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Stress granules ( SGs) and processing bodies ( PBs) comprise the main types of cytoplasmic RNA foci during stress. Our previous data indicate that knockdown of human Tudor staphylococcal nuclease (Tudor-SN) affects the aggregation of SGs. However, the precise molecular mechanism has not been determined fully. In the present study, we demonstrate that Tudor-SN binds and colocalizes with many core components of SGs, such as poly(A)+ mRNA binding protein 1, T-cell internal antigen-1-related protein and poly(A)+ mRNA, and SG/PB sharing proteins Argonaute 1/2, but not PB core proteins, such as decapping enzyme 1 a/b, confirming that Tudor-SN is an SG-specific protein. We also demonstrate that the Tudor-SN granule actively communicates with the nuclear and cytosolic pool under stress conditions. Tudor-SN can regulate the aggregation dynamics of poly(A)+ mRNA-containing SGs and selectively stabilize the SG-associated mRNA during cellular stress. [ABSTRACT FROM AUTHOR]

Objective To construct eukaryotic Flag (DYKDDDDK) expressing recombinant plasmids, pCMV-N-Flag-SND1-No1/2, which contain the coding sequence of human SND1-No1 (from 1st AUG)or SND1-No2 (from 2nd AUG), and perform the cellular localization analysis of Flag-tagged SND1-No1/2 under stress condition to study the function of the two AUG in the SND1 containing stress granules formation. Methods The gene fragments of SND1-No1/2 were amplified by PCR from the whole SND1 transcript and inserted into pCMV-N-Flag expressing vector through BamHI/EcoRI double enzyme digestion and T4 DNA Ligase connection. The recombinant pCMV-N-Flag-SND1-No1/2 plasmids were transfected into HeLa cells and the expression of Flag-SND1-No1/2 fusion proteins was examined by Western blotting assay. Immunofluo-rescence assay was performed to detect the co-localization of Flag-SND1-No1/2 with endogenous SND1 granule. Results The pCMV-N-Flag-SND1-No1/2 were sequenced and digested correctly by restriction single/double enzyme. The Flag-tagged SND1-No1/2 fusion proteins were also detected in transfected HeLa cell by Western blotting assay. Both of them showed the co- localization with endogenous SND1 granule. Conclusion The recombinant eukaryotic plasmids of pCMV-N-Flag-SND1-No1/2 were constructed successfully and expressed effectively. The depletion of 1st AUG failed to affect the formation of SND1 containing stress granules. [ABSTRACT FROM AUTHOR]

Objective To construct eukaryotic enhanced green fluorescent protein (EGFP) expressing recombinant plasmid, pEGFP-C1-hnRNP A1, which contains coding sequence of human hnRNP A1 (heterogeneous nuclear ribonucleo protein A1), and to perform cellular localization analysis of EGFP tagged hnRNP A1 under stress. Methods Total RNA was isolated from HeLa cell used for synthesis of first-strand cDNAs using reverse primers that are specific for the3'-un-translated region of hnRNP A1. hnRNP A1 gene fragments were then amplified by touch-down PCR from those cDNAs and inserted into pEGFP-C1 fluorescent bearing vector through EcoR I/BamH II double enzyme digestion and T4 DNA Ligase connection. The recombinant pEGFP-C1-hnRNP A1 plasmid was transfected into HeLa cells and green fluorescent tagged fusion proteins was examined by Western blot and confocal fluorescence microscopy. Co-localization of EGFP-hnRNP A1 with poly (A)+ mRNA (the marker of the stress granules), or DCP1a (the marker of processome) were detected by RNA fluores cence in situ hybridization and immunofluorescence. Results The pEGFP-C1-hnRNP A1 was sequenced and digested correctly by restriction single/double enzyme. The green fluorescent fusion protein was also detected in transfected HeLa cell by Western blot and confocal fluorescence microscopy. EGFP-hnRNP A1 co-localizes with poly(A)+ mRNA, but not DCP1a. Conclusion Recombinant eukaryotic plasmid of pEGFP-C1-hnRNP A1 was constructed successfully and expressed effectively. EGFP tagged hnRNP A1 takes part in forming stress granules. [ABSTRACT FROM AUTHOR]

