杨树溃疡病生防菌株N6-34抗菌活性物质的分离纯化与结构鉴定

【背景】杨树溃疡病是一种主要由葡萄座腔菌引起的杨树枝干病害,危害严重。前期从杨树中分离到一株内生拮抗细菌N6-34,研究表明该菌株拮抗效果好,对多种植物病原菌均有较强的拮抗作用。【目的】对拮抗细菌N6-34产生的抗菌活性物质进行分离纯化,并鉴定了活性物质组分的结构。【方法】通过硫酸铵盐析、甲醇抽提、分子筛、高效液相色谱等方法分离纯化N6-34菌株的抗菌活性物质,并对其进行结构鉴定。【结果】N6-34菌株发酵液经多步分离纯化,共获得14个组分,其中有13个组分具有抗菌活性,经一级质谱分析,获得了13种抗菌活性组分的分子量;经二级质谱分析,将13种抗菌活性物质鉴定为Fengycin A或Fengycin B的同系物或同分异构体。【结论】从N6-34菌株发酵液中分离获得了13种抗菌成分,为杨树溃疡病的生物防治提供了理论依据。     

COPD association and repeatability of blood biomarkers in the ECLIPSE cohort

Background

There is a need for biomarkers to better characterise individuals with COPD and to aid with the development of therapeutic interventions. A panel of putative blood biomarkers was assessed in a subgroup of the Evaluation of COPD Longitudinally to Identify Surrogate Endpoints (ECLIPSE) cohort.

Methods

Thirty-four blood biomarkers were assessed in 201 subjects with COPD, 37 ex-smoker controls with normal lung function and 37 healthy non-smokers selected from the ECLIPSE cohort. Biomarker repeatability was assessed using baseline and 3-month samples. Intergroup comparisons were made using analysis of variance, repeatability was assessed through Bland-Altman plots, and correlations between biomarkers and clinical characteristics were assessed using Spearman correlation coefficients.

Results

Fifteen biomarkers were significantly different in individuals with COPD when compared to former or non-smoker controls. Some biomarkers, including tumor necrosis factor-α and interferon-γ, were measurable in only a minority of subjects whilst others such as C-reactive protein showed wide variability over the 3-month replication period. Fibrinogen was the most repeatable biomarker and exhibited a weak correlation with 6-minute walk distance, exacerbation rate, BODE index and MRC dyspnoea score in COPD subjects. 33% (66/201) of the COPD subjects reported at least 1 exacerbation over the 3 month study with 18% (36/201) reporting the exacerbation within 30 days of the 3-month visit. CRP, fibrinogen interleukin-6 and surfactant protein-D were significantly elevated in those COPD subjects with exacerbations within 30 days of the 3-month visit compared with those individuals that did not exacerbate or whose exacerbations had resolved.

Conclusions

Only a few of the biomarkers assessed may be useful in diagnosis or management of COPD where the diagnosis is based on airflow obstruction (GOLD). Further analysis of more promising biomarkers may reveal utility in subsets of patients. Fibrinogen in particular has emerged as a potentially useful biomarker from this cohort and requires further investigation.

Trial Registration

SCO104960, clinicaltrials.gov identifier {"type":"clinical-trial","attrs":{"text":"NCT00292552","term_id":"NCT00292552"}}NCT00292552     

"基因电脑克隆"软件SiClone的并行优化研究与实现

生物信息学中,发现、鉴别新基因是承上启下的一步,它既承接了过往如“基因组测序”的工作,又是未来“后基因时代”研究的基石．“基因电脑克隆”是利用计算手段发现、鉴别新基因的方法,SiClone软件实现了“基因电脑克隆”功能．本文对SiClone软件操作的数据库提出并行处理方案,并详述了基于MPI(message passing interface)平台实现的并行优化版本PSiClone．根据已得到的EST数据库,展示了软件并行版PSiClone的运行性能,试验数据库EST序列条数仅仅是NCBI(The National Center for Biotechnology Information)dbEST庞大数据库的很小部分,这也暗示我们软件的并行工作对于大数据库的比较和运算将更有应用前景．     

