笃斯越桔硬枝扦插繁殖技术研究

采用笃斯越桔硬枝扦插,用生根剂处理插穗,并在不同的基质上进行扦插试验,结果表明:笃斯越桔硬枝扦插用河沙作为基质最好,生根剂ABT处理的插穗生根率最高。     

两种一步法RNA提取试剂的比较

RNA的提取在生物医学研究和临床检测上应用广泛。本文分别用进口Invitrogen公司生产的以及我们自己研制的一步法RNA提取试剂，提取大肠杆菌RNA、鸡肌肉组织RNA和尿囊液RNA，通过核酸电泳和荧光定量RT-PCR进行比较。结果表明用我们自己研制的和进口的试剂提取的RNA都不含有DNA；这些RNA都可以作为模板进行RT-PCR检测，并且在数量上相互之间没有明显差异。说明我们自己研制的一步法RNA提取试剂可以取代进口试剂。此外，本文还讨论了此类试剂一些评定方法。     

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.     

基于pGhost4系统保加利亚乳杆菌乙酸激酶敲除载体的构建

保加利亚乳杆菌凭借其微生物特性和效能优异等特点成为当下产乳酸最重要的微生物菌株之一。乙酸是保加利亚乳杆菌代谢乳酸最主要的副产物,它的大量生成降低葡萄糖的代谢利用率,并且消耗了能量。乙酸激酶是控制乙酸生成的关键酶,敲除乙酸激酶基因,理论上可以阻断戊糖磷酸途径中乙酸的生成,从而优化代谢途径提高葡萄糖代谢乳酸的利用率。研究以Lactobacillus delbrueckii subsp.bulgaricus ATCC 11842公布的乙酸激酶基因ack序列设计引物,以保加利亚乳杆菌基因组DNA为模板,PCR克隆出ack上下游片段,利用重叠PCR技术将上下游片段拼接在一起,并连入具有温敏性的p Ghost4载体。     

Early resynchronization protocols for goats: Progestogens can be used prior to an early pregnancy diagnosis without affecting corpus luteum functionality

This study aimed to evaluate the exogenous progesterone (P4) effect on the luteal function from Day 16 to Day 21 of the oestrous cycle in inseminated goats with unknown pregnancy status. A total of 54 does passed through a short progestin-based synchronization protocol and, on Day 16 of the following oestrous cycle, 27 does received a new P4 device which was retained until Day 21. Blood samples were collected daily from all does during this period, as well as on Day 24. Pregnancy diagnoses were performed on Day 30. Serum P4 values from 26 animals (G_NPSP: Group of non-pregnant does with second sponge: n = 8; G_NPNSP: Group of non-pregnant does without second sponge: n = 6; G_PSP: Group of pregnant does with second sponge: n = 5; G_PNSP: Group of pregnant does without second sponge: n = 7) were determined by radioimmunoassay commercial kits. No P4 differences were found between groups (G_NPSP: 3.1 ± 2.8; 1.7 ± 1.8; 0.4 ± 1.0; and 0.0 ± 0.0 vs. G_NPNSP: 4.4 ± 1.8; 3.0 ± 2.2; 0.8 ± 0.8; and 0.0 ± 0.0 or G_PSP: 4.2 ± 1.0; 3.4 ± 0.6; 3.3 ± 1.6; 3.2 ± 0.9; 3.6 ± 1.2; 3.5 ± 1.3; 2.7 ± 1.3 vs. G_PNSP: 4.4 ± 1.6; 3.6 ± 1.5; 3.7 ± 1.5; 3.8 ± 1.4; 3.2 ± 1.2; 3.1 ± 1.2; 3.6 ± 1.1; D16, D17, D18, D19, D20, D21, D24, respectively) or for the interaction of group and time. In conclusion, a second progestogen device had no effect on luteolysis or early pregnancy in the following oestrous cycle.     

土壤交换性铝的三种测定方法比较

为了准确高效地检测土壤交换性铝含量,探寻适宜的检测方法。比较分析了氯化钾交换-中和滴定法、铝试剂法和羊毛铬花青R比色法所测交换性铝的差异性、精密度、准确性和适用性。结果表明,3种方法所测交换性铝无显著性差异。但羊毛铬花青R比色法的精密度优于氯化钾交换-中和滴定法和铝试剂法,羊毛铬花青R比色法平均回收率达99.28%,准确性高于另2种方法。羊毛铬花青R比色法的线性范围在0~0.32 mg/L,对应吸光值范围在0~0.778;铝试剂法的线性范围在0~0.8 mg/L,对应吸光值范围在0.006~0.157;与铝试剂法相比,羊毛铬花青R比色法的线性范围小于铝试剂法,但其吸光值范围大于铝试剂法。羊毛铬花青R比色法显色剂与显色物质吸收峰间隔较远,测定背景干扰小,方法灵敏度较高。羊毛铬花青R比色法检测单个样品的平均用时为4.2 min,检测效率高于另2种方法,且操作简捷,适用性较高。因此,推荐羊毛铬花青R比色法为土壤交换性铝的测定方法。     

不同处理对芹菜种子萌发过程中抗氧化系统及激素水平的影响

为研究芹菜种子的萌发机制,采用不同试剂组合（5 g/L NaOH+10% PEG、10% PEG+500 mg/L壳聚糖、5 g/L NaOH+500 mg/L壳聚糖）和不同温度(18、24、30℃)对芹菜种子进行处理,观察芹菜种子的萌发指标,并探究种子萌发过程中的抗氧化系统及内源激素水平的变化。结果表明,30℃对种子发芽率、发芽势及发芽指数提高具有促进作用,加快种子萌发进程。与蒸馏水对照组相比,试剂组合处理在18、24℃时种子发芽率、发芽势和发芽指数升高,促进种子萌发,18℃时5 g/L NaOH+500 mg/L壳聚糖处理下的发芽势最高,24℃时5 g/L NaOH+10% PEG处理下发芽势达到最大值;30℃时不同试剂组合处理间发芽率、发芽势和发芽指数无明显差异。18、30℃对种子具有一定程度的胁迫作用,种子SOD和CAT活性较24℃条件下有所增加;18℃时MDA含量和脯氨酸含量显著增加;5 g/L NaOH+10% PEG处理能显著降低MDA含量。萌发过程中,芹菜种子内部ABA含量下降,GA₃、ZA含量增加。试剂组合及适当高温有利于提高芹菜种子的发芽率、发芽势和发芽指数,促进种子萌发,提高种子萌发整齐度;芹菜种子萌发过程中抗氧化系统和激素水平会积极响应不同的处理条件。