猪卵巢体外保存温度对卵母细胞染色质构型和成熟能力的影响

Most porcine oocytes used in studies on embryo biotechnology and the in vitro production of embryos are currently obtained from the ovaries of slaughtered gilts. The duration and temperature during ovary transportation and handling might, therefore, affect the recovery of culturable COCs, chromatin configuration and developmental competence of oocytes. The effects of ovary storage temperature on chromatin configuration and in vitro maturation of porcine oocytes were examined in this study. Ovaries collected from a slaughterhouse were stored in vitro for 8 h under different temperatures. The results showed that more culturable COCs were isolated from the ovares stored at 39℃ than that from ovaries stored at 31℃ or 20℃ and before storage. Thirty-one centidegree was the best storage temperature in terms of cumulus expansion, nuclear maturation and morphology of the first polar body after in vitro maturation culture. The ability of cumulus expansion was completely lost in COCs derived from ovaries stored at 39℃ for 8 hours. Ovary storage （at both 31℃ and at 20℃ ） increased the proportion of oocytes with the GVc configuration in which chromatin condensed into a single big clump at the nucleolus and the functional significance of this configuration needs further investigations [ Acta Zoologica Sinica 51 （5）： 919 923, 2005].     

在低蛋白质饲料中补充必需氨基酸对大口黑鲈生长、体组成和免疫指标的影响简

为了研究在低蛋白质饲料中补充晶体必需氨基酸对大口黑鲈生长、体组成和免疫指标的影响,根据鱼体的必需氨基酸组成模式设计了7种等能的试验饲料。其中4种饲料(45CP、40CP、35CP和30CP)的粗蛋白质水平分别为45%、40%、35%和30%,另3种饲料(40AA、35AA和30AA)是在低蛋白质饲料(40CP、35CP和30CP)的基础上添加必需氨基酸,使它们的必需氨基酸含量与45CP(对照组)相一致。用上述饲料对初始体重为(10.13±0.01)g的大口黑鲈进行了89d的饲养试验。饲养试验在室内循环水养殖系统中进行,每种饲料设3个重复,每重复放养30尾鱼。方差分析显示:试验鱼的生长性能、饲料效率、全鱼和肌肉的粗蛋白质含量、成活率以及免疫指标均随着饲料蛋白质含量的降低而显著降低(P0.05)。添加必需氨基酸的35AA和30AA的饲料效率和蛋白质保留率分别显著高于对应的未添加必需氨基酸的35CP和30CP组(P0.05),但仍显著低于45CP组(P0.05)。40AA的试验鱼血清溶菌酶活性和血清补体活性与45CP组差异不显著(P0.05)。35AA和30AA组的头肾白细胞呼吸爆发活性显著高于35CP和30CP组(P0.05)。30AA组的全鱼粗蛋白质含量以及肥满度显著高于30CP(P0.05)。各组试验鱼的水分和灰分均无显著差异(P0.05)。回归分析显示:在低蛋白质饲料中补充晶体必需氨基酸对大口黑鲈幼鱼的生长、饲料效率和蛋白质保留率所产生的影响与其引起增加了的饲料蛋白质水平而不是饲料的必需氨基酸水平正相关。研究表明,在低蛋白质饲料中补充的晶体必需氨基酸对大口黑鲈的生长、体组成和免疫指标产生的有益作用不及等量的以蛋白质为来源的必需氨基酸。     

Phylogeographical Study on Eriophyton wallichii (Labiatae) from Alpine Scree of Qinghai Tibetan Plateau

In this study, we examined the nuclear ribosomal DNA internal transcribed spacer (ITS) sequence variation of Eriophyton wallichii, a perennial endemic to alpine scree of the Qinghai Tibetan Plateau. Nineteen haplotypes were detected by analyzing 187 individuals from 20 populations of E.wallichii. Most populations hold the unique haplotype, which showed different population with different haplotype. An analysis of molecular variance (AMOVA) for populations showed that the genetic variation mainly resided among populations (89.54%), the level of differentiation among populations was very high (GST=0.863; NST=0.957) but didn’t show a significant phylogeographical structure. It is different from foregoing other species reported in Qinghai Tibetan Plateau that the haplotypes of E.wallichii showed a distribution pattern that almost one population hold one unique haplotype, hardly share same halotype among populations.We hypothesized the unique genetic structure of E.wallichii might result from allopatric fragmentation in the “islands of alpine” due to the extremely isolated alpine scree and rough topography of the region, through the Quaternary climate oscillation dramatically caused population expanding or shrinking.     