大鼠主动脉内皮细胞的分离培养和鉴定

目的:探索大鼠主动脉原代内皮细胞体外培养方法,为体外研究提供细胞模型。方法:分离大鼠主动脉,直接贴壁于培养皿中,荧光倒置显微镜观察细胞形态,免疫组化Ⅷ因子相关抗原染色鉴定细胞。结果:约24小时组织块边缘有游离的新生细胞长出,7天即融合成片。消化传代后细胞呈短梭形或三角形,单层生长,铺路石状,Ⅷ因子表达阳性,呈指数增殖。冻存后复苏细胞活性均超过90%。结论:用贴壁法成功建立了大鼠血管内皮细胞体外培养方法,冻存细胞存活率高,为体外研究提供了稳定的模型。     

用杆状病毒-昆虫细胞系统表达、纯化重组的人源化氨基末端LBP

根据已有的资料显示LBP与LPS的结合位点位于其氨基末端的第 91～ 1 0 0个氨基酸残基。在体外构建含有人LBP与LPS结合活性部分的穿梭质粒pBacmidtLBP ,并且带上 6×his的标签 ,用此穿梭质粒转染sf2 1昆虫细胞 ,获得重组病毒。然后用重组病毒液感染对数生长期的sf2 1昆虫细胞 ,72h后收获含有这种截断型人源化氨基末端LBP-(NH-LBP)的培养上清。用金属亲和树脂纯化后 ,采用SDS-PAGE电泳以及Western-Blot鉴定所得到的纯化物。成功获得相对分子质量约为 30 0 0 0的NH-LBP。为进一步研究LBP在介导LPS活化靶细胞中的作用机制奠定了基础。     

八株芽孢杆菌菌株的分类及固氮活性的测定

从8个省市的不同土壤分离了200个菌株,筛选出8株（JR₁,JR₂,JR₅,ZZ₃,ZZ₁₂,ZZ₁₆,M₁,K₄）能在无氮培养基上生长良好的典型的芽抱杆菌,其中6株（JR₁,JR₂,JR₄和ZZ₃,ZZ₁₂,ZZ     

10种草坪及地被植物的同工酶研究

用垂直平板聚丙烯酰胺凝胶电泳技术对２种禾草及８种地被植物的过氧化物酶同工酶及酯酶同工酶酶谱进行了研究。初步结果表明：这种方法可以比较准确地鉴定和分离种与品种及科与属,这对草坪及地被植物的分子水平上的研究和应用具有重要意义。此外,还对所用方法进行了讨论。     

无患子及其制剂的薄层色谱鉴别探讨

目的探讨无患子及其制剂的鉴别方法。方法采用薄层色谱法在不同条件下进行薄层色谱考察,选择适宜的薄层色谱条件。结果最佳薄层色谱条件为:以正丁醇-乙酸乙酯-水-浓氨水(40∶10∶50∶1;V∶V∶V∶V)上层液为展开剂,室内环境中展开,晾干,喷5%磷钼酸乙醇液,105℃加热至斑点清晰,在日光下检视,无患子及其在制剂中的成分分离效果良好,色谱斑点清晰,耐用性好。结论该法操作简便可靠,分离度较好,可作为无患子及其制剂的鉴别方法。     

利用蔗渣半纤维素水解液产油酵母的筛选、鉴定及发酵实验

对已分离到的产油酵母通过木糖摇瓶发酵,从中筛选到2株H2-1和J2-2利用蔗渣半纤维素水解液高产油脂酵母,其利用蔗渣半纤维素水解液油脂得率系数分别为13.49和10.28,均显示出对蔗渣半纤维素水解液的高转化率.根据常规生理生化特征和26S rDNA序列分析结果,确认H2-1为小红酵母(Rhodotorula minuta),J2-2为粘红酵母(Rhodotorula glutinis).     

1株源于人体的双歧杆菌的PCR扩增和鉴定

目的从佳木斯大学健康学生体内分离出1株双歧杆菌。方法利用引物设计软件设计双歧杆菌的引物,通过PCR进行扩增,进行序列分析和鉴定。结果通过BLAST序列比对分析,与GenBank中相应基因同源性为99%。结论确定此菌株为青春双歧杆菌。