血管钠肽、 C型钠尿肽和心房钠尿肽舒血管作用的对比

本实验采用离体血管灌流方法,观察和比较血管钠肽（ＶＮＰ）,Ｃ型钠尿肽（ＣＮＰ）和心房钠尿肽（ＡＮＰ）对大鼠肺动脉,腹主动脉和腹腔静脉的舒张作用。．结果表明,ＶＮＰ,ＣＮＰ和ＡＮＰ对离体大鼠的保留内皮与去内皮的肺动脉,腹主动脉和腹腔静脉均有浓度依赖性舒张作用。     

影响K型杂交小麦粒重潜在因素的遗传分析

1996-1997年采用5×7不完全双列杂交,对影响K型杂交小麦粒重的潜在因素进行分析。结果表明,K型杂交小麦粒重与胚乳细胞数和鲜籽烂一最大体积呈极显正相关。鲜籽粒最大体积对粒重贡献较大;胚乳细胞数的杂种优势较弱,鲜籽粒最大体积的杂种优势十分明显;这两个性状均符合加性-显性遗传模型。以加快效应为主,存在明显的母体效应。     

突触泡蛋白2研究进展

突触泡蛋白2(SV2)是一类跨膜糖蛋白,定位于脊椎动物神经元及内分泌细胞,与神经递质的释放、内分泌泡胞吐作用、突触泡稳态的维持、神经肌肉接头的形成及肾上腺素能受体α2C的定位密切相关。最近还发现SV2是肉毒神经毒素BoNT/A的受体,介导BoNT/A进入神经元。SV2可作为突触泡标记蛋白,广泛应用于生物学研究及肿瘤诊断。此外,SV2还是抗癫痫药物的作用靶标。     

Synthesis of safflomide and its HPLC measurement in mouse plasma after oral administration

Safflomide (N-caffeoyltryptamine) is a compound belonging to a group of phenylpropanoid amides found in plants. In this study, safflomide was chemically synthesized and confirmed by LC-MS, LC-MS/MS and NMR spectroscopic methods, and a high-performance liquid chromatography (HPLC) method was developed for quantifying safflomide in biological samples. The synthesis was simple, and the yield of safflomide was greater than 50%. Using the synthesized safflomide as a standard, HPLC separation was performed on a Nova-Pak C18 column using an isocratic buffer, and the separation was detected using a coulometric electrochemical detector. The detection of safflomide yielded an excellent peak resolution at the retention time of 21 min, and the lower limit of the detection was as little as 100 fmol. Using this HPLC method, the plasma concentrations of safflomide were determined in mouse blood, collected at 5, 10, 15, 20, 25, 30, and 35 min following its oral administrations (1 and 3 mg/30 g body weight). This HPLC method standardized with safflomide is the first reported method able to quantify the compound in standard and plasma samples with good detection limit and consistent reproducibility.     

薄荷油体外抗蠕形螨效果及杀螨机制

采用透明胶带粘贴过夜法获取2种人体蠕形螨,随机分组,观察不同浓度的薄荷油的杀虫效果,并在MoticDMB5图像采集软件系统下拍摄虫体在薄荷油作用下的虫体死亡过程。结果表明,薄荷油有很强的体外杀灭2种人体蠕形螨的作用,尤其对皮脂蠕形螨的杀灭作用显著。随着药物浓度的增加及作用时间的延长,蠕形螨死亡率增高。12.5%、3.125%分别是薄荷油体外杀灭毛囊蠕形螨和皮脂蠕形螨的最适杀螨浓度。薄荷油作用于蠕形螨,可见虫体收缩扭动,活动加剧,消化管剧烈收缩,毛囊蠕形螨和皮脂蠕形螨体壁均有渗出物外溢。虫体表现为兴奋—痉挛—失水—松弛—死亡的典型症状。薄荷油对人体蠕形螨的杀灭机制主要通过神经毒性和直接毒杀作用,造成虫体破裂脱水而死亡。薄荷油是一种极具开发潜能的高效的天然杀螨药。     

