家蝇幼虫血细胞类型及免疫功能的初步研究

目的用不同方法观察家蝇3龄幼虫血细胞的形态,并对血细胞进行分类和免疫功能研究,为昆虫血细胞形态、分类及免疫研究提供实验依据。方法 (1)应用姬氏染色结合相差显微镜及荧光染色方法观察家蝇3龄幼虫血细胞形态,并对家蝇3龄幼虫血细胞进行分类;(2)观察家蝇3龄幼虫感染大肠杆菌后不同时间血细胞总数(THC)、各类血细胞数量(DHC)及形态的变化;(3)应用倒置显微镜观察家蝇3龄幼虫离体血细胞感染大肠杆菌后的形态变化;(4)采用酶细胞化学技术测定感染前后家蝇3龄幼虫血细胞中ACP、POD活性的变化。结果 (1)家蝇3龄幼虫血细胞可分为原血胞、浆血胞、粒血胞、珠血胞、类绛血胞5类,其中浆血胞又分为大核浆血胞和小核浆血胞两种;(2)感染后各时间组血细胞总数、浆血胞和粒血胞数量均显著升高,且浆血胞和粒血胞聚集成堆,出现细胞变形、空泡等形态变化;感染后16h、24h组的珠血胞数显著升高;原血胞和类绛血胞数量和形态无明显变化;(3)家蝇幼虫离体血细胞感染大肠杆菌后粒血胞周围见大量细菌聚集,浆血胞、粒血胞聚集成团将细菌包裹形成包囊,未见原血胞、珠血胞、类绛血胞形态的变化;(4)感染后浆血胞和粒血胞中ACP、POD的活性增强,感染前后原血胞、珠血胞、类绛血胞中均未见ACP、POD的阳性反应物。结论通过3种方法能很好地将家蝇3龄幼虫血细胞分为5类,其中浆血胞和粒血胞是家蝇幼虫参与免疫反应的主要细胞类型,珠血胞不参与感染后的早期细胞免疫反应。     

幼虫密度对草地螟血细胞数量和组成的影响

为了阐明幼虫密度对草地螟Loxostege sticticalis L.（鳞翅目： 螟蛾科）细胞免疫能力的影响, 本研究调查了在活体灰菜植株上1,5,10和20头/瓶（900 mL）4种密度条件下的其5龄幼虫血细胞种类、数量和组成。结果表明： 草地螟幼虫血淋巴中有原血细胞、浆血细胞、 颗粒血细胞、珠血细胞和类绛色血细胞等5种（类）血细胞。血细胞总数、 浆血细胞、颗粒血细胞数量随幼虫密度的增加而显著递增, 但原血细胞、珠血细胞和类绛色血细胞数量在幼虫密度间的差异不明显;各种血细胞所占血细胞总数的比例在4个密度中的排序相同, 但10和20头/瓶密度下的浆血细胞比例显著高于1头/瓶的,其余4种血细胞的比例在不同密度之间无显著差异。可见, 幼虫密度主要是通过影响草地螟幼虫浆血细胞和颗粒血细胞的数量及血细胞总数, 从而影响草地螟的细胞免疫能力。     

阿尔泰蝠蛾幼虫血细胞形态学研究

本研究采用Wright-Giemsa染色方法,利用光学显微镜观察阿尔泰蝠蛾Hepialus altaicola Wang幼虫血细胞.根据鳞翅目昆虫血细胞形态特征对阿尔泰蝠蛾幼虫血细胞进行鉴定,计数并计算各类血细胞所占的比例.发现阿尔泰蝠蛾幼虫血淋巴中存在原血胞、浆血胞、粒血胞、类绛色细胞、珠血胞和囊血胞6种细胞类型,所...     

