人胃粘膜G、D细胞与肠化生关系的研究

用免疫细胞化学和原位杂交技术探讨G、D细胞及胃泌素mRNA与肠化生的关系。标本来自胃镜活检的胃粘膜。结果显示,在与大肠化生区相邻的胃粘膜,G细胞突变消失,假幽门腺化生也缺乏G细胞,而淖肠化生仍保留少数G细胞;D细胞不仅见于小肠化生,而且也出现在假幽门腺化生以及某些大肠化生区。胃泌素mRNA仅限于G细胞分布区,未出现在大肠化生区和假幽门腺化生区,G细胞及胃泌素mRNA在大肠化生区的消失,可能由于局部杯状细胞分泌的硫酸粘蛋白改变了局部的微环境,从而影响了G细胞的分化与发育,至于假幽门腺化生区G细胞及胃泌素mRNA消失的原因还不清楚,应继续研究。     

Met-enkephalin effects on associations formed in utero

Rat fetuses exposed to an odor stimulus and an aversive stimulus in utero showed an aversion to the odor when tested 16 days postnatally. Fetuses that also received 80 μg/kg Met-enkephalin showed a greater aversion to the odor stimulus than those subjects that did not receive the peptide. The difference between these groups was marginally significant. Control subjects did not show an aversion. But, subjects exposed to the odor and Met-enkephalin without the aversive stimulus, when tested, showed a significant preference for the odor over other control groups. These data show that associative learning in rat fetuses at 20 days of gestation may be enhanced by administration of Met-enkephalin.     

mRNA差别显示技术在植物发育研究中的应用

mRNA差别显示技术是近年发展起来的研究基因表达差异的有效方法,它不仅可比较基因表达差异、追踪已知基因的表达情况,而且可作为寻找特异表达基因的重要手段。本文介绍此方法在植物春化作用、光周期反应、花发育、雄性不育、成熟、衰老、种子发育和休眠及根发育等研究领域的应用概况。     

原位缺口平移标记断裂DNA技术及其在检测流行性出血热尸检和实验动物组织中细胞凋亡和早期死亡的应用

原位缺口平移技术（ＩＳＮＴ）已被用于检测细胞核中ＤＮＡ断裂鉴别尸检组织中细胞的凋亡和坏死断裂、流行性出血热（ＥＨＦ）组织中存在散在单个细胞变性死亡和灶性梗死样坏死,前者带有细胞凋亡的特征。本文以ＥＨＦ肝脏和实验性病毒感染鼠脑组织为例,应用缺口平移法,在ＤＮＡ聚合酶或Ｋｌｅｎｏｗ酶的作用下,将地高辛标记的ｄＵＴＰ掺入合成到ＤＮＡ的断裂部位,通过碱性磷酸酶标抗地高辛抗体免疫组化法显示细胞ＤＮＡ的断裂,检测和鉴别细胞的凋亡和坏死。为分析死后解剖时间间隔及组织固定时间对该方法的影响,本文选用死后２～１４０ｈ尸检、经常规固定石蜡包埋后存放１０～３５年的标本和在１０％福尔马林固定了１０～３５年之后再进行常规处理的标本。实验时用蛋白酶Ｋ（ＰＫ）消化前后对比并分别在标记反应液中略去ＤＮＡ聚合酶作为阴性对照,用ＤＮＡ酶消化组织人为制造ＤＮＡ缺日作为阳性对照。结果发现,未经ＰＫ消化的组织,仅灶性肝细胞核ＩＳＮＴ标记阳性,经ＰＫ消化后,散在的带有凋亡特征的肝细胞胞核也出现阳性,灶性肝细胞胞核标记染色增强,但无论是蛋白酶消化与否,明确梗死样坏死的肝细胞均不被标记。结果还发现,死后２～２４ｈ内尸检组织和长时间（１０～３４年）存放的石     

小麦抗叶锈病近等基因系TcLr41 SSH文库构建与分析

以小麦抗叶锈病近等基因系TcLr41和感病亲本Thatcher的叶片cDNA分别作为试验方和驱动方,利用抑制差减杂交技术,构建了一个包含2544个克隆的差减文库。随机提取阳性克隆质粒DNA后经PCR检测,插入片段大部分集中在200～1000bp之间,证明所构建的文库符合要求。在功能已知的基因中,推测过氧化氢酶（catalasc）基因、抗秆锈病基因（rust resistance gene）、铜蓝结合蛋白（blue copper—binding protein）基因、锌指蛋白（ring zinc finger protein）基因、胁迫反应蛋白（stress responsive protein）基因等可能是TcLr41中抗病相关差异表达基因。     

花生ABI3同源基因的定位分析

ABSC ISIC AC ID-INSITIVE3(AB I3)、LEAFY COTYLEDON2(LEC2)和FUSCA3(FUS3)转录因子在种子发育过程中发挥着重要的调控作用。采用Northern杂交技术,用拟南芥AB I3保守的B3结构域部分序列作为探针分别与花生根、茎、叶、子叶RNA进行了杂交,同时也对花生根、茎、叶、子叶(含胚)组织切片进行了原位杂交,结果均显示只有在花生的子叶和胚中有杂交信号出现,表明花生中可能存在AB I3、FUS3和LEC2的同源基因,且它们只分布在花生的子叶和胚中。     

Snake venoms as a source of compounds modulating sperm physiology: Secreted phospholipases A2 from Oxyuranus scutellatus scutellatus impact sperm motility,acrosome reaction and in vitro fertilization in mice

The goal of this study was to identify new compounds from venoms able to modulate sperm physiology and more particularly sperm motility. For this purpose, we screened the effects of 16 snake venoms cleared of molecules higher than 15 kDa on sperm motility. Venoms rich in neurotoxins like those from Oxyuranus scutellatus scutellatus or Daboia russelii, were highly potent inhibitors of sperm motility. In contrast, venoms rich in myotoxins like those from Echis carinatus, Bothrops alternatus and Macrovipera lebetina, were inactive. From the main pharmacologically-active fraction of the Taipan snake O. scutellatus s., a proteomic approach allowed us to identify 16 different proteins, among which OS1 and OS2, two secreted phospholipases A2 (sPLA₂). Purified OS1 and OS2 mimicked the inhibitory effect on sperm motility and were likely responsible for the inhibitory effect of the active fraction. OS1 and OS2 triggered sperm acrosome reaction and induced lipid rearrangements of the plasma membrane. The catalytic activity of OS2 was required to modulate sperm physiology since catalytically inactive mutants had no effect. Finally, sperm treated with OS2 were less competent than control sperm to initiate in vitro normal embryo development. This is the first report characterizing sPLA₂ toxins that modulate in vitro sperm physiology.