Fen1 mutations that specifically disrupt its interaction with PCNA cause aneuploidy-associated cancer

DNA replication and repair are critical processes for all living organisms to ensure faithful duplication and transmission of genetic information. Flap endonuclease 1 (Fen1), a structure-specific nuclease, plays an important role in multiple DNA metabolic pathways and maintenance of genome stability. Human FEN1 mutations that impair its exonuclease activity have been linked to cancer development. FEN1 interacts with multiple proteins, including proliferation cell nuclear antigen (PCNA), to form various functional complexes. Interactions with these proteins are considered to be the key molecular mechanisms mediating FEN1's key biological functions. The current challenge is to experimentally demonstrate the biological consequence of a specific interaction without compromising other functions of a desired protein. To address this issue, we established a mutant mouse model harboring a FEN1 point mutation (F343A/F344A, FFAA), which specifically abolishes the FEN1/PCNA interaction. We show that the FFAA mutation causes defects in RNA primer removal and long-patch base excision repair, even in the heterozygous state, resulting in numerous DNA breaks. These breaks activate the G2/M checkpoint protein, Chk1, and induce near-tetraploid aneuploidy, commonly observed in human cancer, consequently elevating the transformation frequency. Consistent with this, inhibition of aneuploidy formation by a Chk1 inhibitor significantly suppressed the cellular transformation. WT/FFAA FEN1 mutant mice develop aneuploidy-associated cancer at a high frequency. Thus, this study establishes an exemplary case for investigating the biological significance of protein-protein interactions by knock-in of a point mutation rather than knock-out of a whole gene.     

断尾对胎生蜥蜴运动能力和选择体温的影响

尾自切是蜥蜴为了降低被捕食危险而采取的一种反捕食适应策略,但断尾可导致体重减轻、热量收支平衡改变,并影响蜥蜴的运动能力和体温调节.为检验断尾对蜥蜴运动能力和选择体温的影响,于2006年5月选取黑龙江省小兴安岭地区的一个胎生蜥蜴种群进行实验.结果表明:在30 ℃和24℃两个实验温度下,断尾后胎生蜥蜴的运动能力均明显下降,表现在停顿次数增多、最大可持续距离和最大疾跑速度减少等方面;断尾、温度和性别对胎生蜥蝎运动能力的影响在一定程度上是相互独立的,断尾是影响胎生蜥蜴运动能力的主要因素;断尾对胎生蜥蝎的选择体温没有显著影响.     

硅藻纳米技术

硅藻是一类微小的单细胞藻类,具有由无定形氧化硅组成的坚硬细胞壁(硅壳).硅壳具有精致的形态和结构,且随硅藻种类和生长条件不同而千变万化.目前估算的硅藻种类超过200 000种,其独特的纳米结构对光子结构、化学生物传感器、新纳米材料和器件的开发具有启发意义.同时硅藻形态形成学和分子生物学的研究,可以推动硅质材料的仿生合成...     

Imaging long-term fate of intramyocardially implanted mesenchymal stem cells in a porcine myocardial infarction model

The long-term fate of stem cells after intramyocardial delivery is unknown. We used noninvasive, repetitive PET/CT imaging with [(18)F]FEAU to monitor the long-term (up to 5 months) spatial-temporal dynamics of MSCs retrovirally transduced with the sr39HSV1-tk gene (sr39HSV1-tk-MSC) and implanted intramyocardially in pigs with induced acute myocardial infarction. Repetitive [(18)F]FEAU PET/CT revealed a biphasic pattern of sr39HSV1-tk-MSC dynamics; cell proliferation peaked at 33-35 days after injection, in periinfarct regions and the major cardiac lymphatic vessels and lymph nodes. The sr39HSV1-tk-MSC-associated [(18)F]FEAU signals gradually decreased thereafter. Cardiac lymphography studies using PG-Gd-NIRF813 contrast for MRI and near-infrared fluorescence imaging showed rapid clearance of the contrast from the site of intramyocardial injection through the subepicardial lymphatic network into the lymphatic vessels and periaortic lymph nodes. Immunohistochemical analysis of cardiac tissue obtained at 35 and 150 days demonstrated several types of sr39HSV1-tk expressing cells, including fibro-myoblasts, lymphovascular cells, and microvascular and arterial endothelium. In summary, this study demonstrated the feasibility and sensitivity of [(18)F]FEAU PET/CT imaging for long-term, in-vivo monitoring (up to 5 months) of the fate of intramyocardially injected sr39HSV1-tk-MSC cells. Intramyocardially transplanted MSCs appear to integrate into the lymphatic endothelium and may help improve myocardial lymphatic system function after MI.     

