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Find video protocols related to scientific articles indexed in Pubmed.
Role of surfactant proteins A and D in sepsis-induced acute kidney injury.
Shock
PUBLISHED: 09-26-2014
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Sepsis is a major cause of acute kidney injury (AKI) with high rates of morbidity and mortality. Surfactant proteins A and D (SP-A, SP-D) play a critical role in host defense and regulate inflammation during infection. Recent studies indicate SP-A and SP-D are expressed in the kidney. The current study examines the role of SP-A and SP-D in the pathogenesis of sepsis-induced AKI. Wild-type (WT) and SP-A/SP-D double knockout (KO) C57BL/6 mice were treated by cecal ligation and puncture (CLP) or sham surgery. Histological, cellular and molecular indices of kidney injury were investigated in septic mice 6 and 24 h after CLP. 24 h post-CLP, kidney injury was more severe, renal function was decreased, blood creatinine and BUN were higher in septic SP-A/SP-D KO mice (p<0.05, vs septic WT mice). Kidney edema and vascular permeability were increased in septic SP-A/SP-D KO mice (p<0.01, vs septic WT mice). Apoptotic cells increased significantly (p<0.01) in the kidney of septic SP-A/SP-D KO mice compared to septic WT mice. Molecular analysis revealed levels of Bcl-2 (an inhibitor of apoptosis) were lower and levels of caspase-3 (a biomarker of apoptosis) were higher in the kidney of septic SP-A/SP-D KO mice (p<0.01, vs septic WT mice). Furthermore, levels of NF-?B and phosphorylated I?B-? increased significantly in the kidney of septic SP-A/SP-D KO mice than septic WT mice, suggesting SP-A/SP-D KO mice have a more pronounced inflammatory response to sepsis. We conclude SP-A and SP-D attenuate kidney injury by modulating inflammation and apoptosis in sepsis-induced AKI.
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Unique Functional and Structural Properties of the LRRK2 ATP-Binding Pocket.
J. Biol. Chem.
PUBLISHED: 09-16-2014
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Pathogenic mutations in the LRRK2 gene can cause late-onset Parkinson's disease. The most common mutation, G2019S, resides in the kinase domain and enhances activity. LRRK2 possesses the unique property of cis-autophosphorylation of its own GTPase domain. As high-resolution structures of the human LRRK2 kinase domain are not available, we used novel high-throughput assays that measured both cis-auto-phosphorylation and trans-peptide- phosphorylation to probe the ATP-binding pocket. We disclose hundreds of commercially available activity-selective LRRK2 kinase inhibitors. Some compounds inhibit cis-auto-phosphorylation more strongly than trans-peptide phosphorylation, and other compounds inhibit G2019S-LRRK2 more strongly than WT-LRRK2. Through exploitation of structure-activity-relationships revealed through high-throughput analyses, we identified a useful probe inhibitor SRI-29132 (11). SRI-29132 is exquisitely selective for LRRK2 kinase activity and is effective in attenuating pro-inflammatory responses in macrophages and rescuing neurite retraction phenotypes in neurons. Further, the compound demonstrates excellent potency, is highly blood-brain barrier permeant, but suffers from rapid first-pass metabolism. Despite the observed selectivity of SRI-29132, docking models highlighted critical interactions with residues conserved in many protein kinases, implying a unique structural configuration for the LRRK2 ATP-binding pocket. While the human LRRK2 kinase domain is unstable and insoluble, we demonstrate that the LRRK2-homolog from amoeba can be mutated to approximate some aspects of the human LRRK2 ATP-binding pocket. Our results provide a rich resource for LRRK2 small molecule inhibitor development. More broadly, our results provide a precedent for the functional interrogation of ATP-binding pockets when traditional approaches to ascertain structure prove difficult.
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Preparation of camptothecin-loaded targeting nanoparticles and their antitumor effects on hepatocellular carcinoma cell line H22.
