Immunohistochemical study of matrix metalloproteinases‐2 and ‐9, macrophage inflammatory protein‐2 and tissue inhibitors of matrix metalloproteinases‐1 and ‐2 in normal,purulonecrotic and fungal infected equine corneas

Objective Determine the effects of matrix metalloproteinases (MMPs)‐2, ‐9, macrophage inflammatory protein‐2 (MIP‐2), tissue inhibitors of matrix metalloproteinase (TIMP)‐1 and ‐2 by immunohistochemical expression in fungal affected and purulonecrotic corneas. Procedure Paraffin‐embedded equine corneal samples; normal (n = 9), fungal affected (FA; n = 26), and purulonecrotic without fungi (PN; n = 41) were evaluated immunohistochemically for MMP‐2, ‐9, MIP‐2, TIMP‐1 and ‐2. The number of immunoreactive inflammatory cells was counted and statistics analyzed. Western blot was performed to detect MMP‐2, MMP‐9, TIMP‐1 and TIMP‐2 proteins. Results Matrix metalloproteinases‐2, ‐9, MIP‐2, TIMP‐1 and ‐2 immunoreactivity was identified in corneal epithelium of normal corneas, and in corneal epithelium, inflammatory cells, keratocytes, and vascular endothelial cells of both FA and PN samples. Inflammatory cell immunoreactivity was significantly higher in FA and PN samples than in the normal corneas. There was positive correlation between MMP‐2 and MIP‐2, MMP‐9 and MIP‐2, and MMP‐9 and TIMP‐1 in inflammatory cell immunoreactivity in FA samples. There was positive correlation between MMP‐9 and MIP‐2, MMP‐9 and TIMP‐2, MIP‐2 and TIMP‐1, and MIP‐2 and TIMP‐2 in inflammatory cell immunoreactivity in PN samples. Western blot confirmed the presence of all four proteins in equine corneal samples. Conclusion Increased immunoreactivity of MMP‐2 and ‐9 in FA and PN samples is indirectly related to MIP‐2 through its role in neutrophil chemo‐attraction. Tissue inhibitors of matrix metalloproteinase‐1 and TIMP‐2 are up‐regulated in equine purulonecrotic and fungal keratitis secondary to MMP‐2 and MMP‐9 expression. The correlation between MMPs ‐2 and ‐9, MIP‐2, TIMPs ‐1 and ‐2 suggests that these proteins play a specific role in the pathogenesis of equine fungal keratitis.     

寡果糖诱导瘤胃急性酸中毒对山羊瘤胃发酵、蹄组织结构及炎症因子、金属蛋白酶表达的影响

本试验旨在研究寡果糖诱导瘤胃酸中毒对山羊瘤胃发酵、蹄组织结构、蹄部炎症因子及金属蛋白酶表达的影响。采用随机区组设计,将8头健康装有永久性瘤胃瘘管的波杂山羊(波尔山羊×长江三角洲白山羊)随机分为对照组和诱导酸中毒的试验组,每组4头。其中试验组山羊瘤胃寡果糖灌注量为21 g/kg BW。分别于灌注前(0 h)和灌注后4、8、12、24和48 h采集瘤胃液,同时分别在0、4、8、24和48 h通过颈静脉采集血液,灌注后48 h屠宰2组山羊,采集蹄组织。结果表明:与对照组相比,试验组山羊瘤胃液p H、挥发性脂肪酸浓度平均值显著降低(P0.05),血液和瘤胃液中乳酸和脂多糖浓度平均值显著提高(P0.05);组织形态学结果显示,与对照组相比,试验组山羊蹄组织中次级表皮蹄小叶和次级真皮蹄小叶的长度变短,蹄小叶形状不规则;实时定量PCR检测结果显示,较对照组比较,试验组山羊蹄组织中基质金属蛋白酶组织抑制物-1的mRNA的相对表达量显著降低(P0.05),白细胞介素-6、膜型基质金属蛋白酶-1和基质金属蛋白酶-2的mRNA的相对表达量显著升高(P0.05)。结果提示,寡果糖诱导山羊急性瘤胃酸中毒可导致山羊瘤胃发酵紊乱,提高瘤胃液和血液中脂多糖与乳酸浓度,导致山羊蹄组织相关炎症因子与金属蛋白酶表达改变,最终引发山羊急性蹄叶炎。     

