中国东北地区人乳头瘤病毒16型全基因组克隆及HPV16.HaCaT重组细胞模型的构建及初步鉴定

为构建含东北地区人乳头瘤病毒16型(HPV16)全基因组的HPV16.HaCaT细胞模型,收集中国东北地区HPV16单一感染患者宫颈脱落细胞,提取DNA,将HPV16全基因组分成4个区段,通过4对特异性引物对HPV16全基因组进行分段扩增,测序后进行序列拼接及核酸序列分析,克隆HPV16全基因组序列;通过细胞转染,构建含HPV16全基因组的HPV16.HaCaT重组细胞模型;利用聚合酶链式反应(PCR)和细胞免疫荧光法检测重组细胞内HPV16早期基因的表达.成功克隆出中国东北地区HPV16全基因组序列(GenBank登录号:MW320358);构建了东北地区HPV16全基因组的重组质粒及HPV16.HaCaT重组细胞模型;证明了 HPV16早期基因E1-E4、E5、E6和E7在重组细胞模型内均有表达,从而获得中国东北地区HPV16全基因组序列及含有HPV16全基因组的HPV16.HaCaT重组细胞模型.     

Ovule morphogenesis and structure in Menispermaceae,focusing on the development of the single fertile ovule and its systematic significance

Menispermaceae is one of the core groups of Ranunculales. The single fertile ovule in each ovary in Menispermaceae varies greatly in integument number, micropyle formation, and integument lobe. However, data regarding ovule morphogenesis in the family are very limited. In this study, we document ovule development of selected species in the Menispermaceae using scanning electron microscopy and light microscopy. Ovule development in Menispermaceae shows the following characteristics. Two ovules are initiated in a young carpel, one of them degenerates gradually and the other develops into a fertile ovule in subsequent stages. Bitegmic in Sinomenium Diels. and Cocculus DC. and unitegmic in Stephania Lour. The formation of unitegmy is probably due to integumentary shifting. The annularly initiated inner integument is of dermal origin and has 2–3 cell layers in the family, but the semi-annularly initiated outer integument is of both dermal and subdermal origin. Both inner and outer integument are cup-shaped at maturity. The cup-shaped outer integument is formed due to the outer integument's extension to the concave (adaxial) side of the funiculus. The obturator is well developed and consists of 2–3 cell layers in Cocculus or 9–11 cell layers in Stephania. Ovule development of Menispermaceae suggests some common characteristics between Cocculus and Sinomenium, and derived unitegmy supports molecular data that indicate Stephania is one of the late-diverging lineages in the family. Integument lobations are present. The sterile ovule shows variations in the degeneration process. These results will provide evidence for exploring the evolution of ovules in Ranunculales.     

北盘江大峡谷流域浮游生物资源调查及多样性分析

为探究北盘江大峡谷流域内浮游生物群落组成及多样性情况,于2017年10月对该流域进行浮游生物调查及多样性分析.结果表明该流域内浮游植物共有7门40属61种、其中绿藻门、硅藻门和蓝藻门种类分别为21种、19种和12种,分别占到34.43％、31.15％和19.67％,浮游植物平均密度为4 298.33(375～9 500)ind./L,Shannon-Wiener藻类生物多样性指数均处于1和2之间;浮游动物3门12属21种,其中轮虫类、枝角类和桡足类种类分别为11种、5种和5种,各占到52.38％、23.81％和23.81％,浮游动物平均密度为436.11(55～1 437.5)ind./L,Shannon-Wiener生物多样性指数均处于1和2之间.本研究结果该地区水生生物资源积累了本底资料.     

Genomes of 12 fig wasps provide insights into the adaptation of pollinators to fig syconia

Figs and fig pollinators are one of the few classic textbook examples of obligate pollination mutualism. The specific dependence of fig pollinators on the relatively safe living environment with sufficient food sources in the enclosed fig syconia implies that they are vulnerable to habitat changes. However, there is still no extensive genomic evidence to reveal the evolutionary footprint of this long-term mutually beneficial symbiosis in fig pollinators. In fig syconia, there are also non-pollinator species. The non-pollinator species differ in their evolutionary and life histories from pollinators. We conducted comparative analyses on 11 newly sequenced fig wasp genomes and one previously published genome. The pollinators colonized the figs approximately 66.9 million years ago, consistent with the origin of host figs. Compared with nonpollinators, many more genes in pollinators were subject to relaxed selection. Seven genes were absent in pollinators in response to environmental stress and immune activation. Pollinators had more streamlined gene repertoires in the innate immune system, chemosensory toolbox, and detoxification system. Our results provide genomic evidence for the differentiation between pollinators and nonpollinators. The data suggest that owing to the long-term adaptation to the fig, some genes related to functions no longer required are absent in pollinators.     

Coassemble dopamine and GHK tripeptide into fluorescent nanoparticles for pH sensing

Fluorescent nanostructures have been widely applied to biomedical researches and clinical diagnosis such as biolabeling/imaging/sensing and have even acted as therapy reagents. Peptide‐based fluorescent nanostructures attract recent interest from biomedical researchers. Inspired by the natural existence of GHK‐Cu complex with a growth factor‐like effect in human blood, here we have developed a novel approach for designing nanosensors through the co‐assembling of two kinds of biomolecules. By making best use of both π‐π stacking between carbon rings and the easy‐oxidation property of an important transmitter molecule, dopamine (DA), we successfully built up a supersensitive and robust fluorescent pH nanosensor by co‐assembling oxidized DA (DA_ox) with a tripeptide GHK. The GHK‐DA_ox nanostructures have a quantum yield of 20.82%, which might be the brightest one among all the current co‐assembling structures merely through unmodified biomolecules. We envision this approach could open a new avenue for not only hybrid nanostructure construction, but also may inspire the bioengineering of in vivo luminescent probes.     

