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Find video protocols related to scientific articles indexed in Pubmed.
Melatonin prevents cell death and mitochondrial dysfunction via a SIRT1-dependent mechanism during ischemic-stroke in mice.
J. Pineal Res.
PUBLISHED: 11-05-2014
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Silent information regulator 1 (SIRT1), a type of histone deacetylase, is a highly effective therapeutic target for protection against ischemia reperfusion (IR) injury (IRI). Previous studies showed that melatonin preserves SIRT1 expression in neuronal cells of newborn rats after hypoxia-ischemia. However, the definite role of SIRT1 in the protective effect of melatonin against cerebral IRI in adult has not been explored. In this study, the brain of adult mice were subjected to IRI. Prior to this procedure, the mice were given intraperitoneal with or without the SIRT1 inhibitor, EX527. Melatonin conferred a cerebralprotective effect, as shown by reduced infarct volume, lowered brain edema and increased neurological scores. The melatonin-induced up-regulation of SIRT1 was also associated with an increase in the anti-apoptotic factor, Bcl2, and a reduction in the pro-apoptotic factor Bax. Moreover, melatonin resulted in a well-preserved mitochondrial membrane potential (MMP), mitochondria Complex I activity, mitochondrial cytochrome c level while it reduced cytosolic cytochrome c level. However, the melatonin-elevated mitochondrial function was reversed by EX527 treatment. In summary, our results demonstrate that melatonin treatment attenuates cerebral IRI by reducing IR-induced mitochondrial dysfunction through the activation of SIRT1 signaling. This article is protected by copyright. All rights reserved.
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Large tunable optical absorption of CVD graphene under total internal reflection by strain engineering.
Nanotechnology
PUBLISHED: 10-24-2014
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We have developed a method to tune polarization-dependent optical absorption of large-scale chemical vapor deposition (CVD) graphene under total internal reflection (TIR) by strain engineering. Through control of the strain direction, the optical absorption of graphene for transverse magnetic or transverse electric waves can be separately tuned. Strain-induced modulation of the optical absorption has been theoretically expected when light is normally incident through graphene. Under TIR, however, we experimentally observed a significant increase in the strain-induced tunability of optical absorption for CVD graphene, with the modulation efficiency of optical absorption in monolayer graphene increasing by a factor of three times that for normal incidence. We conclude that the strain sensitivity of optical absorption of graphene under TIR offers significant potential for application in many areas such as ultra-thin optical devices and strain sensors.
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Theoretical study of the hydration of atmospheric nucleation precursors with acetic acid.
J Phys Chem A
PUBLISHED: 08-29-2014
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While atmosphere is known to contain a significant fraction of organic substance and the effect of acetic acid to stabilize hydrated sulfuric acids is found to be close that of ammonia, the details about the hydration of (CH3COOH)(H2SO4)2 are poorly understood, especially for the larger clusters with more water molecules. We have investigated structural characteristics and thermodynamics of the hydrates using density functional theory (DFT) at PW91PW91/6-311++G(3df,3pd) level. The phenomena of the structural evolution may exist during the early stage of the clusters formation, and we tentatively proposed a calculation path for the Gibbs free energies of the clusters formation via the structural evolution. The results in this study supply a picture of the first deprotonation of sulfuric acids for a system consisting of two sulfuric acid molecules, an acetic acid molecule, and up to three waters at 0 and 298.15 K, respectively. We also replace one of the sulfuric acids with a bisulfate anion in (CH3COOH)(H2SO4)2 to explore the difference of acid dissociation between two series of clusters and interaction of performance in clusters growth between ion-mediated nucleation and organics-enhanced nucleation.
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A review of melatonin as a suitable antioxidant against myocardial ischemia-reperfusion injury and clinical heart diseases.
J. Pineal Res.
PUBLISHED: 08-16-2014
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Cardiac tissue loss is one of the most important factors leading to the unsatisfactory recovery even after treatment of ischemic heart disease. Melatonin, a circadian molecule with marked antioxidant properties, protects against ischemia-reperfusion (IR) injury. In particular, the myocardial protection of melatonin is substantial. We initially focus on the cardioprotective effects of melatonin in myocardial IR. These studies showed how melatonin preserves the microstructure of the cardiomyocyte and reduces myocardial IR injury. Thereafter, downstream signaling pathways of melatonin were summarized including Janus kinase 2/signal transducers and activators of transcription 3, nitric oxide-synthase, and nuclear factor erythroid 2 related factor 2. Herein, we propose the clinical applications of melatonin in several ischemic heart diseases. Collectively, the information summarized in this review (based on in vitro, animal, and human studies) should serve as a comprehensive reference for the action of melatonin in cardioprotection and hopefully will contribute to the design of future experimental research.
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Novel role of silent information regulator 1 in acute endothelial cell oxidative stress injury.
Biochim. Biophys. Acta
PUBLISHED: 08-14-2014
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Silent information regulator 1 (SIRT1), a class III histone deacetylase, retards aging and plays roles in cellular oxidative stress injury (OSI). However, the biological context in which SIRT1 promotes oxidative injury is not fully understood. Here, we show that SIRT1 essentially mediates hydrogen peroxide (H2O2)-induced cytotoxicity in human umbilical vein endothelial cell (HUVEC). In HUVECs, SIRT1 protein expression was significantly increased in a dose-dependent manner after H2O2 treatment, whereas the acetylation levels of the NF-?B p65 subunit and p53 were decreased. EX527 (a specific SIRT1 inhibitor) conferred protection to the HUVECs against H2O2, as indicated by an improved cell viability, adhesion, an enhanced migratory ability, a decreased apoptotic index, decreased reactive oxygen species (ROS) production and reductions in several biochemical parameters. Immunofluorescence and Western blot analyses demonstrated that H2O2 treatment up-regulated SIRT1, phosphorylated-JNK (p-JNK), p-p38MAPK, and p-ERK expression. EX527 pretreatment reversed these effects on SIRT1, p-JNK, and p-p38MAPK but further increased the p-ERK levels. Similar results were confirmed in SIRT1 siRNA experiments. In summary, SIRT1 signaling pathway inhibition imparts protection against acute endothelial OSI, and modulation of MAPKs (JNK, p38MAPK, and ERK) may be involved in the protective effect of SIRT1 inhibition.
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Intermittently administered parathyroid hormone [1-34] promotes tendon-bone healing in a rat model.
