The influence of progesterone and prostaglandin F2α on decorin and the adhesion of caruncular epithelial cells of bovine placenta at early-mid pregnancy—Part II

One of the most important processes determining the proper course of gestation and its physiological termination in cows is the adhesion of epithelial cells allowing for direct contact of maternal and foetal parts of the placenta. Throughout pregnancy, placental cells are under strict hormonal control, which among others regulates the concentration and activity of specific proteins participating in the extracellular matrix remodelling of foetal membranes. The aim of the study was to evaluate the influence of progesterone and prostaglandin F_2α on the adhesion of epithelial cells at early-mid pregnancy in cows. Additionally, the impact of selected hormones on anti-adhesive properties of decorin was evaluated. Caruncular epithelial cells were isolated from healthy cows during pregnancy, immediately after slaughter. Primary cell cultures derived from the 2nd and 4th month of gestation were used in the experiments. The viability of cells was assessed by MTT assay. The adhesion of cells to fibronectin was measured spectrophotometrically. The activity of metalloproteinases was confirmed by the metalloproteinase assay. Progesterone (10^–5 and 10^–7 mol/L) and prostaglandin F_2α (10^–4_, 10^–5 and 10^–7 mol/L) increased the viability of bovine caruncular epithelial cells in the 2nd month of pregnancy. The treatment with prostaglandin F_2α significantly reduced the number of adherent cells from the 2nd month of gestation at the doses of 10^–4 and 10^–5 mol/L. Both progesterone and prostaglandin F_2α were shown to have an effect of decorin resulting in both a decrease in metalloproteinase activity and an increase in adhesion of cells to fibronectin.     

灵芝发酵产物对感染PCV-2病毒断奶仔猪生长性能和免疫功能的影响

本研究以深层发酵灵芝为原料,评估灵芝发酵产物对感染猪圆环病毒-2(PCV-2)断奶仔猪生长发育及免疫功能的影响。试验1:选择160头21 d断奶的仔猪,随机分为4组,每组4个重复,每个重复10头猪。各组分别在基础日粮中添加0、50、100和150 mg/kg灵芝发酵产物。试验2:将120头21 d断奶的仔猪随机分为2组(4个重复,每个重复15头),分别饲喂基础日粮和基础日粮+50 mg/kg的灵芝发酵产物。两组仔猪在试验当天通过肌肉注射105/mL感染剂量的PCV-2病毒。结果:日粮添加50 mg/kg灵芝发酵产物2周和4周后显著改善了平均日增重(P<0.05),在第4周时150 mg/kg较50 mg/kg灵芝发酵产物组显著降低了平均日增重(P<0.05)。150 mg/kg灵芝发酵产物组平均日采食量较其他组显著降低了8.11%、6.85%和8.11%(P<0.05),日粮添加50 mg/kg灵芝发酵产物显著改善了断奶仔猪的料重比(P<0.05)。日粮添加50~150 mg/kg灵芝发酵产物可显著提高肺泡巨噬细胞的趋化性指数(P<0.05),同时对照组显著提高了28 d伪狂犬病抗体滴度(P<0.05),显著降低了血清葡萄糖水平(P<0.05)。灵芝发酵产物日粮组较对照组显著提高了IL-2、INF-γ和TNF-α相对强度(P<0.05)。结论:日粮添加50 mg/kg灵芝发酵产物可以显著改善断奶仔猪生长性能,在接种伪狂犬病疫苗时可提高肺泡巨噬细胞的趋化能力和抗体滴度。     

