三角帆蚌肝脏瘟病感染期抑制性消减cDNA文库的构建与分析

采用抑制性消减杂交技术构建了三角帆蚌肝脏瘟病感染期抑制性消减cDNA文库。经检验,差异表达基因均被富集了 2¹⁰倍左右,证明构建的 cDNA 消减文库具有很强的消减效率。PCR 鉴定发现,在随机挑取的阳性克隆中,95% 的克隆均含有 0.2～1.0 kb 的插入片段,这些片段可能是三角帆蚌瘟病病毒感染后差异表达基因的cDNA片段。测序共获得 214 个有效 cDNA 序列,分别属于8 大类,共 98 个基因。其中细胞分裂基因 2个、细胞结构与运动基因 9 个、代谢基因 10 个、信号传导基因 7 个、细胞防疫基因 10 个、基因与蛋白表达基因 20 个、未知功能蛋白基因 26 个,GenBank中找不到任何同源序列的基因 14 个。结果说明,构建的差异表达cDNA文库,可较好地反映三角帆蚌瘟病病毒对三角帆蚌影响的基因信息。     

脂肪酸甲酯生物柴油改善低硫柴油的润滑性能

生物柴油可作为改善低硫柴油润滑性能的天然添加剂。该文将豆蔻酸甲酯(C14:0)、棕榈酸甲酯(C16:0)、硬脂酸甲酯(C18:0)、油酸甲酯(C18:1)、亚油酸甲酯(C18:2)、亚麻酸甲酯(C18:3)、蓖麻醇酸甲酯(C18:1 OH)及蓖麻油甲酯和餐饮废油甲酯按照0.5%、1.0%、1.5%和3.0%的体积分数添加到低硫柴油中,在高频往复试验机(high-frequency reciprocating rig,HFRR)上进行润滑性能测试,探究脂肪酸甲酯的碳链长度、不饱和度及含羟基等结构特征对润滑性能的影响。结果表明,长碳链脂肪酸甲酯一般比短链润滑效果好;碳链长度为十八的脂肪酸酯中,不饱和程度即碳碳双键数目越高则润滑性能越好;而在相同碳链长度和不饱和度条件下,含羟基的蓖麻醇酸甲酯的润滑改善效果优于油酸甲酯。由多种脂肪酸酯构成的混合物生物柴油的润滑性能要优于某单一的纯脂肪酸甲酯。在低硫柴油中,当某饱和脂肪酸甲酯的体积分数比例达3.0%时,或不饱和酯的体积分数达到1.5%时,或生物柴油的体积分数达1.0%时,可使低硫柴油的润滑性能指标满足相关标准。研究脂肪酸甲酯的各种结构特征对其润滑性能的影响及作用机制,有助于筛选合适的生物柴油组分及其添加浓度作为低硫柴油的润滑添加剂。     

玉米幼苗对芘污染土壤微生物活性及多样性的影响

通过盆栽土培试验研究了玉米幼苗生长期间对芘污染土壤微生物活性及多样性的影响。结果表明,玉米加快了土壤中芘的降解,提高了芘在土壤中的降解速率。试验期间,根际土中可提取态芘含量显著低于非根际土,根际土微生物生物量碳、微生物熵、多酚氧化酶和脱氢酶活性均高于非根际土,代谢熵低于非根际土。脂肪酸（FAME）分析结果表明,与非根际土相比,芘污染玉米根际土微生物群落结构发生了显著的变化,主要表现在真菌特征脂肪酸以及真菌/细菌的比值显著升高,细菌和GN^-细菌特征脂肪酸显著降低,且这种效应随着培养时间的推移在P〈0.01水平显著。根际土和非根际土中丛枝菌根真菌、GN^＋细菌和放线菌特征脂肪酸差异随着培养时间的延长逐渐加大,45 d时其差异均在P〈0.05水平显著。     

营养强化的轮虫、卤虫对牙鲆仔鱼的成活、生长及体脂肪酸组成的影响

用3种营养强化剂强化的轮虫和卤虫无节幼体投喂牙鲆仔鱼,研究牙鲆仔鱼的生长、成活、体脂肪酸的组成。结果表明:用强化的轮虫和卤虫无节幼体投喂牙鲆仔鱼,成活率、增重均显著高于对照组(p<0 01),其中V号强化剂的效果最好,成活率为29 34%,比对照组提高100%;增重倍数为217 90,比对照组提高68 61%;这是由于V号强化剂强化的卤虫无节幼体体内含有较多的AA的缘故,饵料中AA含量的提高,可以提高牙鲆仔鱼的成活率、促进其生长。牙鲆摄食强化过的轮虫、卤虫无节幼体后,其EPA、DHA、n-3HUFA、PUFA的含量随着饵料中含量的升高而升高,这也是牙鲆仔鱼生长速度和成活率提高的重要因素之一。     

