ET-1、IL-6对肝炎后肝硬化患者预后的评价

目的:探讨肝炎后肝硬化患者血浆活性物质ET-1、IL-6的表达及其对患者预后判断的意义。方法:对70例肝炎后肝硬化患者按照Child-Pugh分级标准分为A级(23例)、B级(22例)和C级(25例)三组,并采用30例健康志愿者作为对照。采用放射免疫(RIA)方法检测ET-1和IL-6水平,对获得的数据进行统计学处理。结果:肝炎后肝硬化组ET-1和IL-6水平较健康组明显升高(P<0.01),且A、B、C三组中的ET-1和IL-6水平比较具有统计学意义。结论:ET-1和IL-6水平可反映肝炎后肝硬化肝脏损害程度,可作为预后判断的重要指标。     

VEGF siRNA降低人肝癌细胞株SMMC7721的致瘤性

本研究应用RNA干扰（RNAi）技术高效抑制VEGF的表达,明显降低了人肝癌细胞株SMMC7721的致瘤性。化学合成针对人VEGF基因的siRNA,体外瞬时转染人肝癌细胞株SMMC7721,RT-PCR和Elisa法检测表明VEGFsiRNA实验组与对照组相比,细胞内VEGFmRNA表达下降了76．16％,VEGF分泌蛋白表达则下降了96．28％,而MTT法结果显示VEGFsiRNA对SMMC7721细胞增殖没有明显作用。体内实验中,不同时间对荷SMMC7721细胞瘤裸小鼠进行siRNA直接瘤内注射,同时测量瘤体积,最后取瘤块进行组织切片观察并进行分子生物学分析,结果显示,与对照组相比,VEGFsiRNA实验组肿瘤体积明显缩小,瘤内出现细胞坏死,同时肿瘤组织中VEGFmRNA和蛋白表达水平均明显降低。     

GST pull-down筛选冠突曲霉MAT的互作蛋白*

目的: 冠突曲霉(Aspergillus cristatus)是一种同宗结合菌,它的产孢受渗透压调控,与构巢曲霉的光调控产孢机制存在较大差异。冠突曲霉的有性生殖主要受MAT1-1-1和MAT1-2-1调控,但MAT基因对该菌有性生殖的调控机制仍不清楚。期望筛选得到冠突曲霉MAT的互作蛋白,为深入研究冠突曲霉有性产孢机制奠定基础。方法: 利用GST pull-down联合液相色谱-串联质谱(LC-MS/MS)技术筛选可能与冠突曲霉MAT1-1-1和MAT1-2-1互作的蛋白,结合ProteinPilot和冠突曲霉基因组注释结果进行互作蛋白的注释及GO分析,其中互作蛋白SI65_00917和SI65_03348利用RT-qPCR探索它们与有性发育的联系,并利用酵母双杂交技术初步验证它们与MAT蛋白的互作关系。结果: 成功构建了GST-MAT1-1-1、GST-MAT1-2-1表达载体,诱导表达纯化出目的诱饵蛋白,分别利用诱饵蛋白捕获冠突曲霉总蛋白中的互作蛋白,经分析、筛选共鉴定出与MAT1-1-1互作的蛋白56个,与MAT1-2-1互作的蛋白413个。GO分析表明,这些蛋白参与翻译调控、代谢过程、蛋白质转运及蛋白结合等生物学过程,具有核苷酸结合活性、催化活性、蛋白结合活性;RT-qPCR结果表明互作蛋白SI65_00917可能与有性发育相关。酵母双杂交结果表明,SI65_00917蛋白具有自激活作用,可能是转录因子;SI65_03348蛋白与MAT1-1-1、MAT1-2-1在酵母中均有互作。结论: MAT通过与其他蛋白直接或间接的相互作用调控其有性发育过程。     

樟子松固沙林林水关系研究进展及对营林实践的指导

中国有着世界上最大面积的人工林, 如何维持人工林的可持续性已成为气候变化背景下需要面对的重大挑战。樟子松(Pinus sylvestris var. mongolica)以其抗旱、抗寒、耐贫瘠等优良特性成为中国北方生态治理中最主要的常绿针叶树种之一, 近70年来发挥了巨大的防风固沙与生态固碳功能。然而, 随着林分的生长与气候变化, 樟子松人工固沙林正经历着越来越严峻的环境胁迫, 部分地区出现了林分“早衰”或死亡的现象, 引起了人们对樟子松固沙林适应与应对气候变化能力的担忧。该文在回溯樟子松基本生物学特征与引种推广历史, 系统总结近年来樟子松林林水关系研究新成果的基础上, 全面分析了樟子松固沙林林水关系存在的主要矛盾, 并提出了基于林水关系相协调的林分经营措施的调整: 由倡导防护功能为主的单一目标向包含林分稳定性、生态固碳功能、可持续发展等多目标平衡方向调整; 由以沙地森林景观培育为主向以良好土壤生境培育为主的方向调整; 由倡导天然更新为主向以人工造林与天然更新相结合的世代更新方向调整。在立足于北方沙地脆弱生境与气候变化客观现实的基础背景下, 应坚持樟子松在固沙林生态系统演化过程中的先锋种与建群种地位, 基于“以水定绿”原则, 采取“隔行带伐+再造林”等方式开展林分密度动态调控, 促进林分向异龄林结构演化, 促进樟子松固沙林生态服务的优质化和生态固碳功能的最大化。     

