基于Pro/E的渐开线直齿轮参数化设计

介绍了一种在Pro／E中创建渐开线直齿轮模型的方法,结合其中的二次开发工具程序和关系,实现了后续制作该类型齿轮时,可通过人机交互方式给定齿轮参数,系统自动生成齿轮模型,避免大量重复的设计工作。     

基于GF-6/WFV卫星遥感的大田冬小麦叶片氮素含量估测

为对大田冬小麦叶片氮素含量（LNC）进行快速、准确及无损监测,通过在江苏省泰州泰兴市、盐城大丰区和南通如皋市布设冬小麦遥感监测大田试验,在获取试验样点冬小麦冠层红光波段反射率（RED_ref）、近红外波段反射率（NIR_ref）和计算的十个光谱指数（RVI、NDVI、DVI、SAVI、OSAVI、MSR、RDVI、EVI2、NLI和SVI）基础上,将12个遥感光谱指标与冬小麦LNC进行相关分析,选出与LNC相关性较好的作为模型输入变量,构建基于BP神经网络的冬小麦LNC估测模型, 并利用GF-6/WFV卫星遥感影像对县域冬小麦LNC的空间分布开展监测。结果表明,12个遥感光谱指标与冬小麦LNC之间存在不同程度的相关性,其中NDVI、RVI、MSR、OSAVI和NLI与冬小麦LNC的相关性较好（相关系数不低于0.65）。将优选的5个遥感光谱指标作为模型输入变量,构建基于BP神经网络的冬小麦LNC估测模型（LNC-BPEM）,模型的估测精度r²=0.866,RMSE=0.246%,ARE=12.9%。将冬小麦LNC-BPEM估测模型和GF-6/WFV影像结合对县域冬小麦LNC的空间信息监测,获得了如皋县域冬小麦LNC的空间分布特征,该区域冬小麦LNC范围在0.9%~2.0%（长势正常）的种植面积为29 693.3 hm²,占冬小麦总种植面积的74%。这说明利用GF-6/WFV卫星的多个遥感光谱指标与神经网络结合建模可有效估测县域大田冬小麦叶片氮素含量。     

副溶血弧菌VP-5的分离鉴定及其与鲈鱼免疫信号通路相互关系研究

为探讨福建省漳州市盛明养殖场鲈鱼的致病菌及其与鲈鱼免疫信号通路的相互关系,从而可以预防和治疗该疾病。解剖患病鲈鱼肝脏组织后划线培养分离纯化了致病菌VP-5。显微镜观察该致病菌VP-5形态为球杆状,弧形,有单鞭毛,运动活跃,革兰阴性菌。在VITEK-32全自动微生物分析仪鉴定后,该致病菌对酪氨酸、葡萄糖、甘露醇等反应阳性,对蔗糖,阿拉伯糖和棉子糖等反应阴性。分析了该致病菌16SrRNA和tdh1基因序列,发现其与副溶血弧菌的亲缘关系非常接近。综合后鉴定致病菌VP-5为副溶血弧菌。并研究了副溶血弧菌VP-5侵染鲈鱼后寄主免疫信号通路基因cd63/cxcr4/il-8的表达水平差异。鲈鱼腹腔注射感染副溶血弧菌VP-5,感染后0-96h解剖鲈鱼肝脏组织和表皮组织。采用Trizol试剂盒抽提鲈鱼总RNA,反转录后生成cDNA。利用实时荧光定量PCR技术对鲈鱼免疫信号通路基因cd63/cxcr4/il-8表达水平进行测定。鲈鱼受到副溶血弧菌VP-5感染后,免疫信号通路基因cd63/cxcr4/il-8在鲈鱼肝脏组织的表达量12h和24h时与对照组相比较差异显著(P<0.05);在表皮组织的表达量在24h和36h时与对照组相比较差异显著(P<0.05)。证实鲈鱼肝脏组织免疫信号通路基因的表达与副溶血弧菌VP-5的感染正相关,相对于表皮组织反应更快。因此鲈鱼的免疫信号通路基因cd63/cxcr4/il-8积极反应于致病菌,这些基因的快速表达会引发鲈鱼自身免疫应答,增强其抵抗致病菌的能力。     

