小鼠输精管内转染法建立人组织型纤溶酶原激活剂(tPA)乳腺定位表达生物反应器的研究

外源基因受精卵内显微注射目前仍然是种常用的建立转基因动物的方法,但此方法操作复杂,成本高。因此利用精子作为载体将外源基因带入卵细胞内建立转基因动物成了极具吸引力的方法,并且已有一些成功的报道。虽然该方法简单,但总体来说可重复性较差。我们在建立乳腺定位表达人组织型纤溶酶原激活剂(tPA)小鼠模型中,对此方法做了改进,即将脂质体包裹的外源基因在小鼠输精管及附睾管内转染精子,而非以往的在体外转染,结果:(1)五只转染小鼠,术后10天内共使10只正常母鼠受精;(2)共繁殖出79只首建者小鼠,存活42只;(3)用PCR检测首建者小鼠外源基因的整合,阳性率为7/42(16.67%);(4)用凝胶平板溶圈试验检测5只PCR阳性的雌性首建者小鼠泌乳期乳汁中的活性tPA,3/5表达,表达量在12—20IU/ml之间(约相当于48—80ng/ml);(5)PCR检测4只子一代小鼠外源基因的遗传情况,阳性率为2/4。     

贵州南部铝土矿的孢粉组合特征及沉积时代讨论*

铝土矿是一种重要的工业矿产资源。目前中国大部分的铝土矿依赖进口。因此, 研究铝土矿的沉积模式对我国的资源安全和矿产开发具有重要意义。铝土矿的成矿受多种因素影响, 包括大地构造背景、古气候、侵蚀速率、母岩、时间、古植被和古地形地貌等。要研究这些成矿因素对铝土矿的影响, 并为下一步的找矿勘察工作提供理论支撑, 首先需要准确评估铝土矿的沉积时代。然而, 由于铝土矿的沉积过程涉及复杂的土壤化作用和再沉积过程, 其中保存的植物根系化石或遗迹化石数量很少, 因此很难确定铝土矿的最终沉积时间。目前, 综合分析铝土矿中保存的孢粉化石组合是确定铝土矿沉积时代更有效的方法。在中国, 已经确定的铝土矿沉积时代主要集中在晚古生代。华南产出的晚古生代铝土矿主要以贵州铝土矿为代表, 然而长期以来对贵州铝土矿的形成时代存在较多争议。为了解决这些争议, 本研究在贵州龙里县龙山镇和花溪区高坡苗族乡的两个铝土矿地层剖面上进行了孢粉地层研究。研究结果显示, 贵州龙里县龙山镇SPM095和花溪区高坡苗族乡SPM097剖面的革老河组、汤耙沟组、丰源层铝土岩和祥摆组底部产出的孢粉组合与西欧密西西比亚纪标准Lycospora pussila (Pu)孢粉组合带基本相当, 表明丰源铝土矿的沉积时代不晚于密西西比亚纪维宪早中期, 这是目前已知贵州地区最早的具有明确化石证据的喀斯特型铝土矿。通过与贵州中部清镇至贵州北部遵义等地铝土矿中发现的孢粉组合进行对比, 发现丰源铝土矿中的孢粉组合比上述地区铝土矿中的孢粉组合大致低一个孢粉组合带 (Perotriletes tessellatus—Schulzosopra camplylotera组合带)或一个牙形刺带(Gnathodus bilineatus带), 时代大概相差420万年。这一发现进一步表明晚古生代大冰期(LPIA)引起的频繁海平面变化导致贵州“南海北陆”的古地理格局由南向北逐次被海水淹没, 并接受沉积, 这是导致贵州不同地区铝土矿呈现由南向北时代梯度递减的主要原因。     

黄土丘陵区草本植物叶片与枯落物生态化学计量学特征

以黄土高原延河流域森林、森林草原和草原3个植被区的4种典型草本植物达乌里胡枝子、长芒草、铁杆蒿、茭蒿的叶片和枯落物为对象,测定叶片和枯落物中碳(C)、氮(N)、磷(P)、钾(K)含量及其比值,分析草本植物在该区域的养分限制及再吸收规律.结果表明: 4种植物叶片N/P均小于14,生长主要受氮元素含量限制;除达乌里胡枝子外,其他3种植物N、P、K的养分再吸收效率平均值分别为48.7%、32.5%、79.9%.茭蒿和铁杆蒿较高的N、P、K养分再吸收效率和叶片K含量有助于其在水分竞争中占据优势.铁杆蒿枯落物的C/N显著低于长芒草和茭蒿的枯落物,因而更易分解和进行养分循环.这可能是铁杆蒿在3个植被区分布广泛的原因之一.     

