通过染色体外同源重组建立乳腺中表达外源基因的转基因小鼠

为研究染色体外重组方法在创建转基因动物中的应用,选取牛asl酪蛋白基因的5′及3′侧翼区和氯霉素乙酰化酶编码区,构建了两个具有3 kb相互重叠的融合基因.将这两个DNA片段末端脱磷酸后,以摩尔比1:1的比例混合,通过显微注射导入小鼠受精原核,最后获得了11个品系的转基因小鼠.对其中10个品系的小鼠的整合分析表明,所注射的两个DNA片段均发生了染色体外同源重组,而且除了 1个品系的小鼠丢失了大约1kb的序列外,其余品系小鼠的重组产物与预想的结构符合.在当代和后代的转基因小鼠乳汁中均可测到氯霉素乙酰化酶的活性.这表明融合基因在转基因小鼠乳腺中得到表达和分泌,也说明显微共注射两个相互重叠的基因片段是建立转基因动物的一个可行途径.     

The direct effect of estrogen on cell viability and apoptosis in human gastric cancer cells

Epidemiology researches indicated that gastric cancer is a male-predominant disease; both expression level of estrogen and expression pattern of estrogen receptors (ERs) influence its carcinogenesis. But the direct effect of estrogen on gastric cancer cells is still unclear. This study aimed to explore the direct effect of β-estradiol (E2) on gastric cancer cells. SGC7901 and BGC823 were treated with a serial of concentrations of E2. The survival rates of both the cell lines were significantly reduced, and the reduction of viability was due to apoptosis triggered by E2 treatment. Caspase 3 was activated in response to the increasing E2 concentration in both SGC7901 and BGC823. Cleaved Caspase 3 fragments were detected, and the expression levels of Bcl-2 and Bcl-xL were reduced. Apoptosis was further confirmed by flow cytometry. The expression level of PEG10, an androgen receptor target gene, was reduced during E2 treatment. Both ERα and ERβ were expressed in these cell lines, and the result of bioinformatics analysis of gastric cancer from GEO datasets indicated that the expression levels of both ERα and ERβ were significantly higher in noncancerous gastric tissues than in gastric cancer tissues. Our research indicated that estrogen can reduce cell viability and promote apoptosis in gastric cancer cells directly; ERs expression level is associated with gastric cancer. Our research will help to understand the mechanism of gender disparity in gastric cancer.     

Analysis of ALS5 and ALS6 allelic variability in a geographically diverse collection of Candida albicans isolates

The Candida albicans ALS (agglutinin-like sequence) gene family encodes eight cell-surface glycoproteins, some of which function in adhesion to host surfaces. ALS genes have a central tandem repeat-encoding domain comprised entirely of head-to-tail copies of a conserved 108-bp sequence. The number of copies of the tandemly repeated sequence varies between C. albicans strains and often between alleles within the same strain. Because ALS alleles can encode different-sized proteins that may have different functional characteristics, defining the range of allelic variability is important. Genomic DNA from C. albicans strains representing the major genetic clades was PCR amplified to determine the number of tandemly repeated sequence copies within the ALS5 and ALS6 central domain. ALS5 alleles had 2-10 tandem repeat sequence copies (mean=4.82 copies) while ALS6 alleles had 2-8 copies (mean=4.00 copies). Despite this variability, tandem repeat copy number was stable in C. albicans strains passaged for 3000 generations. Prevalent alleles and allelic distributions varied among the clades for ALS5 and ALS6. Overall, ALS6 exhibited less variability than ALS5. ALS5 deletions can occur naturally in C. albicans via direct repeats flanking the ALS5 locus. Deletion of both ALS5 alleles was associated particularly with clades III and SA. ALS5 exhibited allelic polymorphisms in the coding region 5' of the tandem repeats; some alleles resembled ALS1, suggesting recombination between these contiguous loci. Natural deletion of ALS5 and the sequence variation within its coding region suggest relaxed selective pressure on this locus, and that Als5p function may be dispensable in C. albicans or redundant within the Als family.     

Chitosan enhanced gene delivery of cationic liposome via non-covalent conjugation

Two new types of stable ternary complexes were formed by mixing chitosan with DOTAP/pDNA lipoplex and DOTAP with chitosan/pDNA polyplex via non-covalent conjugation for the efficient delivery of plasmid DNA. They were characterized by atomic force microscopy, gel retarding, and dynamic light scattering. The DOTAP/CTS/pDNA complexes were in compacted spheroids and irregular lump of larger aggregates in structure, while the short rod- and toroid-like and donut shapes were found in CTS/DOTAP/pDNA complexes. The transfection efficiency of the lipopolyplexes showed higher GFP gene expression than DOTAP/pDNA and CTS/pDNA controls in Hep-2 and Hela cells, and luciferase gene expression 2–3-fold than DOTAP/pDNA control and 70–120-fold than CTS/pDNA control in Hep-2 cells. The intracellular trafficking was examined by confocal laser scanning microscopy. Rapid pDNA delivery to the nucleus enchanced by chitosan was achieved after 4 h transfection.     

