雌激素Beta受体在大鼠脑内表达的免疫组化定位研究

为了探讨雌激素作用于神经系统的机理,采用硫酸镍铵增强显色的免疫组化SP法研究了新的雌激素受体(ER-β)在成年雌雄大鼠脑内的分布。研究证实ER-β免疫阳性物质主要位于神经元的细胞核内,但在个别脑区也可在胞浆甚至突起内检测到。最强的ER-β免疫阳性信号见于前嗅核、大脑皮质、小脑浦肯野细胞、斜角带垂直部、蓝斑和三叉神经运动核等部位;中等强度的染色见于隔内侧核、杏仁外侧核、黑质、中央灰质等部位;较弱的阳性反应见于下丘脑与杏仁复合体的部分核团。在一些部位还存在表达水平甚至细胞内定位模式的性别差异,如前庭上核内的表达只见于雌性;雄性大鼠三叉神经运动核内ER-β蛋白主要表达于胞浆内,细胞核为阴性;而在雌性大鼠该部位ER-β蛋白主要位于细胞核等。以上结果表明ER-β蛋白在大鼠脑内分布广泛并具有一定的性别差异,在与学习记忆有关的脑区如大脑皮质和基底前脑内有很高的表达,提示在脑组织内雌激素可能通过ER-β这一新的信号途径发挥多种重要的调控作用,如学习记忆等。     

大型绿藻浒苔转化表达系统选择标记的筛选

主要研究了条浒苔对抗生素氯霉素和除草剂Basta的敏感性,以确定适合的阳性选择标记基因。应用不同浓度氯霉素(0、25、50、75、100、125μg/ml)和Basta(0、5、12.5、25、37.5、50μg/ml)对不同发育时期条浒苔细胞存活率影响进行了测定。实验结果表明:不同发育时期条浒苔对氯霉素和Basta的敏感性不同。其中最大浓度125μg/ml浓度的氯霉素在15d内对条浒苔孢子和小苗两个不同发育时期的细胞均难以达到全部杀死效果,相对存活率仍分别为1%和20%;而Basta对条浒苔孢子和小苗均具有很强的杀生作用,其中5μg/ml浓度的Basta在3d内可将条浒苔孢子全部杀死,12.5μg/ml浓度下约一周时间可以将浒苔小苗全部致死。本实验结果提示bar基因有可能成为浒苔基因工程较理想的选择标记基因。     

不同土壤水分含量下高寒草地CH4释放的比较研究

2003年6月30日～9月4日,利用密闭箱-气相色谱法,对发育于不同水分状况下的灌丛草甸(GC)、矮嵩草草甸(AC)、藏嵩草草甸(ZC)和季节性湿地(SD)的CH4释放速率进行了比较研究.结果表明,观测期间,季节性湿地处于淹水状态,其它三种土壤平均水分含量分别为39.6%(GC)、38.4%(AC)、65.9%(ZC),而CH4平均释放速率分别为-0.031±0.030(GC)、-0.026±0.018(AC)、1.103±0.240(ZC)和6.922±4.598 mg·m-2·h-1(SD),随着土壤水分含量的增加,高寒草地土壤CH4释放由吸收转为排放,表现出与土壤湿度很好的一致性.矮嵩草草甸不同处理CH4吸收强度AC＜AJ＜AL,它们之间的差异除与土壤水分有关,还可能与处理引起的CH4传输途径不同有关.实验期间,矮嵩草草甸和灌丛草甸土壤-植物系统分别吸收CH438.69和46.13 mg·m-2,是大气温室气体CH4的弱汇,藏嵩草草甸和季节性湿地则是大气温室气体CH4的源,分别排放CH4 1.641和10.30 g·m-2.     

Expression of β-carotene hydroxylase gene (<Emphasis Type="Italic">crtR-B</Emphasis>) from the green alga <Emphasis Type="Italic">Haematococcus pluvialis</Emphasis> in chloroplasts of <Emphasis Type="Italic">Chlamydomonas reinhardtii</Emphasis>

A carotenoid gene (crtR-B) from the green alga Haematococcus pluvialis, encoding β-carotene hydroxylase that was able to catalyze the conversion of β-carotene to zeaxanthin and canthaxanthin to astaxanthin, was cloned into Chlamydomonas reinhardtii chloroplast expression vector p64D to yield plasmid p64DcrtR-B. The vector p64DcrtR-B was transferred to the chloroplast genome of C. reinhardtii using micro-particle bombardment. PCR and Southern blot analyses indicated that crtR-B was integrated into the chloroplast genome of the transformants. RT-PCR assays showed that the H. pluvialis crt R-B gene was expressed in C. reinhardtii transformants. The transformants rapidly synthesized carotenoids in larger quantities than the wild-type upon being transferred from moderate to high-intensity white light. This research provides a foundation for further study to elucidate the possible mechanism of photo-protection by xanthophylls and other carotenoids in high light conditions or through exposure to UV radiation.     

Phage displayed peptides to avian H5N1 virus distinguished the virus from other viruses

The purpose of the current study was to identify potential ligands and develop a novel diagnostic test to highly pathogenic avian influenza A virus (HPAI), subtype H5N1 viruses using phage display technology. The H5N1 viruses were used as an immobilized target in a biopanning process using a 12-mer phage display random peptide library. After five rounds of panning, three phages expressing peptides HAWDPIPARDPF, AAWHLIVALAPN or ATSHLHVRLPSK had a specific binding activity to H5N1 viruses were isolated. Putative binding motifs to H5N1 viruses were identified by DNA sequencing. In terms of the minimum quantity of viruses, the phage-based ELISA was better than antiserum-based ELISA and a manual, semi-quantitative endpoint RT-PCR for detecting H5N1 viruses. More importantly, the selected phages bearing the specific peptides to H5N1 viruses were capable of differentiating this virus from other avian viruses in enzyme-linked immunosorbent assays.     