Int J Mol Sci
PUBLISHED: 08-11-2014
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The objective of this study was to investigate whether intermittent administration of parathyroid hormone [1-34] (PTH[1-34]) promotes tendon-bone healing after anterior cruciate ligament (ACL) reconstruction in vivo. A rat model of ACL reconstruction with autograft was established at the left hind leg. Every day, injections of 60 ?g PTH[1-34]/kg subcutaneously were given to the PTH group rats (n=10) for four weeks, and the controls (n=10) received saline. The tendon-bone healing process was evaluated by micro-CT, biomechanical test, histological and immunohistochemical analyses. The effects of PTH[1-34] on serum chemistry, bone microarchitecture and expression of the PTH receptor (PTH1R) and osteocalcin were determined. Administration of PTH[1-34] significantly increased serum levels of calcium, alkaline phosphatase (AP), osteocalcin and tartrate-resistant acid phosphatase (TRAP). The expression of PTH1R on both osteocytes and chondrocyte-like cells at the tendon-bone interface was increased in the PTH group. PTH[1-34] also enhanced the thickness and microarchitecture of trabecular bone according to the micro-CT analysis. The results imply that systematically intermittent administration of PTH[1-34] promotes tendon-bone healing at an early stage via up-regulated PTH1R. This method may enable a new strategy for the promotion of tendon-bone healing after ACL reconstruction.
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Theoretical study of temperature dependence and Rayleigh scattering properties of chloride hydration clusters.
Phys Chem Chem Phys
PUBLISHED: 08-07-2014
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Cl(-)(H2O)n (n = 5-6) clusters were investigated using a basin hopping (BH) method coupled with density functional theory (DFT). Structures, energetics, thermodynamics, and vibrational frequencies were obtained using high level ab initio calculations. DF-LMP2 (second-order Møller-Plesset perturbation theory using local and density fitting approximations) with an appropriate basis set were employed for final optimization and frequency calculation, which has been benchmarked in a recent study. The global minimum of Cl(-)(H2O)5 was verified and the new competitive local minimum of Cl(-)(H2O)6 was offered. Considering the increasing complexity of the large system and the high flexibility of the hydrogen bonding environment, Boltzmann averaged Gibbs free energy was provided taking into account the contributions of local minima on the potential energy surface. Finally, the temperature dependence of the conformational population for isomers of Cl(-)(H2O)n (n = 5-6) and Rayleigh scattering properties of Cl(-)(H2O)n (n = 1-6) have been investigated systematically for the first time.
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A review of melatonin in hepatic ischemia/reperfusion injury and clinical liver disease.
Ann. Med.
PUBLISHED: 07-18-2014
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Abstract Ischemia/reperfusion injury (IRI) can lead to cellular and, eventually, organ dysfunction, with the liver being one of the most frequently affected organs. Melatonin, a molecule that has notable antioxidant and anti-inflammatory properties, has been shown to protect against hepatic IRI. The purpose of this review is to summarize the protective effects of melatonin on hepatic IRI. The review initially summarizes the antioxidant properties of melatonin. We then discuss the protective effects of melatonin against endothelial and mitochondrial dysfunction. Thereafter, we introduce some information covering melatonin-related signaling pathways, including heme oxygenase-1 (HO-1), toll-like receptor (TLR), c-Jun N-terminal kinase (JNK), and so on. Furthermore, the clinical application of melatonin to hepatic diseases is considered. Finally, the safety of melatonin is evaluated. Taken together, the information compiled in this review will serve as a comprehensive reference regarding the pharmacological benefits of melatonin on hepatic IRI, aid in the design of future experimental research, and promote melatonin as a new therapeutic target.
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Dual mechanisms by which miR-125b represses IRF4 to induce myeloid and B-cell leukemias.
Blood
PUBLISHED: 07-08-2014
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The oncomir microRNA-125b (miR-125b) is upregulated in a variety of human neoplastic blood disorders and constitutive upregulation of miR-125b in mice can promote myeloid and B-cell leukemia. We found that miR-125b promotes myeloid and B-cell neoplasm by inducing tumorigenesis in hematopoietic progenitor cells. Our study demonstrates that miR-125b induces myeloid leukemia by enhancing myeloid progenitor output from stem cells as well as inducing immortality, self-renewal, and tumorigenesis in myeloid progenitors. Through functional and genetic analyses, we demonstrated that miR-125b induces myeloid and B-cell leukemia by inhibiting interferon regulatory factor 4 (IRF4) but through distinct mechanisms; it induces myeloid leukemia through repressing IRF4 at the messenger RNA (mRNA) level without altering the genomic DNA and induces B-cell leukemia via genetic deletion of the gene encoding IRF4.
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Flexible graphene saturable absorber on two-layer structure for tunable mode-locked soliton fiber laser.
Opt Express
PUBLISHED: 06-13-2014
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Using a two-layer structure consisting of polyethylene terephthalate (PET) and polydimethylsiloxane (PDMS) to support graphene grown by chemical vapor deposition (CVD), we demonstrate a flexible integrated graphene saturable absorber (SA) on microfiber for passive mode-locked soliton fiber laser. This method can optimize the light-graphene interaction by using evanescent field in the integration structure. Moreover, the fiber laser with the in-line microfiber-to-graphene SA can realize the tunabilities of both the 3dB bandwidth of output optical spectrum and the pulse width of soliton. This tunable mode-locked soliton laser has potential applications in optical communication, optical microscopy, and so on.
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An overview of the molecular mechanisms and novel roles of Nrf2 in neurodegenerative disorders.
Cytokine Growth Factor Rev.
PUBLISHED: 06-05-2014
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Recently, growing evidence has demonstrated that nuclear factor erythroid 2-related factor 2 (Nrf2) is a pivotal regulator of endogenous defense systems that function via the activation of a set of protective genes, and this is particularly clear in the central nervous system (CNS). Therefore, it is highly useful to summarize the current literature on the molecular mechanisms and role of Nrf2 in the CNS. In this review, we first briefly introduce the molecular features of Nrf2. We then discuss the regulation, cerebral actions, upstream modulators and downstream targets of Nrf2 pathway. Following this background, we expand our discussion to the role of Nrf2 in several major neurodegenerative disorders (NDDs) such as Alzheimer's disease, Parkinson's disease, Huntington's disease, multiple sclerosis and amyotrophic lateral sclerosis. Lastly, we discuss some potential future directions. The information reviewed here may be significant in the design of further experimental research and increase the potential of Nrf2 as a therapeutic target in the future.
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GPS-SUMO: a tool for the prediction of sumoylation sites and SUMO-interaction motifs.
Nucleic Acids Res.
