RG108对肺腺癌A549细胞增殖、凋亡及RASSF1A基因表达的影响

目的探讨DNA甲基转移酶抑制剂RG108对人肺腺癌细胞株A549增殖、凋亡以及对RASSF1A（Ras as-sociation domain proteinfamily1）基因启动子区域甲基化状态、表达的影响。方法用20μmol/L的RG108对A549细胞进行化学干预72h,用MTT法检测细胞生长抑制率;流式细胞术检测细胞周期以及凋亡情况;RT-PCR观察RASSF1A基因mRNA水平变化;Western blot检测RASSF1A蛋白的表达;甲基化特异性PCR（MS-PCR）检测RASSF1A基因启动子区域甲基化状态的改变。结果经RG108干预72h后,A549细胞的抑制率为17.2±0.43%,细胞周期阻滞于G0/G1期,并引起细胞凋亡。RT-PCR和Western blot结果显示在干预组细胞中分别出现RASSF1A基因的DNA条带（329bp）和蛋白质条带（39kD）,而对照组中无相应条带出现。RASSF1A基因启动子区域由甲基化状态转变为非甲基化状态。结论RG108可使RASSF1A基因启动子区域去甲基化,并通过该机制诱导RASSF1A基因在人肺腺癌细胞株A549中表达。     

细菌生物膜的形成与调控机制

细菌通过自身合成的水合多聚物粘附在固体表面,以固着的方式生长从而形成生物膜,细菌生物膜的形成涉及到几个明显的阶段,包括起始的附着、细胞与细胞之间的吸附与增殖、生物膜的成熟、及最后细菌的脱离等四个阶段,生物膜的形成增加了细菌对抗生素的抗性以及帮助细菌逃逸寄主的免疫攻击等,从而引起临床上持续性的慢性感染等各种问题;生物膜结构非常复杂,除了细菌分泌的各种胞外多糖,胞外蛋白质外,最新的研究表明,DNA也是生物膜的一个重要成分.针对近年来的最新文献报道分别对生物膜的形成、结构以及调控机制等进行综述.     

Characterization of electrical penetration graphs of the Asian citrus psyllid, Diaphorina citri, in sweet orange seedlings

Detailed information on probing behavior of the Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae), is critical for understanding the transmission process of phloem‐limited bacteria (Candidatus Liberibacter spp.) associated with citrus ‘huanglongbing’ by this vector. In this study, we investigated stylet penetration activities of D. citri on seedlings of Citrus sinensis (L.) Osbeck cv. Pêra (Rutaceae) by using the electrical penetration graph (EPG‐DC system) technique. EPG waveforms were described based on amplitude, frequency, voltage level, and electrical origin of the observed traces during stylet penetration into plant tissues. The main waveforms were correlated with histological observations of salivary sheath termini in plant tissues, to determine the putative location of stylet tips. The behavioral activities were also inferred based on waveform similarities in relation to other Sternorrhyncha, particularly aphids and whiteflies. In addition, we correlated the occurrence of specific waveforms with the acquisition of the phloem‐limited bacterium Ca. Liberibacter asiaticus by D. citri. The occurrence of a G‐like xylem sap ingestion waveform in starved and unstarved psyllids was also compared. By analyzing 8‐h EPGs of adult females, five waveforms were described: (C) salivary sheath secretion and other stylet pathway activities; (D) first contact with phloem (distinct from other waveforms reported for Sternorrhyncha); (E1) putative salivation in phloem sieve tubes; (E2) phloem sap ingestion; and (G) probably xylem sap ingestion. Diaphorina citri initiates a probe with stylet pathway through epidermis and parenchyma (C). Interestingly, no potential drops were observed during the stylet pathway phase, as are usually recorded in aphids and other Sternorrhyncha. Once in C, D. citri shows a higher propensity to return to non‐probing than to start a phloem or xylem phase. Several probes are usually observed before the phloem phase; waveform D is observed upon phloem contact, always immediately followed by E1. After E1, D. citri either returns to pathway activity (C) or starts phloem sap ingestion, which was the longest activity observed.     

父本剥夺对子代棕色田鼠同性间行为及相关脑区神经元活性的影响

为探究父本剥夺对子代棕色田鼠(Microtus mandarinus)同性间行为及相关脑区神经元活性的影响,本实验观察了父本缺失子代棕色田鼠成年后社会互作实验和相关脑区Fos蛋白的表达情况。社会互作实验结果显示,与雄性对照组相比,雄性父本剥夺组田鼠对另一雄性个体的探究和聚团行为显著减少(P0.01);与雌性对照组相比,雌性父本剥夺组田鼠对另一雌性个体的探究行为有减少的趋势,对视和自饰行为显著减少(P0.05);但父本剥夺组雌雄棕色田鼠静止时间均显著增加(P0.05)。免疫组织化学结果显示,通过短暂社会互作后,与雄性对照组相比,父本剥夺组雄性在终纹床核(bed nucleus of the stria terminalis,BST)、杏仁中央核(central nucleus of the amygdale,Ce)和室旁核(paraventricular nucleus of the hypothalamus,PVN)Fos阳性神经元数量显著增加(P0.01);与雌性对照组相比,父本剥夺组雌性在终纹床核(BST)和室旁核(PVN)阳性神经元数量也极显著增加(P0.01)。综上,父本剥夺可能导致幼仔成年后社会性和活动性下降,并可能增加成年后应激反应相关脑区的神经元活性。     

Engineering large fragment insertions into the chromosome of Escherichia coli

An effective DNA replacement system has been established for engineering large fragment insertions into the chromosome of Escherichia coli. The DNA replacement plasmid, pHybrid I, was first constructed based on the bacterial artificial chromosome (BAC) vector. Two fragments of the E. coli genome, 5.5 and 6.5 kb in length, were introduced into the vector for homologous recombination. In addition to the chloramphenicol gene, a second gene neo was introduced for double marker screening for recombinant clones. By shot-gun cloning and homologous recombination techniques, using our new recombinant vector (pHybrid I), a 20-kb fragment from Lactococcus lactis genomic DNA has been successfully integrated into the chromosome of the E. coli strain J93-140. Plating tests and PCR amplification indicated that the integration remained stable after many generations in cell culture. This system will be especially useful for the chromosome engineering of large heterologous fragment insertions, which is necessary for pathway engineering.     

