小鼠脑组织及培养神经元Neuro-2a 在内质网应激反应时的基因表达谱分析

内质网是真核细胞的重要细胞器。某些细胞内外因素如病原体感染等能引起从内质网到胞浆和胞核的信号传导途径活化,即内质网应激反应。但是,目前国内外尚无针对内质网应激反应的基因表达谱分析报道。本研究中,用3种已报道的内质网应激反应诱导剂,包括蛋白质糖基化抑制剂衣霉素(tunicamycin)、内质网Ca ²⁺-ATPases抑制剂毒胡萝卜素（thapsigargin）和乙脑病毒(Japanese encephalitis virus, JEV),分别处理小鼠颅腔和小鼠脑神经瘤细胞（Neuro-2a）,试剂处理组与未处理组的第二代RNA测序分析发现,衣霉素、毒胡萝卜素和乙脑病毒在体外和体内均引起分子伴侣基因Hsp70表达上调,诱导内质网应激反应。衣霉素、毒胡萝卜素和乙脑病毒体外处理诱导的内质网应激反应信号通路中,基因差异表达相似性高于体内处理组。乙脑病毒和糖基化抑制剂衣霉素体内外处理,主要诱导内质网应激反应的非折叠蛋白质反应信号通路,引起相关基因Atf4、Bip、Edem和Perk等表达上调。内质网Ca ²⁺-ATPases抑制剂毒胡萝卜素主要诱导内质网超负荷反应,激活NF-κB信号通路。乙脑病毒诱导的内质网应激反应相关差异表达基因数量最多,体外与体内合计有40种。乙脑病毒体内外处理上调的基因包括Bax、Casp12、Atf4、Bip、Edem和Perk等,下调的基因包括Sec23/24、Nef、Svip和Jnk等。糖基化抑制剂衣霉素体内外处理上调基因包括Gadd34、Atf4、Ermani和Bip等,下调基因包括Grp94、Atf6、Sec23/24和Nef等。内质网Ca ^-2+-ATPases抑制剂毒胡萝卜素体内外处理上调的基因包括Sec61、Trap和Ask1等。衣霉素、毒胡萝卜素和乙脑病毒体内外处理也通过内质网应激反应,调控与炎症或凋亡相关的MAPK信号通路和P53信号通路。本研究首次通过使用3种内质网应激反应诱导剂分别处理小鼠和细胞,揭示了体内外内质网应激反应引起的基因表达谱变化,为内质网应激反应相关疾病的治疗提供了新思路。     

大猿叶虫卵的颜色与其后代滞育率和繁殖力的关系

大猿叶虫Colaphellus bowringi Baly卵的颜色有浅黄色和深红色2种类型。调查卵的颜色与其后代滞育的发生和繁殖力的关系,结果表明,在25℃、28℃,不同光周期L∶D(11∶13,12∶12,13∶11和15∶9)下,浅黄色卵和深红色卵的后代滞育率没有显著差异;但在L14∶D10下,浅黄色卵的后代滞育率(25℃为58.6%;28℃为33.9%)显著高于深色卵的后代(25℃为45.4%;28℃为11.2%)。卵的颜色与其后代成虫的产卵前期和成虫寿命没有显著相关,但浅黄色卵后代的平均产卵量和平均日产卵量(1663粒和58.6粒)均显著高于深红色卵的后代(1176粒和45.1粒)。     

抗人尿道高分化鳞癌单克隆抗体的研制

用本室建立的人尿道高分化鳞状上皮癌细胞株(Hus-98)免疫Balb/C小鼠,通过细胞杂交技术及ELISA方法进行筛选,SP免疫组织化学方法进行鉴定,获得10株能稳定分泌、选择性强、效价高的杂交瘤细胞株;同时从乳腺癌组织中提出肿瘤相关抗原行免疫印记实验(Western-blot)。结果6株经4次克隆能稳定分泌抗体Ig,免疫组化显示这些抗体是针对来源于上皮的鳞癌、腺癌细胞膜和细胞浆的,而癌旁正常组织及胎儿组织呈阴性反应;通过western-blot结果显示肿瘤相关抗原分子量为46.414kb。     

水稻内生真菌Fusarium moniliform ZSU-1次级代谢产物研究

为了探索内生镰刀菌对稻田病害生物防治的物质基础,对一株来自健康水稻茎部的串珠镰刀菌Fuscrium moniliform ZSU-1进行发酵培养,用色谱技术分别对其菌体提取物、发酵液提取物进行活性次级代谢产物分离,得到4个化合物,通过1D、2D NMR、质谱等波谱分析,分别鉴定为镰刀菌酸(1)、镰刀菌酸·脱氢镰刀菌酸缔合物(1:1)(2)、白僵菌素(3)和甘露醇(4).鉴于镰刀菌酸、镰刀菌酸·脱氢镰刀菌酸缔合物(1:1)有较强的抗菌活性,推测这两个化合物对稻田病害生物防治可能有一定的意义.     

