结直肠腺瘤的微卫星不稳定状态与相关基因表达的研究

应用微切割-聚合酶链反应-单链长度多态性（PCR-SSLP）的方法,检测59例62个结直肠腺瘤,包括散发性腺瘤及家族性腺瘤性息肉病（FAP）腺瘤在BAT26等16个微卫星基因座在结直肠腺瘤标本的微卫星不稳定性（MSI）状态;并应用免疫组织化学SP法检测β-连接素（β-catenin）、TP53、BAX等的表达情况,初步探讨错配修复（MMR）基因在结直肠癌发生的早期即腺瘤阶段的作用及其意义。结果显示：(1)腺瘤16个基因座的总MSI发生率为14.4%;同一病人的不同腺瘤在某些相同的基因座表现出不同的MSI状态;(2)5例FAP病人均表现为MSI-L,其中有3例在hMSH3基因座表现为MSI阳性;(3)β-连接素在腺瘤和腺癌细胞膜阳性率分别为42.9%和11.4%,表达差异有显著统计学意义（P＜0.001）;(4)TP53、D5S346、TCF4(A)9、TGFβRⅡ(GT)3、TGFβRⅡ(A)10等微卫星基因座的MSI改变与相应的免疫组织化学指标TP53、β-连接素、TGFβRⅡ等在腺瘤及腺癌中的阳性表达有密切关系。可以推断：(1)在结直肠癌发生发展的早期即腺瘤阶段即可表现微卫星不稳定性,腺瘤中存在1p染色体的改变、APC基因的改变及TGFβ信号转导途径的异常;(2)随着腺瘤向腺癌的进展,β-连接素的阳性着色由细胞膜转移至细胞内,而且胞浆阳性强度增加;可以推断腺瘤中APC-β-联蛋白-TCF4信号转导途径的异常。 Abstract:In order to understand the role of mismatch repair (MMR) gene in colorectal carcinogenesis,microsatellite instability (MSI) status of 16 microsatellite loci of 62 adenomas from 59 patients,including sporadic and familial adeonmatous polyposis (FAP) adenomas were detected by microdissection-PCR-SSLP,and protein expressions of β -catenin,P53,and BAX,etc.were assayed by immunohistochemistry.Results were as following:(1)The overall MSI alteration rate of the 16 loci was 14.4%.Different adenomas from the same patient showed different microsatellite alterations at the same loci;(2)All of the five FAP patients were MSI-L,three of which showed MSI at the locus of hMSH3;(3)The membrane expression rate of β-catenin in adenomas and accompanied carcinomas was 42.9% and 11.4%,respectively (P＜0.001);⑷Microsatellite alterations of the microsatellite loci of TP53,D5S346,TCF4(A)9,TGFβRⅡ(GT)3 and TGFβRⅡ(A)10 were associated with the changes of their protein expressions.It could be concluded the following:(1)Microsatellite instability existed even in the early stage (adenomas) of colorectal tumorigenesis.The alterations of chromosome 1p,APC genes,and the TGFβ signal transduction pathway could also be deduced;(2)In the progression of adenoma to carcinoma,the staining of β-catenin would be transferred from membrane to cytoplasm and then nucleus,and the cytoplasm stain was stronger in carcinoma than that in adenomas.The abnormality of the signal transduction pathway of APC-β-catenin-TCF4 could be concluded.     

