海洋微生物纤维素酶的研究进展及其应用

海洋极端酶因在极端环境中具有更高的酶活性及更好的稳定性,因而具有重要的理论价值和工业应用前景.随着海洋微生物极端酶的研究开发,海洋微生物纤维素酶的研究也逐渐受到学者们的关注,并取得了较大进展.综述迄今为止分离出的产纤维素酶的海洋微生物种群及其酶学特性,海洋微生物纤维素酶基因克隆与表达的国内外研究现状,分析海洋微生物纤维素酶潜在的应用价值及未来发展前景.     

月季愈伤组织的诱导及植株再生

以可在黑龙江地区露地越冬的5个现代月季（Rosa chinensis）品种为实验材料,分别以其无菌苗的叶片和茎段为外植体,研究了愈伤组织诱导及植株再生方法。实验结果表明：5个寒地月季品种的叶片和茎段均可诱导出愈伤组织,2,4-D诱导愈伤组织的效果较好,高浓度的细胞分裂素不适合用于月季叶片和茎段愈伤组织的诱导;TDZ在月季愈伤组织分化培养过程中具有重要作用,光照培养可促进月季愈伤组织的分化,愈伤组织的分化能力随着继代次数的增加呈下降趋势。该实验成功地从2004-8和2004-9（2个月季品种）愈伤组织中诱导出再生植株,其愈伤组织的分化率分别为45%和38%。     

犬细小病毒VP2基因在枯草芽孢杆菌中的表达及鉴定

从犬细小病毒/犬瘟热二联苗中提取CPV基因组,根据GenBank发表的CPV-VP2基因序列设计一对引物,对VP2基因进行PCR扩增,并克隆至TA Cloning(R) Kit Dual Promoter( pCRⅡ),获得克隆栽体pCRⅡ-VP2.将重组质粒亚克隆到枯草芽孢杆菌表达载体pHT43上,获得重组表达栽体pHT43-VP2.经双酶切鉴定及序列比对分析后,将pHT43-VP2载体转化入枯草芽孢杆菌WB600中进行诱导表达,产物使用PAGE胶蛋白回收法纯化.结果表明,在69 kD处存在目的蛋白,ELISA检测发现,纯化后的目的蛋白与阳性血清存在特异性反应.     

柿ξ-胡萝卜素脱氢酶基因的克隆与序列分析

通过对柿ζ-胡萝卜素脱氢酶(-ζCarotene desaturase,ZDS)基因的克隆及序列分析,旨在为改良柿果实品质奠定基础。根据GenBank中其他植物ZDS基因的保守序列设计了两条PCR扩增引物,以柿果实中所提取的RNA为模板,采用RACE技术扩增出一条约为500bp的条带。经过序列分析,其长578bp,不含内含子,具有部分开放阅读框(177bp)、终止密码子(TGA)和poly(A)尾巴(11bp),共编码58个氨基酸。该序列及其所编码的氨基酸与其他植物ζ-胡萝卜素脱氢酶基因均具有较高的同源性。该基因已提交GenBank数据库,登录号为GU075728。     

果胶酶pelB基因片段克隆及序列分析

根据GenBank上登录的果胶酶基因保守序列设计引物,从本实验室已筛选的一株具有降解果胶质功能的枯草芽孢杆菌S-1中克隆到了pelB基因片段,pelB与pMD20-T载体连接后转化到大肠杆菌JM109中,测序并构建进化树分析.结果表明,其序列与来源于Bacillus sp.果胶裂解酶pel-15和pelA的同源性分别为40.98%和39.20%;与Bacillus Pumilus的pelB一致性为40.32%.推断此pelB为类似果胶酶基因片段.     

亚麻外源基因遗传转化研究进展

亚麻作为重要的经济作物,其遗传转化研究对亚麻新品种的选育工作具有重要意义.分析亚麻遗传转化方法、提高转化效率及转化组织筛选等方面,重点从抗除草剂基因、抗病基因、抗重金属逆性基因、改善亚麻纤维质量基因,以及改善亚麻油质量基因方面综述亚麻遗传转化外源基因的研究和应用进展,并讨论亚麻遗传转化过程中存在的问题和解决策略.     

