Metabolites of PPI-2458, a selective, irreversible inhibitor of methionine aminopeptidase-2: structure determination and In vivo activity

The natural product fumagillin exhibits potent antiproliferative and antiangiogenic properties. The semisynthetic analog PPI-2458, (3R,4S,5S,6R)-5-methoxy-4-(2R,3R)-2-methyl-3-(3-methylbut-2-enyl) oxiran-2-yl]-1-oxaspiro2.5]octan-6-yl] N-(2R)-1-amino-3-methyl-1-oxobutan-2-yl]carbamate, demonstrates rapid inactivation of its molecular target, methionine aminopeptidase-2 (MetAP2), and good efficacy in several rodent models of cancer and inflammation with oral dosing despite low apparent oral bioavailability. To probe the basis of its in vivo efficacy, the metabolism of PPI-2458 was studied in detail. Reaction phenotyping identified CYP3A4/5 as the major source of metabolism in humans. Six metabolites were isolated from liver microsomes and characterized by mass spectrometry and nuclear resonance spectroscopy, and their structures were confirmed by chemical synthesis. The synthetic metabolites showed correlated inhibition of MetAP2 enzymatic activity and vascular endothelial cell growth. In an ex vivo experiment, MetAP2 inhibition in white blood cells, thymus, and lymph nodes in rats after single dosing with PPI-2458 and the isolated metabolites was found to correlate with the in vitro activity of the individual species. In a phase 1 clinical study, PPI-2458 was administered to patients with non-Hodgkin lymphoma. At 15 mg administered orally every other day, MetAP2 in whole blood was 80% inactivated for up to 48 hours, although the exposure of the parent compound was only similar to 10% that of the summed cytochrome P450 metabolites. Taken together, the data confirm the participation of active metabolites in the in vivo efficacy of PPI-2458. The structures define a metabolic pathway for PPI-2458 that is distinct from that of TNP-470 ((3R, 4S, 5S, 6R)-5-methoxy-4-(2R, 3R)-2-methyl-3-(3-methylbut-2-enyl)oxiran-2-yl]-1-oxaspiro2.5]octan-6 -yl] N-(2-chloroacetyl)carbamate). The high level of MetAP2 inhibition achieved in vivo supports the value of fumagillin-derived therapeutics for angiogenic diseases.

Changes in the expression of DNA double strand break repair genes in primordial follicles from immature and aged rats

Oocytes present at birth undergo a progressive process of apoptosis in humans and other mammals as they age. Accepted opinion is that no fresh oocytes are produced other than those present at the time of birth. Studies have shown that DNA repair genes in oocytes of mice and women decline with age, and lack of these genes show higher DNA breaks and increased oocyte death rates. In contrast to the ethical problems associated with monitoring the changes in DNA double-strand breaks in oocytes from young and old humans, it is relatively easy to carry out such a study using a rodent model. In this study, the mRNA levels of DNA repair genes are compared with protein products of some of the genes in the primordial follicles isolated from immature (18-20 days) and aged (400-450 days) female rats. Results revealed a significant decline in mRNA levels of BRAC1 (P < 0.01), RAD51 (P < 0.05), ERCC2 (P < 0.05), and H2AX (P < 0.01) of DNA repair genes and phospho-protein levels of BRAC1 (P < 0.01) and H2AX (P < 0.05) in primordial follicles of aged rats. Impaired DNA repair is confirmed as a mechanism of oocyte ageing. (C) 2014 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.

