非小细胞肺癌患者中EGFR突变、ALK和ROS1融合基因表达变化及其临床病理学意义

目的探讨非小细胞肺癌(non-small cell lung cancer,NSCLC)患者中表皮生长因子受体(epidermic growth factor receptor,EGFR)突变、间变性淋巴瘤激酶(anaplastic lymphoma kinase,ALK)和ROS1融合基因的表达情况及其与临床病理特征的关系。方法应用ARMS法检测379例非小细胞肺癌患者中EGFR突变、ALK和ROS1融合基因的表达情况,并分析其与患者临床病理特征的关系。结果 379例非小细胞肺癌患者组织中,EGFR突变率为36.15%(137/379),19del和L858R突变为其主要突变类型,同时检出L858R和T790双突变4例,L858R和19del双突变2例;EGFR突变人群主要是女性、腺癌、非吸烟患者(P<0.05)。ALK融合基因阳性率为3.43%(13/379),其中ALK-M1融合基因型4例,ALK-M2融合基因型3例,ALK-M3融合基因型3例,ALK-M4融合基因型1例,ALK-M6融合基因型2例。ROS1融合基因阳性率为3.17%(12/379),主要为ROS1-M8融合基因型(8例),存在1例ROS1-M3和ROS1-M8融合基因型双融合。不同性别、年龄、组织学和吸烟状况的NSCLC患者ALK和ROS1基因突变率无统计学差异。结论 EGFR基因在NSCLC患者中存在较高的突变率,而ALK、ROS1融合基因在NSCLC患者中突变率较低,但代表了非小细胞肺癌的特点分子亚型,为指导临床靶向治疗提供依据。     

Reprogramming of Meiotic Chromatin Architecture during Spermatogenesis

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Cosupplementation with a synthetic, lipid-soluble polyphenol and vitamin C inhibits oxidative damage and improves vascular function yet does not inhibit acute renal injury in an animal model of rhabdomyolysis

We investigated whether cosupplementation with synthetic tetra-tert-butyl bisphenol (BP) and vitamin C (Vit C) ameliorated oxidative stress and acute kidney injury (AKI) in an animal model of acute rhabdomyolysis (RM). Rats were divided into groups: Sham and Control (normal chow), and BP (receiving 0.12% w/w BP in the diet; 4 weeks) with or without Vit C (100mg/kg ascorbate in PBS ip at 72, 48, and 24h before RM induction). All animals (except the Sham) were treated with 50% v/v glycerol/PBS (6 mL/kg injected into the hind leg) to induce RM. After 24h, urine, plasma, kidneys, and aortae were harvested. Lipid oxidation (assessed as cholesteryl ester hydroperoxides and hydroxides and F(2)-isoprostanes accumulation) increased in the kidney and plasma and this was coupled with decreased aortic levels of cyclic guanylylmonophosphate (cGMP). In renal tissues, RM stimulated glutathione peroxidase (GPx)-4, superoxide dismutase (SOD)-1/2 and nuclear factor kappa-beta (NFκβ) gene expression and promoted AKI as judged by formation of tubular casts, damaged epithelia, and increased urinary levels of total protein, kidney-injury molecule-1 (KIM-1), and clusterin. Supplementation with BP±Vit C inhibited the two indices of lipid oxidation, down-regulated GPx-4, SOD1/2, and NF-κβ gene responses and restored aortic cGMP, yet renal dysfunction and altered kidney morphology persisted. By contrast, supplementation with Vit C alone inhibited oxidative stress and diminished cast formation and proteinuria, while other plasma and urinary markers of AKI remained elevated. These data indicate that lipid- and water-soluble antioxidants may differ in terms of their therapeutic impact on RM-induced renal dysfunction.     

Revealing genetic diversity of tree peonies at micro-evolution level with hyper-variable chloroplast markers and floral traits

Key message

Highly variable regions of chloroplast genome were found to be useful in the detection of plant genetic diversity at micro-evolution level. Our methodology will improve understanding and conservation of plant diversity.