PUBLISHED: 05-31-2014
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Small ubiquitin-like modifiers (SUMOs) regulate a variety of cellular processes through two distinct mechanisms, including covalent sumoylation and non-covalent SUMO interaction. The complexity of SUMO regulations has greatly hampered the large-scale identification of SUMO substrates or interaction partners on a proteome-wide level. In this work, we developed a new tool called GPS-SUMO for the prediction of both sumoylation sites and SUMO-interaction motifs (SIMs) in proteins. To obtain an accurate performance, a new generation group-based prediction system (GPS) algorithm integrated with Particle Swarm Optimization approach was applied. By critical evaluation and comparison, GPS-SUMO was demonstrated to be substantially superior against other existing tools and methods. With the help of GPS-SUMO, it is now possible to further investigate the relationship between sumoylation and SUMO interaction processes. A web service of GPS-SUMO was implemented in PHP+JavaScript and freely available at http://sumosp.biocuckoo.org.
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Ultrasensitive flow sensing of a single cell using graphene-based optical sensors.
Nano Lett.
PUBLISHED: 05-06-2014
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On the basis of the polarization-dependent absorption of graphene under total internal reflection, we designed a graphene-based optical refractive index sensor with high resolution of 1.7 × 10(-8) and sensitivity of 4.3 × 10(7) mV/RIU, as well as an extensive dynamic range. This highly sensitive graphene optical sensor enables label-free, live-cell, and highly accurate detection of a small quantity of cancer cells among normal cells at the single-cell level and the simultaneous detection and distinction of two cell lines without separation. It provides an accurate statistical distribution of normal and cancer cells with fewer cells. This facile and highly sensitive sensing refractive index may expand the practical applications of the biosensor.
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EZH2 promotes tumor cell migration and invasion via epigenetic repression of E-cadherin in renal cell carcinoma.
BJU Int.
PUBLISHED: 02-13-2014
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To investigate the molecular mechanism and clinical significance for an oncogenic role of EZH2 in renal cell carcinoma (RCC).
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The selective transfer of patterned graphene.
Sci Rep
PUBLISHED: 09-03-2013
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We demonstrate a selective microcleaving graphene (MG) transfer technique for the transfer of graphene patterns and graphene devices onto chosen targets using a bilayer-polymer structure and femtosecond laser microfabrication. In the bilayer-polymer structure, the first layer is used to separate the target graphene from the other flakes, and the second layer transfers the patterned graphene to the chosen targets. This selective transfer technique, which exactly transfers the patterned graphene onto a chosen target, leaving the other flakes on the original substrate, provides an efficient route for the fabrication of MG for microdevices and flexible electronics and the optimization of graphenes performance. This method will facilitate the preparation of van der Waals heterostructures and enable the optimization of the performance of graphene hybrid devices.
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( - )-Arctigenin as a lead compound for anticancer agent.
Nat. Prod. Res.
PUBLISHED: 08-20-2013
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( - )-Arctigenin, an important active constituent of the traditional Chinese herb Fructus Arctii, was found to exhibit various bioactivities, so it can be used as a good lead compound for further structure modification in order to find a safer and more potent medicine. ( - )-Arctigenin derivatives 1-5 of ( - )-arctingen were obtained by modifying with ammonolysis at the lactone ring and sulphonylation at C (6) and C (6?) and O-demethylation at CH3O-C (3), CH3O-C (3?) and CH3O-C (4?), and their anticancer bioactivities were examined.
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Study of Cl(-) (H2 O)n (n = 1-4) using basin-hopping method coupled with density functional theory.
J Comput Chem
PUBLISHED: 07-17-2013
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Cl(-) (H2 O)n (n = 1-4) clusters were investigated using a basin-hopping (BH) algorithm coupled with density functional theory (DFT). Structures, energetics, thermodynamics, vertical detachment energies, and vibrational frequencies were obtained from high-level ab initio calculations. Through comparisons with previous theoretical and experimental data, it was demonstrated that the combination of the BH method and DFT could accurately predict the global and local minima of Cl(-) (H2 O)n (n = 1-4). Additionally, to optimize larger Cl(-) (H2 O)n (n > 4) clusters, several popular density functionals as well as DF-LMP2 (Schütz et al., J. Chem. Phys. 2004, 121, 737) (second-order Møller-Plesset perturbation theory using local and density fitting approximations) were tested with appropriate basis sets through comparisons with MP2 optimized results. DF-LMP2 will be used in future studies because its overall performance in describing the relative binding energies and the geometrical parameters of Cl(-) (H2 O)n (n = 1-4) was outstanding in this study. © 2013 Wiley Periodicals, Inc.
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The telomere-associated homeobox-containing protein TAH1/HMBOX1 participates in telomere maintenance in ALT cells.
J. Cell. Sci.
PUBLISHED: 06-26-2013
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The majority of cancer cells rely on elevated telomerase expression and activity for rapid growth and proliferation. Telomerase-negative cancer cells, by contrast, often employ the alternative lengthening of telomeres (ALT) pathway to maintain telomeres. ALT cells are characterized by long and dynamic telomeres and the presence of ALT-associated promyelocytic leukemia (PML) bodies (APBs). Previous work has shown the importance of APBs to the ALT pathway, but their formation and precise role remain unclear. Here, we demonstrate that a homeobox-containing protein known as HMBOX1 can directly bind telomeric double-stranded DNA and associate with PML nuclear bodies. Hence, we renamed this protein TAH1 for telomere-associated homeobox-containing protein 1. TAH1 knockdown significantly reduced the number of APBs and led to an increase in DNA damage response signals at telomeres. Importantly, TAH1 inhibition also notably reduced the presence of telomere C-circles, indicating altered ALT activity. Our findings point to TAH1 as a novel link between pathways that regulate DNA damage responses, PML nuclear bodies, and telomere homeostasis in ALT cells, and provide insight into how ALT cells may achieve sustained growth and proliferation independent of the telomerase.
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[Establishment of an intrahepatic xenograft tumor model in nude mice and its detection by in vivo fluorescence imaging system].
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi
PUBLISHED: 05-07-2013
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To establish an intrahepatic xenograft tumor model in nude mice and dynamically monitor the tumor growth by in vivo fluorescence imaging system.
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Expression, purification, and immunogenic characterization of Epstein-Barr virus recombinant EBNA1 protein in Pichia pastoris.
Appl. Microbiol. Biotechnol.