Participação do propranolol no reparo tecidual de lesões isquêmicas cutâneas em ratos

Receptores adrenérgicos são amplamente expressos em diferentes tecidos, incluindo na pele. Recentemente foi descoberto que bloqueadores dos receptores β-adrenérgicos são capazes de modular o processo de reparo tecidual. O propranolol é um β-bloqueador não seletivo largamente utilizado na prática clínica para o tratamento de doenças cardiovasculares, parecendo apresentar propriedades antioxidantes. O objetivo desse estudo foi avaliar a resposta de diferentes doses de propranolol durante a lesão isquêmica cutânea em roedores. Ratos Wistar foram tratados com propranolol nas concentrações de 3 e 6 mg/kg. O grupo controle recebeu apenas o veículo e o grupo controle positivo recebeu vitamina E (50 mg/kg/dia). A administração do propranolol iniciou-se no dia da lesão, sendo realizada diariamente até o sacrifício. Incisões bilaterais foram feitas no dorso de cada animal. O flap foi suturado e foram realizadas lesões excisionais totais entre as lesões incisionais. A contração das lesões foi avaliada e os cortes histológicos foram corados com hematoxilina e eosina e vermelho de picrosirius. Foram utilizadas também as seguintes técnicas: Dopplerfluxometria, análises bioquímicas, análise estereológica e expressão de PECAM-1. O grupo tratado com 3 mg de propranolol apresentou uma maior redução na área total da lesão quando comparado ao grupo controle. Da mesma forma, este grupo apresentou um aumento na densidade de vasos sanguíneos, uma maior expressão de PECAM-1 e aumento na perfusão sanguínea na pele isquêmica e no sítio da lesão quando comparado ao grupo controle. Não observamos efeito antioxidante do propranolol neste modelo. Em resumo, nós sugerimos que o propranolol quando administrado na dose de 3 mg/kg/dia foi capaz de modular a angiogênese e melhorar o fechamento da lesão em modelo de roedores

Avaliação da aspirina e do ácido cafeico fenetil estersobre o reparo tecidual cutâneo

Numerosos modelos in vitro e in vivo foram desenvolvidos para estudar o reparo de lesões e identificar os mecanismos chave deste processo. Visando avaliar o processo de cicatrização utilizamos um modelo de lesão excisional total e um modelo de queimadura promovida por escaldamento. No estudo utilizando o modelo de lesão excisional total, abordamos o uso da aspirina (um inibidor não seletivo da COX) e seu efeito diferenciado sobre os sexos na cicatrização cutânea de camundongos. Observamos que os grupos fêmea controle e tratado apresentaram contração atrasada comparado aos grupos macho controle e tratado, respectivamente. Entre os grupos fêmea e macho controles, as fêmeas apresentaram menor atividade da mieloperoxidase e menor quantidade de células MIF-positivas do que os machos controle. Já entre os grupos fêmea e macho tratados, foi observado que nas fêmeas tratadas, a atividade da mieloperoxidase e a quantidade de macrófagos F4/80-positivos estavam maiores do que no grupo macho tratado. Ainda entre os grupos tratados, as fêmeas apresentaram menores níveis de hidroxiprolina e maior expressão proteica de vWF e VEGF comparado aos machos. No estudo das lesões causadas por queimadura, avaliamos as propriedades anti- inflamatórias e antioxidantes do ácido cafeico fenetil ester (CAPE) no reparo destas lesões e observamos que em 7, 14, 21 e 70 dias após a queimadura, o grupo queimado+CAPE apresentou menor área lesada, além de menor atividade da mieloperoxidase e dos níveis de nitrito do que o grupo queimado. Também foi observado que no grupo queimado+CAPE a expressão proteica de CD68 e de PECAM-1 estava reduzida comparada ao grupo queimado. Analisando os parâmetros de dano oxidativo foi observado que os níveis de MDA e de proteínas carboniladas estavam menores no grupo queimado+CAPE do que no grupo queimado, tanto no plasma quanto na lesão. Em suma, nosso estudo avaliou o processo de cicatrização de dois modelos de lesão, em roedores de diferentes espécies e em abordagens distintas. No modelo de lesão excisional total em camundongos observamos que a administração da aspirina prejudicou o processo de cicatrização em camundongos Balb/c fêmeas, mas não afetou esse processo nos machos da mesma linhagem, o que pode ser explicado pela redução dos níveis de estrógenos nas fêmeas. A utilização do CAPE na cicatrização de queimaduras em ratas comprovou sua eficácia anti-inflamatória e antioxidante devido ao grupo queimado+CAPE ter apresentado maior taxa de cicatrização, redução da resposta inflamatória, além da redução do dano oxidativo em lipídeos e proteínas comparado ao grupo queimado.