长非编码RNA THOR在肿瘤中的作用

长非编码RNA（long non-coding RNA,lncRNA）是一类长度超过200 nt并且缺乏蛋白质编码潜能的RNA分子。最初lncRNA被认为是由RNA聚合酶Ⅱ转录的副产物,且无生物学功能。随着转录组测序技术的发展,大规模的lncRNA被鉴定出来。越来越多的证据表明,lncRNA参与多种生物学过程,包括基因印记、基因组重排、染色质修饰、细胞周期调控、转录、剪接、mRNA降解和翻译。lncRNA异常表达与人类多种疾病相关,尤其是增生性疾病,包括胃癌、肝癌和直肠癌等。其中,睾丸相关的高度保守的致癌长非编码RNA（testis-associated highly-conserved oncogenic long non-coding RNA,THOR）是一种非常保守的非编码RNA,在睾丸中特异性表达,并广泛存在于人的多种肿瘤组织中,如肝癌、胃癌、鼻咽癌、肾细胞癌、骨肉瘤、视网膜母细胞瘤、黑色素瘤、非小细胞肺癌和舌鳞状细胞癌中,在其发生和发展过程中发挥重要作用。在鼻咽癌、肾细胞癌、骨肉瘤、黑色素瘤、非小细胞肺癌和舌鳞状细胞癌中,THOR主要通过与胰岛素样生长因子2 mRNA结合蛋白1（insulin-like growth factor 2 mRNA-binding protein 1,IGF2BP1）相互作用,促进肿瘤细胞的增殖。在肝癌中,THOR分别通过PTEN/AKT和β-联蛋白信号促进癌细胞的增殖和肝肿瘤干细胞的扩增。在胃癌和骨肉瘤中,THOR主要通过提高SOX9的表达增强癌细胞的干性。在视网膜母细胞瘤中,THOR主要通过提高c-myc的表达促进癌细胞增殖。在鼻咽癌中,THOR主要通过提高YAP的表达增强癌细胞的干性。     

The long-distance migration of Nilaparvata lugens (Stål) (Delphacidae) in China: radar observations of mass return flight in the autumn

Abstract.

• 1 A case study is presented of the autumn migration of the brown planthopper, Nilaparvata lugens (Stål), in the area of Nanjing in the People's Republic of China. The study was made using a high frequency (8 mm wavelength) radar and a net suspended from a kytoon.
• 2 The observations confirmed that long-distance return migrations occur in China in mid and late September, with N.lugens being carried on the prevailing north-easterly wind towards the autumn infestation and overwintering areas of the species.
• 3 After mass take-off in the late afternoon or at dusk, the migrants flew for several hours during the evening, often in a dense layer which formed at heights between about 400 and 1000m above ground. These layers often had well-defined ceilings corresponding to an air temperature of about 16°C. The migration height was above the top of the surface temperature inversion, i.e. the migrants did not fly at the height of the warmest air.
• 4 The dense layer concentrations overflying the radar were backtracked to source areas up to 240 km away in the north-east of Jiangsu Province. Planthoppers observed emigrating from the Nanjing area would reach areas in south Anhui Province or north Jiangxi Province if they flew for 12 h.
• 5 There was a second period of mass take-off at dawn. Insect layers sometimes formed but did not last longer than 1–2h.
• 6 The present results were strikingly different from those previously observed in the dry season in the Philippines, where migratory flight durations were largely confined to periods of about 30min at dusk and dawn.
• 7 Our observations are discussed in relation to the equator-wards return migrations undertaken in autumn by other insect species, and the importance of these migrations for the maintenance of long-flying genotypes in the overwintering populations is considered.

     

荒漠土壤微生物量碳、氮变化对降水的响应

以腾格里沙漠东南缘的典型荒漠植被为研究对象,通过遮雨棚和滴灌系统设置5个降水梯度,即极端干旱处理、中度干旱处理、对照、增水处理I和增水处理II,研究了荒漠土壤微生物量碳(MBC)、氮(MBN)和微生物碳氮比(MBC/MBN)对季节、降水和土壤深度的响应规律,以期为极端降水事件影响干旱荒漠区土壤微生物量碳、氮及其循环规律的深入研究提供科学依据。结果表明:(1)MBC、MBN和MBC/MBN对降水处理的响应存在差异,三者的变化范围为:230.14—272.87 mg/kg,13.82—17.58 mg/kg,19.78—36.06。其中,降水处理对MBC、MBN的影响显著,对MBC/MBN的影响不显著,在极端干旱处理下,MBC、MBN均显著高于其他降水处理;(2)两年间的MBC、MBN和MBC/MBN差异显著,2017年较2016年MBC、MNB显著减少,MBC/MBN显著增加;(3)MBC、MBN和MBC/MBN变化均表现季节性差异,变化范围分别为:153.31—337.09 mg/kg,7.89—22.29 mg/kg,14.82—46.04,其中MBC、MBN为春季最高、秋季最低,M...