人脐带间充质干细胞通过分泌IL-6介导冈田酸对SH-SY5Y细胞毒性的保护作用

阿尔茨海默病(Alzheimer’s disease,AD)是一种国际公认的难治性神经退行性疾病,是引起痴呆的最常见的病因.其主要的病理学变化是由Aβ过度沉积引起的老年斑(SP),以及Tau蛋白过度磷酸化引起的神经纤维缠结(NFTs).从人脐带华通胶中分离出的间充质干细胞(hUC-MSCs)由于其强大的旁分泌作用,已经被证实对神经系统疾病有治疗效果,其中包括AD,这种治疗机制尚不明确.本研究用冈田酸对SH-SY5Y细胞系进行损伤,建立AD体外模型,然后用种有hUC-MSCs的transwell小室或其条件培养基对模型进行治疗,并发现其分泌的IL-6可能是介导这种修复作用的关键因子.     

PKIB抑制PKA活性调节细胞衰老

人cAMP依赖型蛋白激酶B抑制剂(PKIB)是一种新的耐热蛋白.实验证明,PKIB对PKA催化亚单位具有抑制作用.为研究PKIB在细胞衰老中的功能,以年轻和年老人胚肺二倍体成纤维细胞(2BS细胞株)为实验对象,通过实时 PCR发现,在年老细胞中,PKIB表达高于年轻细胞;用PKA活化剂处理年轻和年老细胞,结果显示,在年老细胞中PKA活性变化较年轻细胞小.通过免疫共沉淀实验证实,在年老细胞中,PKIB与PKA催化亚单位结合较年轻细胞中多;进一步通过在年轻细胞中过表达PKIB,检测细胞PKA活性,发现PKA活性明显降低,进一步证实了在人胚肺二倍体成纤维细胞中PKIB对PKA活性的抑制作用;利用Western实验结果证实,PKA催化亚单位在年轻细胞中的表达高于年老细胞.以上结果证明,在2BS细胞中,PKA活性受PKIB的抑制;这种抑制作用与细胞的衰老有一定的关联作用.     

Transient Receptor Potential Vanilloid-1 (TRPV1) and Ankyrin-1 (TRPA1) Participate in Visceral Hyperalgesia in Chronic Water Avoidance Stress Rat Model

Stressfull life events have powerful influences on visceral perception of certain IBS patients. In the present study, we aimed to examine the involvement of TRPV1 and TRPA1 in the stress-induced visceral hyperalgesia. Rats were exposed to 1-h water avoidance stress (WAS) daily for 10 consecutive days. The abdominal withdrawal reflex (AWR) to colorectal distension was assessed at the end of the 10-day period. Western-blotting analysis was applied to investigate the alterations of TRPV1 and TRPA1 in the colonic afferent dorsal root ganglia (DRG). Compared with control rats, the WAS-treated rats demonstrated a significant increase in the AWR with the pressure ≥40 mm Hg (P < 0.05). Meanwhile, in the WAS-treated rats, western-blotting analysis showed significant upregulation of TRPV1 and TRPA1 in the colonic afferent DRG. The results indicate that WAS could induce the upregulation of TRPV1 and TRPA1 in the colonic afferent DRG, and both receptors may be candidate molecules involved in the stress-induced visceral hyperalgesia in rats.     

