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Abstract Background microRNAs (miRNAs) are a group of regulatory RNAs that regulate gene expression by binding to specific sequences on target mRNAs. However, functional identification of mRNA targets is usually difficult and time consuming. Here we report hybrid-PCR as a new and rapid approach to screen putative mRNA targets in vitro. Results Fifteen putative target mRNAs for human cytomegalovirus (HCMV) miR-UL112-1, including previously confirmed HCMV IE72, were identified from mRNA-derived cDNAs using hybrid-PCR. Moreover, we randomly validated six different target candidates by luciferase reporter assays, and confirmed that their luciferase activities were down-regulated with co-transfection of HCMV miR-UL112-1. Conclusions Our study demonstrated that hybrid-PCR is an effective and rapid approach for screening putative miRNA targets, with much more advantage of simplicity, low cost, and ease of implementation.

To determine the function of miR-206 in epilepsy. Epileptic rat model was established by intra-amygdala injection of kainic acid (KA). Expression levels of miR-206, C–C Motif Chemokine Ligand 2 (CCL2) and interleukin-1β (Il-1β) in hippocampus tissues was measured by reverse transcription-quantitative PCR (RT-qPCR) and western blot. Dual luciferase reporter assay was performed to determine the binding of miR-206 to 3′ untranslated region (UTR) of CCL2. Finally, brain waves were recorded and Hematoxylin and eosin (HE) staining and Nissl's staining were performed on the epileptic rat injected with LPS, miR-206 agomir, adeno-associated virus (AAV) expressed CCL2 alone or in combination. Expression of miR-206 was specially decreased in hippocampus tissues compared to cortex in response to KA induced pathologic brain activity. Enforced expression of miR-206 by injection miR-206 agomir not only decreased seizure activity, but also protected KA-induced neuronal loss. And enforced expression of miR-206 suppressed increase of C–C Motif Chemokine Ligand 2 (CCL2) and interleukin-1β (Il-1β) which were induced by injection of KA or KA combined with lipopolysaccharide (LPS). Further more, results of dual luciferase reporter assay confirmed CCL2 was a target of miR-206. Finally, co-injection adeno-associated virus (AAV) expressed CCL2 with miR-206 agomir abolished the function of miR-206 agomir. Taken together, our results showed that expression of miR-206 could inhibit seizure-induced brain injury by targeting CCL2. Our results showed that expression of miR-206 could inhibit seizure-induced brain injury by targeting CCL2. [ABSTRACT FROM AUTHOR]

Objectives: Neuropathic pain (NP) is one of the main challenges towards NP syndrome treatment. miR-340-5p exhibit different expression levels in NP models. Its effects on NP remained unclear. The objective of this study was to explore the potential regulation mechanisms of miR-340-5p in NP. Methods: Rat model of chronic constriction injury (CCI) was established to induce NP in vivo. NP levels were assessed using mechanical withdrawal threshold (MWT). The inflammation response in CCI rats were determined by HE staining and ELISA assay. The target genes of miR-340-5p were verified by luciferase report assays. Results: In CCI rats, level of miR-340-5p was down-regulated both in spinal cord tissues and isolated microglial cells. Paw withdrawal threshold (PWT) and paw withdrawal latency (PWL) were decreased in CCI rats, which were restored upon miR-340-5p overexpression. miR-340-5p overexpression also decreased inflammation as well as expression levels of COX-2, IL-1β, TNF-α and IL-6 in CCI rats. Luciferase report assays revealed Rap1A was a target gene of miR-340-5p in the experimental model. Elevated miR-340-5p decreased Rap1A expression level in vitro and in vivo. Overexpression of Rap1A protein restored expression levels of COX-2, IL-1β, TNF-α and IL-6, reduced the PWT and PWL and increased inflammation response in CCI rats. Conclusion: miR-340-5p alleviated CCI-induced NP by targeting Rap1A. miR-340-5p and Rap1A may be the potential treatment targets for NP therapeutics. [ABSTRACT FROM AUTHOR]