Drug Deliv
PUBLISHED: 08-22-2014
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Abstract Camptothecin (CPT) is an effective anticancer agent against various cancers but the clinical application is limited because of its poor water solubility, low bioavailability and severe toxic side effects. The aim of the present study was to evaluate the feasibility of using targeted NPs as a high-performance CPT delivery system that targets liver cancer cells through intravenous (i.v.) administration route. CPT was incorporated into biotin-F127-PLA or F127-PLA polymeric nanoparticles (NPs) by a dialysis method. The preparation of the targeting NPs was performed by conjugating biotin-F127-PLA NPs with anti-3A5 antibody. The antitumor effect of the CPT-loaded nanoparticles against H22 cells in vitro was determined using an MTT assay. Tissue distribution and tumor inhibition in vivo were also evaluated. Survivin mRNA expression was assessed by real-time polymerase chain reaction. Results showed that the targeted CPT NPs exhibited regular spherical shapes with a mean diameter of approximately 180?nm. In vitro release of the targeted CPT NPs exhibited an initial burst (40%) within 12?h, followed by a slow release. Cytotoxicity test against H22 cells indicated that the targeted CPT NPs exerted significant antitumor effects. Compared with free CPT and non-targeted CPT NPs, the targeted CPT NPs showed superior inhibition ratio against tumor in vivo, which may be associated with reduced survivin mRNA expression. The results suggested that the new targeted CPT NPs may be a promising injectable delivery system for cancer therapy.
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[Functional analysis of Oct4 promoter in Xuhuai goat].
Yi Chuan
PUBLISHED: 08-22-2014
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The aim of this study was to determine the activity region of Oct4 (octamer-binding transcription factor 4) promoter in Xuhuai goat, and to investigate the effect of TSA (trichostatin A) and VPA(valproicacid) on Oct4 promoter activity. Specific PCR primers of Oct4 promoter including different lengths of fragments were designed by Primer 5.0, then were amplified and cloned into PGL3-Bacic luciferase reporter vector. All the reconstruction vectors were transfected into gEF, P19 and COS7 cells, respectively. After TSA and VPA treatment, the activity of dual-luciferase reporter gene in these three transfected cells was detected. In addition, the CMV promoter of pEGFP-N1 was replaced by the -1516?+30 bp fragment of Oct4 promoter, GFP fluorescence was used to detect the activity of Oct4 promoter. The results indicated that different fragments of Oct4 promoter showed different degrees of activity in gEF, P19 and COS7 cells, and the maximal activity region of Oct4 promoter was -1516?+30 bp, the basal activity region was -238?+30 bp. Positive regulatory domains existed in the region of -1516?-946 bp and -615?-96 bp, while negative regulatory domains existed in the region of -1936?-1516 bp and -946?-615 bp. The optimum induction concentration to enhance the activity of Oct4 promoter was 1 ?mol/L of TSA and 4 mmol/L of VPA. The GFP expression can be started by the fragment of -1516?+30 bp. This study provides an experimental basis for revealing the mechanism of expression and regulation of Oct4 in goat.
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Promotion effect of KMnO ? on the oxidation of As(III) by air in alkaline solution.
J. Hazard. Mater.
PUBLISHED: 08-17-2014
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The mechanism of oxidation of As(III) in alkaline solution by air with promotion effect of KMnO4 was studied. The experimental results indicated that the superstoichiometric oxidation of As(III) by KMnO4 could be attributed to the catalytic effect of reductive product of KMnO4. The XRD and XPS results demonstrated that the catalyst was nascent MnO2 rich in potassium. The results also showed that the mole ratio of Mn/As and the initial pH had significant effects on the oxidation of As(III). The time for the oxidation by air was less than 2h with the mole ratio of Mn/As less than 1/10.5 and the initial pH higher than 13. The kinetics of the catalytic oxidation of arsenic was interpreted using the pseudo first order reaction, and the apparent active energy was about 15.01 kJ/mol. The study suggested that the initial oxidation was firstly dominated by the direct oxidation of KMnO4 followed by the catalytic oxidation with the nascent MnO2.
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Protein phosphatase PP1 negatively regulates the Toll-like receptor- and RIG-I-like receptor-triggered production of type I interferon by inhibiting IRF3 phosphorylation at serines 396 and 385 in macrophage.
Cell. Signal.
PUBLISHED: 08-01-2014
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The production of type I interferon must be tightly regulated, and the aberrant production of this protein is harmful or even fatal to the host. The transcription factor IRF3 phosphorylation is a central regulator of type I interferon meditated antiviral response. Protein phosphatase-1 (PP1) has been reported to be important in many cell functions, including development, differentiation, and tumorigenesis. However, the roles of PP1 in Toll-like receptor (TLR)- or retinoic acid-inducible gene I like receptor (RLR)-triggered IRF-3 activation remain unclear. Here, we show that the activity of PP1 is downregulated in macrophages upon stimulation with TLR or RLR ligands, including lipopolysaccharide, and poly(I:C), or vesicular stomatitis virus (VSV), respectively. The overexpression of PP1 selectively inhibits TLR- and VSV-induced interferon regulatory factor 3 (IRF3) activation but has no substantial effect on TANK-binding kinase 1 (TBK1),?B kinase ? (IKK?) activation. Conversely, RNA interference of PP1 significantly promotes IRF3 activation. Consistently, The overexpression of PP1 inhibits TLR- and VSV-triggered IFN-? production while PP1 knockdown significantly increases the production of IFN-? in macrophages. We further demonstrate that PP1 directly interacts with IRF3 and dephosphorylates IRF3 at Ser385 and Ser396, resulting in the suppression of TLR- and RLR-triggered IFN-? production. Thus, PP1 functions as a negative feedback regulator of TLR- and RLR-triggered antiviral immune responses by acting as an IRF3 phosphatase.
