羽扇豆醇通过抑制RhoA-ROCK1信号通路抑制结肠癌细胞增殖

该文探讨了羽扇豆醇(Lupeol)对人结肠癌HCT116和SW620细胞增殖的影响及相关作用机制。使用不同浓度的Lupeol处理HCT116和SW620细胞后,用MTT法检测细胞活性,CCK8法检测细胞增殖能力,平板克隆实验检测细胞克隆形成能力,流式细胞术检测细胞周期和细胞凋亡,(quantitative real-time PCR,qPCR)和Western blot检测相应mRNA和蛋白表达水平,免疫荧光检测β-Catenin蛋白细胞内分布情况。通过构建shRNA敲低两种结肠癌细胞中RhoA,进一步研究Lupeol影响细胞增殖的分子机制。结果显示,Lupeol处理后,HCT116和SW620细胞增殖能力明显下降,克隆形成能力受到抑制,细胞周期阻滞于G0/G1期,细胞内RhoA、ROCK1、β-Catenin、Cyclin D1 mRNA和蛋白表达水平均显著下降,β-Catenin蛋白胞质和胞膜上分布减少。敲低RhoA后抑制了细胞增殖,同时使得RhoA-ROCK1-β-Catenin信号通路蛋白受到抑制,β-Catenin蛋白胞质和胞膜上分布减少。综上所述,Lupeol可通过抑制RhoA-ROCK1信号通路,抑制β-Catenin蛋白表达,进而抑制HCT116和SW620细胞增殖,Lupeol有望成为临床结肠癌治疗的新药物。     

SIRT3激动剂对乙肝病毒转录和复制的影响

该文探讨了SIRT3激动剂(Honokiol,HKL)对乙肝病毒(Hepatitis B virus,HBV)转录和复制的影响。培养HepG2-NTCP和人原代肝细胞(primary human hepatocytes,PHH),感染HBV颗粒后,用Honokiol(5μmol/L、10μmol/L、20μmol/L)处理细胞后继续培养10天,通过荧光定量PCR检测细胞内HBV DNA、cccDNA和HBV RNAs水平,Southern blot实验进一步检测胞内HBV DNA水平。构建SIRT3-KO细胞,检测敲除SIRT3后,Honokiol对细胞内HBV DNA、cccDNA和HBV RNAs的影响。通过小鼠尾静脉高压注射pCMV-KRAB-Cre质粒和precccDNA质粒构建持续感染小鼠模型,一周后腹腔注射Honokiol持续20天。荧光定量PCR检测小鼠血清中HBV DNA拷贝数,肝组织内HBV DNA、cccDNA和HBV RNAs水平。结果表明,Honokiol浓度依赖性地抑制HepG2-NTCP和PHH细胞内HBV DNA以及HBV RNAs水平,此外,Honokiol可以降低cccDNA的转录活性;敲除SIRT3后,Honokiol不能发挥抗病毒作用;小鼠模型中,Honokiol能够降低血清中HBV DNA和肝组织内HBV DNA拷贝数,以及能够显著抑制肝组织内HBV RNAs水平和cccDNA的转录活性。该研究结果表明,Honokiol能够抑制乙肝病毒转录和复制。     

Three proline rotamases involved in calcium homeostasis play differential roles in stress tolerance,virulence and calcineurin regulation of Beauveria bassiana

FK506‐sensitive proline rotamases (FPRs), also known as FK506‐binding proteins (FKBPs), can mediate immunosuppressive drug resistance in budding yeast but their physiological roles in filamentous fungi remain opaque. Here, we report that three FPRs (cytosolic/nuclear 12.15‐kD Fpr1, membrane‐associated 14.78‐kD Fpr2 and nuclear 50.43‐kD Fpr3) are all equally essential for cellular Ca²⁺ homeostasis and contribute significantly to calcineurin activity at different levels in the insect‐pathogenic fungus Beauveria bassiana although the deletion of fpr1 alone conferred resistance to FK506. Radial growth, conidiation, conidial viability and virulence were less compromised in the absence of fpr1 or fpr2 than in the absence of fpr3, which abolished almost all growth on scant media and reduced growth moderately on rich media. The Δfpr3 mutant was more sensitive to Na⁺, K⁺, Mn²⁺, Ca²⁺, Cu²⁺, metal chelate, heat shock and UVB irradiation than was Δfpr2 while both mutants were equally sensitive to Zn²⁺, Mg²⁺, Fe²⁺, H₂O₂ and cell wall‐perturbing agents. In contrast, the Δfpr1 mutant was less sensitive to fewer stress cues. Most of 32 examined genes involved in DNA damage repair, Na⁺/K⁺ detoxification or osmotolerance and Ca²⁺ homeostasis were downregulated sharply in Δfpr2 and Δfpr3 but rarely so affected in Δfpr1, coinciding well with their phenotypic changes. These findings uncover important, but differential, roles of three FPRs in the fungal adaptation to insect host and environment and provide novel insight into their essential roles in calcium signalling pathway.     