鸡肉、鸡蛋中4种酰胺醇类药物残留的液相色谱-三重四级杆串联质谱(UPLC-MS/MS)分析方法建立

建立了鸡肉、鸡蛋中4种酰胺醇类药物(氯霉素CAP、氟苯尼考FF、甲砜霉素TAP、氟苯尼考胺FFA)残留的超高效液相色谱-三重四级杆串联质谱(UPLC-MS/MS)快速分析方法。样品用80%的乙腈震荡提取,经过PRiME HLB固相萃取柱直接净化,在40℃下氮吹浓缩,最后用10%乙腈复溶。以0.01%甲酸水-0.01%甲酸乙腈溶液作为流动相进行梯度洗脱,采用多反应监测(MRM)模式进行测定,内标法定量。4种酰胺醇类药物在0.5~20 ng/mL的浓度范围内呈现良好的线性关系,R₂^在0.9982~ 0.9988范围内。添加量为0.5、1、5μg/kg时,鸡肉中4种酰胺醇类药物回收率为89.6%~104.2%,鸡蛋中4种酰胺醇类药物回收率为85.9%~105.5%。通过回收率和精密度试验得出该方法具有较好的准确度和重复性。通过试验得出4种酰胺醇类药物检出限为0.05~0.1 μg/kg,定量限为0.2~0.5 μg/kg,本法操作简便、灵敏,准确,适合大批量鸡肉、鸡蛋样品中4种酰胺醇类药物的检测。     

Body weight of newborn and suckling piglets affects their intestinal gene expression

Modern hyperprolific sows must deal with large litters (16–20 piglets) which reduce piglet birthweight with a concomitant increase in the proportion of small and intrauterine growth retarded piglets. However, larger litters do not only have a greater variation of piglet weights, but also a greater variation in colostrum and milk consumption within the litter. To further understand the impact that body weight has on piglets, the present study aimed to evaluate the degree of physiological weakness of the smallest piglets at birth and during the suckling period (20 d) compared to their middle-weight littermates through their jejunal gene expression. At birth, light piglets showed a downregulation of genes related to immune response (FAXDC2, HSPB1, PPARGC1α), antioxidant enzymes (SOD2m), digestive enzymes (ANPEP, IDO1, SI), and nutrient transporter (SLC39A4) (P < 0.05) but also a tendency for a higher mRNA expression of GBP1 (inflammatory regulator) and HSD11β1 (stress hormone) genes compared to their heavier littermates (P < 0.10). Excluding HSD11β1 gene, all these intestinal gene expression differences initially observed at birth between light and middle-weight piglets were stabilized at the end of the suckling period, when others appeared. Genes involved in barrier function (CLDN1), pro-inflammatory response (CXCL2, IL6, IDO1), and stress hormone signaling (HSD11β1) over-expressed compared to their middle-weight littermates (P < 0.05). In conclusion, at birth and at the end of suckling period, light body weight piglets seem to have a compromised gene expression and therefore impaired nutrient absorption, immune and stress responses compared to their heavier littermates.     

2种头孢氨苄片在比格犬体内的药代动力学及生物等效性试验

【目的】研究头孢氨苄片受试制剂和参比制剂在比格犬体内的生物等效性。【方法】采用双周期和双序列交叉设计,将22只健康比格犬随机分成2组,按30 mg/kg BW分别单剂量口服头孢氨苄片受试制剂Trolevis~?300和参比制剂Rilexine~?300,于给药前(0 h)和给药后0.5、1、1.5、2、2.5、3、4、6、8、12、17和24 h从臂头静脉采血。对超高效液相色谱串联质谱(UPLC-MS/MS)方法进行特异性、线性、检测限、准确度、精密度、稳定性等方法学考察。利用建立好的UPLC-MS/MS方法测定血浆中的药物浓度,并用WinNonlin^TM 8.1软件对药代动力学参数进行分析计算。【结果】方法学结果显示,在100～5 000 ng/mL浓度范围内相关性良好,相关系数(R²)≥0.99,标准曲线方程为y=10.6828x-176.481;高、中、低3个浓度的相对回收率平均值分别为105.63%、104.35%和102.40%;日内和日间变异系数均<15%;检测限为50 ng/mL,定量限为100 ng/mL。药代动力学结果...     