Mutations affecting somite formation in the Medaka (Oryzias latipes)

The metameric structure of the vertebrate trunk is generated by repeated formation of somites from the unsegmented presomitic mesoderm (PSM). We report the initial characterization of nine different mutants affecting segmentation that were isolated in a large-scale mutagenesis screen in Medaka (Oryzias latipes). Four mutants were identified that show a complete or partial absence of somites or somite boundaries. In addition, five mutations were found that cause fused somites or somites with irregular sizes and shapes. In situ hybridization analysis using specific markers involved in the segmentation clock and antero-posterior (A-P) polarity of somites revealed that the nine mutants can be compiled into two groups. In group 1, mutants exhibit defects in tailbud formation and PSM prepatterning, whereas A-P identity in the somites is defective in group 2 mutants. Three mutants (planlos, pll; schnelles ende, sne; samidare, sam) have characteristic phenotypes that are similar to those in zebrafish mutants affected in the Delta/Notch signaling pathway. The majority of mutants, however, exhibit somitic phenotypes distinct from those found in zebrafish, such as individually fused somites and irregular somite sizes. Thus, these Medaka mutants can be expected to provide clues to uncovering novel components essential for somitogenesis.     

Mutations affecting retina development in Medaka

In a large scale mutagenesis screen of Medaka we identified 60 recessive zygotic mutations that affect retina development. Based on the onset and type of phenotypic abnormalities, the mutants were grouped into five categories: the first includes 11 mutants that are affected in neural plate and optic vesicle formation. The second group comprises 15 mutants that are impaired in optic vesicle growth. The third group includes 18 mutants that are affected in optic cup development. The fourth group contains 13 mutants with defects in retinal differentiation. 12 of these have smaller eyes, whereas one mutation results in enlarged eyes. The fifth group consists of three mutants with defects in retinal pigmentation. The collection of mutants will be used to address the molecular genetic mechanisms underlying vertebrate eye formation.     

Aminoacetone induces iron-mediated oxidative damage to isolated rat liver mitochondria

Aminoacetone (AA) is a threonine metabolite accumulated in threoninemia, cri-du-chat, and diabetes, where it contributes toward the formation of cytotoxic and genotoxic methylglyoxal (MG). Oxyradicals yielded from iron-catalyzed AA aerobic oxidation to MG are shown here to promote Ca2+ -mediated mitochondrial membrane permeabilization in an AA dose-dependent way. The inhibitory effect of added EGTA, cyclosporin A, Mg2+, and DTT observed in this study suggests the formation of transition pores in the inner mitochondrial membrane by AA, associated with thiol protein aggregation. That the mitochondrial iron pool plays a coadjutant role in the transition of mitochondrial permeability is indicated by the dramatic inhibitory effect of added o-phenanthroline. Iron released from ferritin by AA oxidation products--superoxide anion and AA enolyl radicals--is shown to act as an alternative source of ferrous iron, intensifying the mitochondrial damage. These findings may contribute to clarify the role of accumulated AA and iron overload in the mitochondrial oxidative damage reportedly occurring in diabetes mellitus.     

Quantitative assessment of the associations between MTHFR C677T and A1298C polymorphisms and risk of fractures: a meta-analysis

Many studies have investigated the associations between methylenetetrahydrofolate reductase (MTHFR) C677T and A1298C polymorphisms and risk of fractures, but the impact of MTHFR polymorphisms on fractures risk is unclear owing to the obvious inconsistence among those studies. This study aims to quantify the strength of association between MTHFR C677T and A1298C polymorphisms and risk of fractures. We searched the PubMed, Embase and Wanfang databases for articles relating the association between MTHFR C677T and A1298C polymorphisms and risk of fractures in humans. We estimated summary odds ratios (ORs) with their confidence intervals (CIs) to assess the associations. Meta-analyses suggested MTHFR C677T polymorphism was associated with increased risk of any site fractures (for T vs. C, OR = 1.17, 95 % CI 1.03–1.32; for TT vs. CC, OR = 1. 31, 95 % CI 1.11–1.54; for TT vs. CT, OR = 1.22, 95 % CI 1.04–1.43; for TT vs. CT/CC, OR = 1.31, 95 % CI 1.13–1.51). Besides, MTHFR A1298C polymorphism was also associated with increased risk of any site fractures. Subgroup meta-analyses suggested MTHFR C677T polymorphism was associated with increased risk of vertebral fractures under three genetic contrast modes (for TT vs. CC, OR = 1.43, 95 % CI 1.05–1.95; for TT vs. CT, OR = 1.36, 95 % CI 1.01–1.85; for TT vs. CT/CC, OR = 1.50, 95 % CI 1.17–1.91), but there was no association between MTHFR C677T polymorphism and risk of hip fractures and non-vertebral fractures (all P values were more than 0.05). Thus, individuals with homozygote genotype TT of MTHFR C677T have obviously increased risk of vertebral fractures compared those with heterozygote genotype CT or homozygote genotype CC. There is no association between MTHFR C677T polymorphism and risk of hip fractures and non-vertebral fractures.