罗非鱼TRAF3基因的表达及功能研究

肿瘤坏死因子受体(TNFR)相关因子3(TRAF3)是多种免疫途径中的关键调控因子。从尼罗罗非鱼(Oreochromis niloticus)中克隆获得了TRAF3基因, 命名为OnTRAF3(GeneBank No. MN258118), 该基因包含1个环指结构域、1个锌指结构域、1个卷曲螺旋和MATH结构域。多序列比对表明, OnTRAF3与其他已知的TRAF3蛋白具有高度的相似性, 尤其是MATH结构域。实时荧光定量PCR(qRT-PCR)分析显示, OnTRAF3在各组织中广泛分布, 且在脑、皮肤、肠和鳃中表达量较高。在无乳链球菌诱导后, 多个组织中的OnTRAF3表达量出现了不同程度的上调, 说明OnTRAF3参与了罗非鱼的抗菌免疫应答。亚细胞定位实验显示, OnTRAF3分布在HEK293的细胞质和细胞核中。此外, 荧光素酶报告基因实验结果显示, 野生型(WT)OnTRAF3可显著激活NF-κB信号, 而coiled-coil和MATH 结构域缺失后, 依然能够显著激活NF-κB 活性, 而RING 和Zinc缺失后, 该激活作用则明显减弱, 表明RING 和Zinc 结构域是OnTRAF3在免疫信号通路中行使功能的关键结构域。研究为探索TRAF3在罗非鱼免疫应答中的功能提供了重要的基础。     

长江流域水牛血液蛋白多态性研究

采集长江流域德昌、涪陵、江汉、滨湖和海子5个水牛地方品种共416 头牛的血样。电泳检测血红蛋白、血清运铁蛋白多态性,估算各等位基因频率及基因型频率,根据Hb及Tf两个位点的基因频率进行聚类分析。结果表明,5个水牛地方品种间遗传变异较小,其亲缘关系较近。     

Establishment of HIV-1 model cell line GHOST(3) with stable DRiP78 and NHERF1 knockdown

Chemokine receptors CXCR4 and CCR5 are indispensable co-receptors for HIV-1 entry into host cells. In our previous study, we identified that dopamine receptor-interacting protein 78(DRi P78) and Na+-H+ exchanger regulatory factor 1(NHERF1) are the CXCR4 and CCR5 homo- or hetero-dimerinteracting proteins. DRi P78 and NHERF1 are able to influence the co-receptor internalization and intracellular trafficking. Over-expression of NHERF1 affects the ligands or HIV-1 gp120-induced CCR5 internalization and HIV-1 production. It is reasonable to speculate that DRi P78 and NHERF1, as well as the signaling pathways involved in viral replication, would probably affect HIV-1 replication through regulating the co-receptors. In this present study, we designed two short hairpin RNAs(sh RNAs) targeting the DRi P78 and NHERF1, respectively, and constructed the p Lenti6/BLOCK-i T-DEST lentiviral plasmids expressing DRi P78 or NHERF1 sh RNA. The packaged lentiviruses were used to transduce the widely-applied HIV-1 model cell line GHOST(3). Then, cells with stable knockdown were established through selecting transduced cells with Blasticidin. This study, for the first time, reported the establishment of the GHOST(3) with DRi P78 and NHERF1 knockdown, which is the first stable cell line with HIV-1 co-receptor-interacting molecular defects.     

黄鳝二价染色体上地高辛配基标记随机引物原位DNA合成的研究

在黄鳝二价染色体上, 运用地高辛配基标记的随机引物原位DNA合成技术诱导出了明暗相间、基本稳定且类似于R带的带状结构。本文对该结果进行了较详细的分析和讨论。 Abstract:The dark bands alternating with light bands,relatively stable and R-band-like structure was showed on the pachytene bivalents of Rics-field eel(Monopterus albus Zuiew)by using the random-primed in situ digoxigenin-labelled DNA synthesis technique,and the results were analysed and discussed in detail.     

Revealing the Circadian Rhythm of Taliang Crocodile Newt(Liangshantriton taliangensis) During Breeding Migration Using a Novel Monitoring Technique

Many animals migrate during the breeding season. It is important to study the patterns of breeding migration of wild animals, especially rare and endangered species. However, current data acquisition methods are too coarse and imprecise for investigating the circadian rhythms of migrations in amphibians. Based on the frustrated total reflection image(FTRI), we developed a new device and recorded the precise migration time of an endangered salamander, Liangshantriton taliangensis.During the breed...     

