Effect of 25-hydroxyvitamin D3 on rotavirus replication and gene expressions of RIG-I signalling molecule in porcine rotavirus–infected IPEC-J2 cells

The study evaluated whether a 25-hydroxyvitamin D₃ (25D₃) supplementation decreases the replication of rotavirus by the retinoic acid-inducible gene I (RIG-I) signalling pathway in a porcine small intestinal epithelial cell line (IPEC-J2). The results show that IPEC-J2 cells express high baseline levels of 1α-hydroxylase (CYP27B1), which converts inactive 25D₃ to the active 1,25-dihydroxyvitamin D₃ (1,25D₃). Porcine rotavirus (PRV) infection alone resulted in a significant increase in CYP27B1 mRNA, which augmented the production of active vitamin D. Physiological concentrations of 25D₃ were found to decrease PRV replication in IPEC-J2 cells. RIG-I plays an important role in the recognition of double-stranded RNA virus by host cells. Upon recognition, RIG-I triggers a series of signalling molecules such as interferon-β (IFN-β) promoter stimulator 1 (IPS-1) leading to the expression of type I interferons (IFN-β). Active 25D₃ that was generated by PRV-infected IPEC-J2 cells led to an increased expression of toll-like receptors 3 (TLR3), RIG-I, IPS-1, IFN-β and IFN-stimulated genes 15 (ISG₁₅) with important innate immune functions. Inhibiting CYP27B1 also failed to increase RIG-I, IPS-1, IFN-β and ISG₁₅ mRNA expression. These observations suggest that 25D₃ can directly inhibit PRV in IPEC-J2 cells, which requires this active form of vitamin D. The anti-rotavirus effect of 25D₃ is mediated at least in part by RIG-I signalling pathways in IPEC-J2 cells.     

自发性高血压大鼠肠系膜微静脉白细胞-内皮细胞相互作用和微淋巴管收缩特性的研究

目的:探讨自发性高血压大鼠(Spontaneously Hypertensive Rat,SHR)肠系膜微静脉白细胞-内皮细胞相互作用和微淋巴管收缩的特性。方法:取8周龄Wistar大鼠、8周龄SHR(SHR8W)和13周龄SHR(SHR13W),麻醉、固定并暴露肠系膜后,微循环显微镜下观察肠系膜微循环并录像;回放录像,计算微静脉白细胞滚动数和滚动的白细胞-内皮细胞接触时间(Rolling leukocyte-endothelial contact time,RLECT),用Vas Track自动测量系统对微淋巴管口径进行动态测量,并计算微淋巴管收缩特性指标。结果:SHR13W的白细胞滚动数显著低于Wistar;SHR8W和SHR13W的RLECT均显著低于Wistar,且SHR13W的RLECT显著低于SHR8W;进一步按照管径分级后,三组间白细胞滚动数在10~20μm管径级别下未见差异;各个管径级别下,SHR8W和SHR13W的RLECT均未见差异。SHR13W的淋巴管收缩分数显著低于Wistar和SHR8W;SHR8W及SHR13W的总收缩活性指数均显著低于Wistar;SHR13W的淋巴管动力指数显著低于Wistar。结论:SHR肠系膜微静脉白细胞滚动数及RLECT减少,其中白细胞滚动数在不同管径级别微静脉中的分布不均匀,而RLECT随SHR周龄降低,意味着SHR淋巴管收缩功能降低。     

PNPLA3在实验性非酒精性脂肪性肝病中的表达及其CpG岛甲基化状态的研究

目的:探讨PNPLA3在非酒精性脂肪性肝病中的表达水平及其Cp G岛甲基化状态,探讨PNPLA3在NAFLD发生、发展中的作用。方法:1.采用10μg/m L的油酸作用于人正常肝细胞株L02建立脂肪肝细胞模型,72 h后经油红O染色,观察正常组、脂肪肝模型组及给药组细胞内脂滴形成情况,并用荧光定量PCR检测PNPLA3在三组肝细胞中的表达情况。2.采用专用DNA提取试剂盒分别提取正常组、脂肪肝模型组、给药组细胞中DNA,经亚硫酸转化后行PCR扩增目的基因,并用焦磷酸测序方法检测不同组PNPLA3 Cp G岛中甲基化水平,探讨其在非酒精性脂肪肝中的作用。结果:1、脂肪肝模型组肝细胞PNPLA3 m RNA的表达高于正常组,经姜黄素给药后,其表达降低,差异均有统计学意义(P0.05)。2、在PNPLA3启动子区6个检测位点中,脂肪肝组位点2甲基化水平高于正常组,姜黄素给药后甲基化水平降低,差异均有统计学意义(P0.05),在其余各检测位点中甲基化水平无差异。结论:1、油红O染色后,脂肪肝模型组细胞内发现有大量红色脂滴积聚,而正常组基本上未见脂滴,姜黄素给药组脂滴较模型组减少,表明用10μg/m L的油酸诱导能成功建立体外肝细胞模型。2、PNPLA3 m RNA在肝细胞中高表达及其启动子区甲基化状态异常参与非酒精性脂肪肝发病。3、抑制肝脏PNPLA3活性或降低肝细胞PNPLA3 m RNA的表达水平可能是今后治疗非酒精性脂肪性肝病(NAFLD)的一个新的治疗方法。     