Objective To explore the factors affecting the male sexual quality after PANP rectal cancer radical dissection and provide reference to improve postoperative sexual quality of the male. Methods A total of 134 patients undertaken pelvic autonomic nerve preservation (PANP) rectal cancer radical dissection in this hospital were investigated through questionnaire survey and analyzed by the logistic regression in its influencing factors. Results The total score of postoperative sexual quality of 134 male patients in this study was (136.51±45.96). The quality of daily sleep, the self-cognition to the disease, the attitude to the patients and coordination of their families ranked the first three places successively. The self-cognition to the disease, the attitude to the patients of the families and cooperation were the key factors of sexual quality, which had statistical significance in difference (P<0.05), Conclusion Strengthening the self-cognition to the disease, the attitude to the patients of the families and cooperation are the important factors in the nursing health education after PANP rectal cancer radical dissection. [ABSTRACT FROM AUTHOR]

Electro-Fenton is a commonly used approach for pollutant removal from different wastewater that requires high energy consumption. Coupling electro-Fenton with solar energy could potentially overcome high power consumption. Thus, this study combined photocatalysis with three-dimensional electro-Fenton to treat acrylamide (AM) wastewater. The removal efficiencies of chemical oxygen demand (COD), ammonia nitrogen (NH 3 -N), and total organic carbon (TOC) in photocatalysis-coupled electro-Fenton were studied. The effects of current density, iron powder dosage, plate spacing, and photocatalyst dosage on COD and NH 3 -N removal were also optimised using response surface methodology (RSM). The photocatalysis coupled three-dimensional electro-Fenton reduced energy consumption compared to single photocatalysis or electro-Fenton technology. The COD and NH 3 -N removal rates were 82.444% and 92.810%, respectively, at the current density of 16.000 mA/cm 2 , iron powder dosage of 1.330 g, plate spacing of 16.643 mm, photocatalyst dosage of 0.2 g. This study demonstrated that organic pollutants can be degraded efficiently at a low concentration of catalysts coupled with the electro-Fenton process, offering a low-energy consumption treatment of wastewater.

Stress granules (SGs) are dynamic dense structures in the cytoplasm that form in response to a variety of environmental stress stimuli. Staphylococcal nuclease and Tudor domain containing 1 (SND1) is a type of RNA-binding protein and has been identified as a transcriptional co-activator. Our previous studies have shown that SND1 is a component of the stress granule, which forms under stress conditions. Here, we observed that SND1 granules were often surrounded by ɑ-tubulin-microtubules in 45°C-treated HeLa cells at 15 min or colocalized with microtubules at 30 or 45 min. Furthermore, Nocodazole-mediated microtubule depolymerization could significantly affect the efficient recruitment of SND1 proteins to the SGs during heat shock stress. In addition, the 45°C heat shock mediated the enhancement of eIF2α phosphorylation, which was not affected by treatment with Nocodazole, an agent that disrupts the cytoskeleton. The intact microtubule cytoskeletal tracks are important for the efficient assembly of SND1 granules under heat shock stress and may facilitate SND1 shuttling between cytoplasmic RNA foci. Anat Rec, 300:2192-2199, 2017. © 2017 The Authors The Anatomical Record published by Wiley Periodicals, Inc. on behalf of American Association of Anatomists., (Copyright © 2017 The Authors The Anatomical Record published by Wiley Periodicals, Inc. on behalf of American Association of Anatomists.)

The family of STAT proteins consists of seven members that mediate highly specific functions in cytokine signaling. STAT6 is a critical regulator of transcription for interleukin-4 (IL-4)-induced genes. Activation of gene expression involves recruitment of coactivator proteins that function as bridging factors connecting sequence-specific transcription factors to the basal transcription machinery, and as chromatin-modifying enzymes. In this report, we show that the coacitivator p100 protein can interact with STAT6 through its SN domain both in vivo and in vitro, resulting in enhancement of STAT6-mediated gene transcriptional acitivation. Consistent with our previous reports, we identified intracellular localization of p100 and STAT-6 by confocal microscopy examined in response to IL-4. Moreover, in consideration of STAT molecules sharing significant homology in structure and function, we detected whether p100 can associate with STAT-1. In conclusion, this study found no evidence that p100 functions as a transcriptional coactivator for STAT1-dependent gene regulation.