静原鸡let-7a-5p的靶基因预测及组织表达分析

【目的】了解let-7a-5p的生物学信息,初步探索其在静原鸡肌肉组织中的功能作用。【方法】利用生物信息学方法对转录组测序获得的静原鸡let-7a-5p的成熟序列进行保守性分析,使用miRDB、TargetScan和miRanda在线软件预测let-7a-5p的靶基因,取其交集进行GO功能和KEGG通路富集分析,并利用实时荧光定量PCR检测let-7a-5p在静原鸡公鸡和母鸡各组织中的表达水平。【结果】let-7a-5p的成熟序列在各物种间高度保守,共预测得到38个基因集合。在生物过程、细胞组分和分子功能中靶基因富集较多的GO条目有细胞过程、生物调节、细胞、结合和催化活性;靶基因显著富集于聚酮糖单元生物合成、AGE-RAGE信号通路在糖尿病并发症中的作用、泛素介导的蛋白水解、TGF-β信号通路和加压素调节的水重吸收信号通路,且富集最多的是代谢途径、AGE-RAGE信号通路在糖尿病并发症中的作用、泛素介导的蛋白水解和MAPK信号通路。let-7a-5p-靶基因互作主要通过作用于靶mRNA的3′-UTR抑制表达或使其降解。组织表达发现,let-7a-5p在静原鸡公、母鸡的心脏、肝脏、脾脏、...     

广灵驴FTO基因克隆、序列分析及组织差异表达

【目的】对广灵驴脂肪和肥胖相关基因(fat mass and obesity-associated gene, FTO)进行克隆及序列分析,同时检测其在广灵驴各组织中的表达差异,以探究广灵驴FTO基因结构对其生理代谢功能的影响。【方法】运用RT-PCR技术扩增并克隆广灵驴FTO基因CDS序列,进行基因及蛋白质功能分析,同时运用实时荧光定量PCR方法检测FTO基因在广灵驴7种组织(心脏、肝脏、脾脏、肺脏、肾脏、背最长肌及皮下脂肪)中的差异表达。【结果】广灵驴FTO基因CDS区序列长1 518 bp,编码505个氨基酸,序列提交至NCBI,登录号:MZ169553。广灵驴FTO基因与马、猪、牛、人、羊驼、绵羊和山羊的相似性为99.3%、90.3%、89.5%、90.8%、90.7%、89.2%和89.2%;系统进化树分析发现,广灵驴与马的亲缘关系最近。FTO蛋白分子质量为58.35 ku,理论等电点为5.07,脂肪系数为80.36,不稳定系数为48.82,平均疏水指数为-0.550,为不稳定的酸性亲水蛋白。FTO蛋白无信号肽和跨膜区,主要定位于细胞质中,具有34个磷酸化位点与5个糖基化位点...     

抑制差减杂交法(SSH)及其在植物中的应用

简述了抑制差减杂交法的基本原理、方法及优缺点,并对其在植物中的应用进行了综述。     

Penetration, Development and Emigration of Juveniles of the Nematode Meloidogyne arenaria in Myrobalan Plum (Prunus cerasifera) Clones Bearing the Ma Resistance Genes

Penetration, development and emigration of M. arenaria in the roots of three Myrobalan plum (Prunus cerasifera) clones genetically characterized for their resistance to root-knot nematodes (RKN) were studied during the 10 (penetration) and 15 (emigration) days following the date of inoculation (D) of 2500 juveniles (J2s) per plant into the soil. Miniaturized tests were conducted on the two resistant clones P.2175 (Ma1 gene) and P.1079 (Ma2 gene) and the susceptible clone P.2032 (recessive for both genes), obtained from micropropagated plantlets and grown in mini-containers under controlled conditions at 25°C in a growth chamber. For penetration and development studies, nematodes in the roots were recovered by the acid fuchsin-lactophenol staining technique. Equivalent numbers of J2s were recovered in all the clones at D+1 and D+2. Subsequently, the numbers increased rapidly in P.2032 and were significantly different from those in P.1079 and P.2175 that remained at a low level. No swollen larvae were observed in the resistant clones. In P.2032, the first swollen larvae were observed at D+4, the first females were observed at D+12, whereas the first females with attached egg sacs and the first new-generation J2s were obtained between D+21 and D+28. Our data suggest that the resistance phenomenon does not act on the very early nematode penetration but acts later by preventing feeding-site induction and development into the third-stage. For emigration studies, plants in which J2s had been allowed to penetrate for two days (from D to D+2) were washed free of soil, repotted and then, after various periods of growth, soil-free roots were placed under a mistifier to evaluate the numbers of emigrating individuals. Emigration of J2s from the roots occured mainly from D+2 to D+4 in all the genotypes and was very limited from D+4 to D+10. There was no significant differences in the number of emigrated juveniles between the resistant and susceptible clones, indicating that emigration cannot explain the difference in the numbers of nematodes recovered in the roots.