W86F/W86A突变对细菌视紫红质光循环影响的差异性分析

细菌视紫红质(bacteriorhodopsin,bR)是一种光驱受体蛋白,每个活性单元由3个单体组成,每个单体由一分子视蛋白和一分子的视黄醛发色团共价结合而成,其功能为从胞内向胞外定向传输质子,利用形成的质子浓度梯度将光能转化为化学能.光照后视黄醛发色团构型发生all-trans向13-cis转变,蛋白的构象也随之发生了一系列具有稳定中间态的变化并驱动质子的定向传递.为探讨bR视黄醛键合区保守性氨基酸色氨酸86 (Tryptop-han86,w86)对其光循环中间态和质子泵功能的影响,本研究采用定点突变技术将W86突变为侧链大小不同的F86和A86,通过原位紫外-可见光吸收光谱、闪光光解光谱、pH滴定、固体核磁共振等技术手段,探究W86F和W86A突变对bR光循环及质子泵功能影响差异性的分子机制.结果 表明,W86F和W86A突变均造成了bR暗适应状态下视黄醛顺反异构平衡向顺式构型占优的方向移动,且W86F突变可造成反式构型的完全消失.此外,无论是W86F还是W86A突变都造成了蛋白光循环中间态的减弱且衰减延长,以及质子泵功能减弱,但影响机制各有所不同,这可能与这两个突变对视黄醛多烯链上电子云的分布以及周围残基造成的扰动程度不同有关.     

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.     

传染性单核细胞增多症患者外周血T细胞亚群检测及临床意义

目的检测传染性单核细胞增多症(IM)患者及非本病的发热患者外周血T淋巴细胞亚群,探讨EB病毒感染导致的传染性单核细胞增多症的免疫反应机制及与非本病发热患者的鉴别诊断价值,同时通过检测患者血液中的EBV-DNA载量加以确证。方法选取确诊为传染性单核细胞增多症的患者30例(IM组),非传染性单核细胞增多症的发热患者30例(对照组)。应用流式细胞技术检测患者外周血中淋巴细胞亚群,同时应用荧光定量PCR技术检测血清EBV-DNA载量,采用卡方检验及Mann-Whitney检验进行组间差异分析。结果 IM组与对照组患者T淋巴细胞亚群相比,CD8~+T淋巴细胞所占比例显著升高(Z=1.776,P0.001),CD4~+T淋巴细胞所占比例降低(F=4.008,P0.050),同时CD4~+/CD8~+比例明显下降(Z=6.653,P0.001)。IM组EBV-DNA病毒载量显著高于对照组,差异具有统计学意义(P0.050)。结论传染性单核细胞增多症患者外周血T淋巴细胞亚群主要以CD4~+T淋巴细胞及CD8~+T淋巴细胞的变化为主,T淋巴细胞亚群结合血清EBV-DNA病毒载量的联合检测对IM及非本病的发热患者具有鉴别诊断价值。     

MicroRNA-520c-3p suppresses vascular endothelium dysfunction by targeting RELA and regulating the AKT and NF-κB signaling pathways

Journal of Physiology and Biochemistry - Endothelial injury, which can cause endothelial inflammation and dysfunction, is an important mechanism for the development of atherosclerotic plaque. This...     

中华蜜蜂AcNPC2b的原核表达和鉴定

[目的]中华蜜蜂Apis cerana cerana(简称中蜂),其胞内胆固醇转运体AcNPC2a能够识别并结合几种微生物的细胞壁,AcNPC2存在于该蜂触角,参与嗅觉通讯,关于AcNPC2b的功能尚未见报道.[方法]通过对中华蜜蜂AcNPC2b基因进行原核表达,以纯化的重组蛋白免疫小鼠,制备多克隆抗血清;利用荧光定量PCR和免疫印迹检测AcNPC2b的转录活性及蛋白水平,通过微生物结合测定对AcNPC2b的功能进行探索.[结果]中蜂AcNPC2b含有信号肽和ML结构域,具有3个二硫键,与果蝇Drosophila melanogaster DmNPC2b聚为一个亚枝.荧光定量qRT-PCR检测发现,感染中蜂囊状幼虫病毒(Chinses sacbrood virus,简称CSBV)的幼虫体内AcNPC2b基因的表达呈现出先微弱下调再持续上调的趋势.获得了约16ku的重组蛋白AcNPC2b并制备了多克隆抗体,免疫印迹结果表明,中蜂AcNPC2b蛋白在工蜂头、胸与中肠中的表达量最高,触角、马氏管与脂肪体中次之.AcNPC2b蛋白在正常与感染CSBV病毒的中蜂幼虫中均有表达,且在感染CSBV的中蜂幼虫中有微弱上调.重组蛋白AcNPC2b微生物结合测定实验结果显示中蜂AcNPC2b重组蛋白均能与活的大肠杆菌、金黄色葡萄球菌、芽孢杆菌以及白僵菌结合.[结论]正常与感染CSBV病毒的中蜂幼虫体内AcNPC2b基因转录水平和AcNPC2b蛋白的表达含量存在差异,重组蛋白AcNPC2b具有识别并结合病原菌的能力,为后续深入研究AcNPC2b蛋白的分子功能奠定了一定理论基础.     

