Molecular cloning and functional analysis of the follicle-stimulating hormone (FSH) receptor gene promoter from the Jintang black goat

A 762 bp fragment of the 5’-flanking region of the FSHR gene from the Jintang black goat was cloned. The putative initial transcript site was the A at 681 bp and there were 7 putative cis-acting elements and 3 AT-rich regions. The sequence of the FSHR promoter from the Jintang black goat is 99.34% homology to Capra hircus, 32.38% to Gallus gallus and 38.55% to mouse. It could promote the EGFP, FSHR transcription in HEK293 cells, the fluorescence intensity was weaker than the CMV promoter, but the expressed FSHR could respond to the FSH signaling with signal intensity much higher than that at 24 h. This indicated that the FSHR promoter of the Jintang black goat is a strong promoter and may be a gene-special promoter.Key words: Follicle-stimulating hormone receptor, gene promoter, Jintang black goat, molecular cloning, functional analysis

Dynamic Changes in the MicroRNA Expression Profile Reveal Multiple Regulatory Mechanisms in the Spinal Nerve Ligation Model of Neuropathic Pain

Neuropathic pain resulting from nerve lesions or dysfunction represents one of the most challenging neurological diseases to treat. A better understanding of the molecular mechanisms responsible for causing these maladaptive responses can help develop novel therapeutic strategies and biomarkers for neuropathic pain. We performed a miRNA expression profiling study of dorsal root ganglion (DRG) tissue from rats four weeks post spinal nerve ligation (SNL), a model of neuropathic pain. TaqMan low density arrays identified 63 miRNAs whose level of expression was significantly altered following SNL surgery. Of these, 59 were downregulated and the ipsilateral L4 DRG, not the injured L5 DRG, showed the most significant downregulation suggesting that miRNA changes in the uninjured afferents may underlie the development and maintenance of neuropathic pain. TargetScan was used to predict mRNA targets for these miRNAs and it was found that the transcripts with multiple predicted target sites belong to neurologically important pathways. By employing different bioinformatic approaches we identified neurite remodeling as a significantly regulated biological pathway, and some of these predictions were confirmed by siRNA knockdown for genes that regulate neurite growth in differentiated Neuro2A cells. In vitro validation for predicted target sites in the 3′-UTR of voltage-gated sodium channel Scn11a, alpha 2/delta1 subunit of voltage-dependent Ca-channel, and purinergic receptor P2rx ligand-gated ion channel 4 using luciferase reporter assays showed that identified miRNAs modulated gene expression significantly. Our results suggest the potential for miRNAs to play a direct role in neuropathic pain

Patient-derived xenograft (PDX) models in basic and translational breast cancer research

Patient-derived xenograft (PDX) models of a growing spectrum of cancers are rapidly supplanting long-established traditional cell lines as preferred models for conducting basic and translational preclinical research. In breast cancer, to complement the now curated collection of approximately 45 long-established human breast cancer cell lines, a newly formed consortium of academic laboratories, currently from Europe, Australia, and North America, herein summarizes data on over 500 stably transplantable PDX models representing all three clinical subtypes of breast cancer (ER+, HER2+, and "Triple-negative" (TNBC)). Many of these models are well-characterized with respect to genomic, transcriptomic, and proteomic features, metastatic behavior, and treatment response to a variety of standard-of-care and experimental therapeutics. These stably transplantable PDX lines are generally available for dissemination to laboratories conducting translational research, and contact information for each collection is provided. This review summarizes current experiences related to PDX generation across participating groups, efforts to develop data standards for annotation and dissemination of patient clinical information that does not compromise patient privacy, efforts to develop complementary data standards for annotation of PDX characteristics and biology, and progress toward "credentialing" of PDX models as surrogates to represent individual patients for use in preclinical and co-clinical translational research. In addition, this review highlights important unresolved questions, as well as current limitations, that have hampered more efficient generation of PDX lines and more rapid adoption of PDX use in translational breast cancer research

Hyperacute stabilization (\u3c12 hours) for polytrauma patients with unstable spinal fractures

BACKGROUND: The objective of this study is to validate the safety of hyperacute stabilization. METHODS: Patient demographics, American Spinal Injury Association (ASIA) grade on initial evaluation and serial follow up grades, hospital length of stay, Intensive Care Unit length of stay, intraoperative blood loss, postoperative length of stay, comorbidities, Injury Severity Score and complications of recumbency were recorded. RESULTS: Corroborating previous studies, our study shows polytrauma patients undergoing a hyperacute stabilization of a spinal fracture displayed a trend towards better neurological outcome and decreased hospital stays while having a similar complication rate to those operated on in a delayed fashion. CONCLUSIONS: Hyperacute stabilization can be associated with improved ASIA grades even in complete injuries