开花红哺鸡竹叶片的显微结构与光响应参数研究

为阐释竹子开花过程中的生理功能变化,探寻竹子花后复壮更新的途径,本研究选取红哺鸡竹(Phyllostachys iridescens)竹林换叶后出现开花征兆的竹株异形叶和外观正常叶,并以未开花竹株正常叶为对照,进行了叶片结构观察和光响应测定。结果发现:红哺鸡竹从营养生长向生殖生长转变的过程中,叶片的发育有变小变薄的特征,相邻维管束距离增大,维管束横切面积和气孔密度有显著变小的趋势(p0.05)。开花竹异形叶的叶片厚度、上表皮厚度、维管束横切面积和气孔密度是未开花竹正常叶的71.59%、87.40%、77.79%和73.56%,相邻维管束距离增加了19.34%,这种结构的变化相应地导致了开花竹叶片光合和蒸腾作用的显著降低(p0.05)。开花竹异形叶的光响应参数,如光饱和点、表观量子效率和最大光合速率是未开花竹正常叶的67%、40.5%和8.27%,分别为900、0.015、1.22μmol·m-2·s-1;光补偿点比未开花竹正常叶高出208.5%,达85.11μmol·m-2·s-1,而开花竹外观正常叶的光响应参数介于两者之间。开花竹株的叶片结构弱化和随之产生的生理功能异化可能是多年生竹子无法像树木那样连年开花、持续生长的重要原因之一。     

Expression of gibberellin 2-oxidase 4 from Arabidopsis under the control of a senescence-associated promoter results in a dominant semi-dwarf plant with normal flowering

Gibberellin (GA), a plant hormone, is involved in many aspects of plant growth and development both in vegetative and reproductive phases. GA2-oxidase plays a key role in the GA catabolic pathway to reduce bioactive GAs. We produced transgenic Arabidopsis plants expressing GA2-oxidase 4 (AtGA2ox4) under the control of a senescenceassociated promoter (SEN1). As we hypothesized, transgenic plants (SEN1::AtGA2ox4) exhibited a dominant semi-dwarf phenotype with a decrease of bioactive GAs (e.g., GA4 and GA1) up to two-fold compared to control plants. Application of bioactive GA3 resulted in increased shoot length, indicating that the GA signaling pathway functions normally in the SEN1::AtGA2ox4 plants. Expressions of other members of GA2-oxidase family, such as AtGA2ox1, AtGA2ox3, AtGA2ox6, and AtGA2ox8, were decreased slightly in the flower and silique tissues while GA biosynthetic genes (e.g., AtGA20ox1, AtGA20ox2 and AtGA3ox1) were not significantly changed in the SEN::AtGA2ox4 plants. Using proteome profiling (2-D PAGE followed by MALDI-TOF/MS), we identified 29 protein spots that were increased in the SEN1::AtGA2ox4 plants, but were decreased to wild-type levels by GA3 treatment. The majority were found to be involved in photosynthesis and carbon/energy metabolism. Unlike the previous constitutive over-expression of GA2-oxidases, which frequently led to floral deformity and/or loss of fertility, the SEN1::AtGA2ox4 plants retained normal floral morphology and seed production. Accordingly, the expressions of FT and CO genes remained unchanged in the SEN1::AtGA2ox4 plants. Taken together, our results suggest that the dominant dwarf trait carried by SEN1::AtGA2ox4 plants can be used as an efficient dwarfing tool in plant biotechnological applications.     

