猪链球菌2型Orf207基因缺失菌株的构建及其生物学特性检测

【背景】猪链球菌2型(streptococcus suis type 2, SS2)可引起人、猪的脑膜炎、关节炎及败血症等,不仅给养猪业带来巨大的经济损失,同时严重威胁公共卫生安全。本团队前期通过噬菌体展示文库技术发现Orf207编码蛋白可能参与SS2诱导的脑膜炎发生,然而其在SS2致病过程中的具体作用尚不清楚。【目的】探究Orf207基因对SS2致病性的影响。【方法】采用温敏性自杀质粒介导的同源重组系统,构建SC19 Orf207基因缺失菌株ΔOrf207及其回补菌株CΔOrf207,系统比较缺失菌株与野生株间在生长特性、形态、组织定殖能力、毒力情况、细胞黏附与侵袭及抗巨噬细胞吞噬能力等生物学特性方面的差异。【结果】与野生株相比,缺失菌株链长变短,生长速度略慢;而且Orf207缺失显著增加了小鼠的存活率,降低了细菌在血液、心脏、肝脏、脾脏、肺脏、肾脏、脑组织的定殖能力和对肺组织的病理损伤并显著减弱SS2对HeLa细胞的黏附与侵袭能力及抗巨噬细胞吞噬能力。【结论】Orf207基因可以显著降低SS2对宿主的致病能力,本研究结果不仅丰富了SS2的致病机制,也为SS2疫苗等研发提供了新靶点。     

Enhanced resistance to stripe rust disease in transgenic wheat expressing the rice chitinase gene RC24

Stripe rust is a devastating fungal disease of wheat worldwide which is primarily caused by Puccinia striiformis f. sp tritici. Transgenic wheat (Triticum aestivum L.) expressing rice class chitinase gene RC24 were developed by particle bombardment of immature embryos and tested for resistance to Puccinia striiformis f.sp tritici. under greenhouse and field conditions. Putative transformants were selected on kanamycin-containing media. Polymease chain reaction indicated that RC24 was transferred into 17 transformants obtained from bombardment of 1,684 immature embryos. Integration of RC24 was confirmed by Southern blot with a RC24-labeled probe and expression of RC24 was verified by RT-PCR. Nine transgenic T₁ lines exhibited enhanced resistance to stripe rust infection with lines XN8 and BF4 showing the highest level of resistance. Southern blot hybridization confirmed the stable inheritance of RC24 in transgenic T₁ plants. Resistance to stripe rust in transgenic T₂ and T₃ XN8 and BF4 plants was confirmed over two consecutive years in the field. Increased yield (27–36 %) was recorded for transgenic T₂ and T₃ XN8 and BF4 plants compared to controls. These results suggest that rice class I chitinase RC24 can be used to engineer stripe rust resistance in wheat.     

长江经济带森林生态安全评价及时空演变研究

由于森林生态系统的安全关系到人类的生存与发展,因此本文以长江经济带1107个区县为研究对象,运用熵权法、ArcGIS和GeoDA软件、重心分析模型、空间相关分析来分析长江经济带森林生态安全指数（ESI）,结论如下：（1）森林状态指数中,权重最高的指标为森林火灾受灾率,其次为森林有害生物成灾率和林地面积比率;在森林压力指数中,权重最高的指标为政府林业投入强度,其次为年度造林比例和自然保护区占比。（2）从全域来看,森林ESI值长江上游 > 中游 > 下游,长江南岸高于北岸。长江经济带森林ESI值总体水平较低,但在2000-2015年间总体呈上升趋势。从各省看,云南省森林ESI值最高,上海市森林ESI值最低。在此15年间,湖南省森林ESI值提高幅度最大（19.77%）,江苏省提高幅度最小（0.76%）。（3）各支流流域森林ESI值排序：赣江 > 沅江 > 金沙江 > 乌江 > 湘江 > 汉江 > 嘉陵江 > 岷江。从2000-2015年,八大流域的森林ESI值总体呈上升趋势,其中湘江流域增长幅度最大（20.87%）,而金沙江流域增长幅度最小（3.6%）。（4）森林ESI值的重心先后经历了在从南往西、从西往东北和从东北往南等过程。（5）长江经济带森林ESI值有较为显著的集聚性,森林ESI值High-High集聚区域主要分布在四川省和云南省,Low-Low集聚区县主要分布在上海、江苏和安徽,其次在湖北江汉平原、四川成都平原较为集中。（6）基于以上分析,本文建议：①应注重森林火灾、病虫害防治、林业投资、植树造林等工作。②从全域看,生态修复的重点应放在长江下游。从支流流域来看,岷江、嘉陵江和汉江流域应重点加强森林修复工作。③应在上海、江苏、安徽等Low-Low集聚区域加强植树造林和退耕还林的力度,而在四川、云南、江西和浙江等High-High集聚区域适当发展木材加工和林下种植等产业。     

狂犬病毒鼠脑复壮后的典型形态恢复及感染细胞内的形态发生学

[目的]观察比较鼠脑复壮前后狂犬病毒的形态变化,并观察病毒感染BHK-21细胞后不同时间的形态发生情况.[方法]以保存时间较长的SRV9毒株为原始材料,经乳鼠脑传代复壮后接种BHK-21细胞,浓缩、纯化后观察.[结果](1)未经复壮的病毒中DI粒子占较高比例,典型粒子只占少数,而复壮后典型粒子所占比例升高到病毒粒子总数的90%.(2)感染24h后在细胞浆内可以观察到典型病毒粒子,其数量随着培养时间的延长而增加.带毒传代之后的培养过程中细胞内病毒数量增加不明显.(3)病毒可以在细胞内的空泡膜表面以多种方式成堆出芽.[结论](1)鼠脑复壮可恢复狂犬病毒中典型粒子所占比例.(2)带毒传代1～2次时为狂犬病毒收获的最佳时机.(3)本研究为狂犬病毒的装配机制补充了数据.     

