马立克氏病病毒CV1988株pp24基因的克隆与表达

根据马立克氏病病毒（MDV）国际参考强毒CA株pp24基因序列，设计和合成了一对引物，其两端分别含SalI和EcoRI的酶切位点，以CV1988株基因组DNA为模板，通过PCR技术，扩增其pp24基因。PCR产物经纯化后，按正确的阅读框架定向克隆到表达性载体pGEX-6P-1中谷胱甘肽转移酶（GST）基因的下游，并用序列测定验证。将重组质粒转化的大肠杆菌BL21，在1.0mMIPTG和37℃条件下诱导，GST-pp24基因获得了表达。诱导菌体的裂解物经12%的SDS聚丙烯凝胶电泳（SDS-PAGE）和Westemblot试验验证。得到大小为43kD的融合蛋白，与预期大小一致。将该融合蛋白的凝胶带切下研碎后免疫小鼠三次后，其血清与MDV感染的鸡胚成纤维细胞（CEF）在间接免疫荧光试验中呈阳性反应。     

24-Hour Profiles of Plasma Cortisol and Testosterone in the Male Dog: Absence of Circadian Rhythmicity, Seasonal Influence and Hormonal Interrelationships

Contents: Temporal changes of plasma cortisol and testosterone concentrations were measured in intact adult male beagle dogs during the spring and autumn with natural photoperiods of 16L:BD and 10L:14D, respectively. Each animal was fitted with an indwelling cephalic vein catheter and blood was withdrawn every 30 min for 24 h. Examination of the hormonal profiles revealed that both steroids are secreted episodically characterized by irregular short term fluctuations which were variable among different dogs. In several instances sustained elevations of cortisol and testosterone secretion occurred but were unrelated to photoperiod. Despite individual variability in the secretion patterns the daily means of each hormone were similar and not statistically different between spring and autumn .
When the 24 h were divided into three 8-h periods, namely 8.00–16.00 h (I), 16.00–24.00 h (II) and 24.00–8.00 h (III) overall and baseline mean cortisol and testosterone levels were not significantly different between the individual time periods. It appears that the dog differs from other species in that cortisol secretion was not related to the light-dark cycle. Concentrations of both hormones were not significantly correlated with each other throughout the sampling periods. Results from this study indicate that in intact male dogs cortisol and testosterone are secreted episodically without any evidence of circadian rhythmicity and seasonal variation and that an endocrine interrelationship between the adrenal cortex and the testis is not prominent under normal physiological conditions .     

2,4-表油菜素内酯结合氯化钙处理对采后杏果实黑斑病及苯丙烷代谢的影响

为探究减轻杏果实采后黑斑病的方法,以赛买提杏果实为试验材料,经3.0 mg·L^-1 2,4-表油菜素内酯(EBR)结合2%氯化钙(CaCl₂)减压渗透处理后,于温度1.0±1℃、相对湿度90%~95%条件下放置48 h后,进行交孢链格孢(Alternaria alternata)损伤接种,并进行A. alternata离体抑菌试验。结果表明,3.0 mg·L^-1 EBR结合2% CaCl₂对A. alternata的菌落直径和孢子萌发没有直接抑制作用。EBR结合CaCl₂处理可通过诱导苯丙氨酸解氨酶(PAL)、肉桂酸-4-羟化酶(C4H)、香豆酸辅酶A连接酶(4CL)活性上升,增加总酚、类黄酮和木质素含量来抑制杏果实接种发病率与病斑直径的上升。综上,EBR结合氯化钙处理可通过诱导苯丙烷代谢途径来增强杏果实对黑斑病的抗性。本研究结果可为减轻杏果实采后黑斑病提供技术参考与理论依据。     

Effect of 24-epibrassinolide on growth,yield, antioxidant system and cadmium content of bean (Phaseolus vulgaris L.) plants under salinity and cadmium stress

The plants of Phaseolus vulgaris L. were grown in the presence of NaCl and/or CdCl₂ and were sprayed with 5 μM of 24-epibrassinolide (EBL) at 15 days after transplanting (DAT) and were sampled at 30 DAT and at the end of experiment. The plants exposed to NaCl and/or CdCl₂ exhibited a significant decline in growth, the level of pigment parameters, green pod yield and pod protein. However, the follow up treatment with EBL detoxified the stress generated by NaCl and/or CdCl₂ and significantly improved the above parameters. The NaCl and/or CdCl₂ increased electrolyte leakage, lipid peroxidation and plant Cd²⁺ content, and decreased the membrane stability index (MSI) and relative water content. However, the EBL treatment in absence of the stress improved the MSI and relative water content and minimized plant Cd²⁺ content but could not influence electrolyte leakage and lipid peroxidation. The antioxidative enzymes and the level of proline exhibited a significant increase in response to EBL as well as to NaCl and/or CdCl₂ stress.     

