Novel high-resolution characterization of ancient DNA reveals C > U-type base modification events as the sole cause of post mortem miscoding lesions

Ancient DNA (aDNA) research has long depended on the power of PCR to amplify trace amounts of surviving genetic material from preserved specimens. While PCR permits specific loci to be targeted and amplified, in many ways it can be intrinsically unsuited to damaged and degraded aDNA templates. PCR amplification of aDNA can produce highly-skewed distributions with significant contributions from miscoding lesion damage and non-authentic sequence artefacts. As traditional PCR-based approaches have been unable to fully resolve the molecular nature of aDNA damage over many years, we have developed a novel single primer extension (SPEX)-based approach to generate more accurate sequence information. SPEX targets selected template strands at defined loci and can generate a quantifiable redundancy of coverage; providing new insights into the molecular nature of aDNA damage and fragmentation. SPEX sequence data reveals inherent limitations in both traditional and metagenomic PCR-based approaches to aDNA, which can make current damage analyses and correct genotyping of ancient specimens problematic. In contrast to previous aDNA studies, SPEX provides strong quantitative evidence that C > U-type base modifications are the sole cause of authentic endogenous damage-derived miscoding lesions. This new approach could allow ancient specimens to be genotyped with unprecedented accuracy.     

Resistin induces monocyte-endothelial cell adhesion by increasing ICAM-1 and VCAM-1 expression in endothelial cells via p38MAPK-dependent pathway

Resistin, firstly reported as an adipocyte-specific hormone, is suggested to be an important link between obesity and diabetes. Recent studies have suggested an association between resistin and atherogenic processes. The adhesion of circulating monocytes to endothelial cells is a critical step in the early stages of atherosclerosis. The purpose of the present study was to investigate the effect of resistin on the adhesion of THP-1 monocytes to human umbilical vein endothelial cells (HUVECs) and the underlying mechanism. Our results showed that resistin caused a significant increase in monocyte adhesion. In exploring the underlying mechanisms of resistin action, we found that resistin-induced monocyte adhesion was blocked by inhibition of p38MAPK activation using SB203580 and SB202190. Furthermore, resistin increased the expression of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) by HUVECs and these effects were also p38MAPK-dependent. Resistin-induced monocyte adhesion was also blocked by monoclonal antibodies against ICAM-1 and VCAM-1. Taken together, these results show that resistin increases both the expression of ICAM-1 and VCAM-1 by endothelial cells and monocyte adhesion to HUVECs via p38MAPK-dependent pathways.     

利诺吡啶对豚鼠耳蜗外毛细胞和Deiters细胞钾电流的抑制

为探讨KCNQ家族钾通道在耳蜗外毛细胞和Deiters细胞的功能性表达,我们观察并记录了KCNQ家族钾通道阻滞剂利诺吡啶对豚鼠耳蜗单离外毛细胞(outer hair cells,OHCs)和Deiters细胞总钾电流的影响。采用酶孵育加机械分离法分离豚鼠耳蜗单个OHCs和Deiters细胞：运用膜片钳技术,在全细胞模式下记录正常细胞外液中8个外毛细胞和5个Deiters细胞的总钾电流,并观察100μmol／L和200μmol／L利诺吡啶对外毛细胞和Deiters细胞总钾电流的影响。结果观察到,在正常细胞外液中的单离外毛细胞,可记录到四乙基二乙胺敏感的外向性钾电流和静息膜电位附近激活的内向性钾电流(the K^ current activated at negative potential,IKa)两种钾电流,而在单离Deiters细胞中只记录到外向整流性钾电流。在细胞外液中,加入100μmol／L利诺吡啶后,OHCs中的四乙基二乙胺敏感的钾电流峰电流成分被抑制,稳态电流幅值减小,且电流的失活时问常数明显延长;在细胞外液中加入100μmol／L和200μmol／L利诺吡啶后,OHCs的内向性钾电流IKa被完全抑制;而细胞外液中利诺吡啶终浓度为200μmol／L时,Deiters细胞的外向整流性钾电流幅值无明显变化。由此我们推测,KCNQ家族钾通道存在于豚鼠耳蜗外毛细胞,其介导的钾电流是四乙基二乙胺敏感的钾电流的组成部分,并构成全部的IKn,其功能是介导细胞内K^ 外流和防止细胞过度去极化;KCNQ家族钾通道不存在于豚鼠耳蜗Dciters细胞。     

棉铃虫Helicoverpa armigera的环境友好型诱杀技术研究

2003年和2004年,在山东惠民采用杀虫灯、性诱剂、杨树枝把等措施开展了诱杀棉铃虫Helicoverpa armigera成虫的防治试验.杀虫灯+10支性诱剂,杀虫灯+5支性诱剂+5支杨树枝把,杀虫灯+lO支杨树枝把,杀虫灯4种措施的日均诱蛾量分别为11.4、10.4、8.6和7.0头,性诱剂和杨树枝把均未显著提高对棉铃虫的诱杀效果.不同诱捕方案对棉铃虫累积诱蛾量随日期的变化符合Gompertz方程.普通灯和时控灯对棉铃虫的日均诱蛾量分别为19.9头和17.4头,二者差异不显著;时控灯对异色瓢虫和龟纹瓢虫的日均诱捕量均少于普通灯;减少开灯时间可在不显著降低棉铃虫诱杀量的同时,有效保护捕食性瓢虫.与普通灯相比,设置时间合理的时控灯(夜间19:00～23:00和3:00～5:00开灯)未显著降低棉铃虫的日均诱蛾量,而对异色瓢虫和龟纹瓢虫的日均诱捕量分别为2.0头和4.9头,均显著低于普通灯的3.7头和9.2头,合理设置开关灯控制模式可在不显著降低对棉铃虫诱杀效果的同时,降低近50%的天敌杀伤率,节约用电40%.     