PUBLISHED: 03-06-2013
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Epstein-Barr virus (EBV) is a ubiquitous human herpesvirus associated with the development of both lymphoid and epithelial tumors. EBNA1 is the only viral protein expressed in all EBV-associated malignancies and plays important roles in EBV latency. Thus, EBNA1 is thought to be a promising antigen for immunotherapy of all EBV-associated malignancies. This study was undertaken to produce recombinant EBNA1 protein in Pichia pastoris and evaluate its immunogenicity. The truncated EBNA1 (E1?GA, codons 390-641) was expressed as a secretory protein with an N-terminal histidine tag in the methylotrophic yeast P. pastoris and purified by Ni-NTA affinity chromatography. The purified proteins were then used as antigens to immunize BALB/c mice for production of polyclonal antibodies. Western blot analysis showed that the polyclonal antibodies specifically recognized the EBNA1 protein in B95-8 cell lysates. The recombinant E1?GA also induced strong lymphoproliferative and Th1 cytokine responses in mice. Furthermore, mice immunized with E1?GA developed CD4+ and CD8+ T cell responses. These findings showed that the yeast-expressed E1?GA retained good immunogenicity and might be a promising vaccine candidate against EBV-associated malignancies.
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Crystallization and preliminary crystallographic studies of AAL-2, a novel lectin from Agrocybe aegerita that binds nonreducing terminal N-acetylglucosamine.
Acta Crystallogr. Sect. F Struct. Biol. Cryst. Commun.
PUBLISHED: 03-01-2013
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AAL-2 is a recently discovered lectin from the mushroom Agrocybe aegerita that specifically recognizes nonreducing terminal acetylglucosamine (GlcNAc) and that could be used as a probe in studies of protein O-linked ?-N-acetylglucosamination (O-GlyNAcylation). In order to illustrate the mechanism of how this protein specifically recognizes nonreducing terminal GlcNAc and to evaluate the efficacy of AAL-2 as a macromolecular probe in O-GlyNAcylation studies, expression and crystallization studies of AAL-2 were performed and a diffraction data set was collected to 2.0 Å resolution. Preliminary crystallographic studies revealed that the AAL-2 crystals belonged to space group P2(1)2(1)2(1), with unit-cell parameters a = 52.60, b = 111.70, c = 135.97 Å.
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Potent killing of HBV-related hepatocellular carcinoma by a chimeric protein of anti-HBsAg single-chain antibody and truncated Bid.
Biomaterials
PUBLISHED: 02-05-2013
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Targeted therapy is needed for hepatitis B virus (HBV)-mediated hepatocellular carcinoma (HCC) which shows overexpression of HBV surface antigen (HBsAg). We previously developed scFv15, a human single-chain antibody against HBsAg. Here we tested the strategic feasibility of scFv15-mediated delivery of apoptotic effectors for HBsAg-targeted HCC therapy and application of HA2 motif of influenza hemagglutinin to enhance endosome escape and antitumor effect. A class of HBsAg-targeted immunoproapoptotic molecule was generated by sequentially fusing scFv15, the furin-cleavable motif from diphtheria toxin (Fdt), HA2 and a truncated apoptotic protein Bid (tBid). The resulting scFv15-Fdt-HA2-tBid was prokaryotically expressed and functionally characterized for HBsAg-binding capacity, endosome escape activity and antitumor effect as compared with scFv15-Fdt-tBid. Both scFv15-Fdt-HA2-tBid and scFv15-Fdt-tBid retained affinity and specificity for HBsAg, and bound and selectively killed HBsAg-positive HCC cells via apoptosis. Notably, the IC50 of scFv15-Fdt-HA2-tBid in HBsAg-positive PLC/PRF/5 cells was 10 times lower than that of scFv15-Fdt-tBid. In vivo imaging of antitumor activity demonstrated 95% growth inhibition of orthotopic HCC by scFv15-Fdt-HA2-tBid compared with 75% suppression by scFv15-Fdt-tBid. This study represents an extended application of the immunoproapoptotic strategy in the treatment of HBsAg-positive HCC and shows significant potential of HA2 as a functional enhancer for endosome-encapsulated antibody-conjugates.
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Thermopower as a sensitive probe of electronic nematicity in iron pnictides.
Phys. Rev. Lett.
PUBLISHED: 02-04-2013
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We study the in-plane anisotropy of the thermoelectric power and electrical resistivity on detwinned single crystals of isovalent substituted EuFe(2)(As(1-x)P(x))(2). Compared to the resistivity anisotropy, the thermopower anisotropy is more pronounced and clearly visible already at temperatures much above the structural and magnetic phase transitions. Most remarkably, the thermopower anisotropy changes sign below the structural transition. This is associated with the interplay of two contributions due to anisotropic scattering and orbital polarization, which dominate at high and low temperatures, respectively.
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Should bone scan be performed in Chinese prostate cancer patients at the time of diagnosis?
Urol. Int.
PUBLISHED: 01-17-2013
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Prostate cancer (PCa) is increasingly being diagnosed in China. Early detection of bone metastases (BM) is critical in the management of patients with high-risk PCa. The aim of this study is to establish a screening model to determine if bone scan should be performed for BM in Chinese patients at the time when PCa is diagnosed.
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Transcriptome and proteome exploration to provide a resource for the study of Agrocybe aegerita.
PLoS ONE
PUBLISHED: 01-14-2013
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Agrocybe aegerita, the black poplar mushroom, has been highly valued as a functional food for its medicinal and nutritional benefits. Several bioactive extracts from A. aegerita have been found to exhibit antitumor and antioxidant activities. However, limited genetic resources for A. aegerita have hindered exploration of this species.
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Prognostic Value of EZH2 Expression and Activity in Renal Cell Carcinoma: A Prospective Study.
PLoS ONE
PUBLISHED: 01-01-2013
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Increased expression of EZH2 correlates with aggressive clinical behavior in various malignancies. In this study, we aim to investigate the clinical and prognostic values of EZH2 expression and activity in tumor tissues and improve the risk stratification in patients with renal cell carcinoma after surgery. We analyzed EZH2 expression and its activity as indicated by H3K27me3 levels comprising 373 patients with renal cell carcinoma in our institute. Outcome was assessed as overall survival and disease free survival using Kaplan-Meier analysis. Prognostic values of EZH2 and H3K27me3 expression for clinical outcomes were evaluated by Cox regression analysis. We used receiver operating characteristic to calculate diagnostic accuracy. High EZH2 expression correlates with poor overall survival in all patients, especially in advanced RCC, which is an independent prognostic factor in disease free survival and overall survival. Compared with EZH2, H3K27me3 expression is not an independent prognostic factor. The expressions of H3K27me3 and EZH2 are not completely consistent, which might be due to complicated interaction of Polycomb Repressor Complex 2. A combination of EZH2 expression and TNM stage could have better prognostic value than do TNM stage or EZH2 expression alone in both sets for disease free survival and overall survival. These results imply that evaluating intratumoral EZH2 density might improve prognostic value to the TNM staging system and inform treatment decisions for patients with late-stage renal cell carcinoma.
