Effects of cyhalothrin on the transient outward potassium current in central neurons of Helicoverpa armigera

The effects of cyhalothrin on the transient outward potassium current in central neurons of Helicoverpa armigera were studied by using the patch clamp techniques. The results showed that before using cyhalothrin （10.5 mmol/L）, activation potential was approximately -40 mV, after application of the drug, the activation potential shifted roughly 10 mV to the negative potential direction, so channels can be activated more easily. Before and after cyhalothrin application, the change of current amplitude was insignificant. The value of V1/2 and k of activation curves did not change significantly, however, the V1/2 of the inactivation curves changed significantly. Inactivation curves significantly shifted to a negative direction, so that inactivation of the channels was hastened. It is indicated that there may exit a primary way in which cyhalothrin provides neurotoxicity to the nervous system through the regulation of activation potentials and inactivation state of IA channels.     

Characterization of geraniol synthase from the peltate glands of sweet basil

The monoterpene fraction of the lemon-scented sweet basil (Ocimum basilicum) cv Sweet Dani consists mostly of citral (a mixture of geranial and neral), with lower levels of geraniol and nerol. These compounds are stored in the peltate glands found on the leaf epidermis. Younger leaves, which have a higher density of such glands, also have a higher content of monoterpenes than older leaves. Geraniol synthase (GES) activity, generating geraniol from geranyl diphosphate, was shown to be localized exclusively or almost exclusively to glands. GES activity resides in a homodimeric protein that was purified to near homogeneity. Basil GES requires Mn2+ as a divalent metal cofactor for activity and produces only geraniol from geranyl diphosphate. Km values of 21 and 51 microM were obtained for geranyl diphosphate and Mn2+, respectively. In the presence of 18O-labeled water, GES catalyzed the formation of 18O-geraniol from geranyl diphosphate, indicating that the reaction mechanism of GES is similar to that of other monoterpene synthases and is different from the action of phosphatases. A GES cDNA was isolated based on analysis of a glandular trichome expressed sequence tag database, and the sequence of the protein encoded by this cDNA shows some similarity to sequences of other terpene synthases. The expression of the GES cDNA in Escherichia coli resulted in a protein with enzymatic activity essentially identical to that of plant-purified GES. RNA gel-blot analysis indicated that GES is expressed in glands but not in leaves of basil cv Sweet Dani, whose glands contain geraniol and citral, and not in glands or leaves of another basil variety that makes other monoterpenes but not geraniol or citral.     

Heart rate variability analysis in general medicine

Autonomic nervous system plays an integral role in homeostasis. Autonomic modulation can frequently be altered in patients with cardiac disorders as well as in patients with other critical illnesses or injuries. Assessment of heart rate variability is based on analysis of consecutive normal R-R intervals and may provide quantitative information on the modulation of cardiac vagal and sympathetic nerve input. The hypothesis that depressed heart rate variability may occur over a broad range of illness and injury, and may inversely correlated with disease severity and outcome has been tested in various clinical settings over the last decade. This article reviews recent literature concerning the potential clinical implications and limitations of heart rate variability assessment in general medicine.     

用RAPD技术检测野生鲫鱼的四个金鱼代表品种的基因组DNA多态性

利用随机扩增多态DNA技术检测了两个不同地区的野生鲫鱼(Carassius a uratus L.)和4个金鱼(Carassius auratus Var.)代表品种的基因组DNA的多态性。用26个随机引物对各品种实验鱼的基因组DNA进行扩增,平均每个品种观察到约134个标记,单个引物获得的标记在1-16个之间。实验结果经统计学分析表明,金鱼和鲫鱼的随机扩增多态DNA共亨度高,进一步证实了金鱼由野生鲫鱼演化而来,聚类结果表明,草金鱼形成后,首先演化为文种鱼,然后由文种再形成龙种和蛋种两个品系。 Abstract:Genomic DNA polymorphisms in the wild crucian from rwo different areas and in four representative varieties of goldfish were detected by using random amplified polymorphic DNA(RAPD)technique.The genomic DNA of each variety of fish was amplified with 26 primers.On average,about 134 RAPD markers were observed by each variety.The markers obtained by a single primer varied from 1 to 16.The statistical analysis of the experimental results indicated that random amplified polymorphic DNA of the goldfish and wild crucian have a high proportion of random amplified polymorphic DNA fragment shared.This further verified that goldfish is evolved from wild crucian.The cluster analysis suggested that after emerging,grass goldfish is evolved to wen goldfish,then through wen goldfish evolved to dragoneye goldfish and oval goldfish.     