带鱼下脚料水解物对大鼠血管内皮损伤的影响

通过检测带鱼下脚料蛋白酶水解物的药理作用,为带鱼下脚料的开发利用提供理论依据。用带鱼下脚料三酶混合水解物喂养盐酸肾上腺素造成血管内皮损伤模型的大鼠,检测大鼠血清中血管内皮生长因子(VEGF)和血管性血友病因子(vWF)的含量变化。结果表明:实验不给药组与空白对照组及实验给药组相比,VEGF和vWF的含量存在显著差异。注射盐酸肾上腺素成功制造了血管内皮损伤模型,带鱼下脚料蛋白酶水解物对内皮损伤没有显著影响,但对损伤后的内皮修复有明显的促进作用。     

血浆是引起血沉增快的主要因素

目的:阐明血沉增快的原因是血浆还是红细胞。方法:收集72例血沉异常的抗凝血标本,同时收集血型相对应的72例血沉正常标本,组成血型相同的血沉异常和正常标本72对,每对互换血浆后重新测定血沉,与原始血沉结果比较,并通过多元线性回归分析血沉与血浆蛋白及血脂浓度的关系。结果:血沉异常标本的红细胞加入血沉正常标本的血浆后,72例(100%)血沉均减慢,其中30例血沉下降90%以上,35例下降70%-90%,7例小于70%。血沉正常标本的红细胞加入血沉异常标本的血浆后,67例(93%)血沉加快,其中58例(81%)变为异常(18例血沉加快10倍以上,40例加快5-10倍)。球蛋白、白蛋白和纤维蛋白原与血沉具有线性关系,球蛋白(r=0.420,P<0.001)和纤维蛋白原(r=0.673,P<0.001)与血沉呈正相关,而白蛋白(r=-0.558,P<0.001)与血沉呈负相关。结论:血沉增快主要与血浆因素相关,红细胞对于血沉的影响作用很小。     

膜蛋白ATAD3A在线粒体质量控制中的作用

线粒体质量控制对于线粒体网络的稳态和线粒体功能的正常发挥具有重要意义。三磷酸腺苷酶家族蛋白3A(ATAD3A)是同时参与调节线粒体结构功能、线粒体动力学和线粒体自噬等重要生物学过程的线粒体膜蛋白之一。近期研究表明,ATAD3A既可与Mic60/Mitofilin和线粒体转录因子A (TFAM)等因子相互作用以维持线粒体嵴的形态和氧化磷酸化功能,又能与发动蛋白相关蛋白1 (Drp1)结合而正性/负性调节线粒体分裂,还可作为线粒体外膜转位酶（TOM）复合物和线粒体内膜转位酶（TIM）复合物之间的桥接因子而介导PTEN诱导激酶（PINK1）输入线粒体进行加工,显示出促自噬或抗自噬活性。本文对ATAD3A在调控线粒体质量控制中的作用及其机制进行了综述。     

福尔马林对固定标本DNA提取和扩增的影响

福尔马林被广泛应用于生物标本的长期保存。由于福尔马林可能影响标本DNA的质量,因此需要对福尔马林固定标本DNA的提取和扩增过程进行改进。影响从福尔马林保存标本中提取的DNA质量的主要因素包括福尔马林导致的DNA与蛋白质之间、蛋白质与蛋白质之间、DNA与DNA之间的交联,福尔马林溶液的化学成分、pH值及浓度,标本保存的时间和温度,标本保存部位等。本文总结了目前常用的对标本DNA提取和扩增过程的改进措施及其优点。     

EM复合菌对新疆色素辣椒生长及根际细菌群落的影响

【背景】Effective microorganisms （EM）复合菌在我国农业种植上的应用越来越广泛,但对色素辣椒的促生作用与根际细菌群落结构的影响未见报道。【目的】评估EM复合菌对新疆色素辣椒的促生长作用,并分析其对色素辣椒根际细菌群落组成的影响。【方法】通过随水灌溉方式将EM复合菌接种到色素辣椒根部,在收获期测定辣椒生长指标、土壤养分和酶活活性,明确EM复合菌对辣椒生长和土壤质量的影响;利用16S rRNA基因高通量测序技术测定EM复合菌对辣椒根系细菌群落组成和结构的影响。【结果】与对照组相比,EM复合菌的施用使辣椒株高、鲜重、单个果重和单株结果数分别提高23.89%、85.41%、42.31%和46.04%;土壤中碱解氮和速效磷含量分别提高5.83%和13.39%,土壤中脲酶、蔗糖酶和过氧化物酶的活性分别提高11.47%、9.42%和21.43%;施用EM复合菌显著改变辣椒根际微生物群落的α多样性和β多样性,提高有益菌群变形菌门（Proteobacteria）、酸杆菌门（Acidobacteria）、芽单胞菌门（Gemmatimonadetes）、放线菌门（Actinobacteria）和厚壁菌门（Firmicutes）的相对丰度,其中变形菌门黄单胞菌科（Xanthomonadaceae）的相对丰度增加119.32%;在属的水平上,施用EM复合菌显著增加了藤黄色杆菌属（Luteitalea）、藤黄单胞菌属（Luteimonas）、鞘脂单胞菌属（Sphingobacterium）和盐单胞菌属（Halomonas）的相对丰度,尤其是藤黄单胞菌属的丰度提高244.17%,同时显著降低黄杆菌属（Flavobacterium）的相对丰度。此外,与土壤理化指标呈正相关的微生物菌群相对丰度也显著升高。【结论】EM复合菌能够通过提高土壤营养成分与酶活活性,调控根系微生物群落结构,富集大量在盐碱地生存能力较强的有益菌群,进而起到促进色素辣椒生长的功效。