细叶百合低温解除休眠过程中鳞茎内糖分及相关酶的研究

通过低温处理打破细叶百合鳞茎休眠,探讨细叶百合鳞茎解除休眠过程中的糖类化合物及蔗糖、淀粉代谢酶的变化规律。结果表明,鳞茎休眠解除过程中伴随着旺盛的糖类代谢活动,淀粉含量下降,淀粉酶活性升高。内层鳞片在冷藏60 d时可溶性糖含量达到峰值,贮藏前期,蔗糖含量,蔗糖合成酶(SS)和蔗糖磷酸合成酶(SPS)活性上升,SPS和SS协同控制蔗糖代谢及转运。冷藏条件下鳞片及顶芽内淀粉向可溶性糖方向代谢过程中促进了腺苷二磷酸葡萄糖焦磷酸化酶(AGPase)活性的增加。各种淀粉代谢相关酶在鳞茎冷藏不同时期和不同部位的作用规律不同。鳞茎各个部位总可溶性糖含量与蔗糖、葡萄糖含量及淀粉磷酸化酶(SP)、SS、α-及β-淀粉酶活性都存在显著或极显著的正相关性。糖类代谢成为鳞茎休眠解除的物质基础。     

Ethyl acetate fraction of flavonoids from Polygonum hydropiper L. modulates pseudorabies virus-induced inflammation in RAW264.7 cells via the nuclear factor-kappa B and mitogen-activated protein kinase pathways

Pseudorabies virus (PRV) infection leads to severe inflammatory responses and tissue damage, and many natural herbs exhibit protective effects against viral infection by modulating the inflammatory response. An ethyl acetate fraction of flavonoids from Polygonum hydropiper L. (FEA) was prepared through ethanol extraction and ethyl acetate fractional extraction. An inflammatory model was established in RAW264.7 cells with PRV infection to evaluate the anti-inflammatory activity of FEA by measuring cell viability, nitric oxide (NO) production, reactive oxygen species (ROS) release, and mRNA expression of inflammatory factors, inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2). Its functional mechanism was investigated by analyzing the phosphorylation and nuclear translocation of key proteins in the nuclear factor-kappa B (NF-κB) and mitogen-activated protein kinase (MAPK) signaling pathways. Our findings indicate that PRV induced inflammatory responses in RAW264.7 cells, and the responses were similar to that in lipopolysaccharide (LPS)-stimulated cells. FEA significantly suppressed NO synthesis and down-regulated both expression and secretion of COX-2, iNOS, and inflammatory cytokines (P<0.05 or P<0.01). FEA also reduced NF-κB p65 translocation into the nucleus and decreased MAPK phosphorylation, indicating that the NF-κB/MAPK signaling pathway may be closely related to the inflammatory response during viral infection. The findings suggested the potential pharmaceutical application of FEA as a natural product that can treat viral infections due to its ability to mitigate inflammatory responses.     

RT-PCR快速检测口蹄疫病毒

根据口蹄疫病毒VP1基因的序列，设计了1对引物，建立了检测口蹄疫病毒的RT－PCR方法。对FMDV各毒株检测，结果均为阳性，而对猪病毒性疾病相关病毒进行检测，结果均为阴性；检测19份已知阳性样品，检出率100％；与动物接种试验比较，符合率100％，证明该方法特异，与经典方法动物接种试验比较，其灵敏度提高100倍左右；对O3I3株的细胞毒进行检测，其敏感度达10个TCID50。初步结果表明所建立的RT－PCR技术可用于口蹄疫的诊断和流行病学调查。     

西藏牦牛和黑白花奶牛生长激素基因AluⅠ多态性的比较研究

利用聚合酶链式反应 (PCR)技术 ,测定了西藏牦牛和黑白花奶牛生长激素基因的AluⅠ RFLP (限制性内切酶片段长度多态性 ) ,并比较了2牛群的基因频率。在西藏牦牛和黑白花奶牛中 ,等位基因A的频率分别为 0 935和 0 90 5 ,等位基因B的频率分别为 0 0 65和 0 0 65 ,卡方检验差异不显著 (P >0 0 5)。     

供港活猪4种疫病抗体水平监测及免疫效果分析

为掌握广东惠州地区供港活猪主要疫病的免疫状况,我们对供港活猪主要疫病抗体水平作了一次跟踪监测,并对免疫效果进行了分析.2012-2014年,分5次共采取了900头供港猪血清,采用ELISA方法对猪瘟、猪繁殖与呼吸障碍综合征、伪狂犬病、0型口蹄疫的抗体水平进行检测.结果显示,猪瘟抗体免疫合格率在68.9％ ～78.3％之间,口蹄疫在67.2％～82.8％之间,免疫效果不理想;猪繁殖与呼吸障碍综合征抗体免疫合格率在73.9％～ 92.8％之间,伪狂犬病在86.7％ ～ 93.9％之间,免疫效果较理想.     