六株虫生真菌的生物学特性及其对油茶象甲的致病力

为了筛选防治油茶象甲的虫生真菌菌株,本研究测定了不同绿僵菌(Ma09、Ma25、Ma05、Mac985)和白僵菌(Bb01、Bb02)菌株的生长速度、产孢量以及孢子萌发率,采用孢悬液浸渍法测定了上述菌株对油茶象甲成虫和幼虫的致病力。结果表明,Ma09、Ma05、Bb01、Bb02菌株的菌落直径分别为63.13、62.55、61.99、61.90 mm(25℃、15 d),生长速度显著快于其余菌株(P0.05);Ma25、Bb01、Ma09菌株的产孢量分别为3.04×10~8、2.88×10~8、2.85×10~8孢子/cm~2,显著高于其余菌株(P0.05);Bb01和Ma09菌株的孢子萌发率较高,分别达到97.80%和96.40%。不同菌株对油茶象甲成虫和幼虫致病力差异显著,其中Bb01和Ma09在1×10~7孢子/mL时,对油茶象甲成虫的LT_(50)分别为4.64 d和4.93 d,处理后15 d累计校正死亡率均达到100.00%,僵虫率分别为76.00%和78.67%;对幼虫的LT_(50)分别为2.15 d和3.48 d,10 d累计校正死亡率均达到100.00%,僵虫率分别为81.00%和86.31%。本研究筛选的Bb01和Ma09菌株具有生长快、产孢量大、孢子萌发率高、对油茶象甲致病力强等特性,在油茶象甲的生物防治中具有潜在的应用价值。     

核开关结构、机制及应用新进展

核开关(riboswitch)是Breaker等在2002年发现的一种全新的转录后调节机制．它可以通过小分子与mRNA结合来直接调控基因的表达,不需要任何蛋白质的参与．与常见的经由蛋白质的调控方式相比,riboswitch响应更迅速,对细胞内代谢物的变化更敏感．它的发现为RNA研究展示了新的领域． 目前在这个领域,既有基础研究,如riboswitch晶体结构解析、作用机制和动力学研究,又有前沿应用研究,如基于riboswitch的生物传感器和药物设计．Topp等通过设计riboswitch成功地改变了大肠杆菌的趋化性,这为合成生物学和人工生物网络的设计提供了新思路．目前对于riboswitch结构、机制及动力学的研究为基于riboswitch的合理药物设计奠定了基础,有望针对这一新的机制开发新一代抗菌药物．     

Direct and indirect effects of weather variability in a specialist butterfly

1. Climate change poses serious threats to the long‐term persistence of many animal and plant populations. Species having specific niche requirements, or characterised by highly co‐evolved interactions, will face the greatest challenges. An example is represented by Maculinea alcon (Denis & Schiffermüller), a monophagous and univoltine butterfly species, which lays eggs only on larval host plants which occur inparticular phenological conditions. 2. The present 2‐year study focused on two M. alcon populations, both located at the southern boundaries of the species, but facing different climatic conditions (360 m, low altitude versus 860 m, high altitude). Population vulnerability with respect to direct and indirect effects of climate change was analysed, focusing on two important aspects of butterfly biology, i.e. the flight activity of adults and the degree of synchrony in the larval plant–insect interactions. 3. It was observed that, when positive temperature anomalies are reached, the temperature can exert detrimental effects on adults' activity. At a low altitude, in a hotter than usual year, a temperature threshold was recorded (around 32 °C), above which the activity of butterflies is inhibited. In contrast, at a high altitude, temperature increases maintain the opportunity to enhance butterfly activity. Altitudinal differences were also observed in the phenology of the two interacting species, which generate stronger asynchrony at low altitudes. 4. High‐ and low‐altitude populations represent different conservation units: a global increase in temperature would pose a serious threat to the lowland populations, whereas high‐altitude populations would gain a greater role in assuring the persistence of M. alcon at its southern boundaries.     

临床路径教学法在鼻咽癌临床护理教学中的应用评价

目的 探讨临床路径(CP)在鼻咽癌临床护理教学中的应用效果.方法 将2011年7月~2012年5月在我院放射科实习的96名护生随机分为观察组和对照组,观察组采用CP教学法,对照组采用传统实习带教法,并对两组护生的教学效果分阶段进行综合评价.结果 观察组护生在理论考核、技能考核及教学满意度方面均优于对照组,差异有统计学意义(P＜0.05).结论 CP教学有利于护生形成系统的知识结构,对临床思维及综合能力的培养起到重要作用,临床路径教学模式值得推广.     

