Coordinated mRNA and Protein Expression of Human LAMP-1 in Induction of Melanogenesis After UV-B Exposure and Co-Transfection of Human Tyrosinase and TRP-1 cDNAs

In order to better understand the cascade of melanogenic events in melanocytes, this report has introduced our two recent approaches for the expression of melanogenesis/or melanosome-associated genes and encoded proteins in melanocytes (melanoma cells) after repeated exposure to UV -B and after cotransfection of two human genes, i.e., tyrosinase and tyrosinase-related protein-1 (TRP-1). Repeated exposure of UV B (2.5–5.0 mJ/cm²) caused not only upregulation of tyrosinase and TRP-1 genes but also coordinated increase in the gene and protein synthesis expression of Lamp-1 (lysosome-associated membrane protein-1). When COS-7 kidney cells and amelanotic melanoma (C32 and SKMEL-24) and melanotic melanoma (G361 and SK-MEL-23) cells were exposed to cotransfection of human tyrosinase and TRP-1 cDNAs, there was also an increased expression of Lamp-1 mRNA and protein along with tyrosinase activation and new melanin synthesis. Importantly, single transfectants of human tyrosinase cDNA revealed marked cellular degeneration, whereas this degeneration was not seen in single transfectants of TRP-1 cDNA or cotransfectants of human tyrosinase and TRP-1 cDNAs, indicating that TRP-1 prevented, along with Lamp-1, programmed death of melanocytes after transfection of tyrosinase gene. The coordinated expression of TRP-1 and Lamp-1 was further confirmed by antisense oligodeoxynucleotide hybridization experiment against Lamp-1 gene, showing the decreased expression of TRP-1 as identified by three different types of anti-TRP-1 monoclonal antibodies. We propose therefore that human tyrosinase and TRP-l, when activated or expressed together, will coordinate to upregulate the mRNA expression and protein synthesis of Lamp-1. The Lamp-1 molecules will, in turn, cover the inner surface of melanosomal membrane, together with TRP-1 molecules, thus protecting the melanosomal membrane from toxic melanin intermediates generated during melanogenesis in the presence of active tyrosinase. In contrast, the expression of other lysosome-related proteins, e.g., β-galactosidase and CD63 is not stimulated in new melanogenesis.     

A stage—specific protein factor binding to a CACCC motif in both human β—globin gene promoter and 5‘—HS2 region

The DNaseI hypersensitive site 2 (HS2) of human β-globin locus control region(LCR) is required for the high level expression of human β-globin genes.In the present study,a stage-specific protein factor (LPF-β) was identified in the nuclear extract prepared from mouse fetal liver at d 18 of gestation,which could bind to the HS2 region of human β-globin LCR.We also found that the shift band of LPF-β factor could be competed by human β-globin promoter.However,it couldn‘t be competed by human ε-globin promoter or by human ^Aγ-globin promoter.Furthermore,our data demonstrated that the binding-sequence of LPF-β factor is 5‘CACACCCTA 3‘,which is located at the HS2 region of β-LCR(from-10845 to-10853 bp)and human β-globin promoter(from-92 to -84 bp).We speculated that these regions containing the CACCC box in both the human β-globin promoter and HS2 might function as stage selector elements in the regulation of human β-globin switching and the LPF-β factor might be a stage-specific protein factor involved in the regulation of human β-globin gene expression.     

中国喀斯特地区微生物群落结构和功能的研究进展

频繁的人类生产活动使植被遭到破坏,造成基岩裸露的石漠化现象,严重制约了喀斯特地区自然和社会经济的发展,随着生态修复工程的大力开展,微生物群落的结构和功能在生态修复中逐渐受到重视,因为微生物作为喀斯特生态系统的重要组成部分,不仅在物质循环过程中起着重要作用,也在喀斯特生态系统修复中占有十分重要的地位。所以微生物群落结构和功能的研究可以作为衡量生态系统稳定性的重要指标。就中国喀斯特地区的典型植被恢复的不同阶段、成土过程、不同的土地利用方式、矿山修复过程以及不同水域中的微生物群落结构及功能等方面的研究状况进行系统梳理,结合实例综合论述喀斯特地区生态修复过程中微生物作用的研究进展,以期为喀斯特地区生态修复提供参考。     

利用饵料微藻表达抗菌肽及其初步应用

为了解抗菌肽在饵料微藻中表达后的抗菌特性,构建海洋微拟球藻(Nannochloropsis oceanica)、湖泊微拟球藻(N.limnetica)和三角褐指藻(Phaeodactylum tricornutum)的抗菌肽(源自虹鳟,Cath-1a)表达质粒,分别转化相应的微藻,检测转化子中抗菌肽的表达量和体外抑菌效果,将藻株作为鱼饲料添加剂喂食斑马鱼,初步分析了抗菌肽及藻体自身的岩藻黄素和多不饱和脂肪酸对鱼免疫系统的影响。结果表明,外源抗菌肽在3种微藻中均可以成功表达,体外抑菌试验表明,仅三角褐指藻对水产领域常见致病菌爱德华氏菌(Edwardsiella tarda)有一定的抑菌效果,然而抗菌肽的表达并未使3种藻株的体外抑菌性增加。添加藻粉对斑马鱼的生长无明显影响,通过检测鱼体肝脏中与抗氧化和免疫相关基因的表达水平及丙二醛的含量,表明添加藻粉可增强斑马鱼的抗氧化和抗炎症能力,表达抗菌肽(PtC组)能进一步提高斑马鱼的免疫力。另外,添加Pt6(富含岩藻黄素)藻粉组比添加PtC的抗炎效果更显著,表明三角褐指藻中的岩藻黄素和二十碳五烯酸对增强鱼的抗病能力具有潜在作用。     