Utilization of monounsaturated fatty acid (18:1n-9, oleic acid) by freshwater prawn Macrobrachium rosenbergii (de Man) juveniles

The utilization of oleic acid as an energy source and the effects of oleic acid levels and/or dietary soy bean lecithin (SBL) on oleic acid utilization, growth and survival, and lipid class and fatty acid compositions of freshwater prawn, Macrobrachium rosenbergii (de Man), juveniles were determined.
Increase in levels of dietary oleic acid from 10 to 80 g kg⁻¹ significantly ( P ≤ 0.05) reduced growth, survival and feed conversion efficiency of M. rosenbergii juveniles during the 40-day feeding period. Inclusion of 20 g kg⁻¹ SBL had no significant effect ( P ≥ 0.05) on growth and survival, nor was there any interactive effect between dietary SBL and oleic acid levels.
Body fatty acid profile of prawns reflected dietary fatty acid quantity. The fatty acid composition of prawns fed diets containing 80 g kg⁻¹ oleic acid had tremendously high proportions of oleic acid. Polar lipids, mostly phosphatidylcholine (PC), constituted the bulk of the extracted total lipids. Prawns fed with SBL had significantly ( P ≥ 0.05) higher PC content.
Oleic acid was metabolized for energy by M. rosenbergii juveniles at the same rate regardless of dietary level of SBL and/or oleic acid. Expired ¹⁴CO₂ accounted for half of the ingested radioactivity 48 h after feeding with labelled diets. No significant difference in the amount of ¹⁴CO₂ expired by prawns fed the labelled test diets was found. Per cent radioactivity ingested and absorbed into the body was also not significantly different in prawns of the different dietary treatments.     

南美油藤种子发育过程的代谢组学和转录组学联合分析

【目的】南美油藤种子油中富含不饱和脂肪酸,是一种具有重要经济价值和应用前景的油料植物。目前影响南美油藤种子油脂合成的关键基因、途径及其调控机理尚未清晰。因此,对南美油藤种子的脂质代谢物的动态变化规律及重要脂肪酸代谢相关调控基因的研究,不仅能够为提高其种子油产量和改善油脂品质提供理论基础,也可为其他木本油料植物高效开发利用提供有价值的参考依据。【方法】选用不同生长阶段的南美油藤种子(形成初期、发育初期、中期、后期、成熟期)为研究对象,结合GC-MC代谢组学技术和高通量转录组测序技术,分析种子发育过程中脂质代谢物含量的动态变化规律,并根据种子不同生长阶段的差异表达基因寻找出脂质代谢物生物合成与累积的关联酶基因。【结果】脂质代谢组学分析发现,南美油藤种子中高含量的α-亚麻酸和亚油酸主要在种子成熟阶段合成与累积,是判别南美油藤种子中脂肪酸缓慢累积时期与快速累积阶段的依据。转录组学分析表明,南美油藤种子成熟阶段前后的基因表达存在显著差异。结合代谢组学与转录组学的分析结果显示,与脂肪酸生物合成和累积相关的6个关键酶基因的表达模式与脂肪酸的合成和累积存在显著的相关性,基因家族的不同成员存在生物学功能的差异性和多样性。其中FAD2-3、FAD7、FATA、KAS2、LACS2、LACS8和SAD酶基因表达与脂肪酸总含量及主要脂肪酸含量呈显著正相关,说明7个酶基因表达对南美油藤种子油脂的合成与累积有促进作用;FAD2-2、KCS1、KCS10和LACS1酶基因与脂肪酸总含量及主要脂肪酸含量呈显著负相关,说明4个酶基因的表达对南美油藤种子油脂的合成与累积具有抑制作用。【结论】南美油藤种子成熟前后脂肪酸含量及差异表达基因存在显著差异,可依据种子中α-亚麻酸和亚油酸的含量变化将种子发育过程划分为脂肪酸缓慢累积和快速累积2个时期;南美油藤种子脂肪酸代谢中的6个关键酶基因的表达模式与脂肪酸的合成和累积存在显著的相关性,且同一基因家族的不同成员在脂肪酸的累积过程中可能具有不同的生物学功能。研究结果可为利用分子生物学技术提高南美油藤种子油产量和改变脂肪酸组分提供备选基因和相应的理论基础。     

Bacteria inhabiting artificially inoculated xylem of Picea abies

Bacteria inhabiting the xylem of Norway spruce (Picea abies (L.) H. Karsten) were investigated. The trees had been wounded and artificially inoculated with fungi and bacteria obtained from wounds of naturally infected spruce. One and five growing seasons after inoculation the Gram‐negative bacterial population present in the stem of inoculated trees were analysed.