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TRPP2 and TRPV4 form an EGF-activated calcium permeable channel at the apical membrane of renal collecting duct cells.
PLoS ONE
PUBLISHED: 01-01-2013
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Regulation of apical calcium entry is important for the function of principal cells of the collecting duct. However, the molecular identity and the regulators of the transporter/channel, which is responsible for apical calcium entry and what factors regulate the calcium conduction remain unclear.
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FasL expression on human nucleus pulposus cells contributes to the immune privilege of intervertebral disc by interacting with immunocytes.
Int J Med Sci
PUBLISHED: 01-01-2013
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The mechanisms of immune privilege in human nucleus pulposus (NP) remain unclear. Accumulating evidence indicates that Fas ligand (FasL) might play an important role in the immune privilege of the disc. We aimed for addressing the role of FasL expression in human intervertebral disc degeneration (IDD) and immune privilege in terms of the interaction between NP cells and immunocytes via the FasL-Fas machinery. We collected NP specimens from 20 patients with IDD as degenerative group and 8 normal cadaveric donors as control. FasL expression was detected by qRT-PCR, western blotting and flow cytometry (FCM). We also collected macrophages and CD8(+) T cells from the peripheral blood of patients with IDD for co-cultures with NP cells. And macrophages and CD8(+) T cells were harvested for apoptosis analysis by FCM after 2 days of co-cultures. We found that FasL expression in mRNA, protein and cellular resolutions demonstrated a significant decrease in degenerative group compared with normal control (p<0.05). FCM analysis found that human NP cells with increased FasL expression resulted in significantly increased apoptosis ratio of macrophages and CD8(+) T cells. Our study demonstrated that FasL expression tends to decrease in degenerated discs and FasL plays an important role in human disc immune privilege, which might provide a novel target for the treatment strategies for IDD.
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Superconductivity and ferromagnetism in EuFe?(As(1-x)P(x))?.
J Phys Condens Matter
PUBLISHED: 11-03-2011
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Superconductivity and ferromagnetism are two antagonistic cooperative phenomena, which makes it difficult for them to coexist. Here we demonstrate experimentally that they do coexist in EuFe?(As(1-x)P(x))? with 0.2 ? x ? 0.4, in which superconductivity is associated with Fe 3d electrons and ferromagnetism comes from the long-range ordering of Eu 4f moments via Ruderman-Kittel-Kasuya-Yosida (RKKY) interactions. The coexistence features large saturated ferromagnetic moments, high and comparable superconducting and magnetic transition temperatures, and broad coexistence ranges in temperature and field. We ascribe this unusual phenomenon to the robustness of superconductivity as well as the multi-orbital character of iron pnictides.
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CeNiAsO: an antiferromagnetic dense Kondo lattice.
J Phys Condens Matter
PUBLISHED: 04-08-2011
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A cerium-containing pnictide, CeNiAsO, crystallized in the ZrCuSiAs-type structure, has been investigated by measuring transport and magnetic properties, as well as specific heat. We found that CeNiAsO is an antiferromagnetic dense Kondo lattice metallic compound with Kondo scale T(K) ?15 K and shows an enhanced Sommerfeld coefficient of ?(0) ?203 mJ mol K(-2). While no superconductivity can be observed down to 30 mK, Ce ions exhibit two successive antiferromagnetic (AFM) transitions. We propose that the magnetic moment of the Ce ion could align in the G-type AFM order below the first transition at T(N1)=9.3 K, and it might be modified into the C-type AFM order below a lower transition at T(N2)=7.3 K. Our results indicate that the 3d-4f interlayer Kondo interactions play an important role in Ni-based Ce-containing pnictides.
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Temporal genetic variability and host sources of Escherichia coli associated with fecal pollution from domesticated animals in the shellfish culture environment of Xiangshan Bay, East China Sea.
Environ. Pollut.
PUBLISHED: 04-02-2011
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This study was conducted to analyze the genetic variability of Escherichia coli from domesticated animal wastes for microbial source tracking (MST) application in fecal contaminated shellfish growing waters of Xiangshan Bay, East China Sea. (GTG)(5) primer was used to generate 1363 fingerprints from E. coli isolated from feces of known 9 domesticated animal sources around this shellfish culture area. Jackknife analysis of the complete (GTG)(5)-PCR DNA fingerprint library indicated that isolates were assigned to the correct source groups with an 84.28% average rate of correct classification. Based on one-year source tracking data, the dominant sources of E. coli were swine, chickens, ducks and cows in this water area. Moreover, annual and spatial changes of E. coli concentrations and host sources may affect the level and distribution of zoonotic pathogen species in waters. Our findings will further contribute to preventing fecal pollution in aquatic environments and quality control of shellfish.
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[Relationship among coagulation effect of Al-based coagulant, content and speciation of residual aluminum].
Huan Jing Ke Xue
PUBLISHED: 08-12-2010
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The application of AlCl3, Al2 (SO4)3 and poly-aluminum chloride (PAC) in humic acid-kaolin simulated water was studied in this article. It is intended to discuss the relationship among coagulation effect of Al-based coagulants in humic acid-kaolin simulated water and content and speciation of residual aluminum. It was found that, the turbidity removal efficiency and UV254 removal efficiency could reach about 90% at the tested dosage. At higher dosage, PAC gave better coagulation effect. The residual total aluminum content and residual aluminum ratio of PAC, which was 0.9 mg/L and - 3.0% or so respectively, were greatly lower than those of AlCl3 and Al2 (SO4)3. The residual total dissolved aluminum was the predominant content in the effluent after coagulation and sedimentation by the three Al-based coagulants. For the total dissolved aluminum, the proportion of dissolved organic aluminum was significantly higher than that of other aluminum speciation. With respect to humic acid-kaolin simulated water, the content of residual total aluminum in the effluent after coagulation and sedimentation by PAC decreased obviously compared to AlCl3 and Al2 (SO4)3. PAC could effectively decrease the content of residual dissolved aluminum speciation which has higher toxicity. The content of residual total dissolved aluminum in the effluent after coagulation and sedimentation by PAC was about 0.6 mg/L.
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Opposing developmental functions of Agrocybe aegerita galectin (AAL) during mycelia differentiation.