培肥措施对旱地农田土壤CO2排放和碳库管理指数的影响

阐明长期有机物料施肥下土壤CO₂排放特征及其影响机制以及碳库管理指数对黄土高原旱作农业区固碳减排及施肥模式选择的影响尤为重要。基于2012年设置在陇中黄土高原旱作区的长期定位试验,通过不施肥（CK）、氮肥（NF）、有机肥（OM）、秸秆（ST）、有机肥结合无机肥（OMNF）5个处理,测定并计算了2018年不同施肥措施下全年土壤CO₂排放、作物碳排放效率和碳库管理指数的变化,并运用结构方程模型分析了0~30 cm土壤温度、水分、微生物量碳氮、易氧化有机碳、蔗糖酶、脲酶与土壤CO₂排放速率的关系。结果表明：1）与不施肥相比,秸秆、有机结合无机肥和有机肥处理使生育期土壤CO₂排放平均速率提高了42.72%、30.82%和29.79%,秸秆、有机肥处理分别使生育期土壤CO₂排放量显著提高36.35%、32.45%（P<0.05）,有机结合无机肥处理使碳排放效率显著降低41.10%（P<0.05）;2）有机物料处理均能显著提高0~5 cm土层易氧化有机碳、微生物量碳氮、蔗糖酶活性和碳库管理指数,相比不施肥和氮肥处理,有机结合无机肥处理分别使0~30 cm土壤碳库管理指数提高127.41%,99.33%（P<0.05）;3）结构方程模型表明,环境因子对土壤CO₂排放速率的总解释度为53%,对土壤CO₂排放速率总效应较大的影响因素包括土壤温度（2.36）、微生物量碳（1.59）和土壤水分（1.18）,且均间接地影响着土壤CO₂排放速率,土壤温度促进了微生物量碳和蔗糖酶活性的提高,微生物量碳促进了微生物量氮和易氧化有机碳的增加。综合来看,有机结合无机肥处理可以提升土壤碳库管理指数,保持微生物活性,增加作物产量,降低土壤碳排放效率,是陇中黄土高原旱作农业区比较适宜的农田培肥措施。     

高表达线粒体融合蛋白2(MFN2)可抑制高BHBA活化的奶牛肝细胞NF-κB炎性通路

为研究线粒体融合蛋白2(mitofusin 2,MFN2)对高β-羟丁酸(β-hydroxybutyrate,BHBA)活化NF-κB炎性通路的影响,体外培养奶牛肝细胞,添加不同浓度(0.0,1.2,2.4,4.8 mmol/L)的BHBA,并转染过表达MFN2的腺病毒,运用Western blot和qRT-PCR技术检测NF-κB炎性通路关键分子的基因和蛋白表达。结果显示,随着BHBA浓度的增加,IKBα和NF-κB p65的磷酸化水平以及IL-1β、IL-6和TNFα的mRNA表达均显著升高,MFN2的基因和蛋白表达水平则显著降低;过表达MFN2后,显著抑制了高BHBA诱导的IKBα和NF-κB p65磷酸化水平及IL-1β、IL-6、TNFαmRNA表达水平的升高。结果表明,在奶牛肝细胞中过表达MFN2可以显著抑制高BHBA活化的NF-κB炎性通路。     

黄河源区建植19年人工草地生物结皮CO2通量与叶绿素荧光参数的变化

为了解黄河源人工草地不同类型生物结皮和植被群落CO₂释放和固定的变化特征,并探讨二者的影响因子,本文通过调查黄河源建植19年人工草地不同类型生物结皮CO₂通量和植被土壤特征变化发现：藻和地衣结皮CO₂通量均显著小于苔藓结皮（10.58 μmol·m^-2·s^-1）（P<0.05）;苔藓结皮对应的植被群落CO₂通量（11.55 μmol·m^-2·s^-1）显著低于藻和地衣结皮（P<0.05）;藻和地衣结皮下的土壤CO₂通量显著大于苔藓结皮（约2倍,P<0.05）。苔藓结皮叶绿素初始荧光参数F₀（Minimal Fluorescence）和最大荧光参数F_m（Maximal Fluorescence）均最大;藻和苔藓结皮的PS Ⅱ最大光化学量子产量F_v/F_m（Optimal/Maximal Quantum Yield of PS Ⅱ）均高于地衣结皮。CO₂通量和叶绿素荧光参数的主要影响因子相似,均与禾本科高度、土壤铵态氮含量正相关,与pH负相关。因此,在研究草地碳平衡过程中应充分考虑生物结皮的贡献,同时需注意不同类型的生物结皮的CO₂通量和叶绿素荧光参数的差异变化。     

