外源赤霉素对青花菜茎尖内源激素含量的影响

以不同浓度赤霉素（GA）分别在青花菜花芽分化前20 d（播后40 d）和花芽分化后10 d（播后70 d）叶面各喷施1次,研究外源GA对青花菜茎尖内源激素含量的影响。结果表明,青花菜生长发育过程与植株内源激素含量变化密切相关,在花芽分化期茎尖IAA、GA1/3含量保持较低的水平和适宜的iPAs含量有利于花芽分化,而在现蕾期保持适宜水平的IAA、GA1/3含量及较低水平的iPAs含量则有利于花球的形成。外源GA明显地影响了青花菜茎尖内源激素的平衡,GA处理在花芽分化期和现蕾期均抑制了茎尖IAA合成,降低了茎尖IAA含量;较低浓度的GA处理在花芽分化期可增加茎尖GA1/3含量,浓度升高反而降低其含量,但在现蕾期正好相反;GA处理在花芽分化期促进了iPAs含量的升高,而且浓度越高iPAs含量也越高,但在现蕾期低浓度GA处理可提高茎尖iPAs含量,而浓度过高反而导致iPAs含量下降。     

Effects of bFGF on brain edema,nerve function injury and autophagy related protein in rats with traumatic brain injury

AIM:To study the effects of basic fibroblast growth factor (bFGF) on brain edema, nerve function damage and autophagy related proteins in rats with head injury. METHODS:The rat model of craniocerebral injury (CI) was constructed. The rats were divided into control group, CI group, and low-, middle-and high-dose bFGF groups (n=10). The CI model was established in CI group, while the rats in control group were not given epidural impact. The rats in low-dose, middle-dose and high-dose bFGF groups were given bFGF at 2, 4 and 6 μg, respectively, by intraperitoneal injection after 30 min. The neurological function in the rats was evaluated by improved neurological function scoring. The rat brain tissues were taken, and the water content was detected. The levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and IL-1β in the brain tissue were measured by ELISA. The malondialdehyde (MDA) content was analyzed by thiobarbituric acid method. The activity of superoxide dismutase (SOD) was examined by WST-8 assay. The glutathine peroxidase (GSH-Px) activity was detected by colorimetric method. The protein levels of autophagy related proteins LC3-Ⅱ and beclin-1 in the brain tissues were determined by Western blot. RESULTS:The neurological function score was increased significantly of the rats in CI group. The rat model of craniocerebral injury was successfully constructed. Neurological function scores in the rats in low-dose, middle-dose and high-dose bFGF groups were reduced, the water content of the brain tissue was also reduced (P<0.05). The levels of TNF-α, IL-6 and IL-1 β were decreased in the brain tissues (P<0.05), the content of MDA was declined (P<0.05), the activities of SOD and GSH-Px were increased (P<0.05), the protein levels of LC3-Ⅱ and beclin-1 were decreased, compared with the untreated rats in CI group (P<0.05). CONCLUSION:bFGF improves the nerve function of the rats with craniocerebral injury, reduces the water content of the brain tissue, reduces the expression of autophagic protein LC3-Ⅱ and beclin-1.The mechanism is related to the inhibition of inflammatory reaction and oxidative damage.     

Basic fibroblast growth factor promoting the healing of palatal perforation in rats

AIM: To study the influence of basic fibroblast growth factor(bFGF) on treating ed palatal perforation in rats. METHODS: bFGF was given to the early palatal perforation in rat. The granulation tissues in perforations were grossly and pathologically obserVed. RESULTS: The the of wound healing was significantly in- crease in the bFGF group (P<0.01 ). bFGF could obviously promote the proliferation of the granulation tissues and fibroblasts. The number of AgNORs granules in fibrablasts were 3.73 ±0.52 in the buy group and 2.11 ±0.31 in control group (P < 0.05). CONCLUSION: bFGF could promote the growth of granulation tissues and had a strong promotion on the wound healing in palatal perforation. It was helpful in repairing palatal perforation.     