To investigate the correlation between the air phthalate acid ester (PAE) exposure and serum PAE concentration and the effects of PAE exposure on reproductive health among Chongqing traffic-patrol policemen. In 2013, 32 traffic-patrol policemen working in an area with poor air quality in Chongqing and 28 traffic-patrol policemen working in an area with good air quality were selected. Their blood levels of 14 PAEs and six reproductive hormones were determined. Air samples were collected from four traffic-patrol platforms. The concentrations of 14 PAEs in the air samples were evaluated. All 14 PAEs were detected in the blood samples. The concentrations of seven PAEs in the total suspended particulate, namely, dimethyl phthalate, diethyl phthalate, dibutyl phthalate, bis (2-ethox-yethyl) phthalate, dihexyl phthalate, benzyl butyl phthalate, and bis (2-n-butoxyethyl) phthalate, were positively and significantly associated with the blood levels of these PAEs in the participants. All the sex hormone levels measured here were significantly different between the participants from the two areas. The PAE concentrations in the blood samples were correlated with the reproductive hormone levels in the participants. Air PAE pollution may be a major source of PAE exposure in the traffic-patrol policemen of Chongqing. [ABSTRACT FROM AUTHOR]

Purpose Delivering messages and information to potentially interested users is one of the distinguishing applications of online enterprise social network (ESN). The purpose of this paper is to provide insights to better understand the repost preferences of users and provide personalized information service in enterprise social media marketing.Design/methodology/approach It is accomplished by constructing a target audience identification framework. Repost preference latent Dirichlet allocation (RPLDA) topic model topic model is proposed to understand the mass user online repost preferences toward different contents. A topic-oriented preference metric is proposed to measure the preference degree of individual users. And the function of reposting forecasting is formulated to identify target audience.Findings The empirical research shows the following: a total of 20 percent of the repost users in ESN represent the key active users who are particularly interested in the latent topic of messages in ESN and fits Pareto distribution; and the target audience identification framework can successfully identify different target key users for messages with different latent topics.Practical implications The findings should motivate marketing managers to improve enterprise brand by identifying key target audience in ESN and marketing in a way that truthfully reflects personalized preferences.Originality/value This study runs counter to most current business practices, which tend to use simple popularity to seek important users. Adaptively and dynamically identifying target audience appears to have considerable potential, especially in the rapidly growing area of enterprise social media information service. [ABSTRACT FROM AUTHOR]

Abstract With the revolution of the traditional economic and social pattern based on centralized fossil energy consumption, the regional integrated energy system (RIES) has gained rapid development in recent years for achieving higher energy efficiency. This paper expands the Demand Response (DR) concept to the RIES and presents an optimal operation model of RIES considering the DR mechanism on the energy price. In this paper, a comprehensive DR strategy combining energy price and different loads is firstly developed to exploit the demand flexibility of RIES in the DR model established. Based on the DR modeling of the RIES, an operation optimization model with environmental, economic benefits and energy supply reliability as objective functions has been firstly established in detail. Secondly, a complete scheduling scheme is built based on the energy consumption characteristics and system operation characteristic of the integrated energy system. The model presented could reduce the cost of system without causing a significant amount of environmental pollution, and improve the energy efficiency of the RIES efficiently. Besides, the scheduling strategy proposed could also provide support for the operation strategy of integrated energy system under the energy development. Hopefully, this paper will provide reference for future research and engineering projects on DR programs in the RIES. Highlights • The typical structure of the RIES is introduced. • A DR model suitable for the RIES considering different loads is established. • An optimal operation model of the RIES based on DR is established. • The load in the RIES is affected not only by energy price but also by other loads. [ABSTRACT FROM AUTHOR]