基于“玩课网”对“免疫学”翻转课堂课前教学进行评价的实践

翻转课堂是近年来兴起的一种以学生为主体的教学模式。此前我们以QQ群为平台对"免疫学"教学进行翻转课堂实践,取得了良好效果。但受QQ平台技术的限制,难以对学生的自主学习情况进行有效监督与考核。因此,在原有基础上,以"玩课网"为平台,设计了以学生完成课前自学任务获得积分作为翻转课堂的课前教学评价。实践证明不仅促进了学生高质量完成课前自学任务,也使任课教师及时了解学生对课前自主学习内容的掌握情况,使课堂教学更具有针对性,提高了翻转课堂的教学效果。     

笼养海南(开鸟)的活动时间分配

2003年10月至2004年4月,在对海南[开鸟]进行驯养的基础上,采用全部事件取样法研究了笼养海南[开鸟]的活动时间分配。结果表明：笼养海南[开鸟]一天中除静栖和静立外,其余各行为所占时间比例在白天极少,而在晨（5：00～7：00am）、昏（19：00～21：00pm）和午夜（23：00～1：00am）较大,呈明显的日活动节律;节律变化与季节有关;海南[开鸟]各种行为活动所占比例依次为静栖82．48％、静立8．64％、运动5．36％、取食1．97％、身体护理1．30％、其它0．25％。其时间分配不受季节（春、秋、冬季）影响。     

大批量测定植物(互花米草)叶面积的一种新方法

叶面积的准确测定是评价生态系统能流、碳流和水分利用的重要基础.目前常用的叶面积测定方法都有其局限性,有些方法快捷、准确,但仅适用于小批量测定;大多数方法很难在较短时间内准确进行大批量测定.本文以互花米草叶面积测定为例,探讨了以扫描仪和叶片干重为基础,结合PHOTOSHOP图像处理技术,大批量、快捷、准确测定植物叶面积的方法--扫描-干重法.结果表明:虚拟正方形叶片边长与叶子像素成幂函数曲线关系,公式为419.85x^1.9693,R≈1.00;用此公式验证扫描-干重法,相对误差仅为3.651%.用扫描-干重法测得的结果与方格法相比无显著差异(P=0.473),表明该方法是可行的.     

Aurora-A is a critical regulator of microtubule assembly and nuclear activity in mouse oocytes, fertilized eggs, and early embryos

Aurora-A is a serine/threonine protein kinase that plays a role in cell-cycle regulation. The activity of this kinase has been shown to be required for regulating multiple stages of mitotic progression in somatic cells. In this study, the changes in aurora-;A expression were revealed in mouse oocytes using Western blotting. The subcellular localization of aurora-A during oocyte meiotic maturation, fertilization, and early cleavages as well as after antibody microinjection or microtubule assembly perturbance was studied with confocal microscopy. The quantity of aurora-A protein was high in the germinal vesicle (GV) and metaphase II (MII) oocytes and remained stable during other meiotic maturation stages. Aurora-A concentrated in the GV before meiosis resumption, in the pronuclei of fertilized eggs, and in the nuclei of early embryo blastomeres. Aurora-A was localized to the spindle poles of the meiotic spindle from the metaphase I (MI) stage to metaphase II stage. During early embryo development, aurora-A was found in association with the mitotic spindle poles. Aurora-A was not found in the spindle region when colchicine or staurosporine was used to inhibit microtubule organization, while it accumulated as several dots in the cytoplasm after taxol treatment. Aurora-A antibody microinjection decreased the rate of germinal vesicle breakdown (GVBD) and distorted MI spindle organization. Our results indicate that aurora-A is a critical regulator of cell-cycle progression and microtubule organization during mouse oocyte meiotic maturation, fertilization, and early embryo cleavage.     

