武汉单极虫巢式聚合酶链反应检测方法的建立与优化

应用巢式聚合酶链反应(nested PCR)建立了一种检测异育银鲫武汉单极虫(Thelohanellus wuhanensis)基因组DNA的方法,提取的DNA作为模板进行两次扩增,第一次扩增用单极虫18S rRNA序列通用引物:5'-CTGCGGACGGCTCAG TAAATCAGT-3'和5'-CCAGGACATCTTAGGGCATCACAGA-3',扩增长度为1 584 bp;第二次依据第一次扩增产物中武汉单极虫特有的高度保守区设计特异性引物:5'-ACCCACTTCTGTGGC CTTTC-3'和5'-AATCCGACCTACAACGCTGG-3',扩增长度为853 bp。结果表明,通过PCR反应体系中退火温度、dNTP、Mg2+、延伸时间、扩增循环数和模板浓度的优化,优化后的巢式PCR最低DNA检测量达10 fg,优化后比常规PCR检测的灵敏度高104倍。因此,本文所建立的巢式PCR检测方法适合于水环境或水生动物中武汉单极虫的微量检测,也为异育银鲫武汉单极虫病的诊断和流行病调查提供了一种高灵敏的检测技术。     

复方磺胺甲噁唑在松浦镜鲤体内的药动学及残留消除规律

采用高效液相色谱法,研究了以50 mg/kg (体质量)的剂量在口灌给药条件下,复方磺胺甲噁唑(甲氧苄啶.磺胺甲噁唑=1.5)在松浦镜鲤Cyprinus carpio Songpu体内的药动学与残留消除规律。结果表明：单次给药后,甲氧苄啶在血浆和肌肉中的药时关系符合一级吸收一室开放模型,在肝胰脏和肾中的药时关系符合一级吸收二室开放模型,磺胺甲噁唑在血浆、肌肉、肝胰脏和肾中的药时关系符合一级吸收二室开放模型;每天一次连续5 d给药后,甲氧苄啶于第15天开始低于0·05 mg/kg,磺胺甲噁唑于第9天开始低于0·10 mg/kg。研究表明,在本试验条件下,建议休药期不低于15 d。     

超声波辅助提取鱼脑磷脂的研究

以鲣鱼脑为原料,对鱼脑磷脂超声提取方法进行了研究.以磷脂提取率为评价指标,通过单因素和正交试验优化了鱼脑磷脂提取的条件.结果表明：以正己烷-乙醇（体积比2∶1）为提取溶剂,超声提取时间为40 min,超声温度为35℃,固液比为1∶9 （g∶ mL）,超声提取2次,在此条件下鱼脑磷脂的提取率达到氯仿-甲醇法的87.1％;通过薄层色谱分析,确定鲣鱼脑磷脂中含有磷脂酰乙醇胺、磷脂酰胆碱和神经鞘磷脂3种磷脂;用气相色谱法测定鲣鱼脑磷脂的脂肪酸组成,高不饱和脂肪酸中DHA和EPA的含量分别为21.1％和2.4％.研究表明,用超声法提取鱼脑磷脂具有可行性,鱼脑磷脂中含丰富的不饱和脂肪酸,在食品加工中具有一定的应用前景.     

藜麦叶片黄酮类物质的提取及基因型差异

为了优化藜麦Chenopodium quinoa叶片黄酮的乙醇提取工艺和分析基因型间的差异,为藜麦黄酮的开发和高黄酮的品种筛选提供理论依据,采用3因子3水平正交试验设计,探讨了乙醇体积分数、料液比和浸提时间等因素对藜麦叶片黄酮提取率的影响;并采用最佳提取条件,对10个不同基因型品种藜麦的叶片黄酮得率进行了比较分析。结果表明:藜麦叶片黄酮最佳提取条件为体积分数70%乙醇,1∶40料液比,80℃水浴下回流浸提0.5 h。在优化条件下,1次提取工艺得率达85%以上。各因素对叶片黄酮提取率的影响程度依次为:浸提时间>乙醇体积分数>料液比。比较结果发现:藜麦叶片黄酮得率存在明显基因型差异,其中品种PI814932的叶片黄酮类物质得率最高,达0.933%。所测藜麦品种的叶片黄酮平均得率为0.619%,变异系数为34.44%。研究表明:乙醇回流法适于提取藜麦总黄酮类化合物。     

密度调控对女贞人工林土壤铵态氮和硝态氮季节变化的影响

以江苏扬州市江都区7年生女贞人工林为对象,研究密度调控对其土壤铵态氮（NH4＋-N）和硝态氮（NO3--N）含量剖面分布特征及季节变化的影响。结果表明：不同调控密度下女贞人工林土壤NH4＋-N和NO3--N含量均随着土层深度的增加而逐渐降低,且表现出明显的季节变化;3种密度调控下土壤NH4＋-N 含量均是秋冬两季较高,春季最低,夏季又有明显回升,且3种密度中的2275株/hm2下NH4＋-N含量在4个季节均高于另外2种密度;土壤NO3--N 季节变化与NH4＋-N大体一致,在春季含量最低,但不同密度间变化趋势差异较大;土温、含水量及有机质均与土壤NH4＋-N和NO3--N含量存在显著的相关关系（p＜0.05）,且在夏季均达到极显著相关（p＜0.01）。     

