Evidence of a conserved functional role for DNA methylation in termites

Many organisms are capable of developing distinct phenotypes from the same genotype. This developmental plasticity is particularly prevalent in insects, which can produce alternate adaptive forms in response to distinct environmental cues. The ability to develop divergent phenotypes from the same genotype often relies on epigenetic information, which affects gene function and is transmitted through cell divisions. One of the most important epigenetic marks, DNA methylation, has been lost in several insect lineages, yet its taxonomic distribution and functional conservation remain uninvestigated in many taxa. In the present study, we demonstrate that the signature of high levels of DNA methylation exists in the expressed genes of two termites, Reticulitermes flavipes and Coptotermes formosanus. Further, we show that DNA methylation is preferentially targeted to genes with ubiquitous expression among morphs. Functional associations of DNA methylation are also similar to those observed in other invertebrate taxa with functional DNA methylation systems. Finally, we demonstrate an association between DNA methylation and the long‐term evolutionary conservation of genes. Overall, our findings strongly suggest DNA methylation is present at particularly high levels in termites and may play similar roles to those found in other insects.     

Systemic immune responses to an inactivated,whole H9N2 avian influenza virus vaccine using class B CpG oligonucleotides in chickens

Commercial vaccines against avian influenza viruses (AIV) in chickens consist mainly of inactivated AIV, requiring parenteral administration and co-delivery of an adjuvant. Limitations in T helper 1 or T helper 2 biased responses generated by these vaccines emphasize the need for alternative, more efficacious adjuvants. The Toll-like receptor (TLR) 21 ligand, CpG oligodeoxynucleotides (ODN), has been established as immunomodulatory in chickens. Therefore, the objective of this study was to investigate the adjuvant potential of high (20 μg) and low (2 μg) doses of CpG ODN 2007 (CpG 2007) and CpG ODN 1826 (CpG 1826) when administered to chickens with a formalin-inactivated H9N2 AIV. Antibody responses in sera were evaluated in 90 specific pathogen free (SPF) chickens after intramuscular administration of vaccine formulations at 7 and 21 days post-hatch. Antibody responses were assessed based on haemagglutination inhibition (HI) and virus neutralization (VN) assays; virus-specific IgM and IgY antibody responses were evaluated by ELISA. The results suggest that the vaccine formulation containing low dose CpG 2007 was significantly more effective at generating neutralizing (both HI and VN) responses than formulations with high or low doses of CpG 1826 or high dose CpG 2007. Neutralizing responses elicited by low dose CpG 2007 significantly exceeded those generated by a squalene-based adjuvanted vaccine formulation during peak responses. A significantly higher IgM response was elicited by the formulation containing low dose CpG 2007 compared to high and low doses of 1826. Although the low dose of CpG 2007 elicited a higher IgY response than CpG 1826, the difference was not statistically significant. In conclusion, 2 μg of CpG 2007 is potentially promising as a vaccine adjuvant when delivered intramuscularly with inactivated H9N2 virus to chickens. Future studies may be directed at determining the mucosal antibody responses to the same vaccine formulations.     

表观遗传学和环境因素与子宫内膜异位症

子宫内膜异位症(EMs)是一种雌激素依赖的炎性疾病,发病机制尚不清楚。越来越多的证据表明,一些基因的表观遗传学特性与EMs的发病有关。表观遗传学是指DNA序列不发生变化,但基因表达却发生了可遗传的改变,包括DNA甲基化、组蛋白共价修饰及染色质构象变化,这些改变可能会引发一系列包括肿瘤在内的病理变化。EMs异位组织中芳香化酶基因的Cp G岛序列低甲基化和(或)反式作用因子上调芳香化酶基因的表达;卵巢异位上皮及间质细胞的雌激素受体α(ERα)低表达而ERβ显著高表达;在位细胞的孕激素受体α(PRα)和PRβ均高表达;EMs异位内膜细胞的类固醇生成因子1(SF-1)启动子低甲基化而在位内膜高甲基化。此外,EMs异位组织E-钙黏蛋白(E-cadherin)基因及在位组织同源框基因A10(HOXA10)表达降低也与EMs有关;环境因素可能会影响表观遗传基因导致EMs的发生。综述表观遗传学改变和环境因素在EMs发病中的作用。     

Enteroaggregative Escherichia aoliisolated from Chinese diarrhea patients with high-pathogenicity island of Yersinia''''xs involved in synthesis of siderophore yersiniabactin