FBXW7 suppresses epithelial‐mesenchymal transition and chemo‐resistance of non‐small‐cell lung cancer cells by targeting snai1 for ubiquitin‐dependent degradation

Objectives

FBXW7 acts as a tumour suppressor by targeting at various oncoproteins for ubiquitin‐mediated degradation. However, the clinical significance and the involving regulatory mechanisms of FBXW7 manipulation of NSCLC regeneration and therapy response are not clear.

Materials and Methods

Immunohistochemical staining and qRT‐PCR were applied to detect FBXW7 and Snai1 expression in 100 samples of NSCLC and matched tumour‐adjacent tissues. FBXW7 manipulation of cancer biological functions were studied by using MTT assay, immunoblotting, flow cytometry, transwells, wound healing assay, and sphere‐formation assays. Immunofluorescence and co‐immunoprecipitation were used to analyse the possible interaction between Snai1 and FBXW7.

Results

We detected the decreased FBXW7 expression in majority of the NSCLC tissues, and lower FBXW7 level was correlated with advanced TNM stage. Furthermore, those patients with decreased FBXW7 expression tend to have both poorer 5‐year survival outcomes, and shorter disease‐free survival, comparing to those with higher FBXW7 levels. Functionally, we found that FBXW7 enforcement suppressed NSCLC progression by inducing cell growth arrest, increasing chemo‐sensitivity and inhibiting Epithelial‐mesenchymal Transition (EMT) progress. Results further showed that FBXW7 could interact with Snai1 directly to degrade its expression through ubiquitylating alternation in NSCLC, which could be partially abrogated by restoring Snai1 expression.

Conclusions

FBXW7 conduction of tumour suppression was partly through degrading Snai1 directly for ubiquitylating regulation in NSCLC

     

昆虫体内的天然屏障——围食膜

昆虫围食膜是由昆虫中肠上皮细胞分泌的非细胞薄膜状结构,主要成份是几丁质、蛋白质和多糖,是昆虫抵御外界侵害的第一道天然屏障,能够保护中肠上皮细胞不受机械损伤并且能够抵御病毒、细菌及其他有害物质,防止化学损伤.昆虫病毒增效蛋白、荧光增白剂和几丁质酶等生物防治促进因子通过与围食膜上特异位点的结合,能够破坏围食膜结构,加速病原微生物对害虫的感染进程.就围食膜组分、结构、功能以及与害虫防治的关系等方面的研究进展进行综述,并且论述了以围食膜为害虫生物防治靶标的应用前景.     

Interleukin-33/ST2 signaling promotes production of interleukin-6 and interleukin-8 in systemic inflammation in cigarette smoke-induced chronic obstructive pulmonary disease mice

Interleukin-33 is a newly described member of the interleukin-1 family. Recent research suggests that IL-33 is increased in lungs and plays a critical role in chronic airway inflammation in cigarette smoke-induced chronic obstructive pulmonary disease (COPD) mice. To determine the role of IL-33 in systemic inflammation, we induced COPD mice models by passive cigarette smoking and identified the IL-33 expression in bronchial endothelial cells and peripheral blood mononuclear cells (PBMCs) of them. After isolation, PBMCs were cultured and stimulated in vitro. We measured expressions of interleukin-6 and interleukin-8 in PBMCs in different groups. The expression of IL-33 in bronchial endothelial cells and PBMCs of COPD mice were highly expressed. Stimulated by cigarette smoke extract (CSE), the expression of IL-6 and IL-8 were induced and enhanced by IL-33. PBMCs of COPD mice produced more IL-6 and IL-8 stimulated by CSE and IL-33. Expression of IL-6 and IL-8 were decreased when stimulated by IL-33 together with soluble ST2. The mRNA production of ST2 in IL-33 stimulated PBMCs was increased. Being pretreated with several kinds of MAPK inhibitors, the secretions of IL-6 and IL-8 in PBMCs did not decrease except for the p38 MAPK inhibitor. We found that IL-33 could induce and enhance the expression of IL-6 and IL-8 in PBMCs of COPD mice via p38 MAPK pathway, and it is a promoter of the IL-6 and IL-8 production in systemic inflammation in COPD mice.     

山楂叶螨、苹果全爪螨及其捕食性天敌生态位的研究Ⅰ——时间与空间生态位

统计分析表明两种叶螨间生态位重叠值较大、各自生态位宽度较大。两种叶螨的时动空间生态位与各自种群消长曲线相一致,显示了它们对资源有相似要求以及“逐步扩散整树危害”的取食危害方式。它们的生态位分离表现在叶片正、反面及叶片反面的某些小“领域”上。苹果全爪螨更喜欢分布在树冠上层和外层。统计和模糊分析都表明草蛉和小花螨与两种叶螨的时间同步和空间同域性较强,捕食螨与叶螨的时间生态位重叠较小,而空间生态位重叠较大。天敌与叶螨间的时空生态位重叠以及天敌食谱生态位宽度结合分析有助于合理评价天敌的控制作用。