Development of a Loop-Mediated Isothermal Amplification Assay for Porcine Circovirus Type 2

In this study, the loop-mediated isothermal amplification (LAMP) method was used to develop a rapid and simple detection system for porcine circovirus type 2 (PCV2). According to the PCV2 sequences published in GenBank, multiple LAMP primers were designed targeting conserved sequences of PCV2. Using the DNA extracted from PCV2 isolates HUN-09 and SD-09 as the template, LAMP reactions in a PCV2 LAMP system was performed, the amplification products were detected by adding SYBR Green I and could be observed di...     

A prospective controlled study: Minimally invasive stereotactic puncture therapy versus conventional craniotomy in the treatment of acute intracerebral hemorrhage

Background

Young stroke can have devastating consequences with respect to quality of life, the ability to work, plan or run a family, and participate in social life. Better insight into risk factors and the long-term prognosis is extremely important, especially in young stroke patients with a life expectancy of decades. To date, detailed information on risk factors and the long-term prognosis in young stroke patients, and more specific risk of mortality or recurrent vascular events, remains scarce.

Methods/Design

The FUTURE study is a prospective cohort study on risk factors and prognosis of young ischemic and hemorrhagic stroke among 1006 patients, aged 18-50 years, included in our study database between 1-1-1980 and 1-11-2010. Follow-up visits at our research centre take place from the end of 2009 until the end of 2011. Control subjects will be recruited among the patients' spouses, relatives or social environment. Information on mortality and incident vascular events will be retrieved via structured questionnaires. In addition, participants are invited to the research centre to undergo an extensive sub study including MRI.

Discussion

The FUTURE study has the potential to make an important contribution to increase the knowledge on risk factors and long-term prognosis in young stroke patients. Our study differs from previous studies by having a maximal follow-up of more than 30 years, including not only TIA and ischemic stroke but also hemorrhagic stroke, the addition of healthy controls and prospectively collect data during an extensive follow-up visit. Completion of the FUTURE study may provide better information for treating physicians and patients with respect to the prognosis of young stroke.     

Flow‐injection chemiluminescent determination of estrogen benzoate using the tris(1,10‐phenanthroline) ruthenium(II)–permanganate system

Chemiluminescence (CL) detection for the determination of estrogen benzoate, using the reaction of tris(1,10–phenanthroline)ruthenium(II)–Na₂SO₃–permanganate, is described. This method is based on the CL reaction of estrogen benzoate (EB) with acidic potassium permanganate and tris(1,10–phenanthroline)ruthenium(II). The CL intensity is greatly enhanced when Na₂SO₃ is added. After optimization of the different experimental parameters, a calibration graph for estrogen benzoate is linear in the range 0.05–10 µg/mL. The 3 s limit of detection is 0.024 µg/mL and the relative standard deviation was 1.3% for 1.0 µg/mL estrogen benzoate (n = 11). This proposed method was successfully applied to commercial injection samples and emulsion cosmetics. The mechanism of CL reaction was also studied. Copyright © 2011 John Wiley & Sons, Ltd.     

Diblock glycopolymers promote colloidal stability of polyplexes and effective pDNA and siRNA delivery under physiological salt and serum conditions

A series of glycopolymers composed of 2-deoxy-2-methacrylamido glucopyranose (MAG) and the primary amine-containing N-(2-aminoethyl) methacrylamide (AEMA) were synthesized via aqueous reversible addition-fragmentation chain transfer (RAFT) polymerization. The colloidal stability of the polyplexes formed with three diblock glycopolymers and pDNA was assessed using dynamic light scattering, and the polyplexes were found to be stable against aggregation in the presence of salt and serum over the 4 h time period studied. Delivery experiments were performed in vitro to examine the cellular uptake, transfection efficiency, and cytotoxicity of the glycopolymer/pDNA polyplexes in cultured HeLa cells and the diblock copolymer with the shortest AEMA block was found to be the most effective. Additionally, the ability of the diblock glycopolymers to deliver siRNA to U-87 (glioblastoma) cells was screened, and the diblock copolymer with the longest AEMA block was found to have gene knockdown efficacy similar to Lipofectamine 2000.