Both mammalian PIG-M and PIG-X are required for growth of GPI14-disrupted yeast

GPI mannosyltransferase I (GPI-MT-I) transfers the first mannose to a GPI-anchor precursor, glucosamine-(acyl)phosphatidylinositol [GlcN-(acyl)PI]. Mammalian GPI-MT-I consists of two components, PIG-M and PIG-X, which are homologous to Gpi14p and Pbn1p in Saccharomyces cerevisiae, respectively. In the present study, we disrupted yeast GPI14 and analysed the phenotype of gpi14 yeast. The gpi14 haploid cells were inviable and accumulated GlcN-(acyl)PI. We cloned PIG-M homologues from human, Plasmodium falciparum (PfPIG-M) and Trypanosoma brucei (TbGPI14), and tested whether they could complement gpi14-disrupted yeast. None of them restored GPI-MT-I activity and cell growth in gpi14-disrupted yeast. However, gpi14-disrupted yeast cells with human PIG-M, but not with PfPIG-M or TbGPI14, grew slowly but significantly when they were supplemented with rat PIG-X. This suggests that the association of PIG-X and PIG-M for GPI-MT-I activity is not interchangeable between mammals and the other lower eukaryotes.     

Venom gland of the ichneumonid Diadromus collaris： morphology, ultrastructure and age-related changes

Females of the solitary parasitoid Diadromus collaris （Insecta： Hymenoptera： Ichneumonidae） lay eggs in the pupae of Plutella xylostella （Lepidoptera： Plutellidae）, and the venom is synchronously injected into hosts. The venom apparatus consists of two glandular tubules terminating in a common reservoir, A ductule connects the reservoir with the sting apparatus, by which the reservoir content enters the latter. Secretory units line the two glandular tubules. All secretory cells belong to dermal gland type Ⅲ. Dermal gland cells in glandular tubules are more abundant and developed than those in the reservoir. There are extensive rough endoplasmic reticulum and electrondense vesicles, and the microvilli are well developed. By the cuticle-lined central funnel secretion products of secretory units reach the reservoir. Moreover, the secretory apparatus undergoes age-related changes. The secretory units in the venom gland are better developed and more vigorous 7 days after eclosion than those 1 day after eclosion; autolytic processes occur 15 days after eclosion, and the tissue of the reservoir is more prostrate 15 day after eclosion than those 1 day after eclosion. The ovipostion peak of this parasitoid, about 3-7 days after eclosion, corresponds with the period when the venom gland is highly developed in the life span of the wasp.     

miRNA-711-SP1-collagen-I pathway is involved in the anti-fibrotic effect of pioglitazone in myocardial infarction

Although microRNAs(miRNAs) have been intensively studied in cardiac fibrosis,their roles in drug-mediated anti-fibrotic therapy are still unknown.Previously,Pioglitazone attenuated cardiac fibrosis and increased miR-711 experimentally.We aimed to explore the role and mechanism of miR-711 in pioglitazone-treated myocardial infarction in rats.Our results showed that pioglitazone significantly reduced collagen-I levels and increased miR-711 expression in myocardial infarction heart.Pioglitazone increased the expression of miR-711 in cardiac fibroblasts,and overexpression of miR-711 suppressed collagen-I levels in angiotensin II(Ang II)-treated or untreated cells.Transfection with antagomir-711 correspondingly abolished the pioglitazone-induced reduction in collagen-I levels.Bioinformatics analysis identified SP1,which directly promotes collagen-I synthesis,as the putative target of miR-711.This was confirmed by luciferase assay and western blot analysis.Additionally,increased SP1 expression was attenuated by pioglitazone in myocardial infarction heart.Furthermore,transfection of antagomir-711 attenuated pioglitazone-reduced SP1 expression in cardiac fibroblasts with or without Ang II stimulation.We conclude that pioglitazone up-regulated miR-711 to reduce collagen-I levels in rats with myocardial infarction.The miR-711-SP1-collagen-I pathway may be involved in the anti-fibrotic effects of pioglitazone.Our findings may provide new strategies for miRNA-based anti-fibrotic drug research.