Transcriptional changes associated with breast cancer occur as normal human mammary epithelial cells overcome senescence barriers and become immortalized

Abstract Background Human mammary epithelial cells (HMEC) overcome two well-characterized genetic and epigenetic barriers as they progress from primary cells to fully immortalized cell lines in vitro. Finite lifespan HMEC overcome an Rb-mediated stress-associated senescence barrier (stasis), and a stringent, telomere-length dependent, barrier (agonescence or crisis, depending on p53 status). HMEC that have overcome the second senescence barrier are immortalized. Methods We have characterized pre-stasis, post-selection (post-stasis, with p16 silenced), and fully immortalized HMEC by transcription profiling and RT-PCR. Four pre-stasis and seven post-selection HMEC samples, along with 10 representatives of fully immortalized breast epithelial cell lines, were profiled using Affymetrix U133A/B chips and compared using both supervised and unsupervised clustering. Datasets were validated by RT-PCR for a select set of genes. Quantitative immunofluorescence was used to assess changes in transcriptional regulators associated with the gene expression changes. Results The most dramatic and uniform changes we observed were in a set of about 30 genes that are characterized as a "cancer proliferation cluster," which includes genes expressed during mitosis (CDC2, CDC25, MCM2, PLK1) and following DNA damage. The increased expression of these genes was particularly concordant in the fully immortalized lines. Additional changes were observed in IFN-regulated genes in some post-selection and fully immortalized cultures. Nuclear localization was observed for several transcriptional regulators associated with expression of these genes in post-selection and immortalized HMEC, including Rb, Myc, BRCA1, HDAC3 and SP1. Conclusion Gene expression profiles and cytological changes in related transcriptional regulators indicate that immortalized HMEC resemble non-invasive breast cancers, such as ductal and lobular carcinomas in situ, and are strikingly distinct from finite-lifespan HMEC, particularly with regard to genes involved in proliferation, cell cycle regulation, chromosome structure and the DNA damage response. The comparison of HMEC profiles with lines harboring oncogenic changes (e.g. overexpression of Her-2neu, loss of p53 expression) identifies genes involved in tissue remodeling as well as proinflamatory cytokines and S100 proteins. Studies on carcinogenesis using immortalized cell lines as starting points or "normal" controls need to account for the significant pre-existing genetic and epigenetic changes inherent in such lines before results can be broadly interpreted.</p

Transcriptional changes associated with breast cancer occur as normal human mammary epithelial cells overcome senescence barriers and become immortalized.

BackgroundHuman mammary epithelial cells (HMEC) overcome two well-characterized genetic and epigenetic barriers as they progress from primary cells to fully immortalized cell lines in vitro. Finite lifespan HMEC overcome an Rb-mediated stress-associated senescence barrier (stasis), and a stringent, telomere-length dependent, barrier (agonescence or crisis, depending on p53 status). HMEC that have overcome the second senescence barrier are immortalized.MethodsWe have characterized pre-stasis, post-selection (post-stasis, with p16 silenced), and fully immortalized HMEC by transcription profiling and RT-PCR. Four pre-stasis and seven post-selection HMEC samples, along with 10 representatives of fully immortalized breast epithelial cell lines, were profiled using Affymetrix U133A/B chips and compared using both supervised and unsupervised clustering. Datasets were validated by RT-PCR for a select set of genes. Quantitative immunofluorescence was used to assess changes in transcriptional regulators associated with the gene expression changes.ResultsThe most dramatic and uniform changes we observed were in a set of about 30 genes that are characterized as a "cancer proliferation cluster," which includes genes expressed during mitosis (CDC2, CDC25, MCM2, PLK1) and following DNA damage. The increased expression of these genes was particularly concordant in the fully immortalized lines. Additional changes were observed in IFN-regulated genes in some post-selection and fully immortalized cultures. Nuclear localization was observed for several transcriptional regulators associated with expression of these genes in post-selection and immortalized HMEC, including Rb, Myc, BRCA1, HDAC3 and SP1.ConclusionGene expression profiles and cytological changes in related transcriptional regulators indicate that immortalized HMEC resemble non-invasive breast cancers, such as ductal and lobular carcinomas in situ, and are strikingly distinct from finite-lifespan HMEC, particularly with regard to genes involved in proliferation, cell cycle regulation, chromosome structure and the DNA damage response. The comparison of HMEC profiles with lines harboring oncogenic changes (e.g. overexpression of Her-2neu, loss of p53 expression) identifies genes involved in tissue remodeling as well as proinflamatory cytokines and S100 proteins. Studies on carcinogenesis using immortalized cell lines as starting points or "normal" controls need to account for the significant pre-existing genetic and epigenetic changes inherent in such lines before results can be broadly interpreted

Transcriptional changes associated with breast cancer occur as normal human mammary epithelial cells overcome senescence barriers and become immortalized.

