牛卵丘细胞的生长特征与转基因

目的了解牛卵丘细胞的生长特征及构建的基因载体对卵丘细胞的转染情况和转染后阳性细胞的扩增效率。方法用透明质酸酶消化牛卵丘/卵母细胞复合体(COCs)获得牛卵丘细胞,了解牛卵丘细胞的生长特点,并用分子大小分别为7178 bp和31 085 bp两种带有增强绿色荧光蛋白(EGFP)和新霉素抗性基因(Neor)的质粒载体pMSCV-EGFP-Neor和pGC1-collagen-EGFP-Neor分别转染靶细胞。结果用脂质体法转染对数生长期的细胞,均获得了绿色荧光阳性细胞,但小质粒的转染效率在72 h时是大质粒的6倍(14%vs.2.3%)。在800μg/mLG418筛选浓度下,14 d后分别获得了7个和3个较明显的单克隆细胞群,对它们进行挑选、扩大,都获得了较纯的单克隆细胞系。最后根据EGFP和collagen的已知基因序列对转染pGC1-collagen-EGFP-Neor质粒的阳性细胞进行三个不同区域DNA片段的扩增,结果表明,扩增的基因片段大小正确,数目完整。结论对卵丘细胞进行基因转染,分子大小等于或小于31 kb的基因可以有效转入,但小分子载体比大分子载体具有更高的转染效率。经过筛选都可以获得纯的转基因细胞...     

Overexpression of microRNA-99a Attenuates Cardiac Hypertrophy

Pathological cardiomyocyte hypertrophy is associated with significantly increased risk of heart failure, one of the leading medical causes of mortality worldwide. MicroRNAs are known to be involved in pathological cardiac remodeling. However, whether miR-99a participates in the signaling cascade leading to cardiac hypertrophy is unknown. To evaluate the role of miR-99a in cardiac hypertrophy, we assessed the expression of miR-99a in hypertrophic cardiomyocytes induced by isoprenaline (ISO)/angiotensin-II (Ang II) and in mice model of cardiac hypertrophy induced by transverse aortic constriction (TAC). Expression of miR-99a was evaluated in these hypertrophic cells and hearts. We also found that miR-99a expression was highly correlated with cardiac function of mice with heart failure (8 weeks after TAC surgery). Overexpression of miR-99a attenuated cardiac hypertrophy in TAC mice and cellular hypertrophy in stimuli treated cardiomyocytes through down-regulation of expression of mammalian target of rapamycin (mTOR). These results indicate that miR-99a negatively regulates physiological hypertrophy through mTOR signaling pathway, which may provide a new therapeutic approach for pressure-overload heart failure.     

1954-2010年三江平原土地利用景观格局动态变化及驱动力

受自然因素和人为因素的影响,近60年三江平原土地利用景观格局发生了明显变化。以遥感影像为主要信息源,利用地理信息系统技术和数理统计方法,对1954—2010年三江平原土地利用景观格局动态变化及其驱动力进行分析。结果表明:1954—2010年三江平原耕地、居住建设用地和水域面积呈增长趋势,林地、草地和湿地面积呈减少趋势;土地利用综合动态度呈先增加后降低再增加的趋势;三江平原斑块密度、周长面积比、景观分离度和Simpson's多样性指数都呈先上升后下降的趋势,最小值出现在2010年,斑块密度和周长面积比的最大值出现在1976年,景观分离度和Simpson's多样性指数的最大值出现在1986年;1954—2010年三江平原斑块密度和周长面积比的高值区由东北向西南转移,低值区由中部向东部转移,景观分离度指数呈现先聚集后分散的趋势,Simpson's多样性指数由中间高四周低格局,逐渐转变为南部高北部低的格局。     

PI3K和受体型酪氨酸激酶介导家蚕麻痹肽免疫诱导信号的传递

【目的】普遍存在于鳞翅目昆虫中的ENF肽具有非常相似的结构和生理功能, 对昆虫的发育、 免疫、 应激反应等方面都起着重要的调控作用。有研究报道, 作为ENF肽家族成员之一的家蚕Bombyx mori麻痹肽（paralytic peptide, PP）通过激活MAPK来调控家蚕的免疫应答, 但其完整的分子作用机制尚不明确。本研究旨在解析传递家蚕PP免疫诱导信号的分子通路。【方法】首先通过荧光定量PCR检测了多种昆虫细胞系在PP诱导下抗菌肽基因的转录水平, 并利用Western blot检测p38 MAPK的磷酸化水平, 然后利用不同信号传递分子的抑制剂处理BmE细胞筛选参与传递PP免疫诱导信号的分子。【结果】PI3K抑制剂LY294002和受体型酪酸激酶抑制剂Genistein抑制了PP对BmE细胞抗菌肽基因表达的诱导作用; 且BmE细胞和家蚕血细胞在PP的诱导下, Akt和大小约为70 kDa的细胞膜蛋白均出现磷酸化水平的动态变化。【结论】PI3K/Akt信号通路和Genistein敏感性的受体型酪氨酸激酶介导了PP免疫诱导信号的传递。     

地枫皮营养器官解剖结构特征及其叶片结构的生态适应性

为了明确地枫皮的结构特征及其在石灰岩山顶和山腰疏林间两种环境下生长的叶片解剖结构的差异,本研究采用石蜡切片和半薄切片对地枫皮营养器官进行解剖观察并评价了叶片结构对不同生态环境的响应。结果表明:地枫皮根中次生维管组织发达,木射线和韧皮射线明显。老茎的次生构造中,皮层贮藏物质丰富,内有大的石细胞群,韧皮射线和木射线明显;而髓细胞内含有大量晶簇和少量单晶。在叶片横切面观上,叶为异面叶,表皮细胞一层,上表皮无气孔分布,主脉中薄壁细胞中分散有石细胞。叶片解剖结构显示,随海拔高度上升,地枫皮趋向于旱生植物的特点,其主要表现在叶片表皮细胞外壁角质层加厚,栅海比增加、海绵组织排列由紧密变疏松。另外,根皮、茎皮和叶肉中都分布有大量油细胞。     