Detection of the initial site of Toxoplasma gondii reactivation in brain tissue

Detection of the initial site of Toxoplasma gondii reactivation in brain tissue is difficult because the number of latent cysts is small and reactivation is a transient event. To detect the early stage of reactivation in mouse brain tissue, we constructed a cyst-forming strain of T. gondii in the tachyzoite stage, specifically expressing red fluorescence. The PLK strain of T. gondii was stably transfected with a red fluorescent protein gene, DsRed Express, under the control of a tachyzoite-specific SAG-1 promoter and the resulting parasite was designated as PLK/RED. Tachyzoites of PLK/RED growing in Vero cells showed red fluorescence. When C57BL/6J mice were i.p. infected with tachyzoites of PLK/RED, red fluorescent tachyzoites were detected in their brains at the fourth day p.i. However, red fluorescent tachyzoites were not detected in BALB/c mice latently infected with PLK/RED, although non-fluorescent cysts were detected in their brains. After treatment of latently infected mice with dexamethasone for 1 month, the mice showed neurological symptoms. In mice with symptoms, red fluorescent tachyzoites were again detected in their brains and in other organs. To detect the initial site of reactivation, BALB/c mice latently infected with the strain were treated with dexamethasone for 3 weeks, and brains were excised before any symptoms appeared. Excised brains were examined for red fluorescence-positive sites. By a histological study of red fluorescent-positive sites, we detected a cyst containing red fluorescent zoites, which still had a PAS stain-positive cyst wall. A few red fluorescent zoites breaking away from the cyst were also observed. The stage-specific expression of fluorescent protein facilitates detection of a rare transient event and makes it possible to detect the initial site of reactivation.     

科尔沁沙地东部农牧交错带土地承载力研究--以科尔沁左翼后旗为例

科尔沁左翼后旗位于科尔沁沙地东部农牧交错带,生态环境复杂、脆弱,资源、人口和环境的协调发展至关重要,其土地承载力状况是协调三者关系的重要依据。本文应用系统动力学原理,以1995年为起始年份建立了系统动力学模型(SD模型),包括人口、土地资源、种植业、林业、畜牧业、渔业和消费等7个子系统。选择3种人口增长速度和3种营养水平组成9种方案,通过模型的运行预测了科左后旗未来50年人口的动态变化和最大土地资源承载力,并对1995—2000年实际的人口发展状况和生态状况进行了讨论。研究显示,科左后旗土地承载力46．7l—74．5l万人,不同的方案达到最大承载力的时间不同,指出控制人口增长是实现经济环境协调发展的必要条件,科左后旗的实际人口增长控制较好,现有的环境问题是不合理的开发活动造成的。     

兔抗人NESTIN抗血清的制备与鉴定(简报)

巢蛋白(nestin)属Ⅵ类中等纤维蛋白。最初在神经系统发育早期发现有该蛋白的表达。后在正常神经系统中发现仅在未分化的神经前体细胞中有nestin的短暂表达,分化后在神经元和神经胶质细胞中分别被NF(neurofilament,神经丝)和GFAP(Glial fibrillary acidic protein,神经胶质元纤维酸性蛋白)所代替。所以nestin的表达通常被视为神经前体细胞的标志之一。就我们所知,目前商品化nestin的抗体都是鼠抗鼠(克隆Rat401)的抗体,而人鼠之间该蛋白序列的同源性仅为50%左     

Identifying suitable reference genes for developing and injured mouse CNS tissues

Accurate quantification of gene expression is fundamental for understanding the molecular, genetic and functional bases of tissue development and diseases. Quantitative real‐time PCR (qPCR) is now the most widely used method of quantifying gene expression due to its simplicity, specificity, sensitivity, and wide quantification range. The use of appropriate reference genes to ensure accurate normalization is crucial for the correct quantification of gene expression from the early development, maturation, aging to injury processes in the central nervous system (CNS). In this study, we have determined the expression profiles of 12 candidate housekeeping genes (ACTB, CYC1, HMBS, GAPDH, HPRT1, RPL13A, YWHAZ, PPIA, RPLP0, TFRC, GUS, and 18S rRNA) in developing mouse brain and spinal cord. Throughout development, there was a significant degree of fluctuations in their expression levels, indicating the importance and complexity of finding appropriate reference genes. Three software including BestKeeper, geNorm and NormFinder were used to evaluate the stability of potential reference genes. GUS was the most stable gene and GUS/YWHAZ were the most stable reference gene pair across different developmental stages in different CNS regions, whereas HPRT1 and GAPDH were the most variable genes and thus inappropriate to use as reference genes. Therefore, our results identified GUS and YWHAZ as the best combination of two reference genes for expression data normalization in CNS developmental studies. © 2017 Wiley Periodicals, Inc. Develop Neurobiol 78: 39–50, 2018