Construction of an Immunostimulatory Plasmid, pUCpGs10, and Research on its Immune Adjuvant Effect

In order to overcome the instability of CpG ODN in vivo, sequence diversity, and individual differences, eleven CpG ODN fragments were meticulously selected and linked to form a Multi-CpG, which were repeatedly inserted into the cloning vector pUC19 for constructing the recombinant plasmid pUCpGs10 containing ten of Multi-CpG. Using the multi-genotype HCV E1 and multi-epitope complex HCV-T as immunogens, and plasmid pUCpGs10 as the immune adjuvant, Balb/c mice were immunized through nasal and subcutaneous immunization. Strong-specific humoral and cellular immune response were induced, which can obviously inhibit the growth of homograft expressing HCV antigen. The immune adjuvant effect of pUCpGs10 closely matched that of Freund’s complete adjuvant. The plasmid pUCpGs10 can significantly improve IgA content in serum and different mucosal extract and systematical T-cell response via intranasal immunization. In conclusions, the newly constructed immunostimulatory plasmid pUCpGs10 is able to effectively activate the humoral and cellular immune activity, and possesses activation on mucosal immune response.     

Alkyl Polyglucoside (APG) Amendment for Improving the Phytoremediation of Pb-PAH Contaminated Soil by the AquaticPlant Scirpus triqueter

This study assessed the potential abilities of Scirpus triqueter for phytoremediation of soils contaminated with Pb-PAHs, amended with environment-friendly surfactant alkyl polyglucoside (APG). The effects of APG on the removal of PAHs from soil and the plant uptake and translocation of Pb were tested with plant growth and soil enzymatic activities. Experiments demonstrated that APG has an ability to facilitate PAH degradation and Pb uptake in the plant body at appropriate concentrations (20–40mg L^?1). The highest PAH removal rate was observed in 30 mg L^?1 APG treatment, and the highest accumulation of Pb was detected as 40 mg L^?1 APG. Experiments documented the effects of APG on plant growth, soil enzymatic activity, bioaccumulation and translocation of Pb in Scirpus triqueter. Results indicated that the addition of appropriate APG enhanced PAH removal rate and increased plant uptake and translocation of Pb.     

Mouse arylamine N-acetyltransferase 2 (Nat2) expression during embryogenesis: a potential marker for the developing neuroendocrine system

Arylamine N-acetyltransferase (NAT) genes in humans and in rodents encode polymorphic drug metabolizing enzymes. Human NAT1 (and the murine equivalent mouse Nat2) is found early in embryonic development and is likely to have an endogenous role. We report the detailed expression of the murine gene (Nat2) and encoded protein in mouse embryos, using a transgenic mouse model bearing a lacZ transgene inserted into the coding region of mouse Nat2. In mouse embryos, the transgene was expressed in sensory epithelia, epithelial placodes giving rise to visceral sensory neurons, the developing pituitary gland, sympathetic chain and urogenital ridge. In Nat2^+/+ mice, the presence and activity of Nat2 protein was detected in these tissues and their adult counterparts. Altered expression of the human orthologue in breast tumours, in which there is endocrine signalling, suggests that human NAT1 should be considered as a potential biomarker for neuroendocrine tissues and tumours.     

Molecular Phylogeny of the Butterfly Genus Polytremis (Hesperiidae,Hesperiinae, Baorini) in China

Background

The genus Polytremis, restricted to the continental part of the southeastern Palaearctic and northern Oriental regions, is one of the largest and most diverse lineages of the tribe Baorini. Previous studies on the genus were focused mainly on morphological classification and identification of new species. Due to the lack of effective and homologous traits of morphology, there were many challenges in the traditional classification. In this report, we reconstruct the phylogeny to provide a solid framework for these studies and to test the traditional limits and relationships of taxa.

Methodology and Principal Findings

We sequenced a mitochondrial and three nuclear gene fragments, coupled with an evaluation of traditional morphological characters, to determine the phylogenetic relationships for a total of 15 species representing all major species groups of the Polytremis genus in China, and to elucidate their taxonomic status.

Conclusions and Significance

Analysis of mitochrondial and nuclear DNA showed considerable congruent phylogenetic signal in topology at the inter-species level. We found strong support for the monophyly of Polytremis and some clades were recognized with morphological data. Thus, the COI sequence in our study could be used as a DNA barcode to identify almost all members of the genus. However, incongruences of phylogenetic analyses occurred: in contrast to the phylogenetic trees of mitochondrial COI, it was not possible for nuclear rDNA to discriminate P. gotama from P. caerulescens, suggesting a possible recent separation of these two species. Additionally, P. theca was the only species with a greater intra-specific genetic distance compared to some inter-specific genetic distances in this study and some problems associated with the cryptic diversity of the species are discussed. The results of this study will helpful to reveal the causes of the high degree of diversity of butterflies, and possibly other groups of insects in China.