氟对镍钛拉簧的腐蚀性研究

目的:研究模拟应用氟化物防龋的口腔环境下,不同浓度氟化物对镍钛拉簧的应力衰减的影响。方法:选取两种国内常用品牌0.010 inch镍钛拉簧各40根,随机将每种拉簧平均分入四个浓度组(0.05%、0.10%、0.15%和对照组),将相同浓度组的不同品牌弹簧分别固定于间距为20 mm的四个支架的两端。将四个支架分别放入37℃人工唾液中水浴。将实验组按浓度分组放入不同浓度氟化人工唾液(0.05%、0.10%、0.15%)中处理5 min,每天三次。分别于实验前、实验后1周、2周、3周、4周用拉伸机测量每根弹簧拉伸至20 mm时的应力。对实验结果使用重复测量的方差分析。结果:时间因素对两种镍钛拉簧拉伸应力的衰减的影响有统计学意义,氟化物浓度有辅助效应,二者有交互作用。氟化物作用下不同品牌镍钛拉簧的应力腐蚀变化不同。(P0.05)。结论:1.氟化物的存在可以增加镍钛拉簧的应力衰减。2.在正畸治疗的临床工作中在使用氟化物时应考虑增加镍钛拉簧的更换频率。     

Cotton (Gossypium hirsutum) 14‐3‐3 proteins participate in regulation of fibre initiation and elongation by modulating brassinosteroid signalling

Cotton (Gossypium hirsutum) fibre is an important natural raw material for textile industry in the world. Understanding the molecular mechanism of fibre development is important for the development of future cotton varieties with superior fibre quality. In this study, overexpression of Gh14‐3‐3L in cotton promoted fibre elongation, leading to an increase in mature fibre length. In contrast, suppression of expression of Gh14‐3‐3L, Gh14‐3‐3e and Gh14‐3‐3h in cotton slowed down fibre initiation and elongation. As a result, the mature fibres of the Gh14‐3‐3 RNAi transgenic plants were significantly shorter than those of wild type. This ‘short fibre’ phenotype of the 14‐3‐3 RNAi cotton could be partially rescued by application of 2,4‐epibrassinolide (BL). Expression levels of the BR‐related and fibre‐related genes were altered in the Gh14‐3‐3 transgenic fibres. Furthermore, we identified Gh14‐3‐3 interacting proteins (including GhBZR1) in cotton. Site mutation assay revealed that Ser163 in GhBZR1 and Lys51/56/53 in Gh14‐3‐3L/e/h were required for Gh14‐3‐3‐GhBZR1 interaction. Nuclear localization of GhBZR1 protein was induced by BR, and phosphorylation of GhBZR1 by GhBIN2 kinase was helpful for its binding to Gh14‐3‐3 proteins. Additionally, 14‐3‐3‐regulated GhBZR1 protein may directly bind to GhXTH1 and GhEXP promoters to regulate gene expression for responding rapid fibre elongation. These results suggested that Gh14‐3‐3 proteins may be involved in regulating fibre initiation and elongation through their interacting with GhBZR1 to modulate BR signalling. Thus, our study provides the candidate intrinsic genes for improving fibre yield and quality by genetic manipulation.     

Associations of the uric acid related genetic variants in SLC2A9 and ABCG2 loci with coronary heart disease risk

Background

Multiple studies investigated the associations between serum uric acid and coronary heart disease (CHD) risk. However, further investigations still remain to be carried out to determine whether there exists a causal relationship between them. We aim to explore the associations between genetic variants in uric acid related loci of SLC2A9 and ABCG2 and CHD risk in a Chinese population.

Results

A case–control study including 1,146 CHD cases and 1,146 controls was conducted. Association analysis between two uric acid related variants (SNP rs11722228 in SLC2A9 and rs4148152 in ABCG2) and CHD risk was performed by logistic regression model. Adjusted odds ratios (ORs) with 95% confidence intervals (CIs) were calculated. Compared with subjects with A allele of rs4148152, those with G allele had a decreased CHD risk and the association remained significant in a multivariate model. However, it altered to null when BMI was added into the model. No significant association was observed between rs11722228 and CHD risk. The distribution of CHD risk factors was not significantly different among different genotypes of both SNPs. Among subjects who did not consume alcohol, the G allele of rs4148152 showed a moderate protective effect. However, no significant interactions were observed between SNP by CHD risk factors on CHD risk.

Conclusions

There might be no association between the two uric acid related SNPs with CHD risk. Further studies were warranted to validate these results.

Electronic supplementary material

The online version of this article (doi:10.1186/s12863-015-0162-7) contains supplementary material, which is available to authorized users.