用RACE技术扩增并克隆牛BMP4基因3′端序列

RACE技术是一项扩增基因末端序列的新技术。该研究从牛BMP4基因出发,以牛软骨的RNA为模板,按照不同物种BMP4基因的相似性设计特异引物,运用PCR和RACE技术扩增并获得了特异片段,该片段经PCR、酶切和测序验证,证实所克隆序列为牛BMP4的3′端序列,包含有1170bp组成的开放读码框(ORF),编码389个氨基酸,3′非编码区121bp个核苷酸和poly(A)15。同源性分析结果表明,牛BMP4 cDNA最大开放读码框所推测的氨基酸序列与已知人、小鼠、大鼠、狗、羊和鸡等真核生物BMP4氨基酸序列进行比较,分别有94.5%、93.1%、91.9%、87.4%、94.2%、79%的同源性。这为克隆其他物种的BMP4基因提供了依据,同时牛骨形态发生蛋白的测序为我们更好的理解牛的生骨机理提供帮助。     

RPN2与肝癌患者预后及对肝癌细胞生长的作用

目的:探讨核糖蛋白2(ribophorin II,RPN2)在肝细胞肝癌(HCC)组织中的表达和对HCC患者生存的影响,同时分析RPN2对肝癌HepG2细胞生长和克隆形成的作用。方法:应用免疫组化方法和HCC公共芯片数据,从蛋白和m RNA水平检测HCC组织中RPN2的表达,同时分析RPN2与HCC患者临床参数的关系及预后相关性;进一步利用MTS法和克隆形成实验在肝癌HepG2细胞中检测RPN2对细胞生长的作用。结果:98例肝癌组织中,RPN2阳性表达率88.78%,对应癌旁肝组织中,RPN2阳性表达率74.49%;癌组织中RPN2染色评分为5.80±3.15,癌旁肝组织RPN2染色评分为2.13±1.59,肝癌组织中RPN2表达显著上调(P0.001)。3个肝癌公共芯片数据(共522例肝癌)中RPN2的m RNA表达水平同样显著升高(均P0.001)。98例肝癌患者RPN2表达水平与肿瘤直径(P=0.004)、门脉侵袭(P=0.012)和TNM分期(P=0.009)相关;RPN2高表达的患者总体生存期(OS)和无复发生存期(RFS)较RPN2低表达的患者短(OS:P=0.027;RFS:P=0.036)。肝癌HepG2细胞转染RPN2小干扰RNA后,细胞生长能力显著受抑制。结论:RPN2在肝癌中表达显著升高,RPN2的表达与肝癌的恶性进展有关,RPN2显著促进肝癌细胞生长。     

天童常绿阔叶树种栲树生殖个体大小及其生殖构件特征

对浙江天童木荷-栲树林内的常绿阔叶树种栲树(Castanopsis fargesii Franch.)的生殖个体大小、生殖构件的分布及其动态变化特征进行了研究。结果表明, 该地区栲树生殖个体的胸径在17~50 cm 间, 平均胸径为31. 2±8.0 cm, 平均年龄约36.3±6.6 年;林缘附近的生殖个体小于木荷-林内。相对稳定的群落和比较丰富的土壤养分条件有利于生殖枝数量和花序数量的增多。栲树生殖个体的数量在两年中变化较大, 部分栲树个体可以在连续年份中生殖。从枝系水平分析:在持续生殖的栲树个体上, 生殖枝数量有明显变化, 并非所有的生殖枝在两年中都可开花或结果, 保持连续生殖的枝系约占48.2%。栲树果序枝数量在连续年份有明显差异(p < 0.01), 而且果序枝上的幼蕾数、果实数量及结实率等都有明显差异(p < 0.05)。     

Inductive Capacities for the Dorsal Mesoderm of the Dorsal Marginal Zone and Pharyngeal Endoderm in the Very Early Gastrula of the Newt,and Presumptive Pharyngeal Endoderm as an Initiator of the Organization Center*

The organization center of Cynops pyrrhogaster was divided into Parts 1, 2 and 3 of equal size (0.3×0.4 mm²) with presumptive fates as pharyngeal, pharyngeal+prechordal+trunk notochord, and trunk-tail notochord, respectively. Movements and changes in size and shape of each part were followed through gastrulation. Differentiation tendencies of each part were examined under three conditions: I, isolated; II, sandwiched with presumptive ectoderm; 111, sandwiched with presumptive ectoderm after preculture in isolation for various times. In I, Parts 2 and 3 differentiated into dorsal mesoderm. In II, each part induced dorsal mesoderm and neural tissues, the frequency being highest in Part 2 and lowest in Part 3. In III, Parts 1 and 2 realized their presumptive fates, through changes in inductive capacities from trunk-tail to head. This change progressed rapidly in Part 1, and slowly in Part 2. Part 3 required induction by neighbouring Part 2 to realize its presumptive fate. Changes of inductive capacity of Parts 1 and 2 respectively, were chronologically similar in normal development and in preculture experiments. Lastly, the primary presumptive pharyngeal zone at blastula was proposed to act as an initiator of the organization center, its programmed information being transmitted to Part 2, and then to Part 3.     

