组织培养条件下苹果Ty1-copia类逆转座子的转录活性

研究了苹果基因组中Tyl—copia类逆转座子的转录活性、多样性及其甲基化水平。在培养基MS＋2，4-D2mg／L＋BA0．2mg／L＋NAA0．5mg／L上诱导了1个月的愈伤组织中用RT—PCR方法检测到了270bp的目的条带，而对照、继代组培苗和不含2，4-D的愈伤中均没有目的条带的扩增，说明一定浓度的2，4-D可以诱导苹果中Tyl—copia类逆转座子的表达活性。将目的条带回收、克隆和测序后进行分析：所克隆的21条序列虽都为Tyl-copia类逆转座子逆转录酶保守序列，但存在很大的异质性，21个克隆的核甘酸序列变化范围为196—290bp；翻译成氨基酸后存在缺失、插入、移框和终止密码子突变，7个克隆在氨基酸保守序列‘SLYGLKQ’处有1—2氨基酸位点的突变，说明这些序列对应的逆转录酶多数可能已失去了其原有功能。将21条序列进行聚类分析，除ART20外，其它克隆可聚为3类，并分别与GenBank上某些物种的相应序列同源性很高，表明在不同物种间可能存在逆转座子的横向传递作用。甲基化试验表明苹果基因组内Tyl-copia类逆转座子甲基化水平高。这些都表明Tyl-copia类逆转座子对苹果基因型的多样性及基因组的进化具有重要意义。     

柑橘裂皮病类病毒的一步RT-PCR法检测及其在甜橙体内的分布

柑橘裂皮病是由柑橘裂皮病类病毒(Citrusexocortisviroid,CEVd)引起的一种重要的柑橘病害。分别采用快速微量核酸提取法和SDS-KAc抽提法在表现症状的Etrog香橼中提取核酸,采用一步RT-PCR法对CEVd进行检测,结果显示SDS-KAc法可以消除带毒样品中非特异性条带。从嫁接接毒的铜水72-1锦橙植株的接穗部取嫩叶、老叶、皮,从砧木部茎杆上取老皮以及根皮分别提取总核酸进行RT-PCR检测,分析CEVd在甜橙体内的分布情况。初步检测结果表明在接穗的嫩叶、老叶、皮,砧木部茎杆的老皮和根皮上都可以检测到CEVd,以接穗部叶和皮检出稳定性高。     

猪生殖与呼吸综合征病毒M基因和N基因的表达

将猪生殖与呼吸综合征病毒（PRRSV）的M基因和N基因从重组质粒pMD18-T—M—N中亚克隆至pBV220原核表达栽体上，成功构建了重组表达质粒pBVM—N。将pBVM—N转化大肠埃希氏菌DH5α感受态细胞．重组菌经温度敏感诱导表达，其细菌裂解物经SDS—PAGE可检测到分子质量约为19ku的目的蛋白，与M基因表达产物一致；经蛋白质分析软件Bandscan分析，其表达量可达19．1％；Western-blotting分析表明，该重组蛋白可被兔抗PRRSV血清所识别，与预期的M基因表达产物相一致，而N基因未获表达。     

母猪卧向行为学的观察

本文通过2个试验对母猪在分娩圈和分娩栏中的卧向行为进行了观察,试验一中, 选择10头大白(Yorkshire)经产母猪作为观察对象,试验二中分别选择10头大白(Yorkshire) 经产母猪和10头长白(Landrace)经产母猪作为观察对象。观察采用瞬时记录方法,每周观察 三次,隔日观察,每天上午、下午各观察一次,每次3 h,每次观察间隔5 min。观察中发现,母猪 卧向以向外为主。分娩栏(四周是栏杆)中的母猪以选择卧向外(向北)为主,其次是卧向内(向 南),卧向左和卧向右最少且差异不显著,上栏前和下栏后差异不显著。分娩圈(四周是墙壁)中 母猪以选择卧向外为主,卧向内最少,卧向左和卧向右差异不显著。长白母猪比大白母猪选择 卧向外的多,妊娠阶段比哺乳阶段选择卧向外的多。     

菌根与植被恢复

菌根不仅可促进植物的营养吸收、生长发育、抗病、抗逆,而且在保持良好的土壤结构、控制水土流失方面具有直接的作用。在我国西北许多地区,菌根菌与植物间的共生关系已被中断,要恢复该地区的植被,治本的方法应是重建植物与菌根菌的共生关系,形成健康的生态系统。文中还提出了菌根技术在植被恢复中的应用策略。     