LBX2-AS1 promotes ovarian cancer progression by facilitating E2F2 gene expression via miR-455-5p and miR-491-5p sponging

LBX2-AS1 is a long non-coding RNA that facilitates the development of gastrointestinal cancers and lung cancer, but its participation in ovarian cancer development remained uninvestigated. Clinical data retrieved from TCGA ovarian cancer database and the clinography of 60 ovarian cancer patients who received anti-cancer treatment in our facility were analysed. The overall cell growth, colony formation, migration, invasion, apoptosis and tumour formation on nude mice of ovarian cancer cells were evaluated before and after lentiviral-based LBX2-AS1 knockdown. ENCORI platform was used to explore LBX2-AS1-interacting microRNAs and target genes of the candidate microRNAs. Luciferase reporter gene assay and RNA pulldown assay were used to verify the putative miRNA-RNA interactions. Ovarian cancer tissue specimens showed significant higher LBX2-AS1 expression levels that non-cancerous counterparts. High expression level of LBX2-AS1 was significantly associated with reduced overall survival of patients. LBX2-AS1 knockdown significantly down-regulated the cell growth, colony formation, migration, invasion and tumour formation capacity of ovarian cancer cells and increased their apoptosis in vitro. LBX2-AS1 interacts with and thus inhibits the function of miR-455-5p and miR-491-5p, both of which restrained the expression of E2F2 gene in ovarian cancer cells via mRNA targeting. Transfection of miRNA inhibitors of these two miRNAs or forced expression of E2F2 counteracted the effect of LBX2-AS1 knockdown on ovarian cancer cells. LBX2-AS1 was a novel cancer-promoting lncRNA in ovarian cancer. This lncRNA increased the cell growth, survival, migration, invasion and tumour formation of ovarian cancer cells by inhibiting miR-455-5p and miR-491-5p, thus liberating the expression of E2F2 cancer-promoting gene.     

云南高黎贡山地区蝴蝶群落多样性

位于滇西北的高黎贡山是全球生物多样性研究和保护的热点地区之一, 然而该地区昆虫多样性缺乏系统调查和总结。本研究聚焦蝴蝶类群, 考虑该区域高山峡谷特点, 结合海拔梯度、生境类型和季节变化, 采用样线法调查、分析蝴蝶物种多样性及群落结构变化。结果显示: 共观测记录到蝴蝶2,055只, 隶属于5科85属151种, 在历史记录上新增27种, 使该地区已知蝴蝶种类达488种; 其中蛱蝶科物种多样性最高, 灰蝶科次之, 凤蝶科最低。蝴蝶群落多样性分析结果表明: 中海拔1,000-2,000 m区域种类丰富、多样性指数最高; 低海拔区蝴蝶分布明显聚集, 并且与高海拔地区空间上分离, 少有重叠。该地区不同生境中蝴蝶的种类及数量差异也较大, 物种数及多样性指数在自然保护区最高、边缘交错带居中及农业种植区最低。此外, 蝴蝶的种类和数量也存在季节差异, 春季调查到的个体数少, 夏季观察到的物种数少, 两年秋季调查到的物种丰富度、多样性均高, 但存在季节内变化。总之, 高黎贡山地区不同海拔、生境、季节间和季节内蝴蝶群落组成有自身特点, 共存物种有限, 蝴蝶群落相似性低。综合评估分布于该地区的蝴蝶保护种类, 包括易危种17种、近危种50种, 有国家二级保护蝴蝶3种。本研究弄清了高黎贡山地区蝴蝶的物种本底, 并调查获得其多样性随海拔、生境和季节变化的模式, 为加强区域物种多样性监测、保护生物多样性提供了科学依据。     

小鼠阴道菌群紊乱模型的建立

目的建立小鼠阴道菌群紊乱模型,为调整阴道菌群紊乱的药物研发提供模型参考。方法采用雌激素化的C57BL/6小鼠经阴道以不同处理方式(链霉素50μg/只,加德纳菌50μL/只,链霉素50μg/只+抗链霉素加德纳菌50μL/只,每种处理方式各70只/组)诱导小鼠阴道菌群紊乱模型,采用细菌16S rDNA高通量测序考察造模后第1、第2、第3、第5、第7、第9、第11天内加德纳菌在阴道内的定植情况,以及阴道细菌结构和细菌生物多样性,并通过显微镜观察阴道组织的病理改变,比较3种造模方法的差异。另采用阳性药物(定君生,成分为德氏乳杆菌)对模型进行验证。结果采用链霉素联合抗链霉素加德纳菌能诱导形成典型的阴道菌群紊乱模型,造模后可见加德纳菌在阴道内的定植增多,优势菌以铜绿假单胞菌为主,且阴道细菌生物多样性增加,阴道组织可见明显的炎症浸润和上皮细胞坏死等病理改变。定君生能显著减少加德纳菌在阴道内的定植,减少阴道内的细菌生物多样性,并能显著改善阴道组织病理变化。结论采用链霉素联合抗链霉素加德纳菌诱导的小鼠阴道菌群紊乱模型方法更佳,模型的细菌学和组织病理学改变,以及对药物的反应性与临床有一定相似,具有临床应用价值。