Feedback regulation of coronary artery disease susceptibility gene ADTRP and LDL receptors LDLR/CD36/LOX-1 in endothelia cell functions involved in atherosclerosis

A high level of low-density lipoprotein cholesterol (LDL) is one of the most important risk factors for coronary artery disease (CAD), the leading cause of death worldwide. However, a low concentration of LDL may be protective. Genome-wide association studies revealed that variation in ADTRP gene increased the risk of CAD. In this study, we found that a low concentration of oxidized-LDL induced the expression of ADTRP. Further analyses showed that knockdown of the expression of LDL receptor genes LDLR, CD36, or LOX-1 significantly downregulated ADTRP expression, whereas overexpression of LDLR/CD36/LOX-1 markedly increased ADTRP expression through the NF-κB pathway. Like ADTRP, LDLR, CD36 and LOX-1 were all involved in endothelial cell (EC) functions relevant to the initiation of atherosclerosis. Downregulation of LDLR/CD36/LOX-1 promoted monocyte adhesion to ECs and transendothelial migration of monocytes by increasing expression of ICAM-1, VCAM-1, E-selectin and P-selectin, decreased EC proliferation and migration, and increased EC apoptosis, thereby promoting the initiation of atherosclerosis. Opposite effects were observed with the overexpression of ADTRP and LDLR/CD36/LOX-1 in ECs. Interestingly, through the NF-κB and AKT pathways, overexpression of ADTRP significantly upregulated the expression of LDLR, CD36, and LOX-1, and knockdown of ADTRP expression significantly downregulated the expression of LDLR, CD36, and LOX-1. These data suggest that ADTRP and LDL receptors LDLR/CD36/LOX-1 positively regulate each other, and form a positive regulatory loop that regulates endothelial cell functions, thereby providing a potential protective mechanism against atherosclerosis. Our findings provide a new molecular mechanism by which deregulation of ADTRP and LDLR/CD36/LOX-1 promote the development of atherosclerosis and CAD.     

Divergent DNA methylation contributes to duplicated gene evolution and chilling response in tea plants

The tea plant (Camellia sinensis) is a thermophilic cash crop and contains a highly duplicated and repeat-rich genome. It is still unclear how DNA methylation regulates the evolution of duplicated genes and chilling stress in tea plants. We therefore generated a single-base-resolution DNA methylation map of tea plants under chilling stress. We found that, compared with other plants, the tea plant genome is highly methylated in all three sequence contexts, including CG, CHG and CHH (where H = A, T, or C), which is further proven to be correlated with its repeat content and genome size. We show that DNA methylation in the gene body negatively regulates the gene expression of tea plants, whereas non-CG methylation in the flanking region enables a positive regulation of gene expression. We demonstrate that transposable element-mediated methylation dynamics significantly drives the expression divergence of duplicated genes in tea plants. The DNA methylation and expression divergence of duplicated genes in the tea plant increases with evolutionary age and selective pressure. Moreover, we detect thousands of differentially methylated genes, some of which are functionally associated with chilling stress. We also experimentally reveal that DNA methyltransferase genes of tea plants are significantly downregulated, whereas demethylase genes are upregulated at the initial stage of chilling stress, which is in line with the significant loss of DNA methylation of three well-known cold-responsive genes at their promoter and gene body regions. Overall, our findings underscore the importance of DNA methylation regulation and offer new insights into duplicated gene evolution and chilling tolerance in tea plants.     

Revisiting the phosphotyrosine binding pocket of Fyn SH2 domain led to the identification of novel SH2 superbinders.

Protein engineering through directed evolution is an effective way to obtain proteins with novel functions with the potential applications as tools for diagnosis or therapeutics. Many natural proteins have undergone directed evolution in vitro in the test tubes in the laboratories worldwide, resulting in the numerous protein variants with novel or enhanced functions. we constructed here an SH2 variant library by randomizing 8 variable residues in its phosphotyrosine (pTyr) binding pocket. Selection of this library by a pTyr peptide led to the identification of SH2 variants with enhanced affinities measured by EC50. Fluorescent polarization was then applied to quantify the binding affinities of the newly identified SH2 variants. As a result, three SH2 variants, named V3, V13 and V24, have comparable binding affinities with the previously identified SH2 triple‐mutant superbinder. Biolayer Interferometry assay was employed to disclose the kinetics of the binding of these SH2 superbinders to the phosphotyrosine peptide. The results indicated that all the SH2 superbinders have two‐orders increase of the dissociation rate when binding the pTyr peptide while there was no significant change in their associate rates. Intriguingly, though binding the pTyr peptide with comparable affinity with other SH2 superbinders, the V3 does not bind to the sTyr peptide. However, variant V13 and V24 have cross‐reactivity with both pTyr and sTyr peptides. The newly identified superbinders could be utilized as tools for the identification of pTyr‐containing proteins from tissues under different physiological or pathophysiological conditions and may have the potential in the therapeutics.     