感染苏云金杆菌后印度谷螟血细胞的病态变化

光学显微镜下观察苏云金杆菌感染印度谷螟后幼虫血细胞的病态变化证明:幼虫感病后,原血胞、粒血胞、浆血胞、珠血胞、类绛血胞、囊血胞均受到破坏,多数血胞出现空泡和变形,珠血胞珠粒之间互相脱离,细胞质外流,细胞膜破裂,囊血胞则有细胞外溢现象。这些病态变化均可在感病8小时以后观察到。     

金龟子绿僵菌及其粗毒素对樟巢螟幼虫的致病性

【目的】为了筛选感染樟巢螟Orthaga achatina幼虫的高致病力金龟子绿僵菌Metarhizium anisopliae菌株,并研究绿僵菌粗毒素对幼虫的毒力和取食粗毒素后幼虫的血淋巴细胞免疫反应。【方法】以死亡率 时间几率值法和TDM模型分析绿僵菌及其粗毒素对樟巢螟幼虫的致病力,并显微观察处理幼虫的血淋巴细胞的变化。【结果】绿僵菌菌株Ma1291-2对樟巢螟幼虫有较强的致病力,以浓度(1.0±0.5)×108个孢子/mL的孢子悬液接菌11 d后,幼虫的校正死亡率和僵虫率分别为99.8%±2.6%和86.9%±1.3%,LT₅₀为6.29 d。各菌株产粗毒素水平与其对幼虫的校正死亡率和LT₅₀呈显著相关。通过时间-剂量-死亡率模型参数估算,Ma1291-2菌株及其粗毒素对幼虫致死效应较强的时间段分别为接菌后6-7 d和3-4.5 d。幼虫取食绿僵菌粗毒素后2 d,总血细胞、浆血细胞、珠血细胞和类绛血细胞浓度上升到最高值,而后下降;粒血细胞浓度2 d后开始极显著上升,第3天达到最高值后急速下降;原血细胞前3 d未发现有明显的数量变化,第4天显著下降。幼虫取食粗毒素3~4 d后,浆血细胞和粒血细胞有破裂,珠血细胞和类绛血细胞有黑化现象,原血细胞病态变化不明显。【结论】樟巢螟幼虫取食绿僵菌粗毒素后2-3 d,幼虫血淋巴细胞对粗毒素的免疫反应最强烈,且粗毒素对血细胞有毒害和破坏作用。本研究为该害虫的生物防治提供了一定的理论与应用基础。     

东亚飞蝗血细胞形态学研究

通过Wright's染色和光学显微镜对不同地区东亚飞蝗的血细胞进行了观察,发现东亚飞蝗Locusta migratoria manilensis(Meyen)的血细胞包括原血胞、浆血胞、粒血胞和类绛血胞4种类型.原血胞的细胞核为红色,而其它3种血细胞的细胞核均被Wright's染液染成红色和蓝色2种类型.故认为浆血胞是...     

黑翅痂蝗血细胞形态学的研究

【目的】蝗虫血细胞的研究以及不同血细胞类型间的演化关系,是蝗虫细胞免疫学的依据,也是防蝗治蝗的重要依据。【方法】本研究通过Wright-Giemsa对黑翅痂蝗Bryodema nigroptera Zheng血细胞进行了混合染色,并使用光学显微镜进行了观察。【结果】发现黑翅痂蝗血细胞包括原血胞、浆血胞、粒血胞、类绛色血细胞、囊血胞、脂血胞、蠕形细胞和巨核血细胞8种类型。【结论】在成虫体内,原血胞占细胞总数的1%,浆血胞占32.2%,粒血胞占57.3%,类绛色血细胞占1%,囊血胞占6%,脂血胞占1%,蠕形细胞占1%,巨核细胞占细胞总数的0.5%,粒血胞数量最多,巨核细胞最少。     

小金蝠蛾幼虫血细胞的种类、形态及细胞免疫功能

【目的】本研究旨在鉴定小金蝠蛾Thitarodes xiaojinensis幼虫血细胞类型,统计各类血细胞所占比例并检测其细胞免疫能力。【方法】利用光学相差显微镜观察小金蝠蛾7龄幼虫血细胞,并根据鳞翅目昆虫血细胞特有形态对小金蝠蛾幼虫血细胞进行鉴定;利用96孔板对小金蝠蛾各类血细胞进行计数并计算各类血细胞所占的比例;利用酿酒酵母Saccharomyces cerevisiae以及Congo Red染色的DEAE-Sephadex A-25凝胶珠刺激小金蝠蛾7龄幼虫,检验其细胞免疫反应。【结果】小金蝠蛾幼虫血细胞由粒细胞、浆细胞、珠血细胞和类绛色细胞组成,所占比例分别为62.5%,26.6%,10.1%和0.8%,未发现典型的原血细胞。酿酒酵母刺激结果显示,小金蝠蛾幼虫血细胞可以发生吞噬以及结节反应。Congo Red染色的DEAE-Sephadex A-25凝胶珠刺激结果显示,小金蝠蛾幼虫血细胞可以发生包囊反应。【结论】小金蝠蛾幼虫血细胞组成及其形态与其他鳞翅目昆虫一致,其中粒细胞所占的比例最高。小金蝠蛾幼虫血细胞对酿酒酵母刺激敏感,可发生吞噬和结节反应,对Congo Red染色的DEAE-Sephadex A-25凝胶珠刺激敏感,可发生包囊反应。小金蝠蛾细胞免疫功能完善。     