The Gram‐negative bacteria isolated from the trees were identified on the basis of morphological, biochemical and physiological tests and whole‐cell fatty acid composition. The predominant strains were Enterobacteriaceae fermenter strains (E. agglomerans or E. sakazakii), fluorescent and yellow pigmented Pseudomonas, Acinetobacter and Moraxella spp. All Gram‐positive bacteria were Bacillus species.

The Gram‐negative bacteria of Norway spruce differed from the Gram‐positive species in possessing stronger lipolytic activity and in their ability to utilize pine resins for growth. Gram‐positive bacteria were generally able to utilise cellulose and hemicellulose, whereas among the Gram‐negative bacteria only one xylanolytic (yellow Pseudomonas) strain was found.     

国医大师张学文教授治疗原发性高血压病肝热血瘀证用药规律研究

目的 应用中医传承辅助系统软件,挖掘分析张学文教授治疗原发性高血压病肝热血瘀证的用药规律。方法 收集张学文教授近5年治疗原发性高血压病肝热血瘀证的病例,采用关联规则、系统熵聚类等数据挖掘方法,分析张学文教授治疗原发性高血压病肝热血瘀证的用药经验和用药规律。结果 对筛选出的116首典型处方进行统计分析,得出处方中药物的使用频次、药物之间的关联规则、药物核心组合和新处方。结论 张学文教授诊治原发性高血压病肝热血瘀证,以天麻钩藤饮和（或）脑清通汤为基础方加减,多用平肝熄风、活血化瘀药以治标,再加补肾、安神等药以扶正,标本兼治。     

Effect of catheter size and injection rate of contrast agent on enhancement and image quality for triple‐phase helical computed tomography of the liver in small dogs

Rapid contrast injection is recommended for triple‐phase helical computed tomography (CT) of the liver. However, a large‐gauge catheter is needed for faster contrast injection and this is not practical for small breed dogs or cats. The purpose of this crossover group study was to evaluate applicability of a lower injection rate with a small‐gauge (G) catheter for triple‐phase hepatic CT in small dogs. Triple‐phase CT images were acquired for six beagle dogs using three protocols: an injection rate of 1.5 ml/s with a 24 G catheter, 3.0 ml/s with a 22 G catheter, and 4.5 ml/s with a 20 G catheter. Enhancement of the aorta, portal vein, and hepatic parenchyma was measured in each phase (arterial, portal, and delayed) and image quality was scored subjectively by two observers. Injection duration, time to scan delay, and time to peak enhancement were also recorded. Contrast injection duration decreased with a higher injection rate (n = 6, P ≤ 0.01), but time to peak enhancement and time to scan delay were not significantly affected by injection rates and catheter sizes. Contrast injection rate did not significantly affect aortic, portal, and hepatic enhancement. In addition, separation between each phase and quality of images was subjectively scored as good regardless of injection rate. Findings from the current study supported using an injection rate of 1.5 ml/s with a catheter size of 24 G for triple‐phase hepatic CT in small dogs (weight < 12 kg).     

藏药川西獐牙菜抗小鼠免疫性肝损伤的研究

【目的】探讨川西獐牙菜对卡介苗(BCG)/脂多糖(LPS)所致的免疫性肝损伤小鼠免疫器官脏器指数和血清免疫球蛋白含量的影响。【方法】采用尾静脉注射BCG和LPS构建小鼠(体质量20g)免疫性肝损伤模型。制备川西獐牙菜提取物,按高剂量(5.380mg/只)、中剂量(2.690mg/只)、低剂量(0.538mg/只)灌胃免疫性肝损伤小鼠,连续10d后,取小鼠肝脏、脾脏和胸腺,计算其脏器指数,并取血清,测定其中IgG、IgA、IgM的含量。【结果】川西獐牙菜的各给药组能有效防止BCG/LPS对小鼠各脏器的免疫性损伤,并能影响IgG、IgA、IgM的含量,调节B淋巴细胞网络的紊乱,使异常的免疫状态恢复正常。【结论】川西獐牙菜预防性给药能减轻免疫性肝损伤对小鼠肝脏的破坏,减弱体液免疫功能的亢进,从而起到保肝、抗病、抗损伤的作用。