Abstract

Tree peonies are famous flowers with about 2,000 cultivars in the world, belonging to Paeonia sect. Moutan of the Paeoniaceae. They are traditionally classified based on flower forms and colors. Due to the limited number of DNA and morphological markers, and the existence of synonyms and homonyms, evaluation on genetic diversity of so many cultivars remains a challenge. In most cases, it is difficult and even impossible to discriminate tree peony cultivars when they are not in flower. In this study, single nucleotide polymorphism detected from the hyper-variable regions of chloroplast genome was employed to separate tree peony cultivars into different maternal lineages which can be expressed briefly by a nucleotide molecular formula. Our approach enabled a much higher resolution of cultivar identification and classification that has not been obtained before. The newly developed hyper-variable chloroplast markers, as an independent source of taxonomic characteristics, provided novel evidences and higher resolution ability that are helpful in building an effective classification system for evaluation, conservation, and utilization of the tree peony germplasm resources at cultivar level.     

Thermal tolerance and altitudinal distribution of two Sphenomorphus lizards in Taiwan

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We compared the thermal tolerances of a high-altitude skink, Sphenomorphus taiwanensis with that of the lowland S. incognitus to test whether their thermal tolerances correspond to their altitudinal distributions.     

Modulation of inactivation properties of CaV2.2 channels by 14-3-3 proteins

Inactivation of presynaptic Ca(V)2.2 channels may play a role in regulating short-term synaptic plasticity. Here, we report a direct modulation of Ca(V)2.2 channel inactivation properties by 14-3-3, a family of signaling proteins involved in a wide range of biological processes. The structural elements critical for 14-3-3 binding and channel modulation lie in the carboxyl tail of the pore-forming alpha(1B) subunit, where we have identified two putative 14-3-3 interaction sites, including a phosphoserine-containing motif that directly binds to 14-3-3 and a second region near the EF hand and IQ domain. In transfected tsA 201 cells, 14-3-3 coexpression dramatically slows open-state inactivation and reduces cumulative inactivation of Ca(V)2.2 channels. In hippocampal neurons, interference with 14-3-3 binding accelerates Ca(V)2.2 channel inactivation and enhances short-term synaptic depression. These results demonstrate that 14-3-3 proteins are important regulators of Ca(V)2.2 channel activities and through this mechanism may contribute to their regulation of synaptic transmission and plasticity.     

Alteration of subcellular redox equilibrium and the consequent oxidative modification of nuclear factor kappaB are critical for anticancer cytotoxicity by emodin, a reactive oxygen species-producing agent

We previously found that emodin produced reactive oxygen species (ROS) intracellularly. In various tumor cells at low doses it enhances the cytotoxicity of As₂O₃, and at higher doses it renders cytotoxicity independently in vitro and in vivo. The effects involve redox-mediated inhibition of NF-κB activation. In this study, we focus on the mechanisms by which emodin inhibits NF-κB activation. Results in HeLa cells demonstrated that emodin at high doses or in combination with As₂O₃, via generation of ROS especially in the nucleus, altered subcellular redox equilibrium and thus oxidized the redox-sensitive site on NF-κB and prevented its binding to the target DNA. In vivo study showed that tumors exposed to the arsenic/emodin cotreatment had dramatically smaller sizes and weaker antioxidant capacity, compared with arsenic alone. NF-κB binding and transactivation were inhibited in these tumors. These data help in the understanding of the mechanisms by which manipulation of cellular redox and NF-κB activation may enhance chemotherapy.     

Stable transfection of Eimeria tenella: constitutive expression of the YFP-YFP molecule throughout the life cycle

The obligate intracellular apicomplexan parasite Eimeria tenella, one of seven species of Eimeria that infect chickens, elicits protective cell-mediated immunity against challenge infection. For this reason, recombinant E. tenella parasites could be utilised as an effective vaccine vehicle for expressing foreign antigens and inducing immunity against heterologous intracellular microbes. A stable line of E. tenella expressing foreign genes is a prerequisite, and in this work an in vivo stable transfection system has been developed for this parasite using restriction enzyme-mediated integration (REMI). Two transgenic populations of E. tenella have been obtained that express YFP-YFP constitutively throughout the parasite life cycle. Southern blotting and plasmid rescue analyses show that the introduced exogenous DNA was integrated at random into the parasite genome. Although the life cycle of the transgenic populations was delayed by at least 12 h and the output of oocysts was reduced 4-fold relative to the parental BJ strain of E. tenella, the transgenic parasites were sufficiently immunogenic to protect chickens against challenge with either transgenic or parental parasites. These results are encouraging for the development of transgenic E. tenella as a vaccine vector and for more detailed investigation of the biology of the genus Eimeria.