Ischemic stroke is a major cerebrovascular disease resulting from a transient or permanent local reduction of cerebral blood flow. Angiogenesis plays an important role in cerebral microvascular repair after ischemic stroke. This study aimed at investigating the effect of NF-E2-related factor 2 (Nrf2) on the angiogenesis of mouse cerebral microvascular endothelial bEnd.3 cells in a hypoxic environment. We found that Nrf2 expression was temporarily increased in hypoxia-induced bEnd.3 cells. Knockdown of Nrf2 inhibited the proliferation, migration, as well as tube formation in hypoxia-induced bEnd.3 cells. Meanwhile, vascular endothelial growth factor and PI3K/Akt signaling pathways were identified to be regulated by Nrf2 in hypoxia-induced bEnd.3 cells. It was found that silencing of Nrf2 downregulated the expression levels of NAD(P)H:quinine oxidoreductase-1, vascular endothelial growth factor, p-Akt, and heme oxygenase-1 in hypoxia-induced bEnd.3 cells. Data suggested that hypoxia induced the transient increase of Nrf2, which plays a key role in the angiogenesis of cerebral microangiogenesis, and that Nrf2 regulates the proliferation, migration, as well as tube formation likely through PI3K/Akt signaling pathway in hypoxia-induced bEnd.3 cells. Our study provides proof of concept for the modulation of Nrf2, so as to tilt the balance toward angiogenesis, representing a therapeutic strategy for hypoxia or ischemia disorders such as stroke. [ABSTRACT FROM AUTHOR]

Peroxisome-proliferator activated receptor gamma (PPARG) in placenta play an important role in pregnancy. Our previous study showed that it mediated the effects of phthalates on placental mRNA expression of estrogen synthetases in rats. To assess the effects of phthalate exposure on PPARG placental expression, and the contribution of PPARG to the effects of phthalates in human. 207 healthy pregnant women were recruited and their cord blood and placenta were collected upon delivery. Three phthalates, estrogens in cord blood and protein expression of PPARG in placenta were measured. Linear regression were used to analyze the relationship between phthalates exposure, PPARG expression and hormones. Phthalate levels in cord blood were positively associated with PPARG protein expression in placenta (p < 0.05), whereas estrogens in cord blood were negatively associated with phthalate levels and PPARG expression (p < 0.05). This study shows that PPARG in placenta may mediate the adverse effects of phthalates on pregnancy in human. [ABSTRACT FROM AUTHOR]

Endocrine disrupting chemicals may disrupt developing neuroendocrine systems, especially when the exposure occurs during a critical period. This study aimed to investigate whether prenatal exposure to di-(2-ethylhexyl) phthalate (DEHP), a major component of plasticizers used worldwide, disrupted the development of a network of genes important for neuroendocrine function in male rats. Pregnant rats were treated with corn oil (vehicle control), 2, 10 or 50 mg/kg DEHP by gavage from gestational day 14 to 19. The neuroendocrine gene expressions were quantified using a 48-gene Taqman qPCR array in the whole hypothalamus of neonatal rats (postnatal day 1) and in the anteroventral periventricular nucleus (AVPV), medial preoptic nucleus (MPN) and arcuate nucleus (ARC) of adult rats (postnatal day 70). Immunofluorescent signals of ERα and CYP19 were detected using the confocal microscopy in adult AVPV, MPN and ARC. The results showed that prenatal DEHP exposure perturbed somatic and reproductive development of offspring. Eleven genes were down-regulated in neonatal hypothalamus and showed non-monotonic dose-response relationships, that the 10 mg/kg DEHP dosage was associated with the greatest number of gene expression changes. Different from this, 14 genes were altered in adult AVPV, MPN and ARC and most of alterations were found in the 50 mg/kg DEHP group. Also, 50 mg/kg DEHP reduced ERα expression in the ARC, but no alterations were observed in CYP19 expression. These results indicated that prenatal DEHP exposure may perturb hypothalamic gene programming and the influences are permanent. The effects showed dependence on developmental stages and nuclei region. [ABSTRACT FROM AUTHOR]