Cytological and morphology characteristics of natural microsporogenesis within Camellia oleifera

Camellia oleifera is believed to exhibit a complex intraspecific polyploidy phenomenon. Abnormal microsporogenesis can promote the formation of unreduced gametes in plants and lead to sexual polyploidy, so it is hypothesized that improper meiosis probably results in the formation of natural polyploidy in Camellia oleifera. In this study, based on the cytological observation of meiosis in pollen mother cells (PMCs), we found natural 2n pollen for the first time in Camellia oleifera, which may lead to the formation of natural polyploids by sexual polyploidization. Additionally, abnormal cytological behaviour during meiosis, including univalent chromosomes, extraequatorial chromosomes, early segregation, laggard chromosomes, chromosome stickiness, asynchronous meiosis and deviant cytokinesis (monad, dyads, triads), was observed, which could be the cause of 2n pollen formation. Moreover, we confirmed a relationship among the length–width ratio of flower buds, stylet length and microsporogenesis. This result suggested that we can immediately determine the microsporogenesis stages by phenotypic characteristics, which may be applicable to breeding advanced germplasm in Camellia oleifera.Supplementary InformationThe online version contains supplementary material available at 10.1007/s12298-021-01002-5.     

黄河三角洲石油化工区农田土壤-玉米体系PAHs的分布特征及风险评价

为明确黄河三角洲石油开采区表层土壤和玉米中多环芳烃（PAHs）的含量及其污染水平,采集农田土壤和玉米各71个样品,检测农田土壤和玉米各部位中16种PAHs含量,并采用内梅罗指数法和健康风险评价模型评估了农田土壤中多环芳烃的生态健康风险。结果表明,农田土壤、玉米根、茎和叶中多环芳烃的含量分别为256.6-1936、291.4-680.9、324.9-527.9、289.5-2400 μg/kg。农田土壤中多环芳烃以4-6环为主。多环芳烃在玉米根茎叶富集系数大小排序为：叶 > 茎 > 根。玉米不同组织中PAHs浓度与相应农田土壤中PAHs浓度的进行相关分析结果表明,农田土壤中PAHs含量与玉米根、茎中PAHs含量均存在极显著正相关关系,相关系数分别为0.98（P<0.01）、0.98（P<0.01）,表明玉米根和茎的多环芳烃主要来源于农田土壤中,农田土壤中PAHs的含量影响着PAHs在玉米根茎中的积累和分布。玉米叶中PAHs含量与农田土壤中PAHs含量与玉米根、茎中PAHs含量不存在相关关系,表明玉米叶中多环芳烃并非来自土壤中PAHs的迁移,可能来源于大气。内梅罗指数结果表明,农田土壤PAHs达到了中度污染,其中BaA、Pyr和BbF达到了偏重污染;健康风险评价结果表明,农田土壤PAHs对儿童和成人的平均非致癌风险分别为0.44和0.12（均小于1）,表明农田土壤多环芳烃对成人和儿童的非致癌风险是可接受;农田土壤PAHs对儿童和成人的平均致癌风险分别为3.6×10^-5、9.0×10^-6,没有超过致癌风险水平上限（10^-4）,致癌风险尚在可接受范围内。3种暴露途径中,皮肤接触是土壤PAHs的最主要暴露方式,其次是经口摄食,吸入暴露途径甚微,可忽略不计。PAHs对儿童健康的威胁风险要大于成人,所以应尽可能避免儿童直接接触或误食土壤等其他介质的污染物。     