Microsatellite analysis of genetic diversity and population structure of Chinese mitten crab （Eriocheir sinensis）

Chinese mitten crab (Eriocheir sinensis) has higher commercial value as food source than any other species of Eriocheir in China.To evaluate the germplasm resources and characterize the genetic diversity and population structure of the crabs in different water systems,two stocks and two farming populations were assessed with 25 polymorphic microsallite loci available in public GenBank.Basic statistics showed that the average observed heterozygosity (Ho) amongst populations ranged from 0.5789 to 0.6824.However,a remarkable presence of inbreeding and heterozygote deficiencies were observed.To analyze population structure,pairwise FST coefficients explained only ～10.3% variability from the subdivision of mitten crab populations,the remaining variability stems from the subdivision within subpopulations.Although the four populations had slight differentiation,different allelic frequencies resulted in distinct population structures.Two stocks and one farming population were clustered together to the phylogenetic branch of Yangtze crab,with an approximate membership of 95%.Whereas,another fanning population was clustered singly to the phylogenetic branch of the Liaohe crab,with a membership of 97.1%.The tests for individual admixture showed that Yangtze crab had probably been contaminated with individuals from other water systems.Genetic relationships between populations also supported the conclusion that Yangtze crab and Liaohe crab had different gene pools in spite of the origins of the same species.     

Oxytocin, but not arginine vasopressin is involving in the antinociceptive role of hypothalamic supraoptic nucleus

Our pervious study has demonstrated that the hypothalamic supraoptic nucleus (SON) plays a role in pain modulation. Oxytocin (OXT) and arginine vasopressin (AVP) are the important hormones synthesized and secreted by the SON. The experiment was designed to investigate which hormone was relating with the antinociceptive role of the SON in the rat. The results showed that (1) microinjection of l-glutamate sodium into the SON increased OXT and AVP concentrations in the SON perfusion liquid, (2) pain stimulation induces OXT, but not AVP release in the SON, and (3) intraventricular injection (pre-treatment) with OXT antiserum could inhibit the pain threshold increase induced by SON injection of l-glutamate sodium, but administration of AVP antiserum did not influence the antinociceptive role of SON stimulation. The data suggested that the antinociceptive role of the SON relates to OXT rather than AVP.     

Mitochondrial genomes of two luminous beetles, Rhagophthalmus lufengensis and R. ohbai (Arthropoda, Insecta, Coleoptera)

We determined the mitochondrial DNA (mtDNA) sequences of two luminous beetles (Arthropoda, Insecta, Coleoptera), Rhagophthalmus lufengensis from Yunnan, China and Rhagophthalmus ohbai from Yaeyama Island, Japan. We identified all the 37 mtDNA genes of R. lufengensis (15,982 bp) and the 34 genes of R. ohbai (15,704 bp). R. lufengensis and R. ohbai genomes have higher A + T contents than other coleopteran genomes although the gene arrangements are similar. Interestingly, in a study of the evolutionary relationship among R. lufengensis, R. ohbai and the firefly Pyrocoelia rufa, the phylogenetic tree inferred from lrRNA genes from mitochondrial genomes indicates a biogeographic relationship among the bioluminescent insects in East Asia and the phylogenetic tree inferred from luciferase-related genes from nuclear genomes shows an appropriate relationship among coleopterans, reflecting the evolutionary origin of bioluminescence. Thus, the mtDNAs of luminescent beetles can provide an insight into their evolutionary origin and biogeography.     

水稻幼芽细胞生物膜上的赤霉素结合蛋白的结合特性

在水稻 (Oryza sativa)幼芽中存在膜结合的赤霉素结合蛋白 ,其与 GA3 结合的平衡解离常数(Kd)为 6.5× 1 0 -8mol/ L,总浓度为 0 .3 pmol· mg-1 蛋白质。结合蛋白与 GA3 结合活力在 0℃时比 2 5℃时高 1 4 0 %。它与 GA3 结合的最适 p H为 5。 GA3 与此结合蛋白的结合量随反应时间延长而增加 ,1 h达最大值 ,以后又逐渐下降。 IAA、ABA可与 GA3 竞争赤霉素结合蛋白。     

亲和层析法分离腺苷结合蛋白质

本文报告一种新的腺苷亲和层析凝胶的合成方法。利用这种凝胶可从大鼠心脏、肝脏及小牛主动脉平滑肌的水溶部份分离出几种腺苷结合蛋白质,其亚基分子量(据SDS-PAGE)分别为35,000、37,000、46,000、43,000及15,300Dal。现已证明,35,000Dal蛋白质是乳酸脱氢酶及苹果酸脱氢酶,43,000Dal蛋白质是腺苷激酶,46,000Dal蛋白质可能是S-腺苷同型半胱氨酸水解酶。15,000Dal蛋白质前人未有报道。它对腺苷具有高度特导性和亲和力,推测是腺苷的细胞内受体和/或载体。测定了这种低分子量腺苷结合蛋白质的氨基酸组成及某些物理常数:pI=6.5;沉降系数2.42S,微分比容0.727cm~3/g,与腺苷复合物的解离常数K_D=2.3μM。