遮阴对石笔木苗木生长和保护酶活性的影响

为探讨石笔木Tutcheria championi苗木对遮阴环境的适应性,以全光照(无遮阴覆盖)为对照,研究遮阴30%、50%和70%对2年生石笔木苗生长和生理特性的影响。结果表明,全光照下石笔木苗木生长较为健壮,其苗高、地径生长优于遮阴处理;全光照下石笔木总生物量、根生物量比以及根冠比显著大于遮阴处理(P<0.05);50%遮阴处理苗木叶片叶绿素含量显著高于全光照和70%遮阴处理(P<0.05),叶绿素a/b值在各处理间无显著差异。丙二醛(MDA)含量、超氧化物歧化酶(SOD)活性在各处理间无显著差异(P>0.05);70%遮阴处理的苗木叶片过氧化氢酶(CAT)活性最高,显著高于全光照和50%遮阴。由此表明,充足的光照环境有利于石笔木苗木生长。     

了哥王叶功能性状特征及其对土壤因子的响应

为探究了哥王Wikstroemiaindica的叶功能性状特征及其影响因素,在海岛植被调查的基础上对了哥王叶片进行取样并测定其功能性状指标,利用变异系数法和Pearson相关性分析探讨叶功能性状之间的差异与联系,运用冗余分析研究了哥王叶功能性状对土壤因子的响应。结果表明,了哥王的叶功能性状变异系数介于9.76%～23.73%,其中叶体积变异幅度最大(23.73%),叶干物质含量变异幅度最小(9.76%),整体上了哥王叶功能性状保持相对稳定。了哥王各项叶功能性状之间具有一定的相关性,联系较密切。了哥王叶功能性状主要受土壤中有机质、全氮、碱解氮的影响,土壤中有机质、全氮、碱解氮的含量与比叶面积呈正比,与叶厚度、叶体积成反比。了哥王的叶片可以通过一定的性状变异和组合来适应外部环境的变化,以较好地适应海岛恶劣的环境。该研究结果可为了哥王野生种质资源的保护、利用以及人工栽培提供参考。     

钾通道阻断剂所致的蜗内直流电位改变

本实验采用耳蜗外淋巴灌流技术,观察了四氨基吡啶(4-AP)、四乙基铵(TEA)以及奎宁等不同钾通道阻断剂对豚鼠蜗内直流电位(EP)的影响。发现快钾通道阻断剂4-AP对EP无明显影响,但可改变强噪声所致EP改变的形式。TEA与奎宁则可减少负相EP(N-EP)的绝对值。实验结果提示耳蜗内有不同类型的钾离子通道存在并各具有不同的生理意义。     

缺氧与复氧对不同血管α_1肾上腺素受体收缩效应的影响

本实验观察离体大鼠肾动脉(含α_(1V)亚型)、主动脉(含α_(1B)亚型)和肺动脉(含α_(1V)和α_(1B)亚型)在缺氧与复氧时由α_1肾上腺素受体激动所致收缩效应的改变。结果显示肾动脉在缺氧时pD值增大,最大收缩效应降低,复氧时pD值恢复;主动脉在缺氧时pD值减小,最大收缩效应不变,复氧时pD值不恢复:而肺动脉的改变为上述两者的综合。提示σ_1受体不同亚型在缺氧与复氧时发生的变化不同,这可能是不同血管对缺氧、复氧反应不同的机制之一。     

前庭—交感反应的中枢通路分析

用锋电位触发叠加(STA)技术对大鼠前庭内侧核(NVM)、下段脑干网状结构(RF)和前庭小脑进行了探查,观察了这些结构中对摆动旋转起反应的单位(PPU)与内脏大神经(SN)记录的交感反应之间的关系。发现用NVM的PPU进行STA,在SN可得一阳性反应,其潜伏期为33.28±3.1ms;用下段脑干RF的PPU进行STA,SN的峰反应潜伏期为11.3±0.91ms;用前庭小脑的PPU进行STA,SN的峰反应潜伏期为21.86±1.73ms。本结果提示前庭交感反应的最近的脊髓上中继站是下段脑干RF旁正中区核团,其下行冲动可能是由网状脊髓束中慢速传导的纤维传导的。根据三个脑区PPU引起的SN-STA潜伏期的不同,在前庭交感反应中前庭小脑所处的地位可能在NVM和下段脑干RF核团之间。