亮斑扁角水虻人工饲养条件下适宜温度的研究

亮斑扁角水虻作为一种对环境有益的昆虫正在全球范围内受到关注,特别是能够将易腐垃圾转化为高附加值的昆虫蛋白和油脂,为下游的多个产业(食品、饲料)提供稳定的动物蛋白来源,从而实现低碳环保、循环利用的可持续发展模式。亮斑扁角水虻作为热带起源的变温动物,生长发育依赖于适宜的环境温度,本文的研究发现,亮斑扁角水虻孵化的最佳温度为32℃,孵化时间最短(56 h)、收获的初孵幼虫最多(1.86 g);其它阶段发育的适宜温度为28℃,表现为幼虫体重(19.42 g/100 head)、蛹重(13.64 g/100 head)、幼虫期(9.33 d)、蛹期(11.33 d)、成虫寿命(10.33 d)都达到最优。蛹期具有较好的温度耐受性,但成虫寿命显著受温度的影响,温度越高成虫寿命越短;数据显示亮斑扁角水虻的环境温度与主要发育参数(孵化虫量、幼虫期、蛹期、幼虫体重、蛹重)的相关性能够很好地被二项式拟合,说明亮斑扁角水虻的生长发育存在着一个最适温度区域,有利于规模化人工养殖条件下的环境优化。     

Resource allocation in the submerged plant Vallisneria natans related to sediment type, rather than water-column nutrients

1. Phenotypic plasticity in resource allocation by Vallisneria natans was investigated in a greenhouse experiment, using three types of sediment [sandy loam, clay, and a 50 : 50 (by volume) mixture of the two sediments] and two levels of water‐column nutrient. The clay was collected from a highly eutrophic lake in Jiangsu Province, China, and the N and P concentrations applied in nutrient media were at the upper limits observed in most lakes of China. 2. Growth and biomass allocation were significantly affected by sediment type, rather than water‐column nutrients. Plant growth in clay and the mixture were similar, and 2.4–3.4 times higher than that in sandy loam. Compared with the plants grown in clay or the mixed sediments, the plants grown in sandy loam allocated relatively more biomass to root (11–17% versus 7–8% of total biomass), and relatively less to leaf (76–82% versus 86–87% of total biomass). Plastic variations in root area were induced by sediment type alone (P < 0.05), whereas the impacts of sediment type and water‐column nutrients on leaf area were insignificant (P > 0.05). 3. Plant N and P concentrations were significantly affected by both sediment type and water‐column nutrients. Increased nutrient availability in the water column enhanced plant N concentration by 3.5–20.2%, and plant P concentration by 19.1–25.8%. 4. Biomass accumulation and plant nutrient concentration in plants grown in different sediment types and water‐column nutrients indicate that sediment type had more significant impacts on growth and N and P concentrations of V. natans than did water‐column nutrients. Changes in phenotype are a functional response to nutrient availability in sediment, rather than to water‐column nutrients.     

大米草(Spartina anglica)自然衰退种群对N、P添加的生态响应

通过对外来植物大米草(Spartina anglica)自然衰退种群进行N肥、P肥和N-P复合肥不同梯度水平的添加,分析大米草的生长指标差异及其生理生态响应。结果表明:N、P添加后使大米草种群株高均有不同程度的增加,肥效强弱依次为N肥、P肥、N-P肥;叶片数、主根数及总生物量均显著增加(p<0.05)。除N肥外,其它处理的叶片面积和厚度与对照没有显著差异。3种肥源的添加均显著提高了大米草自然衰退种群的光合速率(p<0.05);N和N-P肥均以高浓度效果最显著,但P肥却以中浓度效果最强;光合速率分别比对照增加19.08μmol·m-2·s-1、11.23μmol·m-2·s-1和15.47μmol·m-2·s-1;14d淡水淹没后,肥源添加使大米草自然衰退种群的SOD和POD酶活性增强;中浓度N和中浓度P添加使SOD活性增加最显著,分别比对照增加320.74unit·g-1和134.54unit·g-1;高浓度N和高浓度N-P肥添加使POD酶活性最显著增加。N肥添加可以显著改善大米草自然衰退种群生长生理特性,可以推断大米草种群的衰退与我国海岸带土壤营养中N素营养的限制有一定的相关性。