Incomplete protein disulphide bond conformation and decreased protein expression result from high cell growth during heterologous protein expression in Pichia pastoris

Previous report has shown that the expression of recombinant human consensus interferon-α mutant (cIFN) in Pichia pastoris in bioreactor is limited with respect to the incorrectly folded cIFN with incomplete disulfide bond, which lead to the degradation and aggregation of cIFN. In this study, the origin of incorrectly folded cIFN is firstly studied. Fed-batch fermentation in bioreactor shows that the incorrectly folded cIFN is formed intramolecularly and secreted to the extracellular environment. Further chemostat cultures indicate that the specific growth rate is the critical factor for the production of incorrect cIFN. In addition, cell shows reduced expression level of cIFN at high specific growth rate. We also demonstrate that the incorrectly folded cIFN could form aggregates intracellularly and these aggregates are non-covalent forms. Taken together, these results suggest that the efficient heterologous expression of cIFN is limited by high cell growth that is unique from expression limitations seen for soluble proteins. A balance has to be found between the increase for high efficient expression of heterologous proteins and requirement of the high cell growth during the expression of recombinant proteins in P. pastoris.     

蒜头果地理分布与水热关系分析

利用温度指数(WI)、湿润指数(HI)等气候因子,研究了蒜头果(Malana oleifera Chun et S.Lee)地理分布与气候因子的关系.结果表明,极端最高温、7月均温、年均温、≥10℃年积温和年均降水量是蒜头果分布的主要限制因子;由半峰宽(PWH)计算法确定蒜头果生长的最适温度为18.0～22.0℃,年降水量为1106.5～1 585.1 mm;水热组合较好的乐业地区是蒜头果的主要分布区.极端高温和干燥的气候可能是蒜头果分布区逐渐缩小并濒危的重要因素.     

玉叶金花种内性状置换的初步研究

通过对茜草科玉叶金花属植物玉叶金花传粉生物学及种内形态分化和分布特点的初步研究,发现该种的两个变种玉叶金花和白花玉叶金花在形态和生境适应上都有一定的规律。研究表明,楠藤与开黄花的玉叶金花原变种的传粉者主要是鳞翅目昆虫,而玉叶金花的白花变种的有效传粉者是膜翅目昆虫。认为变种白花玉叶金花居群的出现与同属植物楠藤的同域分布及避免相互竞争传粉者有关。白花玉叶金花变种的出现属于性状置换现象。     

An integrative approach to the taxonomy of the pigmented European Pseudomonocelis Meixner, 1943 (Platyhelminthes: Proseriata)

Only a few species belonging to the Proseriata (Platyhelminthes) show a parenchymatic pigmentation, which may aid identification. Among these, Pseudomonocelis agilis has a yellowish body and is provided with a reddish–brown girdle in front of the statocyst. The species is known for limited areas of northern Europe and the Mediterranean. The present study was conducted to assess both the taxonomic status of populations attributed to the species across the unusually wide range for an interstitial flatworm, which lacks an obvious means of dispersal, and the levels of genetic variability within and among populations, by employing an integrative approach that included the analyses, on six populations, of three molecular markers (small subunit ribosomal 18S‐like gene, inter‐simple sequence repeat, allozymes), karyotypes, and 11 morphological characters. Furthermore, crossbreeding experiments were carried out on the Mediterranean populations. The results obtained revealed the existence of four highly divergent genotypic clusters, accompanied by karyological differences, with complete intersterility among the clusters tested. The combination of approaches adopted strongly supports the conclusion that the wide‐ranging European pigmented species P. agilis is actually composed of four species: P. agilis in the Baltic area; Pseudomonocelis cetinae in the Adriatic; and Pseudomonocelis sp. nov. A and Pseudomonocelis sp. nov. B in the western and eastern Mediterranean, respectively. The latter two species are morphologically indistinguishable for the parameters essayed. Reconstruction of the phylogenetic relationships of these taxa, including congeneric and consubfamilial outgroups, showed that pigmentation is a plesiomorphic condition for the genus Pseudomonocelis and that Pseudomonocelis sp. nov. A shares a previously undetected, sister‐group relationship with species of the unpigmented P. ophiocephala complex. The present study thus depicts complex speciation processes in a mesopsammic species, which involves allopatric divergence operating on different scales and ecological shifts, and highlights that the contribution of microturbellarians to marine biodiversity may be seriously underestimated. © 2009 The Linnean Society of London, Biological Journal of the Linnean Society, 2009, 98 , 907–922.