中国兰科植物沿经纬度的水平分布格局

对濒危物种在大尺度上地理分布的研究,有助于制定合理的保护规划和保护策略.兰科植物作为一大类急需保护的濒危物种,研究其在中国境内的地理分布格局具有重要的理论和实践意义.通过文献查阅、自然保护区数据整理收集兰科植物在全国范围内的调查数据,利用ArcGIS10.0和SPASS18.0软件对其地理分布进行了分析,结果表明：中国西南地区是兰科植物的分布中心和分化中心;兰科植物丰富度表现出显著的经度和纬度相关性,与经度之间呈单峰关系,在100°E附近出现峰值,但随纬度升高丰富度不断下降.     

基于指数施肥法的一年生油松菌根化容器苗的供N研究

为了探讨一年生菌根化油松容器苗的最适苗木供N量和供N速率,采用4种不同指数施肥量进行了试验.通过测定苗木生物量和N含量,显示总供N量为80 mg·株-1的处理组在生物量积累和苗木N含量上显著优于其它各组（P〈0.05）;在100 d时,该处理可获得0.47 mg·株-1·d-1的N最适添加速率;在该速率下,苗木生物量、N含量分别可以达到845.60 mg·株-1和6.51 mg·株-1.根据生物量和N含量随着供N量增加的回归拟合结果,在107 d的生长过程中,67.96 mg·株-1至84.15 mg·株-1的供N总量可以使得一年生油松幼苗获得较高的生物量积累水平和N含量.     

Proliferation of Colorectal Cancer Is Promoted by Two Signaling Transduction Expression Patterns: ErbB2/ErbB3/AKT and MET/ErbB3/MAPK

One of the recent breakthroughs in cancer research is the identification of activating mutations in various receptor tyrosine kinase(RTK) pathways in many cancers including colorectal cancer(CRC). We hypothesize that, alternative to mutations, overexpression of various oncogenic RTKs may also underpin CRC pathogenesis, and different RTK may couple with distinct downstream signaling pathways in different subtypes of human CRC. By immunohistochemistry, we show here that RTK members ErbB2, ErbB3 and c-Met were in deed differentially overexpressed in colorectal cancer patient samples leading to constitutive activation of RTK signaling pathways. Using ErbB2 specific inhibitor Lapatinib and c-Met specific inhibitor PHA-665752, we further demonstrated that this constitutive activation of RTK signaling is necessary for the survival of colorectal cancer cells. Furthermore, we show that RTK overexpression pattern dictates the use of downstream AKT and/or MAPK pathways. Our data are important additions to current oncogenic mutation models, and further explain the clinical variation in therapeutic responses of colorectal cancer. Our findings advocate for more personalized therapy tailored to individual patients based on their type of RTK expression in addition to their mutation status.     

The ?842G/C Polymorphisms of PIN1 Contributes to Cancer Risk: A Meta-Analysis of 10 Case-Control Studies

Background

Peptidyl-prolyl cis–trans isomerase NIMA-interacting 1 (PIN1) plays an important role in cancer development. The relationship between PIN1 −842G/C (rs2233678) polymorphism and cancer risk was inconclusive according to published literature.

Methodology/Principal Findings

A literature search, up to February 2013, was carried out using PubMed, EMBASE and the China National Knowledge Infrastructure (CNKI) database. A total of 10 case-control studies including 4619 cases and 4661 controls contributed to the quantitative analysis. Odds ratio (OR) and 95% confidence intervals (95% CI) were used to assess the strength of association. Overall, individuals with the variant CG (OR = 0.728, 95% CI: 0.585,0.906; P_{heterogeneity}<0.01) and CG/CC (OR = 0.731, 95% CI: 0.602,0.888; P_{heterogeneity}<0.01) genotypes were associated with a significantly reduced cancer risk compared with those with wild GG genotype. Sub-group analysis revealed that the variant CG (OR = 0.635, 95% CI: 0.548,0.735; P_{heterogeneity} = 0.240) and CG/CC (OR = 0.645, 95% CI: 0.559,0.744, Pheterogeneity = 0.258) genotypes still showed an reduced risk of cancer in Asians; while no significant association was observed in Caucasians (CG vs.GG: OR = 0.926, 95% CI: 0.572,1.499, P_{heterogeneity}<0.01; CG/CC vs. GG: OR = 0.892, 95% CI: 0.589,1.353; P_{heterogeneity}<0.01). Furthermore, sensitivity analysis confirmed the stability of results. Begg''s funnel plot and Egger''s test did not reveal any publication bias.

Conclusions

This meta-analysis suggests that the PIN1 −842G/C polymorphism is associated with a significantly reduced risk of cancer, especially in Asian populations.     