Avaliação das alterações macro-hemodinâmicas, microcirculatórias, gasométricas, metabólicas e inflamatórias secundárias à sedação com dexmedetomidina em um modelo experimental de endotoxemia em hamsters

Pela sua alta incidência, morbidade, mortalidade e custos ao sistema de saúde, a sepse se destaca entre as diversas indicações de internação em unidade de terapia intensiva (UTI). A disfunção da microcirculação tem papel central na gênese e manutenção da síndrome séptica, sendo um marco fisiopatológico desta síndrome. Pacientes críticos invariavelmente estão ansiosos, agitados, confusos, desconfortáveis e/ou com dor. Neste contexto, drogas sedativas são amplamente utilizadas na medicina intensiva. A dexmedetomidina, um agonista potente e altamente seletivo dos receptores alfa-2 adrenérgicos, vem conquistando espaço como o sedativo de escolha nas UTIs por seus efeitos de sedação consciente, redução da duração e incidência de delirium e preservação da ventilação espontânea. Apesar destas possíveis vantagens, a indicação de uso da dexmedetomidina na síndrome séptica ainda carece de conhecimentos sobre seus efeitos na microcirculação e perfusão orgânica. Com o intuito de caracterizar os efeitos microcirculatórios da dexmedetomidina em um modelo murino de endotoxemia que permite estudos in vivo da inflamação e disfunção da perfusão microvascular, hamsters Sírios dourados submetidos à endotoxemia induzida por administração intravenosa de lipopolissacarídeo de Escherichia coli (LPS, 1,0 mg.kg-1) foram sedados com dexmedetomidina (5,0 μg.kg.h-1). A microscopia intravital da preparação experimental (câmara dorsal) permitiu a realização de uma análise quantitativa das variáveis microvasculares e do rolamento e adesão de leucócitos à parede venular. Também foram analisados os parâmetros macro-hemodinâmicos e gasométricos (arterial e venoso portal), as concentrações de lactato arterial e venoso portal, a água pulmonar total e a sobrevivência do animal. Animais não-endotoxêmicos e/ou tratados com solução salina a 0,9% serviram como controles neste experimento. O LPS aumentou o rolamento e a adesão de leucócitos à parede venular, diminuiu a densidade capilar funcional e a velocidade das hemácias nos capilares e induziu acidose metabólica. O tratamento com dexmedetomidina atenuou significativamente estas respostas patológicas (p < 0,05). A frequência de pulso dos animais foi significativamente reduzida pela droga (p < 0,05). Outros resultados não foram tão expressivos (estatisticamente ou clinicamente). Estes resultados indicam que a utilização de dexmedetomidina produz um efeito protetor sobre a microcirculação da câmara dorsal de hamsters endotoxêmicos. Efeitos anti-inflamatórios da dexmedetomidina sobre os leucócitos e o endotélio poderiam melhorar a perfusão capilar e representar o mecanismo in vivo de ação da droga na microcirculação.