独叶草叶片性状表型多样性研究

独叶草是特产于中国的一种珍稀濒危植物.为研究独叶草自然居群的叶片形态变异特征及影响因素,选取分布于陕西、四川、甘肃的9个自然居群,对独叶草叶片的叶面积、叶柄长、总齿数、末级叶脉数、盲脉数、网结脉数等指标进行统计分析.结果表明:(1)独叶草叶片各属性间存在显著相关性,个别性状表现出与地理位置的相关性.(2)主成分分析显示独叶草的末级叶脉数、叶面积、总齿数对变异有较大贡献,而网结脉贡献很小.(3)独叶草叶片性状居群间变异程度大于居群内,其中盲脉数量在各个居群间变异程度最大,而末级叶脉数为最稳定的性状.(4)将本研究中形态变异结果与前期的遗传分析结果对比分析认为独叶草形态变异主要来自生态环境,而非遗传组成.(5)独叶草居群的形态及遗传变异聚类结果都显示独叶草形态变异并不具有地理格局规律,可能由于地质历史变化打断了独叶草原本连续的分布区,遗传保守性使其仍然保持分布区断裂前的遗传特性,但其表型随着生境的变化表现出可塑性.     

凡纳滨对虾MT基因序列及其在卵巢发育和低温胁迫中的表达分析

在低温处理仔虾全长cDNA文库的筛选测序中, 获得凡纳滨对虾(Litopenaeus vannmei)金属硫蛋白基因全长cDNA序列, 该序列含有425个碱基, 包含177 bp开放阅读框, 上游98 bp的非编码区及下游150 bp 的非编码区, 编码58个氨基酸, 其中半胱氨酸含量丰富, 富含金属硫蛋白典型的Cys-X(1-3)-Cys 结构。多序列比对表明, 凡纳滨对虾MT蛋白序列与美洲螯龙虾(Homarus americanus)MT蛋白序列具最高同源性72.4%。Real-time PCR结果表明, 凡纳滨对虾MT基因在卵巢组织中呈优势表达, 在不同发育期的卵巢中的表达量都很高, 在低温处理凡纳滨对虾肝胰腺组织中上调表达。实验所得结果为研究凡纳滨对虾金属硫蛋白基因在生殖发育和低温应激中的功能提供了参考。       

青藏高原东缘常见种长毛风毛菊(Saussurea hieracioides)的繁殖分配

对分布在青藏高原东缘高寒草甸常见种长毛风毛菊的6个海拔梯度(3200～3850m)18个居群的繁殖分配进行了研究,结果表明:长毛风毛菊随海拔的升高个体和营养器官减小,繁殖的投入增加;随着海拔的升高,长毛风毛菊的个体管状小花数目和种子数目减少,管状小花重量和百粒重增大;个体管状小花的数目及重量和个体种子的数目及重量间存在权衡关系;长毛风毛菊的个体大小和繁殖分配存在负相关关系。在资源受限的环境下,长毛风毛菊以减少管状小花数目和种子的数目来增加管状小花重量和百粒重,是保证有性繁殖成功的策略之一。     

杨树胞外蛋白的提取分离及2-D电泳体系的建立

该研究以美洲黑杨杂种优良无性系NL895杨(Populus deltoides×Populus euramericana cv.)组培苗叶片和茎段为研究对象,对NL895杨叶片细胞壁蛋白(cell wall proteins,CWPs)和茎段质外体蛋白(apoplastic proteins,APPs)的提取、分离和双向电泳等技术进行了系统研究。结果表明:10g以上叶片、超声波破碎10min、CaCl2法提取的细胞壁蛋白效果较好,经G-6-PDH酶活性检测,提取的细胞壁蛋白胞质污染率较低;真空渗透法提取的茎段质外体蛋白胞质污染率较前者高,但在允许范围之内。TCA/丙酮沉淀法纯化提取的细胞壁蛋白、pH 4~7的24cm胶条、上样量为500μg的双向电泳体系,其蛋白电泳图谱中的斑点多而清晰,斑点数达550多个,是叶片细胞壁蛋白电泳分析较适合的体系;pH 3~10胶条对茎段质外体蛋白的电泳分离效果较好。该研究初步建立了杨树胞外蛋白的提取、分离及2-DE电泳体系,为木本植物胞外蛋白的研究奠定了基础。