粘虫 Pseudaletia separata (Walk.)血细胞的类型辨识和病态变化

利用相差显微镜,并辅以涂片染色、化学反应方法,将粘虫各虫态虫龄的血细胞分为8个类型。他们是:原血胞、浆血胞、粒血胞、珠血胞、类绛血胞、囊血胞、梭血胞及脂血胞。并统计出各类血细胞在粘虫一生中的数量变化。本文认为足血胞与粒血胞不同之处在于前者的伸延部分含有细胞质。符合这一标准的足血胞在此次工作中看到的不多,因此对粘虫足血胞的存在问题暂作保留。 苏芸金杆菌处理后,粘虫血细胞产生了明显的病态变化,随着病症的进展,非正常血胞自2%上升为30%,50%,83%左右。直到死亡,血淋巴内未发现杆菌出现。     

转cry1Ab基因水稻对二化螟幼虫血细胞的影响

研究了转cry1Ab基因水稻“克螟稻1号”对二化螟Chilo suppressalis幼虫细胞免疫系统的影响。结果表明,转cry1Ab基因水稻对二化螟幼虫的血细胞影响明显,取食转cry1Ab基因水稻后,二化螟幼虫各类血细胞都明显低于取食非转基因水稻“秀水11”的对照组（原血细胞和囊血细胞在取食初期例外）,随取食时间延长,各类血细胞数量及血细胞总数均呈递减的趋势。从各类血细胞所占血细胞总数的百分比来看,原血细胞在取食36 h后锐减,而浆血细胞和粒血细胞则比例增加,其余珠血细胞、囊血细胞的变化不明显。另外,血细胞还出现空泡化、肿胀等病态变化,致使血细胞快速破裂。由此推测转cry1Ab基因水稻自身表达的毒蛋白能严重干扰靶标昆虫二化螟幼虫的细胞免疫系统。     

Effects of increasing the number of players and memory size in the iterated Prisoner's Dilemma: a numerical approach

The Prisoner''s Dilemma has become a paradigm for the evolution of altruistic behaviour. Here we present results of numerical simulations of the infinitely iterated stochastic simultaneous Prisoner''s Dilemma considering players with longer memory, encounters of more than two players as well as different pay-off values. This provides us with a better foundation to compare theoretical results to experimental data. We show that the success of the strategy Pavlov, regardless of its simplicity, is far more general by having an outstanding role in the iterated N-player N-memory Prisoner''s Dilemma. Besides, we study influences of increased memory sizes in the iterated two-player Prisoner''s Dilemma, and present comparisons to results of experiments with first-year students.     

Loss of Yme1L perturbates mitochondrial dynamics

Yme1L is an AAA protease that is embedded in the mitochondrial inner membrane with its catalytic domain facing the mitochondrial inner-membrane space. However, how Yme1L regulates mammalian mitochondrial function is still obscure. We find that endogenous Yme1L locates at punctate structures of mitochondria, and that loss of Yme1L in mouse embryonic fibroblast (MEF) cells results in mitochondrial fragmentation and leads to significant increased ‘kiss-and-run'' type of mitochondrial fusion; however, Yme1L knockdown (shYme1L (short hairpin-mediated RNA interference of Yme1L)) cells still remain normal mitochondrial fusion although shYme1L mitochondria have a little bit less fusion and fission rates, and the shYme1L-induced fragmentation is due to a little bit more mitochondrial fission than fusion in cells. Furthermore, shYme1L-induced mitochondrial fragmentation is independent on optic atrophy 1 (OPA1) S1 or S2 processing, and shYme1L results in the stabilization of OPA1 long form (L-OPA1); in addition, the exogenous expression of OPA1 or L-OPA1 facilitates the shYme1L-induced mitochondrial fragmentation, thus this fragmentation induced by shYme1L appears to be associated with L-OPA1''s stability. ShYme1L also causes a slight increase of mitochondrial dynamics proteins of 49 kDa and mitochondrial fission factor (Mff), which recruit mitochondrial key fission factor dynamin-related protein 1 (Drp1) into mitochondria in MEF cells, and loss of Drp1 or Mff inhibits the shYme1L-induced mitochondrial fragmentation. In addition, there is interaction between SLP-2 with Yme1L and shYme1L cells retain stress-induced mitochondrial hyperfusion. Taken together, our results clarify how Yme1L regulates mitochondrial morphology.     

