Roles of protein phosphatase 4 in down-regulation of eNOS Ser633 phosphorylation induced by palmitic acid in human umbilical vein endotheli?al cells

AIMTo investigate the roles of protein phosphatase 4 (PP4) in down-regulation of endothelial nitric oxide synthase (eNOS) Ser633 phosphorylation induced by palmitic acid (PA). METHODSHuman umbilical vein endothelial cells (HUVECs) were treated with PA at 25 μmol/L, 50 μmol/L, 100 μmol/L and 200μmol/L for 36 h, or treated with PA at 100 μmol/L for 12 h, 24 h, 36 h and 48 h. Protein phosphatase 2A (PP2A) family inhibitor fostriecin (FST, 20 nmol/L) or okadaic acid (OA, 5 nmol/L) was selected to pretreat the HUVECs for 30 min. Protein phosphatase 4 catalytic subunit (PP4c) siRNA or protein phosphatase 2A catalytic subunit (PP2Ac) siRNA was transfected into the HUVECs. The protein expression levels of of eNOS, PP4c and PP2Ac, as well as the level of eNOS Ser633 phosphorylation, were detected by Western blot. The intracellular nitric oxide (NO) content was measured by DAF-FM DA. RESULTS(1) Compared with control group, the levels of eNOS Ser633 phosphorylation were decreased in PA groups in which the HUVECs were treated with 25 μmol/L, 50 μmol/L, 100 μmol/L and 200 μmol/L PA for 36 h (P<0.05) and 100 μmol/L PA for 24 h, 36 h and 48 h (P<0.05). No significant difference in the level of total eNOS protein expression among all the groups was observed. (2) Compared with control group, both FST and OA pretreatment reversed the reduction of eNOS Ser633 phosphorylation (P<0.05) and the decrease in intracellular NO content (P<0.05) induced by PA. No significant difference in the level of total eNOS protein expression among all the groups was observed. (3) Compared with si-Control group, the PP4c protein expression was significantly reduced (P<0.05), while the level of eNOS Ser633 phosphorylation was significantly increased in si-PP4c group (P<0.05). Although the levels of PP2Ac protein expression declined significantly (P<0.05), the level of eNOS Ser633 phosphorylation remained unchanged in si-PP2Ac group. No significant differencein the level of total eNOS protein expression among all the groups was found. CONCLUSION PA significantly reduces the level of eNOS Ser633 phosphorylation and the content of NO in the HUVECs, which may be due to PA inducing the activation of the PP2A family member PP4 rather than PP2A.     

Effects of apelin-13 on oxidative stress induced by high uric acid in adipocytes

AIM: To study the effects of apelin-13 on oxidative stress induced by high uric acid in 3T3-L1 adipocytes and its underlying mechanisms. METHODS: 3T3-L1 adipocytes were stimulated with uric acid at 10 mg/dL for 48 h. Some of the adipocytes were administered with 1 μmol/L apelin-13 in the presence of uric acid at 10 mg/dL. The adipocytes stimulated with 100 μmol/L H₂O₂ were served as positive controls. The intracellular reactive oxygen species (ROS) concentrations were detected by flow cytometry. The biochemical kits were used to measure the activities of superotide dismutase (SOD), glutathione peroxidase (GSH-Px), catalase (CAT) and NADPH oxidase (NOX) activity, and the content of malondialdehyde (MDA) in the cell lysate and the supernatant. The mRNA levels of renin-angiotensin system (RAS) components, including angiotensinogen (AGT), angiotensin-converting enzyrne1 (ACE1), angiotensin II type 1 receptor (AT1R) and AT2R, as well as angiotensin II receptor -like 1 (APJ) were measured by real-time PCR. The concentrations of angiotensin II (AngⅡ) in the cell lysate and the supernatant were measured by ELISA. RESULTS: Adipocytes stimulated with uric acid at 10 mg/dL had lower activities of antioxidant enzymes (SOD, GSH-PX and CAT) and higher levels of NOX activity and MDA content (P < 0.05). Accordingly, the intracellular ROS levels were found to be dramatically increased. However, apelin-13 administration attenuated uric acid-induced oxidative stress in the 3T3-L1 adipocytes. Uric acid at 10 mg/dL upregulated the mRNA expression of local RAS, enhanced AngⅡ concentrations both in the cell lysate and the supernatant, and down-regulated the mRNA level of APJ in the adipocytes (P < 0.05). Conversely, apelin-13 partially reversed these parameters. CONCLUSION: Apelin-13 attenuates oxidative stress induced by uric acid, may be via down-regulation of local RAS expression in the 3T3-L1 adipocytes.     