Staphylococcal cassette chromosome mec (SCCmec) characterization and panton-valentine leukocidin gene occurrence for methicillin-resistant Staphylococcus aureus in Turkey, from 2003 to 2006

Methicillin-resistant Staphylococcus aureus (MRSA) cause serious community-acquired and nosocomial diseases all over the world. We determined the SCCmec types and occurrence of the PVL gene by using TaqMan real-time PCR method, and correlated these with phenotypic antibiotic susceptibility patterns for MRSA strains collected from Gulhane Military Medical Academy Hospital (GMMAH) during 4 years study period. To our knowledge, this is the first report from Turkey of molecular SCCmec typing analysis of MRSA stains. A total of 385 clinical MRSA isolates collected in the clinical and Microbiology Laboratory at GMMAH between 2003 and 2006 were included in the study. Overall, SCCmec types-I, II, III, IV, V, nontypeable and PVL occurrence were detected in 11 (2.8%), 3 (0.8%), 316 (82.1%), 20 (5.1%), 20 (5.1%), 15 (3.9%) and 5 (1.3%) isolates, respectively. A total of 330 (85.5%) were SCCmec-I/II/III and 40 (10.3%) were SCCmec IV/V. SCCmec-I/II/III isolates were recovered more from patients with serious infections in surgical departments especially those with intensive care units than the SCCmec-IV/V isolates (χ² = 13.560, P < 0.001). SCCmec-I/II/III MRSA strains were predominantly recovered from blood stream (53.0%, P = 0.014), while SCCmec-IV/V strains were predominately isolated from skin and soft tissue and abscess (55.0%, P < 0.001). The PVL gene was detected in 10.0% of SCCmec-IV/V isolates in contrast to 0.3% in SCCmec-I/II/III (χ² = 25.164, P < 0.001). SCCmec-I/II/III MRSA strains were more resistant to clindamycin (χ² = 5.078, P = 0.024), amoxicillin-clavulanate (χ² = 84.912, P < 0.001), erythromycin (χ² = 4.651, P = 0.031), gentamicin (χ² = 24.869, P < 0.001), and rifampin (χ² = 18.878, P < 0.001) than SCCmec-IV/V MRSA strains. This data indicates that SCCmec-III MRSA strains that do not carry the PVL gene are the predominant MRSA strains in our hospital setting in Ankara, capital of Turkey and that SCCmec-I/II/III MRSA strains may cause serious infections in surgical departments especially those with intensive care units.     

“三生”空间视角下土地利用转型的生态环境效应及驱动力——以甘肃省为例

中国社会经济快速转型背景下土地利用变化所引发的环境问题被广泛关注。以土地利用现状遥感监测数据和统计资料为数据源,借助ArcGIS和SIMCA-P软件平台,基于"三生"空间视角,通过生态环境质量指数测度了1980—2018年甘肃省土地利用转型的生态环境效应,并运用偏最小二乘法揭示生态环境质量演变的驱动机制。研究发现:(1)"三生"空间结构演化和地域分异显著。生产、生活空间集中分布于河西走廊、陇中和陇东黄土高原并呈扩张态势;生态空间主要分布于陇南山地、甘南高原、祁连山地和河西走廊以北地带且整体呈缩减趋势;生产、生活空间的增长源于对生态空间的侵占。(2)生态环境质量改善和恶化趋势并存,整体上经历了先下降后上升的"U"形演化过程;空间上呈"东南高-西北低"的分布特征,空间格局演化具有明显的集聚性。(3)生态建设、科技进步等对生态环境改善具有明显的促进作用;而人口增加、社会消费水平提升在一定程度上加剧了生态环境压力;对外贸易对生态环境的影响具有双面性。     

中国植被分类系统修订方案

为了推动《中国植被志》研编工作, 该文回顾了中国植被分类系统的发展过程和主要阶段性成果, 提出了作为《中国植被志》研编技术框架组成部分的中国植被分类系统修订方案, 对各植被型组及各植被型进行了简单定义和描述, 并针对中国植被分类系统若干问题, 特别就中国植被分类系统总体框架、混交林的界定以及土壤在植被分类中的重要性等问题进行了讨论。1960年侯学煜在《中国的植被》中首次提出了中国植被分类的原则和系统, 1980年出版的《中国植被》制定了分类等级和划分依据等更加完善的系统, 之后《中国植被及其地理格局——中华人民共和国1:1 000 000植被图说明书》和《中国植物区系与植被地理》以及很多省区的植被专著对该系统进行过修订。2017年宋永昌在《植被生态学》中提出了一个分类等级单位调整的方案。本次提出的中国植被分类系统修订方案基本沿用《中国植被》的植被分类原则、分类单位及系统, 采用“植物群落学-生态学”分类原则, 主要以植物群落特征及其与环境的关系作为分类依据, 包含三级主要分类单位, 即植被型(高级单位)、群系(中级单位)和群丛(低级单位); 在三个主要分类单位之上分别增加辅助单位植被型组、群系组和群丛组, 在植被型和群系之下主要根据群落的生态差异和实际需要可再增加植被亚型或亚群系。修订方案包含了森林、灌丛、草本植被(草地)、荒漠、高山冻原与稀疏植被、沼泽与水生植被(湿地)、农业植被、城市植被和无植被地段9个植被型组, 划分为48个植被型(含30个自然植被型、12个农业植被型、5个城市植被型和无植被地段)。自然植被中有23个植被型进一步划分出了81个植被亚型。     

博尔纳病病毒pEGFP-p24基因重组质粒的构建及鉴定

构建博尔纳病病毒pEGFP-p24基因重组表达质粒。通过PCR方法扩增获得博尔纳病痛毒p24基因的完整序列,将此片段定向克隆到pEGFP-N1载体多克隆位点区,筛选重组阳性菌株,提取重组质粒,利用PCR方法和核酸序列测定验证重组质粒构建的正确性。PCR及核酸序列测定证明博尔纳病病毒pEGFP-p24基因重组表达质粒构建成功。构建的重组质粒将为研究博尔纳病病毒p24基因在真核细胞中的功能和作用提供实验依据。