Hamilton's rule meets the Hamiltonian: kin selection on dynamic characters

Many biological characters of interest are temporal sequences of decisions. The evolution of such characters is often modelled using dynamic optimization methods such as the maximum principle. A quantity central to these analyses is the ''Hamiltonian'' function, named after the mathematician William R. Hamilton. On the other hand, evolutionary models in which individuals interact with relatives are usually based on Hamilton''s rule, named after the evolutionary biologist William D. Hamilton. In this article we present a generalized maximum principle that includes the effects of interactions among relatives and we show that a time-dependent (dynamic) version of Hamilton''s rule holds involving the Hamiltonian. This result brings together the power and generality of both the maximum principle and Hamilton''s rule thereby providing a natural framework for understanding the evolution of ''dynamic'' characters under kin selection.     

Morphological variations in a widely distributed Eastern Asian passerine cannot be consistently explained by ecogeographic rules

Ecogeographic rules that describe quantitative relationships between morphologies and climate might help us predict how morphometrics of animals was shaped by local temperature or humidity. Although the ecogeographic rules had been widely tested in animals of Europe and North America, they had not been fully validated for species in regions that are less studied. Here, we investigate the morphometric variation of a widely distributed East Asian passerine, the vinous‐throated parrotbill (Sinosuthora webbiana), to test whether its morphological variation conforms to the prediction of Bergmann''s rule, Allen''s rules, and Gloger''s rule. We at first described the climatic niche of S. webbiana from occurrence records (n = 7838) and specimen records (n = 290). The results of analysis of covariance (ANCOVA) suggested that the plumage coloration of these parrotbills was darker in wetter/warmer environments following Gloger''s rule. However, their appendage size (culmen length, beak volume, tarsi length) was larger in colder environments, the opposite of the predictions of Allen''s rule. Similarly, their body size (wing length) was larger in warmer environments, the opposite of the predictions of Bergmann''s rule. Such disconformity to both Bergmann''s rule and Allen''s rule suggests that the evolution of morphological variations is likely governed by multiple selection forces rather than dominated by thermoregulation. Our results suggest that these ecogeographic rules should be validated prior to forecasting biological responses to climate change especially for species in less‐studied regions.     

巨噬细胞特异性启动子/抗菌肽PR39基因腺病毒载体的构建及其表达

旨在构建由巨噬细胞特异性启动子调控的抗菌肽PR39基因腺病毒表达载体,检测目的基因在小鼠巨噬细胞RAW264.7中的表达。PCR扩增巨噬细胞启动子/抗菌肽PR39成熟肽基因序列,插入腺病毒穿梭载体pAdTrack中,重组穿梭质粒(pAdTrack-SP/PR39)与腺病毒骨架(pAdEasy-1)共转化大肠杆菌BJ5183,通过细菌内同源重组产生重组腺病毒载体(pAd-SP/PR39)。pAd-SP/PR39经PacI线性化后转染293细胞,包装出重组腺病毒(Ad-SP/PR39)。Ad-SP/PR39感染小鼠巨噬细胞RAW264.7,经RT-PCR和免疫细胞化学检测PR39的靶向表达特异性。成功构建了由巨噬细胞特异性启动子调控的抗菌肽PR39基因腺病毒表达载体。包装出的重组腺病毒仍然能感染小鼠巨噬细胞且高效表达抗菌肽PR39。构建的重组腺病毒能够在巨噬细胞中表达抗菌肽PR39基因,为靶向表达抗菌肽在胞内菌感染治疗中的应用奠定了基础。     