南方根结线虫MiV901基因在拟南芥中的异源表达及其表型分析

为明确南方根结线虫Meloidogyne incognita效应蛋白MiV901在其寄生过程中的生物学功能,通过构建MiV901基因的植物表达载体,利用根癌农杆菌Agrobacterium tumefaciens介导的花序浸染法将其转化拟南芥Arabidopsis thaliana,并采用室内人工接种法测定转基因植株对灰葡萄孢 Botrytis cinerea侵染及南方根结线虫寄生的影响。结果显示：经Southern blot检测,MiV901基因以不同的拷贝数插入到转基因拟南芥株系901-6、901-8和901-12的基因组中,且qPCR检测结果证实 MiV901基因能够正常表达。3个转基因拟南芥株系901-6、901-8和901-12叶部接种灰葡萄孢3 d后,叶片上形成的病斑平均直径分别为1.00、1.06、1.05 cm,比野生型对照扩大了9.9%~16.5%。相比野生型对照,转基因拟南芥株系901-12、901-6和901-8接种南方根结线虫2龄幼虫后根系上产生了更多的雌虫和卵块,雌虫数分别显著增加了45.4%、34.4%和23.7%,卵块数分别显著增加了51.2%、 46.3%和31.7%。表明异源表达MiV901基因能够抑制植物免疫,增加拟南芥对灰葡萄孢和南方根结线虫侵染的敏感性。     

蜜蜂(Apis mellifera)毒的柱层析分离及其主要组份Melittin的抗凝作用

 蜜蜂(Apis mellifera)毒通过葡聚糖凝胶G50分离为七个组份,并进行了酶活力测定和毒性测定,酶测定作了磷酯酶A₂,水解蛋白酶,三磷酸腺甙酶,一磷酸腺甙酶等。对直接溶血组份在血液凝固系统上的作用进行了体内外实验,实验表明:抗血凝固作用是由于抑制了凝血活酶的生成和凝血酶原激活物的生成。     

Low genetic diversity threatens imminent extinction for the Hungarian meadow viper (Vipera ursinii rakosiensis)

Meadow vipers (Vipera ursinii) are small venomous snakes whose range in Hungary has been greatly fragmented by anthropogenic habitat disturbance (especially, agriculture). We obtained DNA from a total of eight Hungarian snakes. Genetic variability at the major histocompatibility (Mhc) class I loci was much lower for these snakes than for specimens from two large Ukrainian populations. Within two Hungarian populations for which we had multiple individuals, band-sharing indices were 100 and 84.6% (versus 63.3 and 57% for the Ukraine populations). The Ukrainian snakes also displayed more RFLP fragments than the Hungarian vipers (mean 13.7 versus 9.0, respectively). In combination with reports of birth deformities, chromosomal abnormalities and low juvenile survival, these data strongly suggest that the Hungarian vipers are experiencing inbreeding depression. Genetic diversity is still present in the Hungarian vipers but among rather than within populations. Given the very low numbers of animals, the only feasible strategy to increase the genetic diversity and to save the Hungarian vipers from extinction is to implement a captive breeding program based on genetically screened animals.     

Novel genes continue to enhance population growth in adders (Vipera berus)

In 1999, we published a report on how an introduction of 20 males into a severely inbred and isolated population of adders (Vipera berus) halted its decline towards extinction, induced a profound change in population genetic variability, resulted in a dramatic increase in offspring viability and thus in a rapid increase in numbers. Since the publication of our paper we have received numerous inquiries as to whether we have continued to monitor this population, as this is the case we herein present population demographic data gathered between 1981 and 2003. The population has continued to increase in numbers and in 2003, we collected 39 adult male adders, more than at any time over the 23-year study. Our continued work, hence, lends strong support to the importance of novel genes in enhancing the viability of inbred populations and supports the importance of preserving genetic variability in order to maintain viable wild populations.     

GA3和KT对蛇瓜种子萌发和幼苗生长的影响

用不同浓度的GA3和KT对蛇瓜种子进行了处理,结果表明:GA3可以提高蛇瓜种子的发芽率、发芽势、发芽指数,对其幼苗生长有明显的促进作用,并可极大地提高其活力指数;而KT也可以提高种子的发芽率、发芽势,但作用不显著。同时KT对幼苗的生长有一定的抑制作用,但可以增加幼苗的地上部和根的鲜重。