辽东栎天然更新

在北京东灵山地区对辽东栎的天然更新进行研究。辽东栎的结实具有大年现象（masting），大年现象十分复杂，在种群水平、个体之间都存在差异。 动物对辽东栎的天然更新有重要的影响。捕食和传播辽东栎坚果的脊椎动物至少包括小型啮齿目动物中的岩松鼠、花鼠、大仓鼠、社鼠、大林姬鼠和棕背鼠平，鸟类中的松鸦。岩松鼠、花鼠和大仓鼠有发达的颊囊，能够将坚果暂时放在颊囊中进行传播，岩松鼠有分散埋藏坚果的习性;松鸦能够将坚果放在素囊中进行长距离传播。成熟坚果在森林地表的消失非常快，少部分被脊椎动物就地消耗，大部分被传播到巢穴中或分散埋藏。成熟坚果的丢失存在时空差异。无脊椎动物中的昆虫对辽东栎去叶有重要影响。去叶与昆虫幼虫的密度以及长度显著相关。幼树去叶在辽东栎种群大树密度最高的地点最高，而在密度最低的地点最低。大树去叶在辽东栎种群总胸径面积最大的地点最大。 辽东栎实生苗在野外十分匮乏，可能是由于东灵山地区小型啮齿目动物缺乏天敌，使它们的活动十分强烈。它们有异常强大的捕食和搬运辽东栎坚果的能力，使能够逃逸捕食的坚果数量很少。现阶段辽东栎的更新几乎完全靠萌生苗进行，大树的生长大多弯曲不直。实生幼苗在小的林窗中的生长比在树冠下的生长好。综合考虑实生个体和萌生个体，50 龄林中种群大小结构稳定，能够长时间生存发展下去;30 龄林种群中个体间的竞争激烈，种群处于衰退状态。 研究结果建议：对辽东栎林的天然更新，应该采取综合治理的方法，充分考虑各种生物和非生物因素的影响，处理好生物因素的非生物因素的关系;人为进行适当干扰，例如在森林中创建适当大小的林窗，以更好地促进辽东栎的天然更新。

