Inhibition study of red rice polyphenols on pancreatic α-amylase activity by kinetic analysis and molecular docking

This study sought to investigate the possible inhibition mechanism of red rice polyphenols (RRP) on pancreatic α-amylase (PA) activity. RRP showed strong inhibition against PA activity and the half-inhibitory concentration (IC₅₀) value was 3.61 μg/mL. The fluorescence quenching of PA by RRP was a combination of static quenching and dynamic quenching. RRP could aggregate with PA and the physiochemical properties of the aggregates were closely related to the concentration of RRP. Kinetic analysis suggested that the inhibition mode of RRP on PA was reversible inhibition, which was a mixing of competitive inhibition and noncompetitive inhibition. Molecular docking speculated that RRP could form hydrogen bonds with PA by binding to the catalytic active sites (ASP197, GLU233 and ASP300) and the microenvironments of TRP58 and TRP59 were altered, thus inhibiting PA activity.     

Effects of the severity and timing of basal leaf removal on the amino acids profiles of Sauvignon Blanc grapes and wines

The effects of the severity and timing of leaf removal(LR) on the amino acids of Sauvignon Blanc grapes and wines were studied during the 2017 growing season. High-performance liquid chromatography(HPLC) was used to analyze the amino acids profiles of grape berries and wines. The basal leaves were removed at three time points(40, 56 and 72 days after flowering, named LR40, LR56 and LR72, respectively) at two severity levels(one at which the first, third, and fifth basal leaves of each shoot were removed(50% level); and another at which the first six basal leaves were removed(100% level)). The results showed that leaf removal had little impact on total soluble solids(°Brix), titratable acidity, pH or berry weight. The LR72-50% treated grapes had higher berry weight, titratable acidity and °Brix than those of the other treatments. The highest concentrations of total amino acids and of total amino acids except proline were detected in LR72-50% treated grapes(2 952.58 and 2 764.36 mg L~(-1), respectively); the lowest were detected in LR72-100% treated grapes(2 172.82 and 2 038.71 mg L~(-1), respectively). LR72-50% treatment significantly promoted the synthesis of aspartic acid, serine, arginine, alanine, aminobutyric acid and proline at both severity levels for grapes, the concentrations of all of these amino acids were increased relative to the control concentrations. The LR72-50%, LR40-100% and LR72-100% treated wines had higher total amino acids concentrations and higher concentrations of some individual amino acids, such as arginine, alanine and serine, than did the control wines. Of all the amino acids studied, glycine, tyrosine, cysteine, methionine and lysine were not significantly influenced by the timing or severity basal defoliation in grapes and wines. The present study reveals the effects of the timing and severity of leaf removal on the amino acids profiles of grapes and wines.     

Reduction in cadmium accumulation in japonica rice grains by CRISPR/Cas9-mediated editing of OsNRAMP5

Cadmium (Cd) intake is harmful to human health and Cd contamination in rice grains represents a severe threat to those consuming rice as a staple food. Knockout of Cd transporters is a promising strategy to reduce Cd accumulation in rice grains. OsNRAMP5 is the major transporter for Cd and manganese (Mn) uptake in rice. Nevertheless, it is uncertain whether knockout of OsNRAMP5 is applicable to produce low Cd rice without affecting plant growth and grain yield. In this study, we adopted CRISPR/Cas9-based gene editing technology to knock out OsNRAMP5 in two japonica varieties. We generated three independent transgene-free osnramp5 mutants and investigated the effect of osnramp5 mutations on Cd accumulation and plant growth. Hydroponic experiments showed that plant growth and chlorophyll content were significantly reduced in osnramp5 mutants at low Mn conditions, and this defective growth in the mutants could be fully rescued by supply of high levels of Mn. Cd and Mn accumulation in both roots and shoots was markedly reduced in the mutants compared to that in wild-type plants. In paddy field experiments, although Cd in flag leaves and grains was greatly reduced in osnramp5 mutants, some agronomic traits including plant height, seed setting rate, and grain number per panicle were affected in the mutants, which ultimately caused a mild reduction in grain yield. The reduced plant growth in the mutants can be attributed to a marked decrease in Mn accumulation. Our results reveal that the manipulation of OsNRAMP5 should be treated with caution: When assessing the applicability of osnramp5 mutants, soil pH and soil water content in paddy fields need to be taken into consideration, since they might affect the levels of available Mn in the soil and consequently determine the effect of the mutation on grain yield.     