利用piggyBac转座子制备牛成体多能干细胞诱导技术研究

通过逆转录病毒等媒介表达核转录因子Oct4、Sox2、Klf4、c-Myc可将体细胞重编程为诱导多能干细胞(induced pluripotent stem cells, iPSc)。时至今日,已经报道了小鼠、人、大鼠、猪、羊、马、牛的iPS细胞,但大动物iPS的多能性特别是嵌合体形成和生殖细胞传代还没有得到确认。与逆转录病毒等不同的是,piggyBac转座子转染效率高且无病毒源性、操作简单,可以在转座酶的存在下被安全切除。首次尝试了采用piggyBac转座子携带鼠源Oct4、Sox2、Klf4、c-Myc、Rarg和Lrh16个核转录因子诱导胎牛成纤维细胞,成功获得牛类iPS细胞,其形态与小鼠胚胎干细胞相似,克隆边界清晰、呈丘状、克隆内细胞致密、核大。RT-PCR与免疫组织化学染色分析均显示牛类iPS细胞表达多能性基因。该类细胞体外诱导分化可形成类胚体EB,且表达3个胚层的基因;体内诱导分化可形成畸胎瘤,苏木精、伊红染色显示瘤体有三胚层的分化。上述结果显示利用piggyBac转座子制备牛多潜能干细胞诱导技术可行,产生的牛类iPS细胞具有潜在多能性。     

结核分枝杆菌生物膜形成相关基因的筛选与鉴定

目的:通过菌落表型变化并结合生物膜生长缺陷筛选并鉴定可能与生物膜形成相关基因.方法:利用带有Himarl转座子的MycoMarT7转座子系统建立结核分枝杆菌H37Ra随机插入突变库;筛选细菌表面结构发生变化和生物膜形成有变化的突变菌株;运用T-A克隆法并结合抗性标记挽救法获得突变菌株的随机插入基因侧翼序列从而鉴定突变基因,并运用生物信息学方法分析预测突变基因的功能.结果:通过菌落形态变化及生物膜缺陷表型筛选出39株突变株,成功鉴定其中16株突变株,涉及16个基因发生突变,其中5个与脂质代谢相关,4个与细胞壁合成相关、2个与中间代谢和呼吸作用相关、1个调节蛋白相关基因,1个毒力相关基因,1个PE/PPE家族基因,还有2个功能未知基因.结合生物膜形成缺陷分析,其中8个基因可能与H37Ra体外生物膜的形成相关.结论:成功构建库容量约为l×104结核分枝杆菌转座子随机插入突变文库,筛选获得生物膜生长受损突变株及可能与结核分枝杆菌生物膜形成相关的基因信息,为后续深入开展生物膜形成机制研究奠定基础.     

Wnt5a抑制Wnt3a促黑素细胞黑素生成的作用探究

目的：研究WntSa对Wnt3a处理过的melan—a细胞分泌黑色素的影响。方法：体外培养黑色素细胞（melan-a细胞）,分别进行GFP、Wnt3a、Wnt3a＋WntSa处理,比较细胞的突起,酪氨酸酶的活性以及黑素合成相关基因（TYR、TRP2、MITF）表达情况。结果：Wnt3a促进黑色素细胞突起的生长和TYR、TRP2、MITF的表达,而Wnt5a逆转了Wnt3a对黑色素细胞的作用。结论：Wnt5a抑制Wnt3a促黑素细胞黑素生成的作用,表明在melan．a黑素细胞中Wnt5a可有效抑制wnt经典通路。     

木薯ftsZ基因分离及在原核生物中功能初步鉴定

ftsZ基因是控制细胞分裂的关键基因,其蛋白能够在分裂位点形成一个环状结构而影响细胞分裂.为了研究木薯质体分裂与木薯淀粉品质形成的关系,根据木薯基因组数据库上的预测序列,设计引物,从木薯基因组中分离了与质体分裂相关的ftsZ家族3个新基因(ftsZ1,ftsZ2,ftsZ3).分别将它们与荧光蛋白基因(GFP)融合,构建了3个原核表达载体pET-fisZ1-GFP、pET-fisZ2-GFP、pET-fisZ3-GFP,并转化大肠杆菌BL21(DE3).通过荧光显微镜观察菌体的表型和分裂,初步鉴定了木薯质体分裂相关基因ftsZ家族对细胞分裂的作用.结果显示:尽管木薯与大肠杆菌的亲缘关系较远,ftsZ基因的同源性较低,但是两者表现出相似的功能,木薯ftsZ基因的表达能严重影响大肠杆菌细胞分裂.这一结果为进一步研究木薯ftsZ家族基因的功能奠定了基础.     

重组人源性脂联素球状结构的表达、纯化与活性检测

目的:利用基因工程方法构建人源性脂联素球状结构(gAd)基因的高效原核表达体系,并对重组蛋白进行诱导表达、纯化、鉴定及活性检测.方法:从正常人脂肪组织里面提取总RNA,反转录合成cDNA,经PCR扩增、酶切后连入pET-22b(+)载体构建重组质粒pET-22b(+)-gAd,重组质粒转化大肠杆菌BL21(DE3)感受态细胞.经诱导剂诱导后目的蛋白以包涵体形式产生,采用强碱促溶包涵体并用丙酮沉淀蛋白的方法进行复性和纯化,得到高纯度的人源性gAd.运用SDS-PAGE、Western blotting对重组蛋白进行鉴定,通过对蛋白激酶(AMPK)的磷酸化水平和对小鼠的心肌缺血再灌注损伤的保护作用来检测纯化蛋白的生物学活性.结果:成功构建了原核表达载体pET-22b(+)-gAd,实现了人源性gAd在原核细胞中的表达,并对形成的包涵体变性、复性和纯化,纯化出的蛋白经过SDS-PAGE和Western分析证实为gAd;通过对AMPK的磷酸化水平的检测和对小鼠的心肌缺血再灌注损伤的保护作用证明纯化出的gAd具有高生物学活性.结论:成功构建、表达和纯化了无标签、高生物学活性的人源性脂联素球状结构(gAd),为其进一步的理论研究、生产开发奠定了基础.