三峡大坝的修建对库区动物的影响

世界上最大的大坝—三峡大坝―于2003年6月正式建成并蓄水，三峡水库的水位由蓄水前的84m上升至蓄水后的139m。三峡大坝的修建与蓄水，已经显著地改变了三峡库区陆地生态系统和水域生态系统的生态环境。三峡大坝的修建将如何影响生活在这一区域（包括陆地和水域）的野生动植物资源，是个亟待解决的问题。 对于陆地生态系统，三峡大坝的修建，使三峡库区的生态环境进一步破碎化（Fragmentation）和岛屿化（Island）。为了了解三峡大坝的修建对三峡库区野生动物所产生的影响，我们选择适应性强且对环境变化敏感的啮齿动物为研究对象，在三峡库区选取两个岛屿（洛碛岛和皇华岛）及其对岸为研究点，在野外调查了啮齿动物的群落组成、种群密度与分布，并用胃容物分析和稳定性同位素分析方法研究了两个研究点优势种啮齿动物的食物来源与组成。结果表明，两个研究点间啮齿动物群落组成有很大差异，这主要是由于啮齿动物分布的地域性差异引起的。在同一个研究点，岛屿上的啮齿动物群落组成与对岸的没有差异，但在岛屿上的啮齿动物种群密度却明显高于其对岸的，这说明随着库区水位的大幅度提升，岛屿的面积急剧减小，使原本栖息在水边的啮齿动物不得不向岛屿的中上部迁移，致使岛屿上的啮齿动物的种群密度迅速增加。啮齿动物的食物来源有四种：C3非豆科植物，C3豆科植物，C4植物及水生生物。不同种啮齿动物的食物种类组成也不同，四川短尾鼩（Anourosorex squamipes）的食物中有水生生物的组成成分，但其它啮齿动物食物中不含水生生物。不同食物来源在啮齿动物的整体食物中所占比例也不同，并呈明显的季节性变化，这种变化与田间的农作物种类密切相关。动物组织的稳定性同位素组成不仅可以示踪动物的食物来源，也反映了动物的生存状况。我们的结果显示：对岸啮齿动物的稳定性同位素组成相对集中，并且不同种啮齿动物间没有重叠。这一方面说明对岸啮齿动物的食物来源相对丰富，啮齿动物可以选择各自喜食食物，在此环境下的啮齿动物种间竞争相对较弱，至少是种间食物竞争较弱。岛屿上啮齿动物的稳定性同位素值分布范围远大于在对岸的，并且种间稳定性同位素组成有明显的重叠部分。稳定性同位素组成的重叠意味着有相似的食物来源，也即栖息生境相似。这说明岛上的啮齿动物种间存在较剧烈的食物竞争关系。在资源限制情况下，多种啮齿动物不得不利用共同的食物来源。换句话说，岛屿化过程将加剧岛屿上啮齿动物的种间竞争。 洛碛岛上的四川短尾鼩因为可以利用水生食物来源，这使得它在岛屿化过程中处于有利地位。随着岛屿化进程的加剧，四川短尾鼩在岛屿上的优势将更加明显。而皇华岛屿上的褐家鼠的食物来源单一，会因岛屿面积的进一步减小，食物来源更加缺乏，它们将不得不改其食物组成或面临消失。而白腹巨鼠的栖息环境靠近江边，将因水位的上升进一步上移，它们的栖息环境与普通田鼠的发生重叠，使得白腹巨鼠与普通田鼠发生栖息地的竞争与食物竞争，并面临更多的人为因素干扰。 对于水域生态系统，长江是我国最大的河流，也是我国淡水鱼类最丰富的区域。三峡大坝的修建已经显著改变了三峡库区的水文特征。为了了解三峡大坝的修建是否会改变三峡库区的有机物组成，并进而影响到三峡库区水生生物的食物来源和组成，我们选择了三个水文特征不同的研究点（洛碛江段、皇华城江段和茅坪江段）调查了三峡库区的常见鱼种类组成，并用稳定性同位素方法研究了三峡库区洪水前后的有机物组成变化与鱼类的食物网模型，用稳定性同位素划分了鱼类食物网结构及鱼类体长与其肌肉δ15N值间的关系。结果表明，三峡大坝的修建，已经显著改变了皇华城和茅坪江段的水文特征，同时也“干扰”了生活在这一区域的鱼类。适应于流水环境的鱼类在库区回水处和大坝附近几乎消失，而喜欢静态环境的其它鱼类却得到极大的发展，例如鲢鱼（H. molitrix）和草鱼（ C. idellus）。 洪水前后，三峡库区的有机物组成成分有明显改变。洪水前，水中有机物主要以河流自身生产力产生的有机物为主（浮游植物、藻类等）;洪水后，水中有机物主要以外来有机物为主（陆地植物、土壤有机物和从上游带来的有机物）。对于三个食物网模型：河流连续体模型（RCC）、脉冲模型（FPC）和河流生产力模型（RPM），河流生产力模型能更好的解释三峡库区的水生生物的食物来源，即三峡库区的水生生物的食物主要来源于河流自身生产力产生的有机物。但外来有机物作为水生生物的一种辅助食物来源，在洪水期间起到不可忽视的作用。 在本研究中，鱼类体长与其δ15N值间的关系与选取鱼的种类有关，比如南方大口鲇（Silurus asotus）的体长与其肌肉的δ15N值呈明显负相关关系（R2≈0.5），而鱖鱼（Siniperca）、铜鱼（Coreius guichenoti）和草鱼（Ctenopharyngodon idellus）的体长与其肌肉的δ15N值却呈正相关关系。 三峡库区常见鱼类主要以杂食性和广普性鱼类为主，食物网结构十分复杂。通过胃容物分析和稳定性同位素分析，三峡库区水生生物营养级间的判别值约为3.1‰。依据鱼类肌肉的δ15N值，三峡库区的常见鱼类可以划分为四个营养级：草食性鱼类（herbivorous fish）、初级杂食性鱼类（1ºomnivorous fish）、次级杂食性鱼类（2ºomnivorous fish）和食鱼性鱼类（piscivorous fish）。营养级间没有明确的分界限，鱼类的营养级从2到4.8，是连续分布的营养级结构，从而更真实的反映了自然界中动物的捕食关系和在食物网中的位置。 自然丰度变化的稳定性同位素(Stable isotope)作为一种天然的示踪物，在动物生态学上已经得到广泛应用。动物的稳定性同位素可以清楚的示踪动物的食物来源、食物组成、栖息地情况和生存状况等多种信息，结合传统的胃容物分析，或其它的粪便分析、储藏物分析等，稳定性同位素技术在动物生态学研究方面必将得到更广泛的应用。

Composition and evolution of the V2r vorneronasal receptor gene repertoire in mice and rats

