丝猴的社群组成、行为、等级序位、繁殖与生育等方面做了综述。     

六种藓类植物茎的比较解剖学研究

通过对6种藓类植物,即褶叶青藓(Brachythecium salebrosum(Web.et Mohr.)B.S.G.)、湿地匐灯藓(Plagiomnium acutum(Lindb.)Kop.)、侧枝匐灯藓(Plagiomnium maximoviczii(Lindb.)Kop.)、大凤尾藓(Fissidensnobilis Griff.)、大羽藓(Thuidium cymbifolium(Doz.et Molk.)B.S.G.)和大灰藓(Hypnum plumaeforme Wils.)嫩茎和老茎的石蜡切片和显微观察发现,同一藓类植株的嫩茎和老茎,茎结构稳定,不同种藓类植物茎横切面具有不同特征.植物体茎横切面形状、表层细胞的层数、细胞大小和细胞壁厚薄、皮层细胞大小和形状、中轴的有无以及比例等特征可以作为藓类植物的分科分类依据之一.     

Photoregulation of seed germination of wild-type and of an aurea-mutant of tomato

Seed germination of an aurea mutant of tomato ( Lycopersicon esculentum Mill.) is promoted by continuous irradiation with red, far-red or long-wavelength far-red (758 nm) light as well as by cyclic irradiations (5 min red or 5 min far-red/25 min darkness). Far-red light applied immediately after each red does not change the germination behaviour. Seed germination of the isogenic wild-type, cv. UC-105, is promoted by continuous and cyclic red light while it is inhibited by continuous and cyclic far-red light and by continious 758 nm irradiation. Far-red irradiation reverses almost completely the promoting effect of red light. The promoting effect (in the aurea mutant) and the inhibitory effect (in the wild-type) of continuous far-red light do not show photon fluence rate dependency above 20 nmol m⁻² s⁻¹. It is concluded that phytochrome controls tomato seed germination throgh low energy responses in both the wild type and the au mutant. The promoting effect of continuous and cyclic far-red light in the au mutant can be attributed to a greater sensitivity to P_fr.     

Cause of Senescence of Nine Sorts of Flowers

The levels of endogenous phytohormones and respiratory rate in nine sorts of flowers such as Cymbidium faberi Rolfe, Nopalxochia ackermannii Kunth and others were investigated both at full bloom and senescence and meanwhile the effect of exogenous phytohormones on prolonging the blossoms and promoting ethylene production were tested. There is a high content of endogenous ethylene in all the long-lived flowere, about 3–16 folds higer than the short-lived ones. There is a high level of ABA at full blooming flowers of short-lived flowers, in which there is no or only some cytokinins in it, but the ratio of CTK (6BA+zeatin)/ABA is smaller(l.7). The endogenous ABA reached a much higher level at senescence in all nine sorts of flowers, so it is reasonable to consider that it is ABA which plays an important role of regulation in controlling flower's senescence. There is a much higher level of GA3 and zeatin in the long-lived flowers which is not demonstrated in the shortlived ones. The respiratory rate is one of the factors controtling the longevity of flowers, but it does not play a decided role. Application of 6BA and zeatin prolongs distinctly orchid’s longevity, however exogenous IAA through the promotive action on ethylene production, evidently extends the longevity of the flowers of the Nopalxochia ackermannii Kunth.     

龙胆科药用植物化学成分的研究现状

龙胆科植物在我国的分布范围很广,且多数为药用植物,其多数种属的药用植物,至今其化学成分尚未被系统研究。综述了目前龙胆科药用植物的化学成分的研究现状及一般提取方法,对近年来发现的环烯醚萜及裂环烯醚萜类化合物进行了总结,为本科药用植物的更深入研究提供了参考。     

Interconnection of parameters of regulation system of lipid peroxidation and of morphophysiological parameters of mouse liver

A complex analysis of seasonal fluctuations of the mean group parameters of the system of regulation of lipid peroxidation has been performed in liver of Balb/c mice. Association of lipid characteristics and morphophysiological parameters is studied in the Balb/c mouse liver. An inter-connection is revealed between the liver index and the amount of lysoforms of phospholipids, the scale and character of the interconnection differing essentially depending on proportion of phos-phatidylcholine in mouse liver phospholipids.     

真菌类遗传学分析的知识结构教学

本文以认知结构理论为指导,讨论了真菌类遗传分析与高等动植物遗传分析的内在联系,认为利用这种内在联系进行教学可收到好的效果并说明了作者的具体教学过程。 Abstract:In the paper, the relationship between genetic analysis of Fungi and genetic analysis of high animal and plant was discussed.A good results were obtained when we adopted this method in the teaching.     

Physiological characteristics of various species of strains of carboxydobacteria

Seven strains of aerobic carbon monoxide-oxidizing bacteria (carboxydebacteria) when growing on CO as sole source of carbon and energy had doubling times which ranged from 12–42 h. The activity profiles obtained after discontinuous sucrose density gradient centrifugation indicated that the CO-oxidizing enzymes are soluble and the hydrogenases are membrane-bound in all strains examined. The CO-oxidizing enzymes of Pseudomonas carboxydohydrogena, Pseudomonas carboxydoflava, Comamonas compransoris, and the so far unidentified strains OM2, OM3, and OM4 had a molecular weight of 230,000; that of Achromobacter carboxydus amounted to 170,000. The molecular weights of the CO-oxidizing and H₂-oxidizing enzymes turned out to be identical. The cell sonicates were shown to catalyze the oxidation of both CO and H₂ with methylene blue, thionine, phenazine methosulfate, toluylene blue, dichlorophenolindophenol, cytochrome c or ferricyanide as electron acceptors. Methyl viologen, benzyl viologen, FAD⁺, FMN⁺, and NAD(P)⁺ were not reduced. The spectrum of electron acceptors was identical for all strains tested. Neither free formate, hydrogen nor oxygen gas were involved in the CO-oxidation reaction. Methylene blue was reduced by CO at a 1:1 molar ratio. The results indicate that CO-oxidation by carboxydobacteria is catalyzed by identical or similar enzymes and that the reaction obeys the equation CO+H₂OCO₂+2H⁺+2e^- as previously shown for Pseudomonas carboxydovorans.Dedicated to Otto Kandler remembering almost three decades of enjoyable cooperation     

Modulation of allostery of pyruvate kinase by shifting of an ensemble of microstates

Since the introduction of the concepts of allostery about four decades ago, much advancement has been made in elucidating the structure-function correlation in allostery. However, there are still a number of issues that remain unresolved. In this review we used mammalian pyruvate kinase (PK) as a model system to understand the role of protein dynamics in modulating cooperativity. PK has a triosephosphate isomerase (TIM)(α/β)₈ barrel structural motif. PK is an ideal system to address basic questions regarding regulatory mechanisms about this common (α/β)₈ structural motif. The simplest model accounting for all of the solution thermodynamic and kinetic data on ligand-enzyme interactions involves two conformational states, inactive E^T and active E^R. These conformational states are represented by domain movements. Further studies provide the first evidence for a differential effect of ligand binding on the dynamics of the structural elements, not major secondary structural changes. These data are consistent with our model that allosteric regulation of PK is the consequence of perturbation of the distribution of an ensemble of states in which the inactive E^T and active E^R represent the two extreme end states. Sequence differences and ligands can modulate the distribution of states leading to alterations of functions. The future work includes: defining the network of functionally connected residues; elucidating the chemical principles governing the sequence differences which affect functions; and probing the nature of mutations on the stability of the secondary structural elements, which in turn modulate allostery.