陕甘宁老区建立生态特区的必要性分析

本文从历史与现状方面分析了陕甘宁老区加强生态环境建设的必要性。阐述了陕甘宁老区历史上范围的变化及其对中国革命和经济建设的贡献。从多方面分析了它的自然条件 ,揭示了其自然环境的脆弱性。说明了该区经济建设取得的成就和存在的问题 ,揭示了其贫困落后的社会经济状况。提出了在该区建立生态特区以促进其生态环境建设的观点。     

关中平原小麦产量对气候变化区域响应的评价模型研究

根据关中地区宝鸡、西安、渭南与咸阳 4地 (市 )的 1 949～ 1 999年的逐年小麦单产记录序列以及 4地 (市 )的气象观测站点自建站以来至 2 0 0 0年近 5 0年的气象记录序列 ,对关中地区小麦产量与年均温、年降水作相关分析 ;探讨了关中地区小麦单产对气候变化区域响应的评价模型。结果发现 :关中平原气候具有暖干化趋势 ;随着气温变暖 ,小麦产量增加幅度减小 ;小麦产量对降水波动的响应比对气温波动的响应显著。     

Protective effect of onychin on the human umbilical vein endothelial cells injured by menadione

AIM: To investigate the protective effect of onychin on the endothelial cells injured by oxidative stress. METHODS: The injured model was established by endothelial cells treated with menadione. The growth inhibitory rate of endothelial cell was determined by MTT assay; NO₂^-／NO₃^- concentration in the medium was determined by nitrate reductase assay; eNOS and caveolin-1 protein levels were determined by Western blot. RESULTS: Onychin significantly decreased the growth inhibitory rate of endothelial cells injured by menadione, increased NO₂^-／NO₃^- concentration in the medium and eNOS activity and up-regulated caveolin-1 expression. CONCLUSION: Onychin possesses a protective effect against endothelial cell injury induced by menadione via caveolin-1/eNOS pathway.     

Selection and amplification of the liver stem cell subset from rat bone marrow cells with a medium containing cholestatic serum in vitro

AIM: To explore the feasibility of direct separat and selective enlargement of the bone marrow-derived liver stem cells (BDLSC) from bone marrow cells with a culture system containing cholestatic serum in vitro. METHODS: Bone marrow cells of rats were cultured with selective media containing 2%, 5%, 7% and 10% cholestatic rat serum, respectively. The BDLSC were then induced to proliferate with the addition of hepatocyte growth factor (HGF) on the firth day. BDLSC were characterized using immunocytochemistry and RT-PCR for lineage markers, glycogen staining and urea synthetic assay for functions 2 weeks later. RESULTS: Bone marrow cells were unble to form colony in the presence of 2% cholestatic serum and apopotosis appeared gradually in 7% or 10% cholestatic serum. The BDLSC survived in the medium containing 5% cholestatic serum while the other types of cells did not. The survival cells proliferated with a high speed during the second week and then formed hepatocyte-like colony-forming units (H-CFU). Cells in the H-CFU expressed the characteristic proteins of fetal hepatocytes. Furthermore, they had glycogen storage and urea synthesis functions, two of the critical features of hepatocytes. CONCLUSION: The selective micro-environment effectively selected BDLSC from the bone marrow cell, and will be a new way to provide an abundant source of donor hepatocytes for clinical cell therapy.     

Effect of mutant β-catenin gene on the proliferation of hepatocytes

AIM:The β-catenin is a key molecule in the Wnt signal pathway, which plays a critical role in normal development and tumorigenesis. However, the mechanisms of the β-catenin on the cell growth control are still not completely defined. The aim of this study was to test the hypothesis that the mutant β-catenin may regulate the hepatocyte proliferation. METHODS: The immortalized murine hepatocyte cell line, AML12, was used for this study. A plasmid that contain mutant β-catenin S33Y was transfected into the AML12 cells and a stable cell line AML12S33Y was established. The cell growth property of this cell line and the parental cell were compared by flow cytometry analysis and direct cell count. The cells were also tested for the ability to form soft agar colonies, and the ability to form tumors in the severe immune deficient mice (SCID). RESULTS:1. The mutant β-catenin containing cell line AML12S33Y has higher proliferating index compared with the parental AML12 cells (P<0.01), suggesting that mutant β-catenin promotes cell growth. 2. The mutant β-catenin cells formed small colonies in soft agar after 4 weeks of culture, but did not generate tumor in SCID mice. CONCLUSION:The mutant β-catenin promotes liver cell growth.