Pheromones are chemicals produced and detected by conspecifics to elicit social/sexual physiological and behavioral responses, and they are perceived primarily by the vomeronasal organ (VNO) in terrestrial vertebrates. Two large superfamilies of G protein-coupled receptors, V1rs and V2rs, have been identified as pheromone receptors in vomeronasal sensory neurons. Based on a computational analysis of the mouse and rat genome sequences, we report the first global draft of the V2r gene repertoire, composed of similar to 200 genes and pseudogenes. Rodent V2rs are subject to rapid gene births/deaths and accelerated amino acid substitutions, likely reflecting the species-specific nature of pheromones. Vertebrate V2rs appear to have originated twice prior to the emergence of the VNO in ancestral tetrapods, explaining seemingly inconsistent observations among different V2rs. The identification of the entire V2r repertoire opens the door to genomic-level studies of the structure, function, and evolution of this diverse group of sensory receptors. (c) 2005 Elsevier Inc. All rights reserved.

Stepwise loss of motilin and its specific receptor genes in rodents

Specific interactions among biomolecules drive virtually all cellular functions and underlie phenotypic complexity and diversity. Biomolecules are not isolated particles, but are elements of integrated interaction networks, and play their roles through specific interactions. Simultaneous emergence or loss of multiple interacting partners is unlikely. If one of the interacting partners is lost, then what are the evolutionary consequences for the retained partner? Taking advantages of the availability of the large number of mammalian genome sequences and knowledge of phylogenetic relationships of the species, we examined the evolutionary fate of the motilin (MLN) hormone gene, after the pseudogenization of its specific receptor, MLN receptor (MLNR), on the rodent lineage. We speculate that the MLNR gene became a pseudogene before the divergence of the squirrel and other rodents about 75 mya. The evolutionary consequences for the MLN gene were diverse. While an intact open reading frame for the MLN gene, which appears functional, was preserved in the kangaroo rat, the MLN gene became inactivated independently on the lineages leading to the guinea pig and the common ancestor of the mouse and rat. Gain and loss of specific interactions among biomolecules through the birth and death of genes for biomolecules point to a general evolutionary dynamic: gene birth and death are widespread phenomena in genome evolution, at the genetic level; thus, once mutations arise, a stepwise process of elaboration and optimization ensues, which gradually integrates and orders mutations into a coherent pattern.

Improving effects of huperzine A on spatial working memory in aged monkeys and young adult monkeys with experimental cognitive impairment

Our previous studies demonstrated that huperzine A, a reversible and selective acetylcholinesterase inhibitor, exerts beneficial effects on memory deficits in various rodent models of amnesia. To extend the antiamnesic action of huperzine A to nonhuman primates, huperzine A was evaluated for its ability to reverse the deficits in spatial memory produced by scopolamine in young adult monkeys or those that are naturally occurring in aged monkeys using a delayed-response task. Scopolamine, a muscarinic receptor antagonist, dose dependently impaired performance with the highest dose (0.03 mg/kg, i.m.) producing a significant reduction in choice accuracy in young adult monkeys. The delayed performance changed from an average of 26.8/30 trials correct on saline control to an average of 20.2/30 trials correct after scopolamine administration. Huperzine A (0.01-0.1 mg/kg, i.m.) significantly reversed deficits induced by scopolamine in young adult monkeys on a delayed-response task; performance after an optimal dose (0.1 mg/kg) averaged 25.0/30 correct. In four aged monkeys, huperzine A (0.001-0.01 mg/kg, i.m.) significantly increased choice accuracy from 20.5/30 on saline control to 25.2/30 at the optimal dose (0.001 mg/kg for two monkeys and 0.01 mg/kg for the other two monkeys). The beneficial effects of huperzine A on delayed-response performance were long lasting; monkeys remained improved for about 24 h after a single injection of huperzine A. This study extended the findings that huperzine A improves the mnemonic performance requiring working memory in monkeys, and suggests that huperzine A may be a promising agent for clinical therapy of cognitive impairments in patients with Alzheimer's disease.