百合花色机理研究进展

百合花色主要由花青苷和类胡萝卜素形成,一般粉色和棕色花中主要成分是花青苷;黄色和橙色花中主要成分是类胡萝卜素;红色是由花青苷和类胡萝卜素共同组成。百合花色的遗传现象比较复杂。目前初步构建了几个群体,通过遗传连锁图谱标记了花色相关位点。百合中花青苷和类胡萝卜素合成代谢途径中的大部分结构基因已经被克隆和研究。花青苷合成途径中的转录因子LhMYB12调控百合花被片中的花青苷的有无,其转录水平调控花青苷的颜色强度和着色类型。本文总结了百合花朵的着色机理研究,为进一步开展百合花色育种提供了理论参考。     

FAD-dependent sulfhydryl oxidase activity of augmenter of liver regeneration protein and its activity motif

AIM:To establish a sensitive and effective method to determine whether augmenter of liver regeneration protein (ALRp) has the activity of sulfhydryl oxidase.METHODS:The cysteine at the position of 65 in CXXC motif contained in ALRp was changed into alanine through site-directed mutation.There were four groups in the experiment:① ALR group;② ALR-FAD group;③Mutated ALR-FAD group;④ Control group.Using GSH as substrate,at different time points,the concentration of thiol in each group was measured.RESULTS:The concentration of thiol groups in the ALR-FAD group was decreased during the time of the experiment.But the thiol in the other groups didn’t show any significant decrease.CONCLUSION:A sensitive and effective method to determine the sulfhydryl oxidase activity of ALRp was established.ALRp is a FAD-dependent sulfhydryl oxidase,and the conserved CXXC motif is indispensable to ALRp.     

黑木耳原生质体制备、再生及单核体荧光鉴定

利用正交试验研究了黑木耳原生质体制备、再生的最佳条件,结果表明,原生质体制备最佳条件是酶解温度31℃、酶浓度1．0％、菌龄5d、酶解时间4h,原生质体再生的最佳条件是酶解温度27℃、酶解时间4h、酶浓度1．5％、菌龄11d。稳渗剂种类对黑木耳原生质体再生率的试验结果表明,采用0．5mol／L蔗糖为稳渗剂,各黑木耳菌株的原生质体再生率最高。HW2号菌株原生质体经核染色其单核化比率最高;在荧光显微镜下观察菌丝核相,单、双核清晰可辨。     

珠美海棠MzWRKY基因家族盐胁迫应答模式研究

对苹果属植物现有的58个WRKY 的EST序列进行分析,得到了37条uniESTs。珠美海棠（Malus zumi Mats）是耐盐的优良苹果砧木,能在土壤全盐含量为0.6%的盐碱地上存活。根据已知的37条uniESTs序列,通过半定量RT-PCR研究了珠美海棠中MzWRKY基因家族的盐应答模式。28个MzWRKY基因能检测到RT-PCR的产物,其中21个基因受盐诱导,1个受盐抑制。根据MzWRKY基因盐应答模式可将这些基因分为两组,基因表达模式的差异说明MzWRKY在珠美海棠的盐应答中角色的多样性。     

子宫内膜中TLR4的定位及pcDNA3.1-TLR4载体的构建

试验旨在研究TLR4(Toll like receptor4)蛋白在子宫内膜组织中具体的分布与表达,从而构建TLR4真核表达载体,为TLR4在细胞水平上的功能研究提供一种技术手段.选择早期妊娠绵羊(9d、13d、17d、21d、25d)子宫组织,采用免疫组织化学和细胞免疫学抗体着色法检测各阶段子宫组织中TLR4蛋白的分布与表达,确定受体蛋白的表达部位.同时将构建的pcDNA3.1-TLR4质粒转染到确定的受体细胞,检测TLR4蛋白的表达水平.结果表明:TLR4蛋白在子宫内膜各部位均有表达,呈一定的动态时空模式,且第17d胚胎附植后主要表达于内膜基质细胞;重组质粒转染到基质细胞后,TLR4蛋白的表达水平明显增强.此研究成功构建pcDNA3.1-TLR4表达载体,为TLR4在生殖细胞免疫学功能的研究奠定了一定的理论基础.     