基于耦合协调度的大理河流域径流和输沙关系分析

深入研究流域水沙关系变化是解决黄河流域高质量发展关键问题的重要科学途径之一。自20世纪50年代以来,黄河中游地区实施了大规模的水土保持工程,径流和输沙显著减少,水沙关系发生变化。该研究利用Mann-Kendall趋势检验法分析了1960—2015年来大理河流域的径流输沙变化趋势,并构建基于耦合协调度理论和Pettitt检验方法的径流输沙关系变异诊断方法,识别大理河流域径流和输沙关系的突变点,并通过Copula方法进行验证。结果表明:1)大理河流域径流输沙在月尺度和年尺度呈现显著的下降趋势;2)大理河流域径流和输沙关系在1996年发生显著突变;3)径流输沙关系发生突变后,径流和输沙均明显减少,输沙的减少幅度比径流的减少幅度高29.19个百分点,径流对输沙贡献程度下降幅度为5.10%,单位径流的输沙能力降低;4)淤地坝建设和植被恢复是造成大理河流域水沙变化的主要因素,修建淤地坝和恢复植被对减少泥沙具有积极作用。研究成果可以为进一步深化理解大理河流域水沙关系变化以及水土流失治理提供依据。     

荔枝FKBP16-2基因启动子的克隆与瞬时表达分析

前期研究中发现了一个果肉低表达而叶片中高表达的荔枝基因FKBP16-2。本文克隆了该基因1 578 bp的启动子片段并对其功能进行了初步分析,结果表明:荔枝FKBP16-2基因启动子序列中含有大量的TATA-box和CAAT-box保守元件,以及TCA-element,ARE,HSE,GCN4_motif,O2-site等各种转录调控相关的顺式作用元件。该启动子能驱动GUS基因在荔枝的花、叶、根、果皮以及种子中表达而在果肉中不表达,表达具有组织特异性。     

硼对平邑甜茶幼苗硝态氮吸收、利用及分配特性的影响

以平邑甜茶幼苗为试材,运用~(15)N同位素示踪和非损伤微测技术,研究了不同供硼(硼酸0、1.0、2.0、3.0、4.0、5.0和6.0 mg·L~(-1))水平对平邑甜茶根系生长及氮素吸收、利用和分配特性的影响。结果表明,3.0 mg·L~(-1)硼酸处理的幼苗根系活力及根系形态指标显著高于其他处理,幼苗的全氮量及~(15)N吸收量增幅最大,分别比对照提高了19.4%和75.0%。随供硼水平的增加,植株氮素利用率呈现先增高后降低的趋势,在3.0 mg·L~(-1)硼酸处理时最大,为14.8%,是对照的1.8倍。施硼处理对幼苗的~(15)N分配率有一定的影响,3.0 mg·L~(-1)硼酸处理的根系~(15)N分配率达到最大,且显著高于对照。非损伤微测结果显示,3.0 mg·L~(-1)硼酸处理时,平邑甜茶根系对NO_3~-有强烈吸收且内流速度达到最大,在缺硼和高硼(硼酸0和6.0 mg·L~(-1))处理时有明显外排趋势。因此,3.0 mg·L~(-1)硼酸处理最有利于平邑甜茶根系的生长、根系活力的提高和氮素的吸收利用,而低硼和过量供硼均会抑制根系生长及氮素的吸收利用。     