Molecular and evolutionary analyses of formyl peptide receptors suggest the absence of VNO-specific FPRs in primates

Formyl peptide receptors (FPRs) were observed to expand in rodents and were recently suggested as candidate vomeronasal chemosensory receptors. Since vomeronasal chemosensory receptors usually underwent positive selection and evolved concordantly with the vomeronasal organ (VNO) morphology, we surveyed FPRs in primates in which VNO morphology is greatly diverse and thus it would provide us a clearer view of VNO-FPRs evolution. By screening available primate genome sequences, we obtained the FPR repertoires in representative primate species. As a result, we did not find FPR family size expansion in primates. Further analyses showed no evolutionary force variance between primates with or without VNO structure, which indicated that there was no functional divergence among primates FPRs. Our results suggest that primates lack the VNO-specific FPRs and the FPR expansion is not a common phenomenon in mammals outside rodent lineage, regardless of VNO complexity.

Effect of plateau zokors on vegetation characteristics and productivity of alpine meadow

This research was conducted on alpine meadow site at Menyuan county, Qinghai Province, People's Republic of China to determine the effects of native, subterranean rodent of Qinghai-Tibet grasslands, the plateau zokors (Myospalax baileyi), on seasonal above-and below-ground plant biomass, plant species diversity and productivity. Both total peaks of above-and below-ground biomass were the greatest (413.600 g/m~2 and 2297.502 g/m~2) in the patch no any plateau zokors colonized by plateau zokors over 10 years in August and October, respectively. Both above-and below-ground biomass were significantly increased in the patches where plateau zokors were removed or the burrow systems were abandoned for five years compared to the patches plateau zokors colonized over 10 years. However, both above-and below-ground biomass in abandoned patches were significantly lower than that in uncolonized patches. Monocotyledonous biomass was reduced greatly, but the non-palatable dicots were significantly increased in colonized patches. The palatable biomass of monocots and dicots were increased in abandoned patches. Total plant species diversity was the greatest in uncolonized patchesand least in abandoned patch. The total net primary production in colonized patches was reduced by 68.98% compared with uncolonized patches. Although the patches were without any plateau zokors disturbance for fives years, the total net primary production just reached 58.69% of the uncolonized patches. The above-ground net primary production in abandoned patches increased 28.74% and the below-ground increased 54.91% compared with the colonized patches. We suggest that plateau zokor-induced changes in plant above- and below-ground biomass and species diversity may lead to further alterations of nutrient cycling and trophic dynamics in this alpine meadow ecosystem.

Genetic diversity of the subterranean Gansu zokor in a semi-natural landscape

We studied the relationship between genetic diversity of the subterranean Gansu zokor Myospalax cansus and habitat variability in the Loess Plateau, Qinghai Province, China. We used a combination of geographic information systems and molecular techniques to assess the impact of habitat composition and human activities on the genetic diversity of zokor populations in this semi-natural landscape. Although they occurred relatively infrequently in the landscape, woodland and high-coverage grassland habitats were the main positive contributors to the genetic diversity of zokor populations. Rural residential land, plain agricultural land and low-coverage grassland had a negative effect on genetic diversity. Hilly agricultural land and middle-coverage grassland had little impact on zokor genetic diversity. There were also interactions between some habitat types, that is, habitat types with relatively better quality together promoted conservation of genetic diversity, while the interaction between (among) bad habitat types made situations worse. Finally, habitat diversity, measured as patch richness and Shannon's diversity index, was positively correlated with the genetic diversity. These results demonstrated that: (1) different habitat types had different effects on the genetic diversity of zokor populations and (2) habitat quality and habitat heterogeneity were important in maintaining genetic diversity. Habitat composition was closely related to land use thus emphasizing the importance of human activities on the genetic diversity of subterranean rodent populations in this semi-natural landscape. Although the Gansu zokor was considered to be a pest species in the Loess Plateau, our study provides insights for the management and conservation of other subterranean rodent species.