Progress in relationship between epithelial sodium channel and related respiratory diseases

Epithelial sodium channel (ENaC) is an ion channel widely distributed in various tissues and organs of human. It is composed of 3 homologous subunits and allows the flow of sodium ion across epithelial cells, maintain?ing water-salt balance in the cells. Recent studies show that abnormal expression or dysfunction of ENaC in the respiratory system affects water-salt balance, fluid transportation and cell mobility, and causes abnormal changes of the airway surface liquid level and impaired clearance. ENaC is closely related to the development of respiratory diseases, such as cystic fibrosis, asthma and chronic obstructive pulmonary disease. This article reviews the progress in ENaC structure, function and roles in related respiratory diseases in order to provide a reference for the treatment of the diseases.     

刺萼龙葵种子中适宜内参基因的筛选

入侵杂草刺萼龙葵Solanum rostratum Dunal传播扩散的主要载体是种子,研究其种子休眠萌发基因的激素调控对于其防除具有重要意义,而选择合适的内参基因可以提高相关基因表达分析的准确性。本研究以赤霉素、脱落酸和水处理的刺萼龙葵种子为材料,利用geNorm、NormFinder、BestKeeper和RefFinder 4种软件对15个候选内参基因进行表达稳定性评价,并通过检测ABI5(abscisic acid-insensitive 5)的表达验证所筛选的内参基因的适用性。结果表明,对于赤霉素、脱落酸和水处理过的种子,最稳定的内参基因分别为eIF(eukaryotic initiation factor)、SAND(SAND protein family)和ACT(β-actin);对所有种子样本而言,PP2Acs(a catalytic subunit of protein phosphatase 2A)是最稳定的内参基因。研究结果将为刺萼龙葵种子休眠萌发的遗传调控研究提供重要参考。     

雀麦花叶病毒外壳蛋白基因原核表达及抗血清制备

正雀麦花叶病毒(brome mosaic virus,BMV)为正义单链RNA病毒。BMV寄主范围广泛,在波兰等地均有发现(Trzmiel et al.,2015),大多危害禾本科、豆科、茄科等植物,感病植株常表现出褪绿、轻微花叶症状。因此,为减轻BMV造成的经济损失和丰富BMV的检测方式,本研究拟构建BMV的外壳蛋白(coat protein,CP)基因的原核表达载体,在大肠杆菌Escherichia coli中表达BMV的外壳蛋白,制备高特异性和高灵敏度的抗血清,以期用于BMV的有     

微波辐照对直接研磨法全麦粉的储藏 稳定性与品质的影响

为了明确微波辐照对全麦粉储藏稳定性及品质的影响,通过对全麦粉进行不同微波功率和不同微波时间处理,分析了全麦粉储藏过程中菌落总数以及脂肪酸值、面团流变学特性和馒头品质的变化。结果表明,随着微波功率的增加和处理时间的延长,全麦粉脂肪酸值和菌落总数增加速率得到抑制;湿面筋含量、面团稳定时间先增加后降低;气体释放曲线的最大高度(H′_m)和产气量(R_1)逐渐增加,气体保留体积(R_2)先增加后降低,发酵性能得到改善;馒头比体积与综合评分先增加后降低。当微波辐照功率为400 W、处理时间为120 s时,全麦粉可以更好储藏,同时全麦粉面团的粉质特性、流变发酵特性及馒头的品质得到改善。     