Fungal Biol
PUBLISHED: 05-07-2010
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Mycelia of basidiomycetes differentiating into fruiting body is a controlled developmental process, however the underlying molecular mechanism remains unknown. In previous work, a novel fungal Agrocybe aegerita galectin (AAL) was isolated from A. aegerita in our laboratory. AAL was shown to promote mycelial differentiation in A. aegerita and Auricularia polytricha, indicating that AAL might function as a conserved fruiting initiator during basidiomycete mycelia development. In the current work, we investigate the role of AAL in mycelia differentiation and fruiting body formation. First, the expression and localization of AAL in mycelia, primordium and fruiting body were assessed by Western blotting and immunohistochemistry. AAL was found to be ubiquitously expressed in the primordium and fruiting body but not in the mycelia. AAL facilitated mycelia congregation and promoted fruiting body production when AAL was applied on mycelia. At the same time, when AAL was spread on potato dextrose agar (PDA) medium prior to mycelia inoculation, mycelia exhibited slowed growth rates, resulting in mycelia cords formation and inhibition of fruiting body formation. The 5 regulatory sequence of aal was cloned by genome walking. Here, we show that aal lack introns in the coding region and the upstream 740 bp sequence was characterized by the existence of core promoter elements, which included: two CCAAT boxes (-535/-280), a GC box (-145), a TATA box (-30) and a fungal leader intron within the 5 UTR. The identification of regulatory expression elements may provide an explanation to the stage-specific and high-level expression of aal during fruiting development.
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La2Co2Se2O3: a quasi-two-dimensional mott insulator with unusual cobalt spin state and possible orbital ordering.
J. Am. Chem. Soc.
PUBLISHED: 04-30-2010
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The new oxyselenide La(2)Co(2)Se(2)O(3), containing Co(2)O square-planar layers, has been successfully synthesized using solid-state reactions under vacuum. The compound crystallizes in space group I4/mmm with lattice parameters a = 4.0697(8) A and c = 18.419(4) A. Magnetic susceptibility measurements indicate an antiferromagnetic transition at approximately 220 K. The magnetic entropy associated with the transition is close to R ln 2, suggesting an unusual low-spin state for the Co(2+) ions. The as-prepared sample shows insulating behavior with room-temperature resistivity of approximately 10(7) ohms cm, which decreases by 4 orders of magnitude under a pressure of 7 GPa. Band structure calculations using the LSDA+U approach reproduce the insulating ground state with low spin for Co and suggest strong orbital polarization for the valence electrons near the Fermi level. It is also revealed that the spin and orbital degrees of freedom in the antiferromagnetic checkerboard spin-lattice are mutually coupled.
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Reconstruction of the corneal epithelium with induced marrow mesenchymal stem cells in rats.
Mol. Vis.
PUBLISHED: 03-31-2010
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To explore the feasibility of bone marrow mesenchymal stem cells (MSCs) transdifferentiating into corneal epithelial cells in a limbal stem cell deficiency (LSCD) model in rats.
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MicroRNA-155 functions as an OncomiR in breast cancer by targeting the suppressor of cytokine signaling 1 gene.
Cancer Res.
PUBLISHED: 03-30-2010
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MicroRNA-155 (miR-155) is overexpressed in many human cancers; however, the mechanisms by which miR-155 functions as a putative oncomiR are largely unknown. Here, we report that the tumor suppressor gene suppressor of cytokine signaling 1 (socs1) is an evolutionarily conserved target of miR-155 in breast cancer cells. We found that mir-155 expression is inversely correlated with socs1 expression in breast cancer cell lines as well as in a subset of primary breast tumors. We also identified a 24A-->G mutation in the miR-155 binding site of the SOCS1 3 untranslated region in a breast tumor that reduced miR-155 repression, implicating a mechanism for miRNA targets to avoid repression. Ectopic expression of miR-155 significantly promoted the proliferation of breast cancer cells, the formation of soft agar foci in vitro, and the development of tumors in nude mice. In breast cancer cells, RNA interference silencing of socs1 recapitulates the oncogenic effects of miR-155, whereas restoration of socs1 expression attenuates the protumorigenesis function of miR-155, suggesting that miR-155 exerts its oncogenic role by negatively regulating socs1. Overexpression of miR-155 in breast cancer cells leads to constitutive activation of signal transducer and activator of transcription 3 (STAT3) through the Janus-activated kinase (JAK) pathway, and stimulation of breast cancer cells by the inflammatory cytokines IFN-gamma and interleukin-6 (IL-6), lipopolysaccharide (LPS), and polyriboinosinic:polyribocytidylic acid [poly(I:C)] significantly upregulates mir-155 expression, suggesting that miR-155 may serve as a bridge between inflammation and cancer. Taken together, our study reveals that miR-155 is an oncomiR in breast cancer and that miR-155 may be a potential target in breast cancer therapy.
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Influence of magnolol on the secretion of alpha-toxin by Staphylococcus aureus.
Molecules
PUBLISHED: 01-18-2010
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In this study we investigated the antimicrobial activity of magnolol on Staphylococcus aureus. The minimal inhibitory concentrations of magnolol against 31 S. aureus strains ranged from 4-32 microg/mL. In addition, hemolysin assays, Western blotting, and real-time RT-PCR were performed to investigate the effect of magnolol on alpha-toxin secretion by both methicillin-sensitive S. aureus (MSSA) and methicillin-resistant S. aureus (MRSA). The results indicated that sub-inhibitory concentrations of magnolol dose-dependently inhibited the transcription of hla (the gene encoding alpha-toxin) in S. aureus, resulting in a reduction of alpha-toxin secretion and, thus, hemolytic activities.
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A nuclear ligand MRG15 involved in the proapoptotic activity of medicinal fungal galectin AAL (Agrocybe aegerita lectin).
Biochim. Biophys. Acta
PUBLISHED: 01-15-2010
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We have previously reported a novel fungal galectin Agrocybe aegerita lectin (AAL) with apoptosis-induced activity and nuclear migration activity. The importance of nuclear localization for AALs apoptosis-induced activity has been established by mutant study. However, the mechanism remains unclear.
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Focal adhesion kinase signaling pathway participates in the formation of choroidal neovascularization and regulates the proliferation and migration of choroidal microvascular endothelial cells by acting through HIF-1 and VEGF expression in RPE cells.
Exp. Eye Res.