南荻MlNAC2基因差异表达与耐盐生理响应的相关性分析

为探究南荻（Miscanthus lutarioriparius）耐盐生理及其与基因表达的相关性,采用0%,0.2%,0.5%,0.8%浓度的NaCl处理奇岗和不同基因型南荻,测量NaCl溶液胁迫下的MlNAC2基因相对表达量和6个耐盐指标,并对二者之间的相关性进行分析。结果显示,南荻MlNAC2基因的表达量变化在不同材料之间差异较大,L2,L6,L9,P1出现10倍以上增加,而L1,L5,L8,L10变化量都在3倍以内。生理指标中叶绿素含量、净光合速率、气孔导度、脯氨酸与无胁迫对照相比均差异显著;丙二醛、超氧化物歧化酶在部分材料中与对照相比无显著差异。相关性分析表明,与MlNAC2基因的表达量变化最相关的生理指标是一系列光合作用相关因子：叶绿素含量、净光合速率、气孔导度。本研究结果可为探究南荻耐盐生理响应与分子机理层面的相互关系提供借鉴。     

Effect of dipeptide on intestinal peptide transporter 1 gene expression: An evaluation using primary cultured chicken intestinal epithelial cells

Peptide transporter 1 (PepT1) is a transporter responsible for absorbing dipeptide and tripeptide in enterocytes and is upregulated by dipeptide in mammals. It has not been certain whether intestinal PepT1 expression is responsive to dipeptides in chickens because of the lack of in vitro study using the cultured enterocytes. This study established a primary culture model of chicken intestinal epithelial cells (IECs) in two-dimensional monolayer culture using collagen gel by which the response of chicken PepT1 gene expression to dipeptide stimuli was evaluated. The cultured chicken IECs showed the epithelial-like morphology attached in a patch-manner and exhibited positive expression of cytokeratin and epithelial cadherin, specific marker proteins of epithelial cells. Moreover, the chicken IECs exhibited the gene expression of intestinal cell type-specific marker, villin1, mucin 2, and chromogranin A, suggesting that the cultured IECs were composed of enterocytes as well as goblet and enteroendocrine cells. PepT1 gene expression was significantly upregulated by synthetic dipeptide, glycyl-l-glutamine, in the cultured IECs. From the results, we herein suggested that dipeptide is a factor upregulating PepT1 gene expression in chicken IECs.     

绵羊BMP15基因B2突变可视化检测技术的建立

骨形态发生蛋白15（bone morphogenetic protein 15,BMP15）基因突变对绵羊排卵率、产仔数以及繁殖力有很大影响,本研究旨在建立快速可视化检测BMP15基因B2突变的技术。将改良的扩增阻滞突变系统（amplification refractory mutation system,ARMS）与核酸染料SYBR Green Ⅰ结合,针对BMP15基因B2突变设计ARMS特异性引物,使引物下游3'端的最后一个碱基为A,并在下游引物3'端的第3位碱基处设计额外的错配,以提高引物的特异性;经ARMS PCR扩增后,向产物中加入SYBR Green Ⅰ,通过肉眼观察PCR管内的颜色变化来检测BMP15基因的B2突变。经测序发现所采集的样本均为野生型,因此,利用重叠延伸PCR构建BMP15基因B2突变型模板,测序结果显示BMP15基因B2突变型模板构建成功。ARMS PCR结果显示,ARMS特异性引物能够只扩增突变型模板,而不会扩增野生型模板,且扩增效果良好。ARMS PCR扩增后加入SYBR Green Ⅰ发现已知野生型模板与阴性对照一致,呈SYBR Green Ⅰ原始颜色橙黄色,而已知突变型模板呈亮绿色,且色差明显,肉眼可辨。用建立的可视化检测BMP15基因B2突变的技术检测50个小尾寒羊基因组DNA （随机向其中20个样本中加入构建的BMP15基因B2突变型模板）,将可视化检测的突变型样本编号与混合样本时记录的编号对比,结果表明该技术能够准确检测绵羊BMP15基因的B2突变,且准确率高达100%。将构建的BMP15基因B2突变型的模板进行梯度稀释,对本试验可视化检测体系的灵敏度进行检测,发现ARMS可视化检测技术的灵敏度达到0.006 ng/μL。因此,本研究建立的技术能够可视化地检测绵羊BMP15基因B2突变,且准确率高,有望为高繁殖力绵羊的选育提供技术支持。