天津大神堂海洋特别保护区生境修复初步评价

2012年4月、9月和2013年5月、9月份对渤海湾天津大神堂海洋特别保护区分别进行了大型底栖动物调查,用丰度/生物量比较曲线法分析了6个调查站位的大型底栖动物群落投礁前后受污染扰动的变化状况。结果表明,在调查期间保护区大型底栖动物群落表现出从最初的受到中等程度干扰→趋向于受到干扰→群落状况趋向正常,保护区生态状况已经大有好转,人工生态修复效果初显。     

虾夷扇贝C型凝集素母源传递与抑菌作用的初步研究

为研究虾夷扇贝C型凝集素的母源传递及其抑菌作用,实验运用qRT-PCR技术检测了经鳗弧菌刺激后虾夷扇贝卵巢中C型凝集素的表达模式,比较分析了C型凝集素基因在正常虾夷扇贝和鳗弧菌刺激的虾夷扇贝所产的卵及其胚胎发育前期的存在与变化;通过抑菌实验研究了卵胞浆中C型凝集素抑制细菌存活的作用.结果表明,鳗弧菌刺激能够诱导虾夷扇贝卵巢中的C型凝集素mRNA表达量显著变化,最高表达量出现在刺激8h后,为对照组的6.2倍;母体中C型凝集素可以传递给卵和胚胎,刺激组表达量极显著高于正常组,且表达量都随胚胎发育逐渐降低,至受精36 h时分别为正常对照卵的0.3和0.2倍;蛋白终浓度为200和400 μg/mL的卵无细胞体系都具有一定的抑菌作用,与C型凝集素家族抗体反应后,细菌存活率显著上升,说明母源C型凝集素在抑制细菌存活方面发挥了重要的作用.     

逆流运动及葵花籽油替代鱼油对吉富罗非鱼生长和体成分的影响

选取初始体质量（73．81±9．02）g的雄性吉富罗非鱼（GIFFOreochromisniloticus）,分别投喂含100％鱼油（A）、50％葵花籽油＋50％鱼油（B）和100％葵花籽油（C）的饲料,每种饲料设置2个流速运动水平（0BL·s-1、1．5BL·s-1）（BL,体长）,共6个试验组合（0A、1．5A、0B、1．5B、0C、1．5C）,饲养42d后测定试验鱼生长和体成分变化。结果显示各组合吉富罗非鱼的成活率没有显著性差异（P〉0．05）。1．5B组合的特定生长率（4．96±0．02）％·d-1显著高于其他各组合（P〈0．05）。逆流运动后红肌和白肌蛋白质含量分别最高增加10．25％和7．74％（P〈0．05）。逆流运动对全鱼和肌肉多不饱和脂肪酸（PUFA）含量没有显著影响（P〉0．05）,但饲料B组和c组鱼体红肌PUFA含量显著高于A组（P〈0．05）,而B组和c组之间则没有显著差异（P〉0．05）。研究结果表明,葵花籽油替代鱼油对吉富罗非鱼的生长和体成分没有显著影响,而逆流运动能够显著提高肌肉蛋白质含量。     

双须骨舌鱼抗缪勒氏管激素基因amh的克隆、组织表达分析和原核表达

为探究双须骨舌鱼抗缪勒氏管激素基因amh的作用和表达模式,本研究通过RACE技术克隆了双须骨舌鱼amh基因全长c DNA序列,发现存在amh1、amh2两个亚型(Gen Bank登录号KU378662、KU378663)。序列分析结果显示,双须骨舌鱼amh1基因的c DNA全长2277 bp,编码623个氨基酸,amh2基因c DNA全长2181 bp,编码355个氨基酸。同源性分析显示,amh基因保守性较低,双须骨舌鱼与同科的美丽硬仆骨舌鱼相似度最高,为85.64%,与不同科鱼类的相似度均低于42%。系统进化分析显示,该基因与骨舌鱼目聚为一支,与鲱形目、鲤形目等较低等的硬骨鱼类亲缘关系较近,与双须骨舌鱼进化地位相符。用荧光定量PCR检测的结果显示,amh基因在双须骨舌鱼不同组织中均有表达,其中amh1亚型在精巢中表达量最高,且明显高于卵巢和其他组织;amh2亚型也在精巢中表达量最高,且明显高于其他组织,但远远低于amh1在精巢中的表达。构建了原核表达载体p ColdⅠ-amh1和p ColdⅠ-amh2并在大肠杆菌中成功诱导出大小分别为68和48 ku的融合蛋白,为进一步研究amh在双须骨舌鱼体内的生物功能奠定了基础。     

尼罗罗非鱼鼠李糖凝集素 (RBL-1)在非特异性细胞防御病原菌感染中的功能

为了探究鼠李糖凝集素（Rhamnose-binding lectin, RBL）在硬骨鱼非特异性细胞防御病原菌感染中的作用,本研究以尼罗罗非鱼为研究模型,首先通过分离头肾单核/巨噬细胞进行体外菌应激实验,发现在罗非鱼两种重要的致病菌-无乳链球菌（Streptococcus agalactiae）和嗜水气单胞菌（Aeromonas hydrophila）应激后,尼罗罗非鱼L-鼠李糖凝集素1（OnRBL-1）的表达量显著上调。然后,通过荧光定量PCR（qRT-PCR）检测发现OnRBL-1重组蛋白能够调节病原菌诱导细胞炎症因子的表达,包括显著抑制菌诱导的IL-6、IL-8和TNF-α的表达,和促进IL-10和TGF-β的表达。此外,通过流式细胞术检测证实OnRBL-1具有促进单核/巨噬细胞的吞噬作用,同时增强呼吸爆发水平和上调活性氧的释放。以上研究结果表明,OnRBL-1在罗非鱼单核/巨噬细胞非特异性细胞防御中发挥重要的调节作用。本研究为探讨RBL-1在硬骨鱼宿主防御病原菌感染中的功能提供了参考,并有助于完善和丰富硬骨鱼类RBL的功能和在抗菌免疫应答中的基础理论体系,具有重要的科学意义。     

