大黄鱼肿瘤抑制因子cylindromatosis (LcCYLD)的序列与免疫反应特征及其功能

为研究大黄鱼肿瘤抑制因子cylindromatosis（CYLD）在免疫反应中的作用,本实验克隆了大黄鱼CYLD的全长cDNA（命名为LcCYLD）并对其进行序列分析;采用荧光定量PCR的方法对大黄鱼各组织及免疫刺激后的大黄鱼肾细胞系中的LcCY LD表达变化进行检测;构建了重组表达载体pTurboGFP-CYLD及pcDNA3.1-CYLD,分别用于亚细胞定位实验及过表达实验;在HEK293T细胞系中过表达LcCYLD后采用双荧光素酶报告系统检测了 NF-κB、TNF-α及IL-1β 启动子活性的变化。结果显示,LcCYLD的ORF包含2 754 bp,编码917个氨基酸,推测具有保守的3个N端的CAP-GLY结构域,1个磷酸化区域和1个C端的UCH结构域,多序列比对结果显示各物种CYLD间高度保守;系统进化分析显示,LcCYLD与来源于其他硬骨鱼的CYLD聚为一支,其中与条纹鲈的CYLD关系最近;转录水平表达分析发现LcCYLD在大黄鱼各组织均有表达,其中在脑中表达量最高;LPS及poly I:C刺激能够显著诱导LcCYLD的表达;亚细胞定位实验表明LcCYLD在细胞质及细胞核中均有表达;过表达LcCYLD能够显著抑制NF-κB及促炎细胞因子TNF-α及IL-1β的转录激活。以上研究结果表明大黄鱼CYLD能够抑制NF-κB的转录激活,为深入了解LcCYLD在大黄鱼先天免疫信号转导中的作用奠定基础。     

Build up of patches caused by Rhizoctonia solani

Rhizoctonia solani is a complex species that is composed of different anastomosis groups (AG). Although these different AGs show differences in their host ranges, generally R. solani is a phytopathogenic species with a wide spectrum of hosts. It has the ability to grow as a saprotroph, which further complicates its behaviour as a parasite. The losses caused by R. solani are very important and need a sustainable management strategy. The patchy appearance of the disease caused by this pathogen is well-known. The patches show within and between season dynamics. The factors which affect the spread of the disease can be grouped into three main categories: host plant, pathogen and environment. However, each of the categories in its detail may depend on or react with the other categories. There are a number of factors that may be involved in dynamics of patches. These potential mechanisms are discussed. It is essential to know about the mechanisms involved to develop an effective control strategy. Although more work is needed to investigate different mechanisms of parasitism deployed by different AGs in different hosts, it seems that many mechanisms external to the host are operating at the same time which necessitates an integrative research approach to study and control the diseases caused by R. solani.     

Suppression of clubroot (Plasmodiophora brassicae) development in Arabidopsis thaliana by the endophytic fungus Acremonium alternatum

This study investigated the ability of an endophytic fungus Acremonium alternatum to reduce clubroot formation in the model plant Arabidopsis thaliana, which is highly susceptible to Plasmodiophora brassicae . Quantitative PCR demonstrated that A. alternatum colonized the P. brassicae -infected roots and shoots of the host plant. When Arabidopsis plants were co-inoculated with P. brassicae and A. alternatum , gall formation was reduced as shown by the reduction of the disease index (DI) by up to 50% compared to plants only infected with P. brassicae, whereas the infection rate was lowered by about 20% only in several, but not all, experiments. Clubroot was similarly suppressed when plants were inoculated with autoclaved A. alternatum spores or spore extracts, showing that viable spores were not needed. However, A. alternatum spores did not inhibit P. brassicae resting spore germination. Compared to the normal root galls, the smaller root galls on A. alternatum -inoculated plants contained fewer resting spores of the clubroot pathogen. It was thus hypothesized that inoculation with A. alternatum delayed the development of P. brassicae . Using quantitative RT-PCR to monitor the expression of P. brassicae genes differentially expressed during the development of the disease, a delayed pathogen development was corroborated. Furthermore, greenhouse experiments identified a time window in which the endophyte had to be administered, where the latest effective time point was 5 days before inoculation with P. brassicae and the optimum treatment was to administer A. alternatum and P. brassicae at the same time. These results indicate that A. alternatum and perhaps similar endophytes could be useful for the management of clubroot disease.     

