2-硝基-1-芳乙烯类化合物的设计合成及其对稻瘟病菌的抑制活性

为了筛选得到对稻瘟病菌Magnaporthe grisea具有较高抑菌活性的新型化合物,根据稻瘟病菌中1,3,8-三羟基萘酚还原酶（3HNR）的结构信息,设计合成了系列2-硝基-1-芳乙烯（2a~2e）和2-溴-2-硝基-1-芳乙烯（3a,3b）目标化合物,并测试了其对3HNR和稻瘟病菌的抑制活性,同时运用分子对接方法对目标化合物与3HNR可能的结合模式进行了分析。结果表明:大部分目标化合物对3HNR都有很好的抑制作用（IC503的抑制活性最好,IC50值为0.53 μmol/L。在50 μg/mL下,目标化合物对稻瘟病菌的生长具有不同程度的抑制作用,其中2e、3a和3b的抑制率高于96%;3a和3b的EC50值分别为16.4和11.6 μg/mL。分子对接方法分析结果表明,硝基苯乙烯骨架结构与稻瘟病菌的3HNR活性空腔的氨基酸残基有较好的相互作用,其中化合物3中的溴原子可与3HNR中Tyr223和Tyr178的羟基形成氢键,从而解释了化合物3对3HNR有较好抑制作用的原因。     

茶树GGPS基因家族的克隆、生物信息学和表达分析

牻牛儿基牻牛儿基焦磷酸合酶(Geranylgeranyl diphosphate synthase,GGPS)是萜类合成途径的结构酶,对植物生长发育具有重要意义。本研究通过RACE和RT-PCR方法克隆得到5条潜在的茶树GGPS序列,分别命名为CsGGPS1-4和CsGGPS9,其中CsGGPS9存在3条等位基因,分别是CsGGPS9-1、CsGGPS9-2和CsGGPS9-3,在系统进化树上与其他基因分成两支。蛋白质序列分析表明,茶树GGPS家族成员都具有polyprenyl_synt结构域,不存在信号肽序列。亚细胞定位预测结果显示,CsGGPS1、CsGGPS2和CsGGPS4定位在叶绿体上,CsGGPS3和CsGGPS9定位在线粒体上。通过Swiss Model进行三维建模,结合"three-floor"模型对茶树GGPS家族成员的功能进行预测,预测结果显示,CsGGPS1、CsGGPS2和CsGGPS4是GGPS;CsGGPS3是异源二聚体形式的牻牛儿基焦磷酸合酶的小亚基;CsGGPS9的催化主产物是碳链数大于30的异戊烯基焦磷酸。q RT-PCR分析表明,CsGGPS1整体表达丰度较低,仅在一芽二叶中表达量稍高;CsGGPS2在茶树各个组织中均有表达,在花中表达量最高,且花发育过程中表达量先上升后下降;CsGGPS3在叶和幼根中的表达量高于花,花发育过程中表达平稳;CsGGPS4在茶树各个组织中表达量数值相近,在花发育过程中表达量变化趋势与CsGGPS2相同;CsGGPS9的表达量在成熟叶中显著低于幼嫩叶片。     