有机污染物芘胁迫下白菜生理特性变化规律

为探究多环芳烃芘对白菜生理特性的影响,通过盆栽实验对华北地区常见的11种白菜进行不同浓度芘胁迫下的培养,研究白菜生理指标的变化规律。结果表明：白菜的种类和芘的浓度对白菜生理指标（生物量、丙二醛、叶绿素）均有显著影响,且随芘浓度变化,不同种类白菜的生理指标呈现不同的变化规律。白菜的生物量随芘的添加呈现两种变化：一是随芘添加浓度增加白菜生物量逐渐下降;二是在中低浓度芘（5、15、45 mg/kg）处理白菜生物量升高,高浓度（135、405 mg/kg）胁迫下生物量下降。白菜的丙二醛（MDA）含量随芘浓度的增加,表现出3种不同规律：各浓度之间均无显著性差异,MDA含量逐渐升高,以及显著低于对照组。叶绿素含量随芘浓度的增加,主要呈现出先下降后上升和逐渐上升2种变化趋势。京春白（JCB）和中白50（ZB-50）对芘的胁迫具有良好的耐受性,生理指标变化较小;中浓度芘胁迫下京翠60（JC-60）和京春绿（JCL）较为敏感,高浓度下京秋65（JQ-65）和吉红308（JH-308）生理指标变化显著,可以作为中低浓度和高浓度芘污染土壤指示作物。     

酿酒酵母代谢工程技术

自20世纪90年代初期诞生以来,代谢工程历经了30年的快速发展。作为代谢工程的首选底盘细胞之一,酿酒酵母细胞工厂已被广泛应用于大量大宗化学品和新型高附加值生物活性物质的生物制造,在能源、医药和环境等领域取得了巨大的突破。近年来,合成生物学、生物信息学以及机器学习等相关技术也极大地促进了代谢工程的技术发展和应用。文中回顾了近30年来酿酒酵母代谢工程重要的技术发展,首先总结了经典代谢工程的常用方法和策略,以及在此基础上发展而来的系统代谢工程和合成生物学驱动的代谢工程技术。最后结合最新技术发展趋势,展望了未来酿酒酵母代谢工程发展的新方向。     

普通烟草NtODB基因的电子克隆、表达及生物信息学分析

ODB基因在植物同源重组依赖性的DNA双键断裂修复过程中起重要作用,对植物诱变育种具有潜在的应用价值。克隆烟草NtODB基因并分析其表达特征为丰富ODB基因在同源重组DNA修复过程中的作用提供证据。为得到烟草NtODB基因序列,采用电子克隆技术获得该基因cDNA序列并克隆验证。进一步使用生物信息学方法分析该基因表达特征,对预测蛋白的理化性质、信号肽、高级结构等进行预测。生物信息学分析结果表明,NtODB基因开放阅读框包含579个碱基,蛋白含192个氨基酸残基,NtODB蛋白具有碱性和亲水性,主要定位于细胞质内;实时荧光定量PCR检测结果显示NtODB基因在不同组织中呈现组成型表达特征;亚细胞定位检测提示NtODB主要表达于细胞膜和叶绿体。NtODB基因的克隆与表达分析及其蛋白高级结构和理化性质的预测,可为进一步丰富ODB基因在同源重组依赖的DNA修复系统中的作用机制提供证据。     

改性聚二甲基硅氧烷 (iPDMS) 蛋白芯片在肿瘤相关抗原自身抗体筛查中的应用

肿瘤标记的快速筛查是临床早期诊断的难题。利用化学发光蛋白质芯片技术,对低丰度的肿瘤相关抗原的自身抗体进行高灵敏度的筛选,是一种有益的尝试。本研究首先将带烯烃末端的、引发聚合反应的表面引发剂加入到常规聚二甲基硅氧烷材料中,再通过热交联反应固定到聚二甲基硅氧烷的三维结构中,形成改性聚二甲基硅氧烷 (iPDMS)。为了使iPDMS材料具有抗蛋白质非特异性吸附的特性,在活性引发位点处通过表面引发的原子转移自由基聚合反应合成poly(OEGMA) 高分子刷。最后将20种肿瘤相关的抗原利用高通量喷点打印技术打印到芯片的特定区域,并组装成iPDMS芯片的48孔检测微孔板。对临床上常见的8种肿瘤患者血清进行分析,发现VEGFR1和VEGF121自身抗体对常见的8种肿瘤具有检测价值,有望成为肿瘤快速筛查的检测指标。     

膳食脂肪、肠道微生物与宿主健康的研究进展

作为三大主要营养物质之一,膳食脂肪为人体提供能量和营养。膳食脂肪摄入不当会破坏肠道微生物的稳态,影响宿主的代谢状况,增加慢性疾病发生的风险。建立疾病动物模型是研究肠道微生物与宿主健康的重要手段。文中综述了膳食脂质的数量和种类、肠道微生物和宿主代谢之间的相互作用及其可能的作用机制,阐述了基于不同的疾病动物模型,膳食脂质影响肠道微生物的结构和功能,以及对宿主代谢的调节,为深入了解膳食脂质、肠道微生态和宿主健康三者之间的关系提供了依据。