犬贾第虫包囊排出规律的研究

对自然感染贾第虫的比格犬进行粪检调查 ,结果表明犬贾第虫包囊排出呈间歇性 ,其间歇期为 7～ 8d ,排囊后 1 2～ 1 4d达到排囊高峰期 ,峰期持续 2～ 3d,排囊持续时间为 2 5～ 2 7d。用 2只幼犬进行免疫抑制接种犬贾第虫 ,接种后第 7天出现排包囊 ,排囊后 6～ 7d达到排囊高峰期 ,峰期持续 4～ 5d。受染动物出现腹泻 ,体重减轻     

Determination of the oxidative burst chemiluminescent response of avian and murine-derived macrophages versus corresponding cell lines in relation to stimulation with Salmonella serotypes.

In contrast to mammalian systems, avian species lack a resident or harvestable macrophage population in the abdominal exudate. Peritoneal macrophages in the chicken can be elicited if an inflammatory agent such as sephadex is injected. This study examines the kinetics of different macrophage populations, derived by different methods of isolation and from different hosts, with respect to the elicited oxidative burst upon infection with host-adapted Salmonella serotypes.

The nature of the oxidative burst elicited by murine and avian-derived and cell line macrophages was determined after stimulation with phorbol myristate (PMA), zymosan A, and Salmonella serotypes. Both murine and chicken peritoneal macrophages, chicken blood monocytes and corresponding cell lines, J774A.1 and HD-11, were unable to produce a detectable chemiluminescent (CL) response after interaction with Salmonella using the luminescent probe luminol. However, both PMA and zymosan A induced a CL response in all cell types, with PMA eliciting a higher and earlier peak response (pkH) than zymosan A. Lucigenin-enhanced CL in both murine and chicken macrophages was achieved with PMA, zymosan A and Salmonella serotypes. In this case, zymosan A induced higher responses than PMA. In the peritoneal macrophages of both hosts, there were no significant differences in the oxidative burst induced by the different Salmonella serotypes. However, the J774A.1 (murine) cells demonstrated significant differences, with S. enterica serotype Choleraesuis (S. choleraesuis and S. gallinarum producing the highest response. In the HD-11 (chicken) cells, S. choleraesuis and S. dublin elicited the higher CL. With both cell lines, S. abortusovis failed to induce an appreciable CL response.

In these experiments it was demonstrated that oxidative burst was not detectable in monocytes/macrophage populations using luminol, which suggests a link to the lack of a myeloperoxidase system in these cells. Lucigenin-enhanced CL appeared independent from the myeloperoxidase system, indicating production of another oxidative species compared with luminol. No discernable effect of host specificity with regard to Salmonella serotype and respective host was seen in host-derived or cell line macrophages, and cell line macrophages displayed altered functional characteristics with regard to oxidative burst in comparison with their primary counterparts.     

BP神经网络在隧道岩爆预测中的应用

西南某水电站拟修建四条引水隧洞以及两条平行交通隧道,但工程区一般埋深1000-2000 m,最大埋深2525m,地处高应力区,目前已开挖的2000m洞段中均发现轻微至中等的岩爆现象。因此,在深埋、长大的隧道以后开挖过程中岩爆烈度如何显得十分敏感,利用BP人工神经网络,建立了以强度脆性系数、应力系数等8指标为要素的岩爆预测模型,对12200m未开挖的辅助洞段进行了预报,结果表明,该方法的预报效果较好。与地质分析判断相吻合,可用于进一步指导实践。     

Improving quality of composted biowaste to enhance disease suppressiveness of compost-amended, peat-based potting mixes

Biowaste can be converted into compost by composting or by a combination of anaerobic digestion and composting. Currently, waste management systems are primarily focused on the increase of the turnover rate of waste streams whereas optimisation of product quality receives less attention. This results in low quality composts that can only be sold on bulk markets at low prices. A new market for quality compost could be potting mixes for horticultural container-grown crops to partially replace non-renewable peat and increase the disease suppressiveness of potting mixes. We report here on the effect of wetsieving biowaste prior to composting on compost quality and on disease suppressiveness against the plant pathogen Pythium ultimum of peat mixes amended with this compost. The increased organic matter and decreased salt content of the compost allow for significantly higher substitution rates of peat by compost. In this study up to 60% v/v compost peat replacement did not affect cucumber growth. However, disease suppressiveness of the potting mixes strongly increased from 31 to 94% when the compost amendment rate was increased from 20 to 60%. It was shown that general disease suppression for P. ultimum can only be effective when the basal respiration rate is sufficiently high to support microbial activity. In addition, organic matter of the compost should reach a sufficient stability level to turn from disease conducive to disease suppressive. Increasing the compost addition from 20 to 60% did not significantly affect plant yield, yield variation were due to differences in nutrient levels. It can be concluded that compost from wetsieved biowaste has high potential to replace peat in growing media for the professional market.