离体条件下TDZ对解除北五味子种子休眠的影响

以深度休眠的北五味子种子为材料,研究了离体条件下,种皮、胚乳因素对北五味子种子休眠的影响;建立了离体条件下,使用TDZ有效解除北五味子种子休眠的方法;并对北五味子种子解除休眠过程中过氧化物酶和过氧化氢酶活性变化规律进行分析.结果表明,(1)种皮和胚乳因素是引起北五味子种子休眠的主要因素,种皮和胚乳对北五味子种子的萌发具有不同程度的抑制作用;(2)MS培养基中添加0.02 mg·L-1 TDZ能有效解除北五味子种子休眠,可使第30天的发芽率达到90.0%,显著缩短北五味子种子休眠时间;(3)随着北五味子种子休眠被解除,过氧化物酶和过氧化氢酶活性均呈明显的增加趋势.     

黄鼬在吉林环长白山旅游公路路域活动的调查研究

2008年11～12月,采用样线法对环长白山旅游公路改扩建工程全线路域200 m范围内黄鼬活动情况进行了3次调查.调查结果显示:K10～K45路段黄鼬穿越公路较多;黄鼬穿越公路频率与路域植被类型和雪被深度有密切的关系;黄鼬已经开始利用涵洞穿越公路;公路对黄鼬具有吸引效应,影响域范围约为50 m.为此提出了相应的保护对策.     

荧光假单胞菌M18的rpoS基因克隆及其功能分析

从荧光假单胞菌 (Pseudomonasfluorescentsp .)M1 8基因组中克隆了RNA聚合酶的稳定期σs 因子编码基因rpoS ,推测其氨基酸序列与铜绿假单胞菌、荧光假单胞菌和恶臭假单胞菌的同源性分别为 99 1 %、87 35 %和87 8%。利用体外定点插入突变和同源重组技术 ,构建了M1 8的rpoS突变株M1 8R- 。对突变株M1 8R- 合成抗生素吩嗪 1 羧酸 (PCA)和藤黄绿菌素 (Plt)的动力学分析结果表明 ,在KB或PPM培养基中 ,突变株合成PCA的能力比野生型分别提高了 2 5或 5 78倍 ,但Plt的积累量不受影响。与野生型相比 ,突变株对碳源饥饿的耐性下降。同时 ,在碳源饥饿条件下对过氧化氢、乙醇和和氯化钠等环境胁迫的交叉保护性减小 ,存活率显著降低     

Ras/MAPK pathway confers basement membrane dependence upon endoderm differentiation of embryonic carcinoma cells

The formation of extraembryonic endoderm is one of the earliest steps in the differentiation of pluripotent cells of the inner cell mass during the early stages of embryonic development. The primitive endoderm cells and the derived parietal and visceral endoderm cells gain the capacity to produce collagen IV and laminin. The deposition of these components results in the formation of basement membrane and epithelium of the endoderm, with polarized cells covering the inner surface of the blastocoels. We used retinoic acid-induced endoderm differentiation of stem cell-like F9 embryonic carcinoma cells to study the role of the Ras pathway and its regulation in the formation of the visceral endoderm. Upon endoderm differentiation of F9 cells induced by retinoic acid, c-Fos expression, the downstream target of the Ras pathway, is suppressed by uncoupling Elk-1 phosphorylation/activation to MAPK activity. However, attachment to matrix gel greatly enhances the activation of MAPK in endoderm cells but not in undifferentiated F9 cells. Enhanced MAPK activation as a result of contact with basement membrane is able to compensate for reduced Elk-1 phosphorylation and c-Fos expression. We conclude that endoderm differentiation renders the activation of the Ras pathway basement membrane dependent, contributing to the epithelial organization of the visceral endoderm.     

Sulfated glycosaminoglycans in guinea pig basophils studied by means of cationic colloidal gold

 Bone marrow embedding in the hydrophilic resin, Lowicryl K4M, followed by cationic colloidal gold (CCG, pH 1.0) staining was used to study the sulfated glycosaminoglycans (GAGs) and their sites of sulfation ultrastructurally in various maturational stages of both basophil granulocytes and basophil granules in the guinea pig. CCG at pH 1.0 is specific for sulfated GAG staining. Basophil granulocytes and granules reacted positively to CCG with a variety of staining according to the stage of maturation. The formation of basophil granules takes place throughout the myelocyte stage. Early basophil myelocytes contain a large Golgi apparatus with active granulogenesis, while late myelocytes contain a small and less active Golgi apparatus as judged by CCG staining. All the immature granules and some of the granules with characteristic ultrastructure stained positively. However, some of the mature granules had lost their affinity for CCG upon maturation. Interestingly, strongly positive CCG staining was also observed in the trans to transmost Golgi apparatus. This indicates that sulfation of GAGs occurs in the trans to transmost Golgi apparatus in all maturational stages of basophil granulocytes. Treatment with chondroitinase ABC or heparinase I abolished the majority of CCG staining. Accepted: 17 July 1997     