Purification, crystallization and preliminary crystallographic investigations of selenium-containing phycocyanin from selenium-rich algae ( Spirulina platensis )

The selenium-containing phycocyanin from the selenium-rich algae (Spirulina platensis) has been crystallized in two crystal forms by the hanging-drop vapor diffusion techniques. A chromatographic procedure of gel filtration and anion exchange was used for purification. Form I crystal with space group P2₁ and cell parameters a =108.0 ?, b= 117.0 ?, c = 184.0?, β= 90.2° and 12(αβ) units in the asymmetric unit was obtained by using (NH₄)₂SO₄ as precipitant. These crystals diffract up to 2.8 ?. Form II crystal obtained by using PEG4000 as precipitant belongs to space group P6₃ with unit cell constants a = 155.0 ?, c = 40.3 ?, γ =120.0° and one(αβ) unit in the asymmetric unit. The crystals diffract beyond 2.9 ?. The possible stacking forms of phycocyanin molecules in the first crystal form were discussed.     

葡萄糖对杜氏盐藻3-磷酸甘油脱氢酶活性的影响

杜氏盐藻是一种抗渗透能力强的单细胞绿藻,甘油在其渗透调节过程中具有重要作用。葡萄糖对杜氏盐藻细胞数量的增加效果不明显,但对盐藻细胞内甘油积累有显著促进作用,在0～15g/L范围内葡萄糖的浓度与胞内甘油积累显著相关(R2=0.9604,P=0.01);葡萄糖浓度达到15g/L时,胞内甘油积累量达到最高值7.80pg/cell,是对照的1.88倍,胞内甘油积累量与葡萄糖的消耗量极显著相关(R2=0.9982,P=0.01)。葡萄糖对盐藻细胞内总蛋白、3-磷酸甘油脱氢酶(GPDH)酶活和比活都有显著影响,在15g/L葡萄糖时这3个值达到最大值,分别是对照的1.354、4.384、3.229倍。数据显示葡萄糖浓度在15g/L时细胞内蛋白质含量增加不多,但GPDH酶活和比活却大幅度增加;葡萄糖导致的渗透压的变化可能诱导新的同功酶的合成。     

中华绒螯蟹白化症、正常肝胰腺组织结构及脂肪酸组成的比较研究

采用气相色谱、组织学及电镜技术分别对中华绒螯蟹雄蟹白化症肝胰腺与正常肝胰腺脂类和脂肪酸组成、显微及超微结构进行了比较研究,结果表明：正常肝胰腺的总脂含量（31.54%肝胰腺湿重）显著高于白化症肝胰腺（19.07%肝胰腺湿重）,两种蟹肝胰腺的脂类组成没有显著差异,但脂肪酸组成存在一定的差异,白化症肝胰腺中C18∶1的含量显著高于正常肝胰腺,中性脂和极性脂HUFA正常蟹肝胰腺含量都高于白化症蟹,但没有显著差异。组织学结果和超微结构观察表明：白化症肝胰腺肝小管的上皮细胞壁结构紊乱,微绒毛及微绒毛黏膜大部分脱落,细胞内仅见有零星的线粒体、脂肪滴分布,R细胞呈现空泡化。     

小麦幼苗根系形态与反复干旱存活率的关系

以35个不同栽培类型的小麦品种（系）作为试验材料,根据其6叶幼苗的根系形态性状进行聚类分析,供试材料的根系类型分为3种：大根系、小根系和中间型根系。具有中间型根系的材料反复干旱存活率最高,这些材料的根系特点是单株根数7－8．5条,最大根长20-22cm,根总干重44－48mg,其中10cm以下根干重占36％－45％,根冠比范围在0.22-0.24。一些水地栽培的育成品种苗期抗旱性较强,旱地栽培的育成品种苗期抗旱性差异较大,个别旱地栽培的地方品种在土壤水分胁迫条件下反而比在正常水分条件下的根系发育更好,可能是长期适用干旱条件的结果。     

0.21nm分辨率BO-(L-精氨酸)牛胰岛素晶体结构研究

应用差值Fourier技术,立体化学最小二乘技术和XPLOR程序并辅以电子密度图的人工拟合,以三方二锌猪胰岛素模型为起点,解析了0.21nm分辨率BO-(L-精氨酸)牛胰岛素的晶体结构,最终R因子为18.2%,与标准键长与键角的均方根偏差分别为0.0022nm和 4.3°.从电子密度图与模型的拟合来看,独立区两个分子中B链N端加长的L-精氨酸清晰可见.相对于三方二锌猪胰岛素分子,B链N端晶体学微环境发生了变化.