BackgroundHuman mammary epithelial cells (HMEC) overcome two well-characterized genetic and epigenetic barriers as they progress from primary cells to fully immortalized cell lines in vitro. Finite lifespan HMEC overcome an Rb-mediated stress-associated senescence barrier (stasis), and a stringent, telomere-length dependent, barrier (agonescence or crisis, depending on p53 status). HMEC that have overcome the second senescence barrier are immortalized.MethodsWe have characterized pre-stasis, post-selection (post-stasis, with p16 silenced), and fully immortalized HMEC by transcription profiling and RT-PCR. Four pre-stasis and seven post-selection HMEC samples, along with 10 representatives of fully immortalized breast epithelial cell lines, were profiled using Affymetrix U133A/B chips and compared using both supervised and unsupervised clustering. Datasets were validated by RT-PCR for a select set of genes. Quantitative immunofluorescence was used to assess changes in transcriptional regulators associated with the gene expression changes.ResultsThe most dramatic and uniform changes we observed were in a set of about 30 genes that are characterized as a "cancer proliferation cluster," which includes genes expressed during mitosis (CDC2, CDC25, MCM2, PLK1) and following DNA damage. The increased expression of these genes was particularly concordant in the fully immortalized lines. Additional changes were observed in IFN-regulated genes in some post-selection and fully immortalized cultures. Nuclear localization was observed for several transcriptional regulators associated with expression of these genes in post-selection and immortalized HMEC, including Rb, Myc, BRCA1, HDAC3 and SP1.ConclusionGene expression profiles and cytological changes in related transcriptional regulators indicate that immortalized HMEC resemble non-invasive breast cancers, such as ductal and lobular carcinomas in situ, and are strikingly distinct from finite-lifespan HMEC, particularly with regard to genes involved in proliferation, cell cycle regulation, chromosome structure and the DNA damage response. The comparison of HMEC profiles with lines harboring oncogenic changes (e.g. overexpression of Her-2neu, loss of p53 expression) identifies genes involved in tissue remodeling as well as proinflamatory cytokines and S100 proteins. Studies on carcinogenesis using immortalized cell lines as starting points or "normal" controls need to account for the significant pre-existing genetic and epigenetic changes inherent in such lines before results can be broadly interpreted

Flexible silver grid/PEDOT:PSS hybrid electrodes for large area inverted polymer solar cells