Isolation and Identification of Variant HeNZK-2014 of Pseudorabies Virus and Sequence Analysis of gE Gene

In order to learn the situation of pig pseudorabies virus (PRV) variant in this study, tissue samples such as lymph nodes which were collected from clinical pigs with suspected PRV infection were identified by PCR. PRV positive sample were inoculated on PK-15 cells after grinding and degerming, with further experiment including virus isolation, plague purification, PCR and IFA identification, TCID₅₀ confirmed by Reed-Muench method, inoculation test and observation of clinical symptoms in mice. The gE gene of purified PRV and brain tissue samples of dead mice were identified by sequencing analysis. The results showed that the virus grown on PK-15 cells could produce typical cytopathic effect (CPE) after 24 h; After three rounds of plaque purification,the isolate was PRV positive identified by PCR and IFA, and nominated as HeNZK-2014; The TCID₅₀ of the isolate was 10^-9.77/0.1 mL; The virus in 1×10⁸ TCID₅₀ inoculation was able to cause itching, tearing, death in infected mice, and PRV could be detected in tissues of dead mice; The molecular genetic variation analysis of gE gene by PCR amplification and clone sequencing indicated that the gE gene from brain tissue of infected mice shared 100.0% homology with HeNZK-2014, and located in a relatively independent branch with newly pandemic isolates in recent years after 2011, but was far from the classical strains before 2011, and both had two insertion of aspartic acid (D) at sites of 48 and 496 amino acids, which were considered to be the typical characteristics of PRV variants. This study successfully isolated a PRV variant, which laid a foundation for further research on vaccine development, prevention and control against PRV variants.     

猪附红细胞体PCR-微孔板杂交-酶免疫检测方法的研究

以猪附红细胞体和多种血营养菌16S rRNA基因的保守区设计合成引物HM-f/HM-r,在反向引物的5′端用生物素标记,以猪附红细胞体广东株16S rRNA基因的可变区序列设计种特异性寡核苷酸探针,探针的5′端用地高辛标记,成功地建立了猪附红细胞体PCR-微孔板杂交-酶免疫检测方法。通过测定不同浓度PCR产物对应的微孔板杂交-酶反应显色OD值,用SPSS统计软件进行回归分析,得到回归方程为y=2.1012-0.4492×logx,其相关系数R为0.977,显示对未知样品可进行定量检测。该方法与猪的多种细菌性病原、猫血巴尔通氏体CA株和E.wenyonii无交叉反应,其敏感性比常规PCR琼脂糖电泳检测提高了近100倍。用建立的方法对38份临床样品进行检测,结果有12份检测结果为阳性。     

基于单时相MODIS数据的草地退化遥感监测研究

以草地NPP减少的相对百分数（由实际NPP和潜在NPP计算）为主要监测指标，提出了一个基于单时相MODIS数据的草地退化遥感监测方法。介绍了利用MODIS数据反演实际NPP的方法，并提出了一个根据实际NPP和草地类型分布图确定潜在NPP的方法。对2003年中国北方草地的监测结果表明，中国北方草地退化严重，退化程度的空间差异性较大，草地退化程度与草地类型密切相关。     

皖西南地区春播牧草的适应性研究

对皖西南地区春播的9个牧草品种的适应性,草产量和营养成分进行分析,结果表明:墨西哥玉米、菊苣、杂交狼尾草、甜高粱产量高,干草产量和鲜草产量均在95 t/hm2和15 t/hm2以上,适应性强,品质优良,适合在皖西南地区大面积推广种植。苦荬菜和籽粒苋产量中等,蛋白质含量高达22%以上,青绿多汁,适口性好,可以做为优质牧草在皖西南地区大面积推广种植。饲用玉米、皖草2号、苏丹草产量较低,鲜草产量均在70 t/hm2以下,饲用玉米无再生性,皖草2号、苏丹草在高温条件下病虫害严重,种植时应该注意病虫害防治。     

家蚕微孢子虫单克隆抗体的制备及鉴定

用家蚕微孢子虫(Nosema bombycis)总蛋白为抗原,采用长程免疫法免疫BALB/c小鼠,取4次免疫后的脾细胞与SP2/0骨髓瘤细胞用PEG-1500进行细胞融合,第1次融合率为13.3%,第2次融合率为55.4%。建立间接酶联免疫吸附测定法(enzyme-linked immunosorbent assay,ELISA),用于检测杂交瘤细胞上清中的单克隆抗体。初检阳性率分别为4.68%和5.26%。经3次亚克隆筛选获得了2株能稳定分泌抗家蚕微孢子虫的单克隆抗体的杂交瘤细胞株,命名为2G10和2B10。间接免疫荧光试验(indirect immunofluorescence test,IFAT)和Western-bloting分析证实所获得的单克隆抗体是特异针对家蚕微孢子虫蛋白的,将在孢子孢壁蛋白的研究中发挥重要作用。