miR-186-5p与α-MSH互作对绵羊黑素细胞黑色素生成的影响

黑色素合成受多种基因和miRNAs的调控,为了研究miR-186-5p与α-MSH协同作用对绵羊黑素细胞黑色素生成的影响。本研究在绵羊黑素细胞中转染miR-186-5p表达载体,同时添加α-MSH,通过实时荧光PCR（quantitative real time polymerase chain reaction,qRT-PCR）和蛋白质免疫印迹（Western blotting）检测MITF、TYR、TYRP1和TYRP2 4种可能与之相关的基因表达情况;利用免疫组化检测MITF蛋白在黑素细胞中的表达和亚细胞定位;通过分光光度法测定黑色素含量变化;利用划痕试验测定细胞迁移情况,试验分为miR-186-5p转染组、miR-186-5p+α-MSH互作组和对照组,每组3个重复。结果显示,miR-186-5p能够抑制绵羊黑素细胞中MITF、TYR、TYRP1和TYRP2的表达,并最终抑制黑色素的生成。而添加α-MSH能缓解miR-186-5p对MITF、TYR、TYRP1和TYRP2表达的抑制作用,进而在一定程度上恢复黑色素的含量。另外,miR-186-5p还能抑制细胞分裂,并阻碍细胞的迁移,而α-MSH无法抵消miR-186-5p产生的这种负面影响。上述结果表明,在绵羊黑素细胞中,miR-186-5p和α-MSH均参与调控黑色素合成途径,其中,miR-186-5p主要起负调控作用,而α-MSH主要参与相关位点的正调控,并能部分恢复miR-186-5p导致的黑色素含量下降。值得注意的是,miR-186-5p还参与调控细胞的增殖和迁移,而α-MSH不参与相关调控。     

绒山羊皮肤毛囊兴盛前期组织形态、chi-miR-107-3p与RHBDF2基因表达分析

为了探究chi-miR-107-3p和菱形家族蛋白2（rhomboid family member 2,RHBDF2）基因在不同品种（系）和光控增绒绒山羊兴盛前期（5~7月份）皮肤毛囊重建时的表达差异,本试验在7月份采集内蒙古阿拉善型绒山羊、敏盖绒山羊和光控增绒技术处理的阿尔巴斯型绒山羊体侧皮肤毛囊组织,采用组织切片技术对组织形态比较分析,实时荧光定量PCR法检测chi-miR-107-3p和RHBDF2基因的表达量。结果显示,在皮肤毛囊兴盛前期,阿拉善型绒山羊次级毛囊正处于重建阶段,敏盖绒山羊和阿尔巴斯型绒山羊（光控增绒）次级毛囊重建已基本完成;在阿拉善型绒山羊皮肤组织中chi-miR-107-3p表达量极显著高于敏盖绒山羊和阿尔巴斯型绒山羊（光控增绒）（P<0.01）,而RHBDF2基因表达量极显著低于其他两品种（系）（P<0.01）,且RHBDF2基因表达量在敏盖绒山羊皮肤组织中显著低于阿尔巴斯型绒山羊（P<0.05）。不同品种（系）和光控增绒绒山羊皮肤毛囊兴盛前期组织显微结构与chi-miR-107-3p、RHBDF2基因在皮肤组织中的表达差异分析结果相一致,次级毛囊重建初期chi-miR-107-3p表达量较高,而RHBDF2基因表达量极低,随着次级毛囊重建完成,chi-miR-107-3p表达量明显降低,RHBDF2基因表达量逐渐增高,且在不同品种（系）内蒙古绒山羊绒毛生长发育过程中的表达机制基本相同。因此,chi-miR-107-3p和RHBDF2基因是绒山羊皮肤毛囊生长发育的重要调控因子。     

美国军事司法禁止强制性自证其罪规则解读

美国军事司法禁止强制性自证其罪规则,与"米兰达规则"有着本质区别。尽管二者在法源上相同,但建立的规则不同。美国军事司法禁止强制性自证其罪规则确立了禁止刑讯逼供原则、预先告知原则和非法证据排除规则,但虐囚事件充分暴露了美国军事司法禁止强制性自证其罪规则的本质。     

食品安全与食品低温流通中的温度监控

食品的温度监控是一种有效的保证食品安全及保持冷藏链完整的手段,对冷冻冷藏食品在低温流通过程中实行温度监控是非常有必要的。目前,我国冷藏运输中的温度管理情况并不乐观,消费者获得的货物通常不能达到预计的要求。要解决这一问题,必须要借助于时间-温度指示器等来对其整个运输过程进行温度监控。本文对温度立法的必然性和前景,温度监控的重要性、原理以及在冷藏运输中的应用作了较详尽的介绍,并列举了几种国外时间-温度指示器的工作原理。