The lose of juvenility elicits adventitious rooting recalcitrance in apple rootstocks

For perennial woody plants, softwood cutting is an efficient technique for larger scale propagation and adventitious rooting of cuttings is one of the most crucial steps. To evaluate the significance of juvenility on adventitious rooting, rooting rates was compared between softwood cuttings collected from apomictic seedlings (juvenile), in vitro cultured plants (rejuvenated), suckers (juvenile like) and canopy shoots (adult) of reproductively mature trees in Malus xiaojinensis. After pre-treatment with indole-3-butytric acid (IBA) (3,000 mg L^?1) + H₂O₂ (50 mM), rooting rates in cutting from juvenile, juvenile like and rejuvenated donor plants were significantly higher (>90 %) than that from adult trees. The effects of IBA on adventitious rooting were enhanced significantly by exogenous H₂O₂. After 15 passages of in vitro subculture, the micro-shoots from adult phase explants were rejuvenated successfully, marked by the elevated expression of miR156 in the leaflets of the micro-shoots. But the rooting ability of rejuvenated micro-shoots was recovered delayed at the 18th or 21st passage of subculture. During the process of rejuvenation, the leaf indole-3-acetic acid contents and the expressions of rooting related genes CKI1, ARRO-1, ARF7 and ARF19 increased significantly. In contrary, the leaf abscisic acid contents decreased. A lack of juvenility is the most important limiting factor governing adventitious rooting of softwood cuttings in apple rootstocks.     

Live Cell Imaging of Primary Rat Neonatal Cardiomyocytes Following Adenoviral and Lentiviral Transduction Using Confocal Spinning Disk Microscopy

Primary rat neonatal cardiomyocytes are useful in basic in vitro cardiovascular research because they can be easily isolated in large numbers in a single procedure. Due to advances in microscope technology it is relatively easy to capture live cell images for the purpose of investigating cellular events in real time with minimal concern regarding phototoxicity to the cells. This protocol describes how to take live cell timelapse images of primary rat neonatal cardiomyocytes using a confocal spinning disk microscope following lentiviral and adenoviral transduction to modulate properties of the cell. The application of two different types of viruses makes it easier to achieve an appropriate transduction rate and expression levels for two different genes. Well focused live cell images can be obtained using the microscope’s autofocus system, which maintains stable focus for long time periods. Applying this method, the functions of exogenously engineered proteins expressed in cultured primary cells can be analyzed. Additionally, this system can be used to examine the functions of genes through the use of siRNAs as well as of chemical modulators.     

Activation of poly(ADP-ribose) polymerase in the rat hippocampus may contribute to cellular recovery following sublethal transient global ischemia

We have investigated the role of poly(ADP-ribose) polymerase (PARP) activation in rat brain in a model of sublethal transient global ischemia. Adult male rats were subjected to 15 min of ischemia with brain temperature reduced to 34 degrees C, followed by 1, 2, 4, 8, 16, 24, and 72 h of reperfusion. PARP mRNA expression was examined in the hippocampus using quantitative RT-PCR, northern blot analysis, and in situ hybridization. Protein expression was assessed using western blot analysis. PARP enzymatic activity was investigated by measuring nuclear [3H]NAD incorporation. The presence of poly(ADP-ribose) polymers was assessed immunocytochemically. Although PARP mRNA and protein expressions were not altered after ischemia, enzymatic activity was increased 4.37-fold at 1 h (p < 0.05 vs. sham) and 1.73-fold (p < 0.05 vs. sham) at 24 h of reperfusion. Immunostaining demonstrated the presence of poly(ADP-ribose) polymers in CA1 neurons. Cellular NAD+ levels were not significantly altered at any time point. Furthermore, systemic administration of 3-aminobenzamide (30 mg/kg), a PARP inhibitor, prevented the increase in PARP activity at 1 and 24 h of reperfusion, significantly decreased the number of surviving neurons in the hippocampal CA1 region 72 h after ischemia (p < 0.01 vs. sham), and increased DNA single-strand breaks assessed as DNA polymerase I-mediated biotin-dATP nick-translation (PANT)-positive cells (p < 0.01 vs. sham). Furthermore, using an in vitro DNA repair assay, 3-aminobenzamide (30 mg/kg) was shown to block DNA base excision repair activity. These data suggest that the activation of PARP, without subsequent NAD+ depletion, following mild transient ischemia may be neuroprotective in the brain.