大肠杆菌代谢工程改造合成L-高丝氨酸及其衍生物研究进展

l-高丝氨酸及其衍生物(O-琥珀酰-l-高丝氨酸和O-乙酰-l-高丝氨酸)是生物合成l-甲硫氨酸的前体,同时也是合成多种C4化合物(异丁醇、g-丁内酯、1,4-丁二醇、2,4-二羟基丁酸等)和l-草铵膦等的平台化合物。因此,发酵法生产l-高丝氨酸及其衍生物成为近年内研究的热点。然而,利用生物法合成l-高丝氨酸及其衍生物仍存在一些不足之处,如发酵产量不高或糖酸转化率过低等。此外,对l-高丝氨酸及其衍生物合成的总体代谢和调控机制鲜有报道。本文综述了大肠杆菌代谢工程改造合成l-高丝氨酸及其衍生物O-琥珀酰-l-高丝氨酸和O-乙酰-l-高丝氨酸的研究进展,从底物摄取、关键节点碳流分配改造、辅酶NADPH的循环供应以及目标产物的外运输出等方面,系统分析了大肠杆菌全发酵法生产l-高丝氨酸及其衍生物的代谢途径及改造策略,为其后续代谢改造及生物法生产提供一定的研究思路。     

中国真猛犸象和披毛犀化石14C年代研究新进展

真猛犸象（Mammuthus primigenius）和披毛犀（Coelodonta antiquitatis）是北半球高纬度地区晚更新世动物群的主要成员,其消亡的年代和原因一直是国际学术界关注的热点科学问题。本文对黑龙江青冈县英贤村最新出土的5个真猛犸象和5个披毛犀化石进行了AMS¹⁴C年代测定,结果均大于4万年,部分化石可能已经超出了目前¹⁴C的测定范围。通过整理并对比已公开发表的中国境内两种动物化石的¹⁴C年代学数据,本文认为早期常规¹⁴C测年方法所获得的年代值需要重新考虑其准确性。埋藏地层与最新的AMS¹⁴C测年数据显示,我国真猛犸象化石年代主要集中于MIS3阶段;披毛犀在我国消亡的时间很可能晚于真猛犸象,至少延续到末次冰消期。中国猛犸象-披毛犀动物群化石仍然需要开展更多的年代学研究。     

Split-enzyme fragment as a single affinity tag that enables protein expression,purification, and functional assays

Expressing, isolating, and characterizing recombinant proteins is crucial to many disciplines within the biological sciences. Different molecular tagging technologies have been developed to enable each individual step of protein production, from expression through purification and characterization. Monitoring the entire production process requires multiple tags or molecular interactions, because no individual tag has provided the comprehensive breadth of utility. An ideal molecular tag is small and does not interrupt expression, solubility, folding or function of the protein being purified and can be used throughout the production process. We adapted and integrated a split-luciferase system (NanoBiT®, Promega ®) to perform the range of techniques essential to protein production. We developed a simple method to monitor protein expression in real time to optimize expression conditions. We constructed a novel affinity chromatography system using the split-luciferase system to enable purification. We adapted western blot analysis, enzyme-linked immunosorbent assay, and cell-based bioassay to characterize the expressed proteins. Our results demonstrate that a single-tag can fulfill all aspects needed throughout protein production.     

Karyometric image analysis for intraepithelial and invasive cervical lesions

OBJECTIVE: To derive an objective, numeric measure for the progression of intraepithelial and invasive squamous cell cervical lesions. STUDY DESIGN: Thin-layer cervical cytology preparations from colposcopically confirmed normal cervix, low grade squamous intraepithelial lesions, high grade squamous intraepithelial lesions and carcinoma were identified from a cross-sectional study. Fifty-nine cases representing 4 diagnostic categories were selected, and 2,375 nuclei from epithelial cells representative of the diagnostic category were randomly selected for imaging and measurement from these cases. Additionally, 1,378 visually normal appearing intermediate cells from low and high grade squamous intraepithelial lesions, as well as from carcinoma cases, were identified for analysis. The nuclei were quantitatively characterized, and discriminant analyses were performed to derive a progression curve from normal cytology to carcinoma. RESULTS: The lesion signatures show a clear increase in nuclear abnormality with increasing progression. A progression curve was derived based on mean discriminant function scores for each diagnostic category and on the mean nuclear abnormality values for the nuclei in each category, as expressed by their deviation in feature values from normal reference nuclei. CONCLUSION: A numeric assessment of lesion progression for cervical precancerous and cancerous lesions based on karyometric measurements is possible and may provide an objective, precise characterization of each lesion as well as a basis for improved performance in automated cytology-based cervical cancer screening.