广东鸟屎榄果实挥发性成分的HS-SPME-GC-MS分析

采用顶空固相微萃取–气相色谱–质谱联用法(HS-SPME-GC-MS)分析鸟屎榄(Canarium pimela)果实的挥发性成分,并通过面积归一法计算各成分的相对含量。结果显示,从鸟屎榄中鉴定出48种挥发性成分,占挥发性成分总量的93.30%,主要有α-蒎烯(30.17%)、对伞花烃(11.84%)、α-水芹烯(8.72%)和β-石竹烯(8.02%)等。对挥发性成分的香味物质分析发现,该果实共涵盖22种香型,其中松柏香、柑橘香和木香等香味载荷大,是构成鸟屎榄果实的主要香韵。     

发状念珠藻对盐胁迫的响应

探讨了发状念珠藻(NostocflagelliformeBornetFlah)对盐胁迫的耐受适应机制,采用含不同浓度NaCl(0、01、02、04、06、08、10mol/L)的BG110培养液处理具有正常生理活性的丝状体,25±05℃,40μmol/m2/s下照光培养12h,测定藻体光合作用、呼吸作用等生理活性以及体内一些物质的含量,结果表明:随培养液中NaCl浓度的升高藻体光合作用、呼吸作用以及PSⅡ活性(Fv/Fm)降低;质膜透性不断增大,丙二醛含量升高,自由水含量、自由水/束缚水比值下降,类胡萝卜素、可溶性糖含量增加,脯氨酸含量变化不大。由此可知,盐胁迫下发状念珠藻正常生理活性受到抑制而表现出一定的抗逆能力;该藻对盐胁迫具有一定的耐受能力,类胡萝卜素的增加有助于清除藻体内的氧自由基,可溶性糖可能是其主要渗透调节物质之一,脯氨酸在盐胁迫中的渗透调节作用不大。       

基于3种基因序列对百合属Leucolirion组的系统发育与种间关系研究

中国拥有丰富的野生百合种质资源,但有关百合属的系统发育与进化关系研究相对较少。该研究基于ITS、cpDNA(trnV-ndhC、petL-psbE)及单拷贝核基因AKT等3个基因片段,选取特有分布于西南地区的百合属6个近缘类群共48份样本及GenBank获得的数据,对百合属Leucolirion组进行了系统发育与种间关系研究。结果表明:(1)ITS结果支持前人的处理,即将Leucolirion组细分为2个亚组,subsect.Leucolirion6a和subsect.Leucolirion6b。建议将湖北百合(L.henryi)和南川百合(L.rosthornii)归入到subsect.Leucolirion 6a,与泸定百合(L.sargentiae)、淡黄花百合(L.sulphureum)、宜昌百合(L.leucanthum)和岷江百合(L.regale)等4个种一起组成Leucolirion组;原本属于Archelirion组的野百合(L.brownii)与subsect.Leucolirion6b聚为一支,故将它与台湾百合(L.longiflorm)、麝香百合(L.formosanum)等组成一个新组,命名为百合组(sect.Lilium)。(2)组间关系上,Leucolirion组与百合组的系统发育关系较远,前者与分布于日本的Archelirion组形成姐妹类群,而后者与Sinomartagon组的亲缘关系更近。(3)基于叶绿体联合基因和单拷贝核基因AKT的6个近缘种的遗传距离显示,湖北百合与南川百合明显区别于Leucolirion组内拥有白色管状花的4个类群。叶绿体联合基因树和单拷贝核基因树结果表明,泸定百合、淡黄花百合、岷江百合和宜昌百合在遗传水平上表现出了极高的相似性,遗传关系十分相近,这可能与百合属植物特殊的繁育系统有关,影响了物种间遗传变异与分化。