USP18分子通过Ⅰ型干扰素通路影响猪瘟病毒复制的研究

猪瘟病毒(CSFV)感染无特定病原(SPF)猪后,分离猪外周血单个核细胞进行转录组分析,数据显示病毒感染后泛素特异性蛋白酶18(USP18)基因的转录水平明显上升。为研究USP18分子对CSFV复制的影响及其作用机制,本研究通过构建过表达USP18的细胞系及应用特异性小干扰RNA下调表达USP18的方法,采用荧光定量PCR验证USP18对CSFV增殖的影响,结果显示USP18分子能够促进CSFV的复制;通过检测IFN-β、NF-κB及ISRE的启动子活性及I型干扰素(IFN-I)信号通路的下游分子m RNA的转录水平,采用荧光定量PCR和双荧光酶报答试验分析USP18对IFN-I信号通路的影响,结果显示USP18分子抑制IFN-I信号通路。以上结果表明CSFV通过诱导USP18分子的上调表达,抑制了IFN-I途径,从而逃避天然免疫防御,进而促进自身的复制。本研究为明确CSFV与宿主之间的相互作用及CSFV致病机制提供参考依据和奠定基础。     

小鼠Wip1基因的表达及其对受精能力的影响

本研究旨在探讨Wip1基因的表达及其对精子受精能力的影响,为阐明Wip1基因对雄性繁殖的影响提供新的着眼点。选取相同饲养环境、同一遗传背景的8周龄左右的野生型雄鼠,利用实时荧光定量PCR技术检测Wip1基因在心脏、肝脏、脾脏、肺脏、肾脏、脑组织及雄性生殖器官中的表达情况,利用免疫荧光技术检测Wip1蛋白在睾丸组织及精子细胞中的分布情况。以Wip1敲除型雄鼠为试验组,野生型雄鼠为对照组,利用ELISA试剂盒检测两组小鼠血清中睾酮水平;通过体外受精试验比较两组的2-细胞率及囊胚率。结果显示,Wip1基因mRNA在野生型雄鼠各组织中广泛表达,且在雄性生殖器官中表达量较高,其中睾丸中表达量最高,附睾体、附睾头、附睾尾次之;Wip1蛋白主要定位于睾丸组织中的长形精子细胞及附睾成熟精子细胞的头部。ELISA检测结果发现,与野生型雄鼠相比,Wip1敲除型雄鼠血清内睾酮含量极显著下降（P<0.01）。体外受精结果表明,Wip1敲除后2-细胞率及囊胚率均极显著下降（P<0.01）。结果表明,Wip1基因敲除能够引起雄性小鼠受精能力降低,这可能与Wip1基因在精子发生过程中发挥作用相关。     

新乡地区部分鸡场鸡大肠杆菌分离鉴定和药敏试验

鸡大肠杆菌病现今在我国养殖业内非常普遍,且高死亡率给养殖业带来了巨大的损失,由于抗生素使用不当,导致细菌耐药性问题形势严峻。试验采集新乡地区部分鸡场疑似大肠杆菌病鸡样品26份,检测结果均阳性。随机选择9株菌株进行耐药性检测,结果显示此地区大肠杆菌对卡那霉素、多粘菌素、庆大霉素、磺胺甲噁唑耐药率均为100%,对其余8种药物均表现出高敏感性。试验为新乡地区养殖鸡时的临床用药提供了理论依据,提高了疗效,减少了耐药性的产生。     

板蓝根多糖对鸡新城疫疫苗免疫效果的影响

150只14日龄蛋公鸡随机均分成3组,用新城疫传染性支气管炎二联(Newcastle disease-infectious bronchi-tis,ND-IB)弱毒苗点眼滴鼻免疫的同时,2个试验组分别肌肉注射高、低剂量的板蓝根多糖(Isatis root polysaccha-ride,IRPS)溶液,对照组注射生理盐水,每天注射1次,连续注射3次。分别于免疫后7、14、21、28、354、2 d各组随机抽取8只鸡翼下静脉采血,分离血清,用微量法测定新城疫血凝抑制(HI)抗体效价;于免疫后10、20、30、40、50 d每组随机抽取5只鸡心脏采血,分离淋巴细胞,用流式细胞仪双染色法检测CD3+CD4+和CD3+CD8+T淋巴细胞含量,并放血致死称取体质量和胸腺、脾脏、法氏囊的质量,计算器官指数。结果表明,板蓝根多糖高、低剂量组的HI抗体效价、CD4+/CD8+比值和免疫器官指数均明显高于对照组;提示板蓝根多糖对新城疫疫苗的免疫效果有明显的增强作用。     

江南地区墨西哥饲用玉米特性特征及栽培技术

根据多年的试验研究成果,系统介绍了墨西哥饲用玉米（Euchlaena mexicana Schrad）在江南地区的植物学、生物学特征及栽培技术要点等,以利推广种植。     