AIM: To investigate the distribution of 12 high-pathogenicity island(HPI) genes and the relation between HPI genes and expression of yersiniabactin(Ybt) in enteroaggregative E.coli(EAggEC) isolated from Chinese diarrhea patients. METHODS: The distribution of 12 HPI genes was investigated by PCR and DNA hybridization in two prototype strains of EAggEC, EAggEC 17-2, EAggEC 042, and 6 clinical EAggEC isolates from China. The production of siderophore Ybt in HPI-positive strains was detected by reporter gene bioassay to determine the relation between HPI genes and expression of Ybt. Flow cytometry was used to detect fluorescent signal of the reporter strain that could designate production of Ybt. RESULTS: Seven strains were HPI-positive and one strain was HPI-negative. Six of the seven HPI-positive strains were inserted into asnT-tRNA site. Moreover, seven EAggEC HPI-positive strains revealed enhanced fluorescence signal but the EAggEC HPI-negative strain did not. However, there was a difference in Ybt expression condition and level among these seven EAggEC HPI-positive strains. Although UFT073 strain, the prototype strain of uropathogenic E.coli(UPEC), carried the complete HPI core part, we did not detect the expression of Ybt in it. CONCLUSION: EAggEC HPI-positive strains can express the Ybt system, but the presence of HPI core part does not mean the functional expression of Ybt.     

插入CpG序列的乙肝病毒核心抗原基因真核表达质粒的构建及在树突状细胞中的表达

目的：构建pEGFP—N1／CpG-HBcAg（ISS）真核表达载体，探讨乙肝病毒核心抗原（HBcAg）在树突状细胞（DC）中的表达，为研制乙肝治疗性疫苗奠定基础。方法：根据HBcAg基因序列，设计合成两对引物，在引物中引入针对人敏感的CpG基序和不合CpG的片段，用PCR方法从慢性乙型肝炎患者血清HBVDNA中扩增出HBcAg基因片段，将扩增产物与pEGFP—N1连接，构建重组体pEGFP—N1／CpG-HBcAg，进行酶切、PCR及测序鉴定；分离人外周血单个核细胞（PBMC），体外诱导分化为DC，通过脂质体将重组质粒pEGFP—N1／CpG-HBcAg和空载体分别转染DC，Western blot检测HBcAg在DC的表达。结果：HBcAg基因体外扩增产物大小为530bp。所构建的pEGFP-N1／CpG—HBcAg经双酶切及PCR鉴定，与预期片段的大小相符。测序结果与GenBank中收录的HBcAg全长序列一致，表明pEGFP—NI／CpG-HBcAg真核表达体构建正确；PBMC体外成功刺激分化为DC，HBcAg可在DC中表达。结论：成功构建了真核重组表达载体pEGFP—N1／CpG—HBcAg，且可在DC中表达，为CpG的功能研究和乙型肝炎治疗性疫苗的研制奠定基础。     

应用锥形束CT对颌骨特发性骨硬化症的调查研究

目的运用锥形束CT(CBCT)分析惠州市颌骨特发性骨硬化症(IO)的影像学特征。方法回顾性分析427例口腔疾病患者的CBCT影像学资料,观察颌骨IO的发病情况、分布情况、好发部位以及类型。结果 427例口腔疾病患者经CBCT发现36例颌骨IO,发病率为8.43%。所有颌骨IO皆为单发,男19例(52.78%),女17例(47.22%)。颌骨IO多发生于50~59岁,发病率为27.78%,不同年龄段患者颌骨IO发病率比较,差异无统计学意义(P>0.05)。颌骨IO发生于下颌34例,发生于上颌2例,好发于下颌前磨牙区,发生率为30.56%。颌骨IO的分类中V型、中央硬化型、根尖-骨皮质型均较高。结论 CBCT更能详细反映颌骨IO的形态及周围结构关系,一般无需治疗,但应该定期复查,一旦发现牙根吸收或对周围结构产生挤压等需进行干预。     

Genetic polymorphisms of 15 STR loci in the population of the island of Cres (Croatia)

Background: The population of the island of Cres presents one of the few persisting Eastern Adriatic isolates and is thereby suitable for human population differentiation analyses.

Aim: The aim of this study was to analyse the genetic structure of the island of Cres with respect to its eight sub-populations and to compare the genetic variation of the island of Cres with other Eastern Adriatic islands and the Croatian mainland.

Subjects and methods: Fifteen AmpFlSTR identifiler loci were analysed in a sample group of 122 unrelated autochthonous individuals from the island of Cres, Croatia.

Results: Analysis of STR polymorphisms revealed genetic homogeneity among sub-populations of the island of Cres and small but significant levels of genetic heterogeneity among geographically distant Eastern Adriatic islands.

Conclusion: Despite a considerable degree of genetic homogeneity among the studied Eastern Adriatic islands, small but significant differentiation between distant islands indicates geographic sub-structuring which follows the isolation by distance model. This study is supportive of the notion that STR markers are useful for genetic differentiation between larger and geographically more distant regions.