Summer diet of two sympatric species of raptors Upland Buzzard (Buteo hemilasius) and Eurasian Eagle Owl (Bubo bubo) in alpine meadow: problem of coexistence

Summer diets of two sympatric raptors Upland Buzzards (Buteo hemilasius Temminck et Schlegel) and Eurasian Eagle Owls (Bubo bubo L. subsp. Hemachalana Hume) were studied in an alpine meadow (3250 m a.s.l.) on Qinghai-Tibet Plateau, China. Root voles Microtus oeconomus Pallas, plateau pikas Ochotona curzoniae Hodgson, Gansu pikas O. cansus Lyon and plateau zokors Myospalax baileyi Thomas were the main diet components of Upland Buzzards as identified through the pellets analysis with the frequency of 57, 20, 19 and 4%, respectively. The four rodent species also were the main diet components of Eurasian Eagle Owls basing on the pellets and prey leftovers analysis with the frequency of 53, 26, 13 and 5%, respectively. The food niche breadth indexes of Upland Buzzards and Eurasian Eagle Owls were 1.60 and 1.77 respectively (higher value of the index means the food niche of the raptor is broader), and the diet overlap index of the two raptors was larger (C-ue = 0.90) (the index range from 0 - no overlap - to I - complete overlap). It means that the diets of Upland Buzzards and Eurasian Eagle Owls were similar (Two Related Samples Test, Z = -0.752, P = 0.452). The classical resource partitioning theory can not explain the coexistence of Upland Buzzards and Eurasian Eagle Owls in alpine meadows of Qinghai-Tibet Plateau. However, differences in body size, predation mode and activity rhythm between Upland Buzzards and Eurasian Eagle Owls may explain the coexistence of these two sympatric raptors.

Analysis of diets of upland buzzards using stable carbon and nitrogen isotopes

We measured the stable carbon and nitrogen isotope ratios for muscles of the upland buzzards (Buteo hemilasius) and their potential food sources, plateau pikas (Ochotona curzoniae), Qinghai voles (Lasiopodomys fuscus), plateau zokors (Myospalax fontanierii), and several passerine bird species at the alpine meadow in Maduo county, Guoluo prefecture of Qinghai province, People's Republic of China, to provide diet information of upland buzzards, highlighting different diet composition of upland buzzards exposed to different locations. The results demonstrated that stable carbon isotope ratios of upland buzzards, passerine birds, plateau pikas, plateau zokors, and Qinghai voles were -24.42 +/- 0.25parts per thousand, -22.89 +/- 1.48parts per thousand, -25.30 +/- 1.47parts per thousand, -25.78 +/- 0.22parts per thousand, and -25.41 +/- 0.01parts per thousand, respectively, and stable nitrogen isotope ratios were 7.89 +/- 0.38parts per thousand, 8.37 +/- 2.05parts per thousand, 5.83 +/- 1.10parts per thousand, 5.23 +/- 0.34parts per thousand, and 8.86 +/- 0.06parts per thousand, respectively. Fractionation of stable carbon and nitrogen isotope ratios between upland buzzards and their food were 1.03parts per thousand and 2.11parts per thousand, respectively. Based on mass balance principle of stable isotopes and the Euclidean distance mixing model, upland buzzards depended mainly on plateau pikas as food (74.56%). Plateau zokors, Qinghai voles, and passerine birds only contributed a small proportion (25.44%) to diets of upland buzzards. The results were closely accordant with analyses of stomach contents and food pellets, which firmly supported the feasibility of using stable carbon and nitrogen isotope ratios to investigate diet information of upland buzzards. Another study based on stable carbon isotopes showed that upland buzzards living in the Haibei prefecture (another prefecture located in the southeast Qinghai province) mainly preyed on passerine birds (64.96% or more) as food supply. We were alarmed by the preliminary results that widespread poisoning activities of small mammals could reshape the food composition of upland buzzards, influencing the stability and sustainability of the alpine meadow. Bio-control on rodent pests should be carried out rather than the chemical measures.