脉红螺附着变态与食性转换的研究

为实现脉红螺苗种的规模化生产,本实验通过对脉红螺幼虫培育、采苗、稚螺培育等方面的系统研究,探索了脉红螺苗种繁育过程中幼虫附着变态率低和饵料转换难的问题。结果显示:脉红螺浮游幼虫前期(0~12 d)生长较快,壳高平均日增长21.8μm;中期(13~18 d)生长较慢,壳高平均日增长3μm;后期(19~30 d)生长加快,壳高平均日增长46.8μm。用取自沉淀池中的新鲜海水对脉红螺浮游幼虫进行附着变态诱导,诱导第1天脉红螺浮游幼虫开始出现附着变态,第4天时,附着变态活动基本结束,附着变态率为40%。为变态后的稚螺投喂刚附着的长牡蛎和魁蚶稚贝,第1~8天,稚螺壳高平均日增长65.6μm;第9天开始,稚螺生长速度加快;第9~12天,稚螺壳高平均日增长136.4μm;第13~16天,稚螺壳高平均日增长307.3μm;第17~20天,稚螺壳高平均日增长445.8μm。研究表明,用取自沉淀池中的新鲜海水诱导脉红螺浮游幼虫的附着变态效果好,刚附着的长牡蛎稚贝和魁蚶稚贝可以作为脉红螺稚螺培育阶段的适宜饵料。     

甘露糖修饰的壳聚糖聚乳酸-羟基乙酸共聚物纳米微球作为口蹄疫病毒核酸疫苗递送载体的特性评价

旨在对制备的甘露糖修饰的壳聚糖聚乳酸-羟基乙酸共聚物[poly （D,L-lactide-co-glycolide）,PLGA]纳米微球作为口蹄疫病毒（foot-and-mouth disease virus,FMDV）核酸疫苗递送载体进行评价。采用西佛碱反应和元素分析制备具有一定取代度的甘露糖修饰的壳聚糖衍生物（mannose modified chitosan,MCS）,然后,经双重乳化挥发法制备得到甘露糖修饰的壳聚糖PLGA纳米微球（MCS-PLGA-NPs）。采用纳米粒径仪检测MCS-PLGA-NPs粒径分布和表面电势（zeta）、扫描电镜考察其形态、琼脂糖凝胶电泳观察其对质粒的吸附和吸附质粒后抵抗核酸酶降解能力、CCK-8法检测MCS-PLGA-NPs的细胞毒性、激光共聚焦观察巨噬细胞对MCS-PLGA-NPs-质粒DNA复合物的摄取、荧光显微镜和Western blot验证MCS-PLGA-NPs加载质粒DNA在细胞中的表达。元素分析结果表明,成功制备了取代度为5%~10%的MCS。纳米粒径测定和扫描电镜结果表明,MCS-PLGA-NPs的zeta为正值、粒径分布均匀且形态规则呈球形。琼脂糖凝胶电泳结果显示,MCS-PLGA-NPs吸附质粒的能力随着其质量的增加而增强并且可以在一定程度上抵抗核酸酶降解质粒DNA。在细胞毒性试验中,不同浓度的MCS-PLGA-NPs与RAW264.7细胞共孵育24 h后,细胞存活率仍在85%以上。在细胞摄取试验中,用激光共聚焦显微镜可以明显观察到质粒DNA结合到纳米微球表面被RAW264.7细胞摄取。荧光显微镜和Western blot试验证明MCS-PLGA-NPs加载质粒DNA可以在细胞中进行表达。综上表明,本研究成功制备了MCS以及具有递送核酸疫苗能力的MCS-PLGA-NPs,为FMDV核酸疫苗的递送研究提供了新的方向和见解,也为该递送载体携带特定抗原靶向抗原递呈细胞表面甘露糖受体以及应用于动物免疫的研究奠定基础。