Assessment of affecting factors in measuring activating platelets with the method of flow cytometry

AIM:To investigate the affecting factors of detecting platelet activation by flow cytometry (FCM).METHODS:Using decoagulant of natrium citricum, anticoagnlated peripheral venous bloods from 6 healthy donors were labeled with the method of three-colour immunofluorescence assay.Platelet activation markers fibrinogen receptor (Fib-R, PAC-1) and P-selectin (CD62P) were measured.In the same time, the reproducibility of FCM was assessed.RESULTS:The platelet activation markers PAC-1 and CD62P at each time point showed significant difference(P<0.05).The ratio was increased with time extending.The positive ratio of PAC-1 and CD62P immediately measured (within 10 min) was 2.7% and 3.5% less than those at time point of 30 min.The results were measured several times under different activation levels.The coefficient of variation was less than 5%.CONCLUSION:In room temperature and with decoagulant of natrium citricum, if the measurements of PAC-1 and CD62P are finished within 30 min after sampling, good reproducibility should be achieved.     

新疆龙蒿开花习性及花芽分化研究

通过对新疆龙蒿整个生长周期的动态观测,发现新疆龙蒿在5月下旬～7月初进行花芽分化,其过程可分为花轴分化期、花序分化期和花器官分化期;顶生花芽和侧生花芽均由营养锥分化而成;舌状花为雌花,管状花为两性花.     

Polyamines enhance chilling tolerance of cucumber (Cucumis sativus L.) through modulating antioxidative system

Cucumber chilling-resistant cultivar Changchun mici and -sensitive cultivar Beijing jietou were used in this study to investigate the effects of exogenous PAs on protection against chilling injury as well as on changes of physiological features, and the fluctuation of free PAs content in the leaves under chilling stress. Upon chilling treatment, free spermidine (Spd), spermine (Spm) and putrescine (Put) were remarkably induced in the leaves of cv. Changchun mici 1 day after treatment. The induction of Put declined thereafter, whereas Spd and Spm levels increased steadily. In the leaves of cv. Beijing jietou, Put content was increased only at 1 day after chilling while Spd content decreased significantly upon chilling treatment. Chilling reduced soluble protein content, and decreased the activities of antioxidant enzymes, including superoxidase dismutase (SOD), peroxidase (POD), catalase (CAT) and ascorbate peroxidase (APX) only in Beijing jietou. However, these changes could be renovated by exogenous application of Put and Spd. It was also found that pretreatment with Put and Spd diminished the increased electrolyte leakage and malondialdehyde (MDA) content caused by chilling in the leaves of both cultivars. Pretreatment of methyglyoxal-bis-(guanylhydrazone) (MGBG), the PAs biosynthetic inhibitor cancelled the effects of PAs in most of the treatments. Moreover, histochemical staining and quantitative measurements showed that exogenous application of Put and Spd eliminated but MGBG exaggerated the hydrogen peroxide (H₂O₂) accumulation caused by chilling stress, especially in leaves of Beijing jietou. Interestingly, Changchun mici was found to contain higher endogenous free PAs contents compared to Beijing jietou. While no significant difference of SOD, POD and CAT activities was found between non-chilling Changchun mici and Beijing jietou seedlings, the former exhibited higher APX activity than the latter. These results suggest that PAs play important roles in the tolerance of cucumber against chilling stress, which is most likely achieved by acting as oxidative machinery against chilling injury.     

异源精子诱导栉孔扇贝雌核发育二倍体的微管动态变化

采用紫外线遗传灭活的长牡蛎（Crassostrea gigas）精子激活栉孔扇贝（Chlamys farreri）卵子,并用6-DMAP诱导染色体加倍的方法,获得第二极体抑制型雌核发育二倍体早期胚胎。多聚甲醛固定、免疫荧光染色后,运用荧光显微镜观察栉孔扇贝正常发育卵子和异源精子诱导的雌核发育卵子,对其受精过程中微管的动态变化过程进行观察和分析。结果表明：（1）正常发育组受精卵内以微管为基础的纺锤体能够顺利地组装并引导卵细胞进行减数分裂和第一、第二极体的排放,以及雌、雄原核融合和第一次卵裂;（2）异源精子诱导雌核发育的卵子经6-DMAP处理后,部分微管变得模糊或消失,纺锤体受到破坏导致染色体的分离无法进行,第二极体的形成受到抑制并使雌核染色体二倍化;去除6-DMAP作用后,微管重新组装,雌核分裂重新启动继而进行卵裂;精核保持固浓缩状态或轻微膨胀形成雄性原核,但卵裂后期则以致密的染色质体（DCB）形式存在于分裂沟上或进入一个卵裂球中。结果证实,长牡蛎精子经紫外线遗传灭活后可顺利进入并激活栉孔扇贝成熟卵子但不参与合子核的形成,6-DMAP也可有效抑制第二极体的排放而获得雌核发育二倍体胚胎。本实验结果为研究异源精子诱导栉孔扇贝雌核发育的可行性提供了细胞学依据。     

城中村改造模式比较研究

近些年随着城市的扩张,许多农村被包围在城市中,形成了所谓的”城中村”,城中村的存在严重制约着城市土地集约利用和城市的可持续发展。本文对现有的城中村改造模式展开了广泛的探讨,结合实际情况对现行模式进行对比分析,提出合理化建议,以便更好地推动城中村改造的顺利进行。