维生素K_3对五龙鹅生长性能、屠宰性能及养分表观利用率的影响

本试验旨在研究饲粮中维生素K3添加水平对五龙鹅生长性能、屠宰性能及养分表观利用率的影响。试验分为2个阶段。1~4周龄阶段,选用1日龄五龙鹅360只,随机分为6个组,每个组6个重复,每个重复10只鹅。Ⅰ组为对照组,饲喂基础饲粮(维生素K3含量1.23 mg/kg),Ⅱ~Ⅵ组在基础饲粮中分别添加1、2、4、8、16 mg/kg的维生素K3。5~16周龄阶段,选用28日龄五龙鹅324只,随机分为6个组,每个组6个重复,每个重复9只鹅。A组为对照组,饲喂基础饲粮(维生素K3含量1.18 mg/kg),B~F组在基础饲粮中分别添加2、4、8、16、32 mg/kg的维生素K3。试验期16周。结果表明:1)经回归分析得出,1~4周龄阶段五龙鹅饲粮中添加4.81 mg/kg维生素K3,平均日增重最大;5~16周龄阶段饲粮中添加11.59 mg/kg维生素K3,平均日增重最大。2)与对照组相比,1~4周龄阶段,饲粮中添加4 mg/kg维生素K3能显著或极显著提高鹅的全净膛率和腹脂率(P0.05或P0.01);5~16周龄阶段,饲粮中添加8 mg/kg维生素K3能显著或极显著提高屠宰率、半净膛率、全净膛率和腿肌率(P0.05或P0.01)。3)与对照组相比,5~16周龄阶段,饲粮中添加8 mg/kg维生素K3能极显著提高钙的表观利用率(P0.01),显著提高干物质、粗蛋白质、粗脂肪、中性洗涤纤维、酸性洗涤纤维和磷的表观利用率(P0.05)。由此可见,本试验条件下,1~4周龄和5~16周龄五龙鹅饲粮中维生素K3适宜添加水平分别为4.81和11.59 mg/kg。     

Increased humoral antibody response of foot-and-mouth disease virus vaccine in growing pigs pre-treated with poly-γ-glutamic acid

This study was conducted to determine if humoral antibody response of foot-and-mouth disease (FMD) vaccine improved in 8-week-old growing pigs born to well-vaccinated sows pre-treated with 60 mg of poly-γ-glutamic acid (γ-PGA) three days before vaccination. Antibody against FMD virus serotype O was measured 0, 2, 4 and 6 weeks post-vaccination, using a PrioCHECK FMDV type O ELISA kit. The results showed that positive antibody reactions against FMDV serotype O antigen among a component of the vaccine significantly increased in response to pre-injection with γ-PGA.     

Comparative analysis of MAPK and PI3K/AKT pathway activation and inhibition in human and canine melanoma

The lack of advanced animal models of human cancers is considered a barrier to developing effective therapeutics. Canine and human melanomas are histologically disparate but show similar disease progression and response to therapies. The purpose of these studies was to compare human and canine melanoma tumours and cell lines regarding MAPK and PI3K/AKT signalling dysregulation, and response to select molecularly targeted agents. Pathway activation was investigated via microarray and mutational analysis. Growth inhibition and cell cycle effects were assessed for pathway inhibitors AZD6244 (MAPK) and rapamycin (PI3K/AKT) in human and canine melanoma cells. Human and canine melanoma share similar differential gene expression patterns within the MAPK and PI3K/AKT pathways. Constitutive pathway activation and similar sensitivity to AZD6244 and rapamycin was observed in human and canine cells. These results show that human and canine melanoma share activation and sensitivity to inhibition of cancer‐related signalling pathways despite differences in activating mutations.     

Temperature modulates liver lipid accumulation in Atlantic salmon (Salmo salar L.) fed low dietary levels of long‐chain n‐3 fatty acids

Atlantic salmon (Salmo salar) were fed five graded levels of eicosapentaenoic acid (EPA, 20:5n‐3) and docosahexaenoic acid (DHA, 22:6n‐3), from 1.4 to 5.2% of total fatty acids (FA, 5–17 mg kg^?1 feed), and grew from ~160 g to ~3000 g, with the period from 1450 g onwards conducted both at 6 °C and at 12 °C. All fish appeared healthy, and there were no diet‐related differences in haematological or plasma parameters, as well as intestinal histological or gut microbiota analysis. Fish reared at 6 °C had higher accumulation of storage lipids in the liver compared to fish reared at 12 °C. Liver lipids also increased with decreasing dietary EPA + DHA at 6 °C, while there was no such relationship at 12 °C. Gene expression of SREBP1 and 2, LXR, FAS and CPT1 could not explain the differences in liver lipid accumulation. In liver polar lipids, DHA was found to be reduced when dietary EPA + DHA was <2.7% of FAs, while the level of EPA in the membranes was not affected. In conclusion, reducing dietary EPA + DHA from 5.2 to 1.4% of total FAs had a minor impact on fish health. Temperature was the factor that most affected the liver lipid accumulation, but there was also an interaction with dietary components.     