<span style="color: rgb(51, 51, 51); font-family: arial, helvetica, sans-serif; font-size: 13px; line-height: 22px; background-color: rgb(248, 248, 248);">Bulk-heterojunction (BHJ) organic&nbsp;</span><span class="hitHilite" style="margin: 0px; list-style: none; padding: 0px; color: rgb(51, 51, 51); font-family: arial, helvetica, sans-serif; font-size: 13px; line-height: 22px; background-color: rgb(255, 255, 0);">solar</span><span style="color: rgb(51, 51, 51); font-family: arial, helvetica, sans-serif; font-size: 13px; line-height: 22px; background-color: rgb(248, 248, 248);">&nbsp;cell (OSC) has become a major thrust in&nbsp;</span><span class="hitHilite" style="margin: 0px; list-style: none; padding: 0px; color: rgb(51, 51, 51); font-family: arial, helvetica, sans-serif; font-size: 13px; line-height: 22px; background-color: rgb(255, 255, 0);">solar</span><span style="color: rgb(51, 51, 51); font-family: arial, helvetica, sans-serif; font-size: 13px; line-height: 22px; background-color: rgb(248, 248, 248);">&nbsp;energy research. It is highly critical to fabricate&nbsp;</span><span class="hitHilite" style="margin: 0px; list-style: none; padding: 0px; color: rgb(51, 51, 51); font-family: arial, helvetica, sans-serif; font-size: 13px; line-height: 22px; background-color: rgb(255, 255, 0);">large</span><span style="color: rgb(51, 51, 51); font-family: arial, helvetica, sans-serif; font-size: 13px; line-height: 22px; background-color: rgb(248, 248, 248);">-</span><span class="hitHilite" style="margin: 0px; list-style: none; padding: 0px; color: rgb(51, 51, 51); font-family: arial, helvetica, sans-serif; font-size: 13px; line-height: 22px; background-color: rgb(255, 255, 0);">area</span><span style="color: rgb(51, 51, 51); font-family: arial, helvetica, sans-serif; font-size: 13px; line-height: 22px; background-color: rgb(248, 248, 248);">&nbsp;high-efficiency OSCs in order to maximize the fraction of active&nbsp;</span><span class="hitHilite" style="margin: 0px; list-style: none; padding: 0px; color: rgb(51, 51, 51); font-family: arial, helvetica, sans-serif; font-size: 13px; line-height: 22px; background-color: rgb(255, 255, 0);">area</span><span style="color: rgb(51, 51, 51); font-family: arial, helvetica, sans-serif; font-size: 13px; line-height: 22px; background-color: rgb(248, 248, 248);">&nbsp;and hence their practical application. In this paper, we have fabricated a promising&nbsp;</span><span class="hitHilite" style="margin: 0px; list-style: none; padding: 0px; color: rgb(51, 51, 51); font-family: arial, helvetica, sans-serif; font-size: 13px; line-height: 22px; background-color: rgb(255, 255, 0);">hybrid</span><span style="color: rgb(51, 51, 51); font-family: arial, helvetica, sans-serif; font-size: 13px; line-height: 22px; background-color: rgb(248, 248, 248);">transparent electrode consisting of high resolution embedded&nbsp;</span><span class="hitHilite" style="margin: 0px; list-style: none; padding: 0px; color: rgb(51, 51, 51); font-family: arial, helvetica, sans-serif; font-size: 13px; line-height: 22px; background-color: rgb(255, 255, 0);">silver</span><span style="color: rgb(51, 51, 51); font-family: arial, helvetica, sans-serif; font-size: 13px; line-height: 22px; background-color: rgb(248, 248, 248);">&nbsp;</span><span class="hitHilite" style="margin: 0px; list-style: none; padding: 0px; color: rgb(51, 51, 51); font-family: arial, helvetica, sans-serif; font-size: 13px; line-height: 22px; background-color: rgb(255, 255, 0);">grid</span><span style="color: rgb(51, 51, 51); font-family: arial, helvetica, sans-serif; font-size: 13px; line-height: 22px; background-color: rgb(248, 248, 248);">&nbsp;(Ag-</span><span class="hitHilite" style="margin: 0px; list-style: none; padding: 0px; color: rgb(51, 51, 51); font-family: arial, helvetica, sans-serif; font-size: 13px; line-height: 22px; background-color: rgb(255, 255, 0);">grid</span><span style="color: rgb(51, 51, 51); font-family: arial, helvetica, sans-serif; font-size: 13px; line-height: 22px; background-color: rgb(248, 248, 248);">) in hybridization with high conductance poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate) (</span><span class="hitHilite" style="margin: 0px; list-style: none; padding: 0px; color: rgb(51, 51, 51); font-family: arial, helvetica, sans-serif; font-size: 13px; line-height: 22px; background-color: rgb(255, 255, 0);">PEDOT</span><span style="color: rgb(51, 51, 51); font-family: arial, helvetica, sans-serif; font-size: 13px; line-height: 22px; background-color: rgb(248, 248, 248);">:</span><span class="hitHilite" style="margin: 0px; list-style: none; padding: 0px; color: rgb(51, 51, 51); font-family: arial, helvetica, sans-serif; font-size: 13px; line-height: 22px; background-color: rgb(255, 255, 0);">PSS</span><span style="color: rgb(51, 51, 51); font-family: arial, helvetica, sans-serif; font-size: 