利用mRNA差别显示技术分析枸杞体细胞胚发生早期基因的差别表达

本研究选用枸杞体细胞胚发生体系中的继代愈伤组织(对照)、胚性愈伤组织和早期胚体为实验材料,提取细胞总RNA,在12种锚定真核生物mRNA3'末端的OligodT12VN中,随机选用OligodT12GA为引物合成了以上三种材料的cDNA第一链,以此cDNA为模板,用随机引物进行PCR扩增,选择差别表达的片段。我们选用了OPA、OPH、OPK和OPB四组的60个随机引物对所得的c DNA进行了PCR扩增,得到了三个在体细胞胚发生早期组织中基因特异表达的片段。结果表明,在体细胞胚发生早期有胚胎发生特异性基因的表达,而且这种特异表达的基因在继代愈伤组织中没有表达,说明植物的体细胞胚发生过程就是细胞内基因差别表达的结果。 Abstract:Embryogenic calli and early embryo can be obtained from both auxin and auxin-free medium.The analysis of differential gene expression in early somatic embryogenesis has been hindered by above-mentioned material.The modifications of the recently described mRNA differential display method were reported and differential gene expression in early slmatic embryogenesis was analyzed.We have obtained three differential bands of cDNA in early somatic embryogenesis.The results indicate that gene expression has temperal and spalil order in early somatic embryogenesis of Lycium barbarum L.Plant somatic embryogenesis is the results of differential gene expression in cell.     

AFLP分析中多态性扩增产物的回收、克隆及鉴定

本研究在摸索和优化了水稻AFLP分析体系的基础上,发展了多态性AFLP产物的高效克隆方法。特异AFLP扩增产物直接从变性聚丙烯酰胺凝胶上分离纯化,再经过一至二轮PCR扩增,即可高效地克隆于pGEM-Teasy vector系统中。本实验利用该方法成功地克隆了水稻温敏核不育等位突变系546 0S和5460F间的4个多态性AFLP产物,Southern bloting分析证明其中3个产物在水稻基因组中为单拷贝序列,另一个为低拷贝序列。AFLP技术强有力的多态性检出能力再结合多态性扩增产物的高效克隆方法,为寻找与目标基因紧密连锁的分子标记提供了有力工具。 Abstracts:An efficient method for cloning DNA fragment from denaturing polyacrylamide gels was developed to allow the isolation of specific bands obtained from amplified fragment length polymorphism(AFLP)products.After isolation and purification from the thin denaturing polyacrylamide gels,specific AFLP products were successfully cloned after one or two rounds of PCR reamplification.Using this method 4 polymorphic AFLP products between a pair of rice allelic lines differing for thermo-sensitive genic male sterile(TGMS)ene were cloned and it was confirmed that 3 of the AFLP products represented single copy sequences and the other 1 represented low copy sequence in rice genome.     

应用大规模测序技术和生物信息学研究造血干/祖细胞的基因表达及新基因 的识别和克隆

从新生儿脐血和成人骨髓中分选出造血干/祖细胞(HSC/HPC),构建成cDNA文库,对其进行大规模表达序列标签(EST)测序,通过生物信息学等手段分析基因表达谱,并进行新基因的全长cDNA克隆。在所测的10512条可分析E ST序列中,有9866条来自脐血CD34+|细胞,其中4697条(47.6%)为已知基因,2603条(26.4%)为已知EST,1415条(14.3%)代表未知EST。在已知基因中,8.2%基因与造血相关,22.7%涉及细胞代谢、结构和迁移,13.0%与细胞分裂和防御相关,26.2%与RNA、蛋白质的合成相关,10.6%和细胞信号传递有关。对一些已知和未知的EST,综合测序、生物信息学等方法,进行全长克隆,已获得23个新基因的全长cDNA。 Abstract:Hematopoietic stem/progenitor cells were isolated from umbilical cord blood and adult bone marrow,and subject to cDNA library construction.The gene expression pattern in CD34+ cells and the identification and cloning of novel genes were performed by sequencing ESTs and analyzing them with the tools of bioinformatics.Among the obtained 10 512 ESTs which could be further analyzed,9,866 were from umbilical cord blood where 4 697(67.6%)were known genes,2 603(26.4%)were known ESTs and 1415(14.3%)represented novel ESTs.Within the identified genes,8.2% was involved in hematopoiesis,22.7% was associated with cell metabolism,structure and mobility,13.0% was linked to cell division and defence,26.2% was related to RNA protein synthesis and 10.6% was related with cell signal transduction.In parallel,we developed an efficient working system combining sequencing,bioinformatics,etc.and obtained 23 full-length cDNAs from both known and novel ESTs identified in this work.     

基因枪转移Bar基因获得抗除草剂(Basta)的糜子蔗克隆再生植株

用糜子蔗(sorghum×sugarcane)×millet sorghum茎尖愈伤组织为受体,通过基因枪轰击将Bar基因转入糜子蔗细胞;用除草剂Basta含量分别为1、2.5、5、10mg/L的8114诱导与分化培养基上选择抗性愈伤组织;在Bast a含量为1与1.5mg/L筛选培养基上,选择出3个快速增殖并大量再生绿苗的体细胞无性系(Clone),如BarC1、BarC2、BarC3;BarC1、C2再生植株用Basta含量为1、2.5-5mg/L的溶液进行抗性检测,证明这些小植株具有极强的抗性。 Abstract:The Bar genes were introduced into〔(sorghum×sugarcane) ×millet sorghum〕callus cells using particle bombardment.The callus were cultured on the 8114 media with Basta,the Bastas concentration is 1,2.5 and 5,10mg/L.3 Clones with regenerated capacity were screened out on the media with Basta 1,2.5mg/L,for example BarC1,BarC2,BarC3.The regenerated plant from BarC1、BarC2 were cultured on the solution with Basta 1.5,2.5,and 5mg/L,high resistance were obtained in these regenerated plants.