基于图形处理器的三维表面质量快速评定技术

针对三维表面粗糙度评定中表面测量数据量大且计算复杂度高,常用处理方法不能满足实时测量要求的问题,提出了一种基于图形处理器(GPU)的三维表面形貌测量数据快速评定方法,应用CUDA平台开发了基于GPU的并行处理算法,在此基础上实现了三维表面质量的快速评定。实验结果表明,该方法在保证计算精度的同时,运算效率提高了60倍以上,可满足三维表面质量在线评定的要求。     

甘薯褪绿矮化病毒RNase3蛋白的原核表达及抗血清制备

以本实验室保存的重组质粒为模板,利用PCR方法扩增获得了甘薯褪绿矮化病毒SPCSV的RNase3基因,RNase3基因由690个核苷酸组成,编码229个氨基酸。将RNase3基因克隆到原核表达载体pET-28a(+),转化大肠杆菌BL21(DE3),经IPTG诱导,对诱导产物进行SDS-PAGE分析。结果表明,RNase3在大肠杆菌中能高效表达,融合蛋白分子量约为26.5kD。以表达的融合蛋白为抗原,免疫家兔,制备了SPCSV-RNase3的特异性抗血清。ACP-ELISA检测结果表明,制备的抗血清对RNase3融合蛋白的效价达10万倍。     

大头鲤、黑龙江鲤、德国镜鲤及其杂种F3肌肉品质的比较

对大头鲤、黑龙江鲤、德国镜鲤及杂种F3背部肌肉的肉色、嫩度、pH、失水率、系水力和肌纤维性状进行研究,评价杂种F3的肌肉品质.结果表明:(1)杂种F3肌肉的亮度为47.37,红度为1.85,黄度3.27,嫩度为1.65 kg·f,pH 6.62,失水率为22.54%,系水力为97.30%,肌纤维的横截面积为128.76 μm2,纤维的密度为每平方毫米183.38根,肌纤维面积的比例为55.55%.(2)杂种F3与亲本肌肉品质的比较,肉色亮度与黑龙江鲤相近,高于大头鲤和德国镜鲤;红度与大头鲤和黑龙江鲤相近,低于德国镜鲤;嫩度与大头鲤相近,小于黑龙江鲤和德国镜鲤;肌纤维的横截面积与大头鲤相近,小于黑龙江鲤和德国镜鲤;肌纤维密度与大头鲤相近,大于黑龙江鲤和德国镜鲤;pH、失水率、系水力、肉色的黄度和肌纤维面积的比例与3亲本差异不显著.研究表明杂种F3具有与大头鲤相似的肌肉品质,经进一步选育可以育成一个优质的鲤新品系.     

A three‐dimensional cell culture system as an in vitro canine mammary carcinoma model for the expression of connective tissue modulators

In this study, derived complex carcinoma (CC) and simple carcinoma (SC) cell lines were established and cultured under two‐dimensional (2D) and three‐dimensional (3D) conditions. The 3D was performed in six‐well AlgiMatrix? (LifeTechnologies®, Carlsbad, CA, USA) scaffolds, resulting in spheroids sized 50–125 µm for CC and 175–200 µm for SC. Cell viability was demonstrated up to 14 days for both models. Epidermal growth factor receptor (EGFR) was expressed in CC and SC in both systems. However, higher mRNA and protein levels were observed in SC 2D and 3D systems when compared with CC (P < 0.005). The connective tissue modulators, metalloproteinases‐1, ‐2, ‐9 and ‐13 (MMPs), relaxin receptors 1 and 2 (RXR1 and RXR2) and E‐cadherin (CDH1) were quantitated. All were upregulated similarly when canine mammary tumour (CMT)‐derived cell lines were cultured under 3D AlgiMatrix, except CDH1 that was downregulated (P < 0.005). These results are promising towards the used of 3D system to increase a high throughput in vitro canine tumour model.