云南蔷薇属部分种质资源的SSR遗传多样性研究

利用简单重复序列SSR标记技术对蔷薇属（Rosa L．）13份野生种、变种、变型及29份栽培品种的遗传多样性进行了研究。用筛选出的18对SSR引物对42份材料DNA进行PCR扩增,在18个位点共检测到148个等位基因,每一位点的等位基因变幅为6～14个,平均8．2个。材料问遗传相似系数为0．282—0．892,表明在分子水平上具有丰富的遗传多样性。在相似系数为0．456时,基于SSR标记的聚类分析将13个蔷薇野生种分为5个组,这与植物形态学分类结果大体一致。在遗传相似系数为0．43水平上,聚类分析将42份供试材料分为5大组群。初步探讨了野生种之间以及野生种与栽培品种之间的亲缘关系。     

Mitral annulus velocities and time intervals for evaluation of global left ventricular diastolic function in patients with coronary artery disease

AIM: To detect and compare the longitudinal mitral annulus diastolic velocity and time interval changes by pulsed Doppler tissue imaging (DTI) in patients with angina pectoris (AP) and myocardial infarction (MI), and to explore the value of mitral annulus diastolic velocities and time intervals for evaluation of global left ventricular diastolic dysfunction. METHODS: Fifty patients with established coronary artery disease were divided into AP group (16 cases) and MI group (34 cases). Sixteen age-matched healthy individuals served as the control group. The septum, lateral, anterior and inferior walls of the mitral annulus were displayed, and selected for DTI spectrum sampling. Peak early and late diastolic velocities and their ratio, time to the onset and peak of the early diastolic wave, and regional isovolumic relaxation time were measured, and the average values of the four mitral annular sites were calculated and presented as Em, Am, Em/Am, QEm, TEm and IVRTm, respectively. RESULTS: Compared with the control group, Em and Em/Am were significantly lower in both the AP and the MI groups (P<0.01). Em was even lower in the MI group than that in the AP group (P<0.01). QEm, TEm and IVRTm were significantly longer in the AP and the MI groups than those in control group (P<0.01 or P<0.05). IVRTm was even longer in the MI group than that in AP group (P<0.01). IVRTm had significantly negative correlation with Em (r=-0.64, P<0.01). CONCLUSION: Em, Em/Am, QEm, TEm and IVRTm as measured by pulsed DTI may be promising indexes for quantitative assessment of global left ventricular diastolic dysfunction in patients with coronary artery disease. Em and IVRTm may indicate the severity of ischemic myocardial damage.     

DIM enhances effect of cis-diamminedichloroplatinum on growth inhibition and apoptosis in human prostate cancer cell PC-3

AIM: To study the effects of the combination of cis-diamminedichloroplatinum(DDP) and 3, 3-diindolylmethane (DIM) on the growth and apoptosis of human prostate cancer cell PC-3. METHODS: MTT method was applied to detect the cell growth inhibitory rate. The cell apoptosis was measured by the flow cytometry and acridine orange staining method. The expression of the anti-oncogene p21 was detected by RT-PCR technique. RESULTS: The combination of 60 μmol·L^-1 DIM and 0.4 mg·L^-1 DDP effectively inhibited the growth and induced apoptosis in PC-3 cells. This result was the same as the effect of using 4 mg·L^-1 DDP only. The cell growth inhibitory and apoptosis rates for the combination of DIM and DDP were much higher than those for the individual effect. Both the combination and the single effect of these two medicines (i.e., DIM and DDP) all strengthen p21 mRNA expression significantly, and the effect of combination was more significant. CONCLUSION: DIM significantly enhances the effects of DDP on the growth inhibition and apoptosis induction in PC-3 cells.     

基于形态数据的大白菜核心种质构建方法的研究

 以国家蔬菜种质资源中期库收集保存的1 651份大白菜种质为试材,按照大白菜分类系统和生态型,采用分层分组法,将所有种质分为6组。基于43个形态性状的数据,比较了4种组内取样比例法、6种总体取样规模和2种取样方法在构建大白菜核心种质中的作用和效果。结果表明：组内以多样性比例法更能使各组的取样份数或比例趋于平衡,并较好地保持原始收集品的变异度。当总体取样规模为15%时,多样性比例法所构建的核心种质的遗传多样性指数达到最大,表型保留比例亦能达到98％左右;而当总体取样规模增加到20%以上时,虽然表型保留比例接近100％,但核心种质遗传多样性指数迅速降低。因此,认为15%总体取样规模较为合适。在一定的组内取样比例法和总体取样规模下,聚类取样构建的核心种质的遗传多样性指数（I）、表型保留比例（RPR）和变异系数（CV）均比随机取样的高。根据获得的优化方案最终在表型水平建立了包含248份种质的中国大白菜核心种质。     

茉莉花ISSR-PCR反应体系的建立

以茉莉花为材料,研究了PCR反应体系的主要成分及退火温度对茉莉花ISSR扩增结果的影响.结果表明:在20μL的反应体中,Mg2 的用量为1.2～1.6 mmol/L,dNTPs 浓度为0.15 mmol/L,引物的浓度为0.5μmol/L,Taq DNA 聚合酶的用量为1U,模板DNA的用量为40～70 ng,在48～50℃的退火温度下40个循环,能得到清晰、多态性高的ISSR带谱.