避暗反应测定大鼠学习记忆功能方法的探讨

目的 为了提高避暗反应测定动物学习记忆功能的敏感性,本实验对三种实验及统计方法进行了评价。方法 采用IBO致基底前脑胆碱能神经元损伤模型和东莨菪碱所致的记忆障碍模型,对单次简单测试法、多次简单测试法和多次记分测试法的敏感性进行了比较。结果 对于基底前脑损伤模型,单次简单测试法的潜伏期学习记忆指标无法显示出模型组与对照组之间的差异。多次简单测试法两种指标均能显示出显著性差异,且较多次记分测试法的指标差异的显著性高。对于东莨菪碱致痴呆模型,也有同样的效果。结论 多次简单测试法更能敏感地反映动物的学习记忆功能。     

Disabled-2 is essential for endodermal cell positioning and structure formation during mouse embryogenesis

The signal transduction adapter protein Disabled-2 (Dab2) is one of the two mammalian orthologs of the Drosophila Disabled. The brain-specific Disabled-1 (Dab1) functions in positional organization of brain cells during development. Dab2 is widely distributed and is highly expressed in many epithelial cell types. The dab2 gene was interrupted by in-frame insertion of beta-galactosidase (LacZ) in embryonic stem cells and transgenic mice were produced. Dab2 expression was first observed in the primitive endoderm at E4.5, immediately following implantation. The homozygous Dab2-deficient mutant is embryonic lethal (earlier than E6.5) due to defective cell positioning and structure formation of the visceral endoderm. In E5.5 dab2 (-/-) conceptus, visceral endoderm-like cells are present in the deformed primitive egg cylinder; however, the visceral endoderm cells are not organized, the cells of the epiblast have not expanded, and the proamniotic cavity fails to form. Disorganization of the visceral endodermal layer is evident, as cells with positive visceral endoderm markers are scattered throughout the dab2 (-/-) conceptus. Only degenerated remains were observed at E6.5 for dab2 (-/-) embryos, and by E7.5, the defective embryos were completely reabsorbed. In blastocyst in vitro culture, initially cells with characteristics of endoderm, trophectoderm, and inner cell mass were observed in the outgrowth of the hatched dab2 (-/-) blastocysts. However, the dab2 (-/-) endodermal cells are much more dispersed and disorganized than those from wild-type blastocysts, the inner cell mass fails to expand, and the outgrowth degenerates by day 7. Thus, Dab2 is required for visceral endodermal cell organization during early mouse development. The absence of an organized visceral endoderm in Dab2-deficient conceptus leads to the growth failure of the inner cell mass. We suggest that Dab2 functions in a signal pathway to regulate endodermal cell organization using endocytosis of ligands from the blastocoel cavity as a positioning cue.     

大剂量地塞米松快速高效诱导巨噬细胞凋亡

运用透射电镜、DNA琼脂糖凝胶电泳、流式细胞术、原位末端标记法(TUNEL)染色等技术,检测了大剂量地塞米松处理小鼠腹腔巨噬细胞的各种变化.结果显示,大剂量地塞米松处理的巨噬细胞发生胞体皱缩、染色质凝聚、胞质浓缩;TUNEL染色呈阳性;0.5 h后DNA凝胶电泳即呈梯状条带;流式细胞术作周期分析出现凋亡峰等明显的凋亡特征.并且随处理时间延长凋亡率升高.结果表明,大剂量地塞米松快速、高效诱导小鼠腹腔巨噬细胞凋亡.     

二斑叶螨滞育特性的初步研究

诱导滞育实验表明 :短日照和低温是诱导二斑叶螨发生滞育的主要因子。测定我国天水种群 ,在 15℃条件下 ,诱导其滞育的临界日照长度为 9小时 4 2分 ,在每日 8小时光照条件下 ,诱导其滞育的临界温度为 15.5℃。解除滞育实验表明 :在每日 13小时的光照条件下 ,温度越高 ,解除越冬雌成螨滞育的速度越快 ;低温处理滞育雌成螨的时间越长 ,解除其滞育的速度也越快。     

牛白细胞粘附缺陷病(BLAD)的调查

牛白细胞粘附缺陷病(BLAD)是一种常染色体单基因隐性遗传疾病, 病因为白细胞表面整合素CD18亚单位基因突变所致。为了解我国奶牛群中BLAD的携带和发生情况, 本研究采用RT-PCR扩增CD18基因片段后, 用TaqⅠ内切酶对扩增产物进行酶切的方法检测了1 000头1~6岁的奶牛, 结果有19头奶牛为BLAD携带牛, 即BLAD携带率为1.9%; 1头为BLAD病牛。