Immunohistochemical analysis of pseudorabies virus spread through neurons innervating the eyeball

Pseudorabies virus (PRV) was inoculated intraocularly into BALB/c mice, and the distribution pattern of cells positive for several neurotransmitters and viral antigens in the eyeball, trigeminal nerve ganglia, and superior cervical ganglia was examined immunohistochemically to clarify the neural route of the virus spread. In the eyeball, substance P (SP)- and calcitonin gene-related peptide (CGRP)-positive cells were detected in the ipsilateral iris and ciliary body, neuropeptide tyrosine (NPY)-positive cells were detected in the choloid membrane, and tyrosine hydroxylase (TH)-positive cells were detected in the ipsilateral inner nuclear layer of the retina; all these cells contained viral antigens. In the superior cervical ganglia, viral antigen-positive cells containing TH or NPY were found at bilateral sites. In the trigeminal nerve ganglia, viral antigen-positive cells containing SP or CGRP were found at bilateral sites. These findings indicated that the SP- and CGRP-positive ganglion cells of the trigeminal nerve ganglia innervating the iris or ciliary body, and the NPY-positive ganglion cells of the superior cervical ganglia innervating the iris, ciliary body, and choroid membrane served as the route for the virus spread. These findings also suggested that dopaminergic neurons, such as the TH-positive retinal cells and TH-positive ganglion cells of the superior cervical ganglia, served as the route for virus spread.     

Developmental changes in pituitary–thyroid axis, and formation of gonads in leptocephali and glass eels of Anguilla spp.

SUMMARY: Pituitary, thyroid gland and gonads in leptocephali of Japanese eel Anguilla japonica (19.8–32.6 mm in total length), A. obscura (45.0 mm), and A. bicolor pacifica (49.5 mm) and those in glass eels of the Japanese eel were histologically and immunohistochemically examined in order to observe the developmental changes of these endocrine organs in the Anguillidae. The pituitary, consisting of adenohypophysis and neurohypophysis in Japanese eel leptocephali over 22.5 mm, did not contain thyroid stimulating hormone (TSH) immunoreactive cells. Such cells were, however, detectable in the more developed pituitaries of leptocephali of A. obscura and A. bicolor pacifica and in those of glass eels. Conversely, thyroxine (T₄)-immunoreactive thyroid follicles could be detected in all specimens, both leptocephalic and glass eel. Only in glass eels, gonads were found in the body cavity, and these gonads harbored one or two primordial germ cells (PGC) per cross-section. Our results indicate that thyroid hormones (TH) production started prior to TSH production, and that TSH and TH are both secreted during the metamorphosis from leptocephalus to glass eel. Therefore, it is plausible that the TSH–TH axis is involved in the metamorphosis from leptocephalus to glass eel, but not in the early growth from preleptocephalus to leptocephalus.     

播期效应对大粒型玉米杂交种籽粒生长性状稳定性影响的研究

以七个籽粒大小不同的玉米杂交种为材料,在三个播期中评价籽粒生长性状的稳定性。结果表明,终结体积的基因型与播期互作不显著,其余性状均存在显著的基因型与播期互作。对于最大鲜体积、终结体积和粒重,只有基因型与播期互作中的线性部分显著。多数大粒型组合的终结体积、最大鲜体积、粒重和有效灌浆期表现较稳定。大粒型杂交种多数性状比小粒型具有较大的均数和回归系数,粒重的稳定性参数与终结体积、有效灌浆期和实际灌浆期的稳定性参数存在一定相关。     

Immune dysregulation in flea allergy dermatitis--a model for the immunopathogenesis of allergic dermatitis

BACKGROUND: Flea allergy dermatitis (FAD) is a common skin disease in dogs and can be induced experimentally. It often coexists with other allergic conditions. So far no studies have investigated the quantitative production of cytokine mRNA in skin biopsies and peripheral blood mononuclear cells (PBMC) in flea allergic dogs. OBJECTIVE: The aim of our study was to improve the understanding of the immunopathogenesis of allergic dermatitis as a response to fleabites. MATERIAL AND METHODS: Allergic and non-allergic dogs were exposed to fleas. Before and after 4 days of flea exposure mRNA was isolated from biopsies and PBMC. Production of chymase, tryptase, IL-4, IL-5, IL-13, TNF-alpha and IFN-gamma mRNA was measured by real-time RT-PCR. The inflammatory infiltrate in the skin was scored semi-quantitatively. The number of eosinophils, mast cells (MC) and IgE+ cells/mm2 was evaluated to complete the picture. RESULTS: FAD was associated with a higher number of MC before flea exposure and with a significant increase of eosinophils after flea exposure as compared to non-allergic dogs. The number of IgE+ cells was higher in allergic dogs before and after flea exposure. In allergic dogs mRNA for most cytokines and proteases tested was higher before flea exposure than after flea exposure. After exposure to fleas an increased mRNA production was only observed in non-allergic dogs. In vitro stimulation with flea antigen resulted in a decreased expression of most cytokines in allergic dogs before flea exposure. In contrast, in PBMC, only increased levels of IL-4 and IL-5 mRNA were observed in allergic dogs before flea exposure. However, after flea exposure and additional stimulation with flea antigen the production of mRNA for all cytokines tested was significantly increased in allergic dogs. CONCLUSION: We demonstrated that the response in biopsies and PBMC is different and that FAD is associated with a TH2 response.     

神经干细胞定向诱导分化条件研究

通过模拟体内分化诱导条件,在体外对神经干细胞(NSCs)进行定向诱导.结果表明,由添加2% B27 100 pg·mL-1 IL-1 0.05 g·L-1 Vc 5 u·mL-1 EPO的DMEM/F12的诱导液A和1份含2% B27的DMEM/F12 1份纹状体提取液混合组成的诱导液B均可以将中脑和大脑皮质的NSCs诱导分化为多巴胺能神经元,TH阳性细胞表达率随诱导时间的增加而提高,中脑来源的NSCs的TH阳性细胞诱导率极显著地高于大脑皮质来源的NSCs,诱导液A的诱导效果优于诱导液B.     

牙鲆变态中甲状腺激素调控的靶基因

本研究旨在采用新的方法找出甲状腺激素受体（TRs）介导的甲状腺激素（TH）调控的靶基因。以牙鲆（）为实验样本,表达并纯化了牙鲆p3×Flag-TRαA和p3×Flag-TRβ融合蛋白,采用一种目的基因循环扩增与筛选的方法,寻找TH直接调控的靶基因,最终筛选得到TRαA的候选靶基因11个,TRβ的候选靶基因12个,其中TRαA和TRβ存在4个相同的候选靶基因：）和MYC相关因子X（）。利用荧光定量PCR技术检验了这4个基因在牙鲆不同组织、不同变态发育期以及外源性甲状腺激素和硫脲处理后各个变态发育时期的表达情况,发现在TH含量升高时,靶基因表达量降低;TH含量降低时,靶基因的表达量上升,从而验证靶基因受到TH的负向调控,证实TH在促进牙鲆变态中也存在抑制靶基因转录的情况。本研究旨为研究TH调控牙鲆变态的发育网络提供基础资料。