Mutations in MIR396e and MIR396f increase grain size and modulate shoot architecture in rice

Grain size and plant architecture are critical factors determining crop productivity. Here, we performed gene editing of the MIR396 gene family in rice and found that MIR396e and MIR396f are two important regulators of grain size and plant architecture. mir396ef mutations can increase grain yield by increasing grain size. In addition, mir396ef mutations resulted in an altered plant architecture, with lengthened leaves but shortened internodes, especially the uppermost internode. Our research suggests that mir396ef mutations promote leaf elongation by increasing the level of a gibberellin (GA) precursor, mevalonic acid, which subsequently promotes GA biosynthesis. However, internode elongation in mir396ef mutants appears to be suppressed via reduced CYP96B4 expression but not via the GA pathway. This research provides candidate gene‐editing targets to breed elite rice varieties.     

Leaf gall polymorphism and molecular phylogeny of a new Bruggmanniella species (Diptera: Cecidomyiidae: Asphondyliini) associated with Litsea acuminata (Lauraceae) in Taiwan,with ecological comparisons and a species description

Two types of cecidomyiid leaf galls, cup‐shaped and umbrella‐shaped, occur on Litsea acuminata (Lauraceae) in Taiwan. Based on the concept of gall shapes as “extended phenotypes” of gall inducers, these two types could be induced by different gall midge species. However, galls with intermediate shapes between the two types were recently discovered, which implies that possible genetic exchanges occur between the gall inducers of both types. To clarify the taxonomic status of gall midges responsible for the two types of galls on L. acuminata, we undertook taxonomic, molecular phylogenetic and ecological studies. Our findings show that the two gall types are induced by the same Bruggmanniella species and the species is new to science. We describe the species forming this range of galls as Bruggmanniella litseae sp. n. , and compare their geographical distribution, galling position and morphometry. Based on our results, a possible evolutionary scenario of B. litseae sp. n. is discussed.     

Preparation of co‐Co3O4/carbon nanotube/carbon foam for glucose sensor

Co‐Co₃O₄/carbon nanotube/carbon foam (Co‐Co₃O₄/CNT/CF) nanocomposites were prepared by soaking melamine foam into a solution of Co(NO₃)₂·6H₂O, followed by calcination in N₂ and air in sequence. The obtained Co‐Co₃O₄/CNT/CF nanocomposites were characterized with scanning electron microscopy and cyclic voltammetry. It was found that Co₃O₄ nanoparticles were grown on the external of CF successfully, while CNTs were grown on the surfaces of CF in a large amount, which further improved the electrical conductivity of the. The prepared Co‐Co₃O₄/CNT/CF nanocomposites were then used to construct nonenzymatic sensor to detect glucose in alkaline solution. The sensor showed detection range from 1.2 μM to 2.29 mM with a detection limit of 0.4 μM (S/N =3) and a high sensitivity of 637.5 μA^?1 cm^?2. The developed sensor also showed an instant response, favorable reproducibility, and high selectivity. The results attest that Co‐Co₃O₄/CNT/CF composites have great potential in the development of nonenzymatic sensors for glucose.     

白及蔗糖合成酶基因的克隆及表达分析

为揭示白及蔗糖合成酶基因与生长发育的关系,该研究以白及为材料,利用RT-PCR技术同源克隆白及蔗糖合成酶的关键基因SuSy,对SuSy基因的生物学特性及表达特征进行了分析,并利用实时荧光定量PCR检测SuSy基因在不同组织中的表达规律。结果表明:(1)白及SuSy基因长度为2 215 bp,编码737个氨基酸,与铁皮石斛、文心兰和蝴蝶兰的蛋白质氨基酸序列的相似性分别为97%、92%和95%。(2)生物信息学分析表明,SuSy蛋白质序列具有较高的亲水性,与拟南芥SuSy蛋白质氨基酸三级结构一致性为75.2%;系统进化树分析发现,白及SuSy蛋白与铁皮石斛处于同一个分支上。(3) qRT-PCR结果表明,SuSy基因在叶片中的表达量最高,块茎中的表达量最低;成熟叶片的表达量高于未成熟叶片的表达量;数据差异性分析显示,SuSy基因在根、块茎中表达量具有极显著性差异,但在一年生叶和二年生叶中的表达量无显著性差异,幼苗叶和一、二年生叶中表达量具有极显著性差异。由此推测,SuSy基因可能受生长发育的诱导,是调控白及生长发育关键基因。     

水分驱动下茵陈蒿(Artemisia capillaris Thunb.)地上生物量模型与异速生长特征

构建生物量预估模型,探究生物量在各器官中的分配策略和异速生长关系及其对环境因子的响应,对理解植物群落结构、功能、碳储存和分配机制具有重要意义。本研究以内蒙古荒漠草原常见种茵陈蒿(Artemisia capillaris Thunb.)为对象,在不同水分处理下,利用易测指标,如株高、基径、分枝数、冠幅和生物量等参数建立生物量模型,采用标准化主轴分析法分析其异速生长关系。结果表明:在不同水分处理下,茵陈蒿的最佳生物量预估模型的变量选择不同;不同水分处理下茵陈蒿各器官间、各器官与地上生物量间的异速生长关系不同,但相对于自然降水量,增水和减水50%下均为等速生长,这说明在不同水分条件下茵陈蒿对各器官间的资源配置存在权衡策略,符合最优分配假说;而在极端气候条件下,各器官对资源的竞争会变弱;在荒漠草原中,对草本植物进行生物量模拟,选择预测变量和方程模型时,应考虑生长季降水量。本研究可为荒漠草原草本植物生物量预估模型的建立和异速生长关系对环境因子适应的理解等提供方法支持及理论依据。