13px; line-height: 22px; background-color: rgb(248, 248, 248);">) (PH1000), which gives both high transparency and low sheet resistance. We further demonstrate its application in</span><span class="hitHilite" style="margin: 0px; list-style: none; padding: 0px; color: rgb(51, 51, 51); font-family: arial, helvetica, sans-serif; font-size: 13px; line-height: 22px; background-color: rgb(255, 255, 0);">large</span><span style="color: rgb(51, 51, 51); font-family: arial, helvetica, sans-serif; font-size: 13px; line-height: 22px; background-color: rgb(248, 248, 248);">-</span><span class="hitHilite" style="margin: 0px; list-style: none; padding: 0px; color: rgb(51, 51, 51); font-family: arial, helvetica, sans-serif; font-size: 13px; line-height: 22px; background-color: rgb(255, 255, 0);">area</span><span style="color: rgb(51, 51, 51); font-family: arial, helvetica, sans-serif; font-size: 13px; line-height: 22px; background-color: rgb(248, 248, 248);">&nbsp;high efficiency OSCs. By carefully tuning the properties of the Ag-</span><span class="hitHilite" style="margin: 0px; list-style: none; padding: 0px; color: rgb(51, 51, 51); font-family: arial, helvetica, sans-serif; font-size: 13px; line-height: 22px; background-color: rgb(255, 255, 0);">grid</span><span style="color: rgb(51, 51, 51); font-family: arial, helvetica, sans-serif; font-size: 13px; line-height: 22px; background-color: rgb(248, 248, 248);">&nbsp;based&nbsp;</span><span class="hitHilite" style="margin: 0px; list-style: none; padding: 0px; color: rgb(51, 51, 51); font-family: arial, helvetica, sans-serif; font-size: 13px; line-height: 22px; background-color: rgb(255, 255, 0);">hybrid</span><span class="hitHilite" style="margin: 0px; list-style: none; padding: 0px; color: rgb(51, 51, 51); font-family: arial, helvetica, sans-serif; font-size: 13px; line-height: 22px; background-color: rgb(255, 255, 0);">electrodes</span><span style="color: rgb(51, 51, 51); font-family: arial, helvetica, sans-serif; font-size: 13px; line-height: 22px; background-color: rgb(248, 248, 248);">, the sheet resistance is further reduced to as low as 1.2 Omega sq(-1).&nbsp;</span><span class="hitHilite" style="margin: 0px; list-style: none; padding: 0px; color: rgb(51, 51, 51); font-family: arial, helvetica, sans-serif; font-size: 13px; line-height: 22px; background-color: rgb(255, 255, 0);">Inverted</span><span style="color: rgb(51, 51, 51); font-family: arial, helvetica, sans-serif; font-size: 13px; line-height: 22px; background-color: rgb(248, 248, 248);">&nbsp;OSCs with device&nbsp;</span><span class="hitHilite" style="margin: 0px; list-style: none; padding: 0px; color: rgb(51, 51, 51); font-family: arial, helvetica, sans-serif; font-size: 13px; line-height: 22px; background-color: rgb(255, 255, 0);">area</span><span style="color: rgb(51, 51, 51); font-family: arial, helvetica, sans-serif; font-size: 13px; line-height: 22px; background-color: rgb(248, 248, 248);">&nbsp;of 1.21 cm(2) exhibited a record PCE of 3.36%&nbsp;</span><span class="hitHilite" style="margin: 0px; list-style: none; padding: 0px; color: rgb(51, 51, 51); font-family: arial, helvetica, sans-serif; font-size: 13px; line-height: 22px; background-color: rgb(255, 255, 0);">for</span><span style="color: rgb(51, 51, 51); font-family: arial, helvetica, sans-serif; font-size: 13px; line-height: 22px; background-color: rgb(248, 248, 248);">&nbsp;poly(3-hexylthiophene):[6,6]-phenyl-C-61 butyric acid methyl ester (P3HT:PC61BM) blend film as the active layer; and the PCE of PTB7:PC71BM devices reached 5.85%. To the best of our knowledge, the PCE of 5.85% is the highest in&nbsp;</span><span class="hitHilite" style="margin: 0px; list-style: none; padding: 0px; color: rgb(51, 51, 51); font-family: arial, helvetica, sans-serif; font-size: 13px; line-height: 22px; background-color: rgb(255, 255, 0);">large</span><span style="color: rgb(51, 51, 51); font-family: arial, helvetica, sans-serif; font-size: 13px; line-height: 22px; background-color: rgb(248, 248, 248);">-</span><span class="hitHilite" style="margin: 0px; list-style: none; padding: 0px; color: rgb(51, 51, 51); font-family: arial, helvetica, sans-serif; font-size: 13px; line-height: 22px; background-color: rgb(255, 255, 0);">area</span><span style="color: rgb(51, 51, 51); font-family: arial, helvetica, sans-serif; font-size: 13px; line-height: 22px; background-color: rgb(248, 248, 248);">&nbsp;</span><span class="hitHilite" style="margin: 0px; list-style: none; padding: 0px; color: rgb(51, 51, 51); font-family: arial, helvetica, sans-serif; font-size: 13px; line-height: 22px; background-color: rgb(255, 255, 0);">flexible</span><span style="color: rgb(51, 51, 51); font-family: arial, helvetica, sans-serif; font-size: 13px; line-height: 22px; background-color: rgb(248, 248, 248);">&nbsp;OSCs (&gt;1 cm(2)) till date. (C) 2014 Elsevier Ltd. All rights reserved.</span