CMY-39型AmpC酶新基因亚型的序列分析与原核表达

目的对弗劳地枸橼酸杆菌所产CMY-39型AmpC酶新基因亚型进行基因克隆和重组表达。方法以产CMY-39型AmpC酶新基因亚型的弗劳地枸橼酸杆菌总基因组DNA为模板,PCR扩增CMY-39,将其克隆入pGEM-T载体后测定该核苷酸序列,再将CMY-39基因克隆到pET-32 a（＋）系统进行重组,重组菌在大肠埃希菌BL21中表达,SDS-PAGE电泳鉴定酶蛋白的表达。结果 PCR扩增出大小为1 146 bp的基因片段,与GenBank上CMY-39的基因序列同源性为99%。大肠埃希菌BL21转化pET-32 a（＋）/CMY-39重组质粒后,AmpC酶三维试验为阳性。此基因能在大肠埃希菌中大量表达,SDS-PAGE电泳显示,蛋白分子质量大约为60 kD。结论此基因为CMY-39新基因亚型,登陆号为HM565135;成功表达重组的CMY-39型酶,为进一步做酶动力学及酶的其他分子生物学特性研究奠定了基础。     

斜纹夜蛾核多角体病毒VP39-GST融合蛋白的原核表达

用PCR的方法扩增得到斜纹夜蛾核多角体病毒（Spodoptera litura multicapsid nucleopolyhedrovirus,SpltMNPV）vp39全长基因。将其克隆至原核表达载体pGEX-4T-1上,构建重组表达质粒pGEX-4T-vp39,转化大肠杆菌BL21（DE3）,在1 mmol/L异丙基硫代-β-D-半乳糖苷（IPTG）诱导下超量表达了与理论预测值相符的一个约60 kD的VP39-GST融合蛋白。VP39-GST融合蛋白的成功表达为进一步研究VP39在病毒侵染过程中与宿主细胞成分或病毒粒子蛋白间的相互作用奠定了基础。     

PR-39,一种多功能的抗菌肽

PR－39（proline-arginine-rich)是哺乳动物体内的一种含有39个氨基酸残基的小分子抗菌肽,因其富含脯氨酸而得名。分子量小（4719．7）,结构稳定,具有广谱抗菌活性。近年来许多实验研究表明,PR－39在抗肿瘤,组织修复等方面同样具有生物学作用。     

斜纹夜蛾核多角体病毒(SpltNPV)基因组EcoR I-G片段的序列分析

斜纹夜蛾核多角体病毒(SpltNPV)基因组EcoR I-G片段全长7 450bp,包括7个开放读码框vp39、lef-4、cg30、p91、vp33、tlp20、AcMNPV ORF81同源基因和一个同源区(hr).基因组排列比较分析发现,这些基因在基因组中的排列,在所有已知序列的杆状病毒中都比较保守.     

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高速的城市化进程带来一系列生态破坏和环境污染等可持续发展挑战,需要在城市规划阶段尽量规避这些风险。生态基础设施建设倡导生态系统修复和环境协同治理的理念,是新兴城市化背景下指导城市规划的一种有效手段。目前,国内外生态基础设施建设在宏观尺度研究比较系统,而在小尺度的研究相对欠缺。我国社区等小尺度生态基础设施建设,由于规模较小,缺乏网络化构建,导致了建设后雨水内涝、面源污染问题仍层出不穷。构建了一种城市小尺度生态基础设施的设计方法和技术流程,以雄安新区启动区为研究区开展生态基础设施设计：（1）基于实际调研与理论研究分析场地现状,综合考虑自然要素、物理感知、心理感知、生态过程等相关因素构建"廊道为骨,斑块为节"的生态基础设施体系;（2）辨识区域主要动物活动、迁徙以及保护植物多样性等功能生态斑块,构建串联全城提供多种功能的系统性廊道、增加城市内部生态系统服务的结构性廊道、将生态系统服务渗入城市肌理的局部功能性廊道;（3）从景感满意度和年径流总量控制率2个方面进行生态基础设施建设预期效果评估。研究结果有助于缓解雄安新区建设所面临生态环境问题,保障人居生活环境品质,并为今后城市建设工作提供参考。