水葫芦生防真菌Cercosporasp.FJ24的分离、鉴定及其分子生物学研究

从福建省得到被致病菌强烈感染的水葫芦植株,从中分离到致病菌菌株FJ24。形态学鉴定表明,该致病菌属于尾孢属。对该菌株的ITS区、EF-1、β-Tub、His-H3基因进行PCR扩增,测序结果显示其与目前已经分离、发现的多种尾孢属水葫芦致病菌存在着99%以上的序列相似性,可以认为其属于半知菌类（无性类）、丝孢纲、丝孢目、淡丛梗孢科、尾孢属的Cercospora piaropi。     

智慧图书馆24小时自助服务架构及其实现研究

[目的/意义]智慧图书馆被认为是未来图书馆发展的主导模式和最高形态,其提供的智慧服务本身具有开展自助服务的天然需求,读者在24小时“无人值守”的情况下可获取到更加高效、便捷、智能的服务。[方法/过程]从技术、经济、操作3个层面分析了智慧图书馆开展24小时自助服务的可行性,从管理系统、人员管理、读者管理、空间改造、服务创新5个方面来实现智慧图书馆的24小时自助服务,并分析智慧图书馆24小时自助服务普及的制约因素。[结果/结论]提出构建智慧空间、鼓励和支持社会力量参与建设民间图书馆、借助乡村振兴和文化扶贫等战略推进公共文化服务均等化、科学选址、精准服务等若干建议。     

Effects of 24-Epibrassinolide on Antioxidant System in Cucumber Seedling Roots Under Hypoxia Stress

This article aims to study the effects of exogenous 24-epibrassinolide (EBR) on the changes in ROS, activities of antioxidative enzymes and antioxidants in cucumber (Cucumis sativus L.) seedling roots under hypoxia stress. Seedlings of a hypoxianormoxic or hypoxic nutrient solutions that were added or not added with 10-3 mg L-1 EBR. Under hypoxia stress, the ROS levels and the lipid peroxidation were significantly increased in the roots upon exposure to hypoxia stress, which were inhibited by EBR application. The EBR treatment significantly increased the seedlings growth and SOD, APX, GR activities, and contents of AsA and GSH under hypoxia stress. From the results obtained in this study, it can be concluded that oxidative damage on seedling roots by hypoxia stress can be considerably alleviated and the tolerance of plants was elevated.     

MicroRNA-24 regulates apoptosis of cardiomyocytes after myocardial infarction

AIM：To evaluate the expression of miR-24 in infarcted myocardial tissues and to investigate the function of miR-24 during cardiomyocyte apoptosis in vitro and in vivo. METHODS：The mouse model of myocardial infarcton (MI) was established. The expression of miR-24 in the sections of infarcted myocardial tissues was measured by qRT-PCR. The expression of miR-24 was modified by transfecting oligonucleotide mimic and inhibitor of miR-24 into cardiomyocytes, or injecting lentiviral vectors intramyocardially. The apoptosis of cardiomyocytes was detected by Caspase-Glo 3/7 Assay System. The heart functions were determined by echocardiography and the scar size in MI model was observed with Masson trichrome staining. The apoptosis of infarcted myocardial tissues was detected by TUNEL method. Microarray and bioinformatic analysis were also used to predict the targets of miR-24. RESULTS：The expression of miR-24 in the infarct and border areas was down-regulated after MI. Overexpression of miR-24 in cardiomyocytes reduced the apoptosis induced by hypoxia. miR-24 transfection resulted in reduction of the scar size, and improved left ventricular fractional shortening (LVFS) and left ventricular ejection fraction (LVEF) of the mice in treatment group after 2 weeks. Furthermore, miR-24 reduced cell apoptosis in the infarct region. BCL2L11, ANK3 and SGPL1 may be the targets of miR-24 during the process of anti-apoptosis. CONCLUSION：miR-24 is down-regulated in the infarcted myocardial tissues. In vitro, miR-24 reduces apoptosis of cardiomyocytes induced by hypoxia. In vivo, miR-24 attenuates cell apoptosis in the infarct and border areas of the heart 2 weeks after MI, and ultimately improves heart functions.     

Pulsatilla saponin A affects proliferation and radiosensitivity of breast cancer cells by regulating miR-24-3p/RNF2 expression

AIMTo investigate the effect of Pulsatilla saponin A on proliferation and radiosensitivity of breast cancer cells and its mechanism. METHODSHuman breast cancer MCF-7 cells were treated with Pulsatilla saponin A at concentrations of 0, 5, 10, 15 and 20 mg/L and transfected with microRNA-24-3p (miR-24-3p) over-expression vector or inhibitory expression vector. The proliferation and radiosensitivity of the MCF-7 cells were measured by MTT assay and colony formation assay. The miR-24-3p expression and ring finger protein 2 （RNF2） mRNA level were detected by RT-qPCR. The protein expression of RNF2 was determined by Western blot. The luciferase reporter assay was used to detect the targeting relationship between miR-24-3p and RNF2. RESULTSCompared with control group (0 mg/L), the proliferation inhibitory rate of the MCF-7 cells was significantly increased in 5, 10, 15 and 20 mg/L Pulsatilla saponin A groups (P<0.05). The survival score of the MCF-7 cells treated with Pulsatilla saponin A was significantly decreased after irradiation, and the expression of RNF2 was significantly decreased (P<0.05). miR-24-3p targeted RNF2 and negatively regulated its expression. When the MCF-7 cells were simultaneously treated with Pulsatilla saponin A and miR-24-3p, the cell survival curve significantly shifted down. Inhibition of miR-24-3p expression reversed the proliferation-inhibiting and radiation-sensitizing effects of Pulsatilla saponin A on the MCF-7 cells. CONCLUSION Pulsatilla saponin A may affect the proliferation and radiosensitivity of breast cancer cells through miR-24-3p/RNF2 signaling pathway.     

一种基于模板DNA的PCR扩增难易预测方法

借助赋予四种碱基A,G,C,T四个三维空间中的点(0,0,1),(0,1,0),(-1,-1,0),(1,0,0),将一条DNA序列转化为一个二十四维向量。在此基础上利用Fisher线性判别法对人类基因组中189条基因片段进行PCR扩增难易预测,识别率达到了90%。