碾皮处理对全麦粉及馒头品质的影响

利用碾皮设备对小麦进行碾皮处理,对比普通小麦粉和不同碾皮时间下的全麦粉农药残留量、呕吐毒素含量、含砂量、干湿面筋含量、粉质特性、流变发酵特性、淀粉糊化特性以及馒头的品质指标来研究碾皮技术对全麦粉及馒头品质的影响。结果表明,随着碾皮时间的延长,小麦碾皮率逐渐升高,全麦粉中的农药残留量、呕吐毒素含量和含砂量呈现下降趋势;全麦粉精度和亮度增大,干湿面筋含量逐渐降低,但始终高于普通小麦粉,弱化度总体呈现下降趋势,全麦粉面团产气量和持气量呈现先增加后减小趋势,并在碾皮时间为20 s时达到最大,淀粉糊化特性总体呈增加趋势;全麦馒头的亮度、比容随着碾皮时间的延长逐渐升高,其感官评分在碾皮时间为20 s时最高。综上所述,碾皮处理在一定程度上提升了全麦粉和馒头的品质,碾皮时间为20 s时,全麦粉及其馒头的各项安全指标和品质特性均处于最佳状态。     

高炭基肥配施氮肥对牡丹江烤烟生长及产量和质量的影响

采用田间试验,设置4个处理[CK(常规施氮量,不施高炭基肥)、T1(常规施氮量,900 kg·hm~(-2)高炭基肥)、T2(85%常规施氮量,900 kg·hm~(-2)高炭基肥)、T3(70%常规施氮量,900 kg·hm~(-2)高炭基肥)],研究了减氮配施高炭基肥对牡丹江烤烟生长和产量、质量的影响。结果表明,T1处理烤烟各生育时期的农艺性状均有所提高,T3处理对烤烟的生长产生了负面效应;T1处理显著提高了中上部叶总氮含量(1.59%,1.65%)、烟碱含量(1.71%,1.99%),T1和T2处理显著提高了中部叶钾含量(1.68%,1.69%);与CK相比,T1处理中上部叶感官评吸总得分均有显著提高,增幅分别为4.06%和5.92%,其烟叶产量、产值及中上等烟比例均有所提高。在牡丹江烟区白浆土条件下,控制总施氮量为45 kg·hm~(-2)、高炭基肥料施用量为900 kg·hm~(-2)时,有利于促进烤烟生长、提高经济效益、改善烟叶品质。     

219份甜瓜种质资源的遗传多样性分析

以来自国内外的219份甜瓜种质资源为材料,对其进行形态学标记及分子标记的研究。结果发现,果面覆纹形状、果皮底色、果面覆纹颜色等7个质量性状和果实质量、果实长度、果实宽度等14个数量性状遗传多样性指数均≥1,果肉厚度、果肉硬度、种子宽度等7个数量性状变异系数≥50%,表明本研究所用材料间具有广泛的多样性。利用22对核心SSR标记对甜瓜材料的指纹图谱分析显示,每个标记平均可以检测到5.91个基因位点,平均Shannon′s值为1.07,通过分子水平的聚类与群体结构分析,219份材料被明显划分为厚皮、薄皮和野生3种类型。     

荸荠去皮机设计与试验

针对荸荠(Eleocharis dulcis(Burm. f.) Trin.)外形不规则以及去皮损失大、去不净的问题,采用平刀去顶芽、斜刀去底部、圆弧刀去侧皮的组合切削方式,设计1种荸荠去皮机。通过对荸荠外形参数的测量与建模,拟合得到荸荠外轮廓曲线,确定刀具的结构参数范围。基于3D打印技术,采用单因素试验,确定了切削速度、底面斜刀倾角与长度、侧面圆弧刀弦长对去皮损失率与去净率的影响,优化得到了适宜的结构参数与运动参数,在转速为40 r/min、底面斜刀倾角为82°、底面斜刀长度为14 mm、侧面圆弧刀弦长为22 mm(直径42.75 mm)的条件下,去净率达到93%,损失率为24.4%,显著高于人工去皮与机械摩擦去皮。