PUBLISHED: 10-09-2009
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Choroidal neovascularization (CNV) is one of the most frequent causes of severe and progressive vision loss, while its pathogenesis is still poorly understood. Focal adhesion kinase (FAK), a non-receptor tyrosine kinase, plays a crucial role in linking signals initiated by both the extracellular matrix (ECM) and soluble signaling factors and controls essential cellular processes. Extensive evidence has shown that FAK is activated in angiogenic response. This study aims to investigate the effect of FAK on CNV formation. The Brown-Norway (BN) rats underwent laser rupture of Bruchs membrane to induce CNV and were then killed at 1, 3, 7, and 14 days following laser injury. Immunofluorescence and Western blot were processed to detect FAK protein. Retinal pigment epithelial (RPE) cells were cultured under hypoxia and RNA interference (RNAi) technique was used to knock down the FAK gene in RPE cells. Expression of hypoxia inducible factor-1 (HIF-1alpha) and vascular endothelial growth factor (VEGF) in RPE cells were investigated by RT-PCR and Western blot. Two kinds of coculture models were used to observe the effects of specific blockade of FAK in RPE cells on the proliferation and migration of choroidal microvascular endothelial cells (CECs), respectively. FAK was highly expressed in the rat RPE-choroid tissue after photocoagulation. In vitro experiment showed that FAK was involved in hypoxia signaling in cultured RPE cells. The absence of FAK effectively reduced the expression of hypoxia-induced HIF-1alpha and VEGF in RPE cells, resulting in the inhibition of proliferation and migration of CECs. Our results suggest that FAK pathway activation plays a role in the development of CNV, and regulates the proliferation and migration of CECs by acting through HIF-1 and then up-regulating the expression of the angiogenic factor VEGF in RPE cells. It is reasonable to propose that FAK siRNA will potentially provides a means to attenuate the strong stimuli for neovascularization in CNV-dependent disorders, which could present a therapeutically relevant strategy for the inhibition of CNV.
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Superconductivity up to 30 K in the vicinity of the quantum critical point in BaFe(2)(As(1-x)P(x))(2).
J Phys Condens Matter
PUBLISHED: 08-24-2009
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We report bulk superconductivity induced by an isovalent doping of phosphorus in BaFe(2)(As(1-x)P(x))(2). The P-for-As substitution results in shrinkage of the lattice, especially for the FeAs block layers. The resistivity anomaly associated with the spin-density-wave (SDW) transition in the undoped compound is gradually suppressed by the P doping. Superconductivity with a maximum T(c) of 30 K emerges at x = 0.32, coinciding with a magnetic quantum critical point (QCP) which is shown by the disappearance of SDW order and the linear temperature-dependent resistivity in the normal state. The T(c) values were found to decrease with further P doping and no superconductivity was observed down to 2 K for x?0.77. The appearance of superconductivity in the vicinity of QCP hints at the superconductivity mechanism in iron-based arsenides.
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Immune responses of Fenneropenaeus chinensis against white spot syndrome virus after oral delivery of VP28 using Bacillus subtilis as vehicles.
Fish Shellfish Immunol.
PUBLISHED: 07-25-2009
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The protective efficacy of oral administration of VP28 using Bacillus subtilis as vehicles (rVP28-bs) in shrimp, Fenneropenaeus chinensis, upon challenge with white spot syndrome virus (WSSV) was investigated. The calculated relative percent survival (RPS) value of rVP28-bs fed shrimp was 83.3% when challenged on the 14th day post-administration, which is significantly higher (p < 0.001) than that of the group administered recombinant Escherichia coli over-expressing rVP28 (rVP28-e21). After immunization, activities of phenoloxidase (PO), superoxide dismutase (SOD) and inducible nitric oxide synthase (iNOS) in hemolymph were analyzed. It was found that the supplementation of rVP28-bs into shrimp food pellets resulted in the most pronounced increase of iNOS activity (p < 0.001), but had the least influence on activities of PO and SOD. Besides, in the shrimp orally administered with rVP28-bs, the caspase-3 activity was one-fifth that of the control, though the signs of apoptosis (chromatin margination, nuclear fragmentation and apoptotic bodies) could not be observed by transmission electron microscope (TEM). These results suggest that by oral delivery of rVP28-bs, shrimp showed significant resistance to WSSV and an effect on the innate immune system of shrimp. The remarkably enhanced level of iNOS after rVP28-bs administration might be responsible for antiviral defense in shrimp.
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Importance of nuclear localization for the apoptosis-induced activity of a fungal galectin AAL (Agrocybe aegerita lectin).
Biochem. Biophys. Res. Commun.
PUBLISHED: 05-31-2009
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Agrocybe aegerita lectin (AAL) was identified previously in our group as a novel galectin from medicinal fungi Agrocybe aegerita, and has been shown to effectively induce cancer cell cycle arrest and apoptosis in vitro and tumor regression in vivo. Here, AAL was observed to translocate into the HeLa cell nucleus and induce cell apoptosis when it was predominantly in the nucleus. The N-terminus and C-terminus of AAL were required for nuclear localization. Site mutated proteins were generated based on AAL structure. Dimer interface mutant I25G, carbohydrate recognition domain (CRD) mutant R63H, and loop region mutant L33A could not enter the nucleus and lost the ability to induce apoptosis. CRD mutant H59Q and loop region mutant I144G maintained nuclear localization activity, and H59Q retained residual bioability but I144G had no activity, indicating that nuclear localization is important but not sufficient for AAL to become apoptotically active. Our findings provide a novel antitumor mechanism of fungal galectin.
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Superconductivity induced by phosphorus doping and its coexistence with ferromagnetism in EuFe2(As0.7P0.3)(2).
Phys. Rev. Lett.
PUBLISHED: 01-19-2009
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We have studied EuFe2(As0.7P0.3)(2) by the measurements of x-ray diffraction, electrical resistivity, thermopower, magnetic susceptibility, magnetoresistance, and specific heat. Partial substitution of As with P results in the shrinkage of lattice, which generates chemical pressure to the system. It is found that EuFe2(As0.7P0.3)(2) undergoes a superconducting transition at 26 K, followed by ferromagnetic ordering of Eu2+ moments at 20 K. This finding is the first observation of superconductivity stabilized by internal chemical pressure, and supplies a rare example showing the coexistence of superconductivity and ferromagnetism in the ferroarsenide family.
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[A new method for purification and identification of hepatocellular carcinoma stem cell of SMMC-7721].
Zhonghua Yi Xue Za Zhi
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To explore a new efficient purification method of hepatocellular carcinoma (HCC) stem cells and identify their features.
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Genome-wide transcriptome and proteome analysis on different developmental stages of Cordyceps militaris.
PLoS ONE
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Cordyceps militaris, an ascomycete caterpillar fungus, has been used as a traditional Chinese medicine for many years owing to its anticancer and immunomodulatory activities. Currently, artificial culturing of this beneficial fungus has been widely used and can meet the market, but systematic molecular studies on the developmental stages of cultured C. militaris at transcriptional and translational levels have not been determined.