Anatomical Study on the Multi-Ovule Development and Abortion of Hanfu Apple

The terminal flower buds of 6-yr-old Hanfu apple were used to study the ovule development, ovular characteristics, cell death of abortive ovules, and dynamic change of starch grain quantity in the embryo sac with paraffin slices and terminal deoxynucleotidyl transferase-mediated fluorescein deoxyuridine triphosphate nick-end labeling （TUNEL） system. Four anatropous ovules in each ventricle could be observed before flowering. With the developing of floral organ, the bulk of normal ovules enlarged in each ventricle, the mature embryo sac differentiated into nucellus, and the egg cell developed into zygote by double fertilization. A large number of starch grains were observed during pollen tube growth and double fertilization, which guaranteed basic nutrient supply in the normal development of ovules. Moreover, abortion phenomenon of runtish ovules emerged at the stages of mature embryo sac, double fertilization and zygote development. The abortion characteristics included deformity of ovule development, degradation of nucellus tissue, separation between funiculus and ovule, abnormality of four-nucleate embryo sac, as well as development interruption of mature embryo sac. TUNEL analysis proved that ovule abortion was programmed cell death.     

结球菊苣品种比较试验

以3个结球菊苣品种为试验材料,进行品种比较试验,结果表明,红叶菊苣叶球形态呈长椭圆形,单株重、单位面积产量最高,抗逆性和抗病性最强,抽薹率低,适合吉林地区栽培,可作为结球菊苣推广品种在吉林地区推广.     

Detection of Penaeus monodon-type baculovirus (MBV) infection in Penaeus monodon Fabricius by in situ hybridization

Abstract. A digoxigenin-labelled DNA probe was used for in situ detection of the Penaeus monodon -type baculovirus (MBV) derived from cloned MBV polyhedrin genome in cultured Penaeus monodon Fabricius. First, the specificity of the probe against MBV DNA with dot blot hybridization analysis was verified. This probe indicated that cloned MBV polyhedrin fragment can be used as an MBV-specific probe. This was then used to microscopically examine sections of MBV-infected tissues for a blue-purple precipitate indicative of a positive reaction for MBV. MBV-positive cells were located only in the epithelium of the hepatopancreatic tubules and of the midgut. Furthermore, comparison of the susceptibility to MBV infection among several life-stages of the shrimp showed that the MBV genome was found in the zoea, mysis, post-larva, and adult stages, whereas MBV DNA was not detected in either eggs or nauplii. The results were quantified from in situ hybridization with an image analyser to compare the degree of cell infection among groups of cultured P. monodon collected from various farms in Taiwan.     

大白菜突变体库的构建及M2叶片表型变异的研究

 利用不同浓度甲基磺酸乙酯（EMS）诱变大白菜自交系‘A03’种子,根据M₁代植株发芽率、成活率、育性及M₂代成活率,筛选出适宜的EMS诱变浓度为0.4%。采取EMS种子诱变1次、EMS种子诱变2次及EMS花蕾诱变结合小孢子培养的方法构建了含有4 253个家系及相应自交M₂代种子的大白菜突变体库。在该突变体库M₂群体中分别对苗期6 404株幼苗及莲座期7 756个单株的叶片性状进行了形态学调查,总的表型变异频率高达37.62%和26.33%。     

Effects of maternal limb ischemic preconditioning on fetal hippocampal neuron apoptosis induced by intrauterine distress-reoxygenation in rats

AIM: To evaluate the influence of maternal limb ischemic preconditioning (LIP) on the apoptosis of fetal hippocampal neurons induced by intrauterine distress-reoxygenation in rats. METHODS: Intrauterine ischemia was induced by clamping the uterine and uterine branch of the ovarian blood vessels with aneurysm clamps for a period of 15 min followed by removal of the clamps to permit reperfusion. Sprague-Dawley (SD) rats (n=12) were randomly divided into 4 groups on the 19th pregnant day: sham (S) group, LIP group, fetal distress (FD) group and LIP+FD group. The cesarean birth occurred on embryonic day 21 to obtain 12 fetal rats alive in each group. The fetal rats were decapitated and the pyramidal cells in CA1 hippocampus were observed under light microscope. The neuronal apoptosis was detected by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) staining and the apoptotic rate was calculated. The expression of Bcl-2 and Bax were detected by immunohistochemical method and Western blotting. RESULTS: The rates of neuronal apoptosis in FD group and LIP+FD group were significantly higher than that in S group (P<0.05), while no significant difference was observed between S group and LIP group (P>0.05). The expression of Bcl-2 and Bax in FD group and LIP+FD group was significantly higher than that in S group (P<0.05), while no significant difference was observed between S group and LIP group (P>0.05). The ratio of Bcl-2/Bax was lower in FD group than that in S group. Compared with FD group, the rate of neuronal apoptosis was significantly lower (P<0.05), while the ratio of Bcl-2/Bax was significantly higher (P<0.05) in LIP+FD group. CONCLUSION: Maternal limb ischemic preconditioning attenuates the apoptosis of fetal hippocampal neurons induced by intrauterine distress-reoxygenation in rats, which may be associated with the up-regulation of Bcl-2 expression.