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Recombinant VP1 protein expressed in Pichia pastoris induces protective immune responses against EV71 in mice.
Biochem. Biophys. Res. Commun.
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Human enterovirus 71 (EV71) is one of the major causative agents of hand, foot and mouth disease and is also associated with serious neurological diseases in children. Currently, there are no effective antiviral drugs or vaccines against EV71 infection. VP1, one of the major immunogenic capsid proteins of EV71, is widely considered to be the candidate antigen for an EV71 vaccine. In this study, VP1 of EV71 was expressed as a secretory protein with an N-terminal histidine tag in the methylotrophic yeast Pichia pastoris, and purified by Ni-NTA affinity chromatography. Immunogenicity and vaccine efficacy of the recombinant VP1 were assessed in mouse models. The results showed that the recombinant VP1 could efficiently induce anti-VP1 antibodies in BALB/c mice, which were able to neutralize EV71 viruses in an in vitro neutralization assay. Passive protection of neonatal mice further confirmed the prophylactic efficacy of the antisera from VP1 vaccinated mice. Furthermore, VP1 vaccination induced strong lymphoproliferative and Th1 cytokine responses. Taken together, our study demonstrated that the yeast-expressed VP1 protein retained good immunogenicity and was a potent EV71 vaccine candidate.
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Purification and characterization of an antitumor protein with deoxyribonuclease activity from edible mushroom Agrocybe aegerita.
Mol Nutr Food Res
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Mushrooms are well known for their nutritional and medicinal value. Agrocybe aegerita has been used as a nutritious food around the world and for its herbal medicinal properties in Asia. In recent years, several antitumor proteins have been identified from A. aegerita. The objective of this study was to purify a novel antitumor protein from A. aegerita.
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Deep insight into the Ganoderma lucidum by comprehensive analysis of its transcriptome.
PLoS ONE
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Ganoderma lucidum is a basidiomycete white rot fungus and is of medicinal importance in China, Japan and other countries in the Asiatic region. To date, much research has been performed in identifying the medicinal ingredients in Ganoderma lucidum. Despite its important therapeutic effects in disease, little is known about Ganoderma lucidum at the genomic level. In order to gain a molecular understanding of this fungus, we utilized Illumina high-throughput technology to sequence and analyze the transcriptome of Ganoderma lucidum.
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Mycophenolate mofetil inhibits macrophage infiltration and kidney fibrosis in long-term ischemia-reperfusion injury.
Eur. J. Pharmacol.
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Immunosuppressants have been widely used in renal transplantation, in which ischemia-reperfusion injury is inevitable. Mycophenolate mofetil (MMF) is a relative novel immunosuppressant and also attenuates ischemia-reperfusion injury in the acute phase, but its long-term effects are still obscure. Unilateral renal ischemia-reperfusion injury model was established in Sprague-Dawley rats and 30 mg/kg/day MMF or natural saline was administered a day before the surgery. Renal function was monitored, and histological changes and fibrosis in the kidney were evaluated in both short and long terms. TGF-?1 secretion and MCP-1 expression were determined by immunohistochemistry and real-time PCR respectively. The infiltration of macrophages in renal tissues was also assessed by fluorescence activated cell sorting (FACS). MMF treatment significantly improved renal function in ischemia-reperfusion injury rats in the short and long-term and also effectively prevented interstitial fibrosis. TGF-?1 secretion and MCP-1 expression in the renal tissue of MMF-treated rats were much lower than those in natural saline-treated rats, with much less macrophage infiltration as well. MMF treatment effectively prevented the deterioration of renal function and interstitial fibrosis in ischemia-reperfusion injury rats, which may be associated with decreased TGF-?1, MCP-1 and macrophages. These results provide evidence for the choice of MMF in the renal transplant patients not only for acute renal injury but also for long-term survival of renal allograft.
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A novel miR-155/miR-143 cascade controls glycolysis by regulating hexokinase 2 in breast cancer cells.
EMBO J.
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Cancer cells preferentially metabolize glucose through aerobic glycolysis. This phenomenon, known as the Warburg effect, is an anomalous characteristic of glucose metabolism in cancer cells. Chronic inflammation is a key promoting factor of tumourigenesis. It remains, however, largely unexplored whether and how pro-tumourigenic inflammation regulates glucose metabolism in cancer cells. Here, we show that pro-inflammatory cytokines promote glycolysis in breast cancer cells, and that the inflammation-induced miR-155 functions as an important mediator in this process. We further show that miR-155 acts to upregulate hexokinase 2 (hk2), through two distinct mechanisms. First, miR-155 promotes hk2 transcription by activation of signal transducer and activator of transcription 3 (STAT3), a transcriptional activator for hk2. Second, via targeting C/EBP? (a transcriptional activator for mir-143), miR-155 represses mir-143, a negative regulator of hk2, thus resulting in upregulation of hk2 expression at the post-transcriptional level. The miR-155-mediated hk2 upregulation also appears to operate in other types of cancer cells examined. We suggest that the miR-155/miR-143/HK2 axis may represent a common mechanism linking inflammation to the altered metabolism in cancer cells.
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A novel lectin from Agrocybe aegerita shows high binding selectivity for terminal N-acetylglucosamine.
Biochem. J.
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A novel lectin was isolated from the mushroom Agrocybe aegerita (designated AAL-2) by affinity chromatography with GlcNAc (N-acetylglucosamine)-coupled Sepharose 6B after ammonium sulfate precipitation. The AAL-2 coding sequence (1224 bp) was identified by performing a homologous search of the five tryptic peptides identified by MS against the translated transcriptome of A. aegerita. The molecular mass of AAL-2 was calculated to be 43.175 kDa from MS, which was consistent with the data calculated from the amino acid sequence. To analyse the carbohydrate-binding properties of AAL-2, a glycan array composed of 465 glycan candidates was employed, and the result showed that AAL-2 bound with high selectivity to terminal non-reducing GlcNAc residues, and further analysis revealed that AAL-2 bound to terminal non-reducing GlcNAc residues with higher affinity than previously well-known GlcNAc-binding lectins such as WGA (wheatgerm agglutinin) and GSL-II (Griffonia simplicifolia lectin-II). ITC (isothermal titration calorimetry) showed further that GlcNAc bound to AAL-2 in a sequential manner with moderate affinity. In the present study, we also evaluated the anti-tumour activity of AAL-2. The results showed that AAL-2 could bind to the surface of hepatoma cells, leading to induced cell apoptosis in vitro. Furthermore, AAL-2 exerted an anti-hepatoma effect via inhibition of tumour growth and prolongation of survival time of tumour-bearing mice in vivo.
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