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[Transanal endoscopic microsurgery by transanal glove port combined with colonoscopy for excision of rectal tumors].
Zhonghua Wei Chang Wai Ke Za Zhi
PUBLISHED: 05-27-2014
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To evaluate the feasibility and efficacy of transanal endoscopic microsurgery (TEM) by transanal glove port combined with colonoscopy for excision of rectal tumors.
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Comparative genetic mapping and genomic region collinearity analysis of the powdery mildew resistance gene Pm41.
Theor. Appl. Genet.
PUBLISHED: 05-20-2014
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By applying comparative genomics analyses, a high-density genetic linkage map narrowed the powdery mildew resistance gene Pm41 originating from wild emmer in a sub-centimorgan genetic interval. Wheat powdery mildew, caused by Blumeria graminis f. sp. tritici, results in large yield losses worldwide. A high-density genetic linkage map of the powdery mildew resistance gene Pm41, originating from wild emmer (Triticum turgidum var. dicoccoides) and previously mapped to the distal region of chromosome 3BL bin 0.63-1.00, was constructed using an F5:6 recombinant inbred line population derived from a cross of durum wheat cultivar Langdon and wild emmer accession IW2. By applying comparative genomics analyses, 19 polymorphic sequence-tagged site markers were developed and integrated into the Pm41 genetic linkage map. Ultimately, Pm41 was mapped in a 0.6 cM genetic interval flanked by markers XWGGC1505 and XWGGC1507, which correspond to 11.7, 19.2, and 24.9 kb orthologous genomic regions in Brachypodium, rice, and sorghum, respectively. The XWGGC1506 marker co-segregated with Pm41 and could be served as a starting point for chromosome landing and map-based cloning as well as marker-assisted selection of Pm41. Detailed comparative genomics analysis of the markers flanking the Pm41 locus in wheat and the putative orthologous genes in Brachypodium, rice, and sorghum suggests that the gene order is highly conserved between rice and sorghum. However, intra-chromosome inversions and re-arrangements are evident in the wheat and Brachypodium genomic regions, and gene duplications are also present in the orthologous genomic regions of Pm41 in wheat, indicating that the Brachypodium gene model can provide more useful information for wheat marker development.
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Arachnoid cysts with subdural hematoma or intracystic hemorrhage in children.
Pediatr Emerg Care
PUBLISHED: 05-03-2014
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Arachnoid cyst (AC) is a common congenital intracranial lesion in children. It may be complicated by subdural hematoma (SDH) and intracystic hemorrhage (ICH) to cause intracranial hypertension after minor head injury or spontaneously. However, because most bleeding after trauma is delayed, it is often overlooked. At the same time, it remains controversial for treatment of ACs complicated with SDH and ICH. So far, it lacks review, especially for pediatric patients who have ACs with SDH or ICH. Here, we report 3 pediatric cases in our department from 2010 to 2011. At the same time, we review 41 pediatric patients reported in the last 20 years. We conclude that a child with AC should be regularly followed up after minor head injury, and that therapy for children with complicated ACs should be more aggressive than for adults. The cyst wall should be resected, and communication between cyst and cerebral cistern should be established.
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Solution-processable functionalized graphene oxide as an efficient hole transport layer in organic photovoltaics.
J Nanosci Nanotechnol
PUBLISHED: 04-17-2014
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Solution-processable functionalized graphene oxide (SPFGO) thin films as the hole transport layer in organic photovoltaics (OPVs) are fabricated. The performance of the device has been improved by the incorporation of SPFGO. The best results of the open-circuit voltage (V(oc)), short-circuit current density (J(sc)), fill factor (FF) and power conversion efficiency (PCE) were 0.57 V, 7.7 mA/cm2 0.65, and 2.90%, respectively, which are achieved when the thickness of SPFGO film is 3 nm. The dramatic increase in the PCE of OPVs are comparable to those devices fabricated with PEDOT:PSS as the hole transport layers. Therefore, we suggest that SPFGO thin films could be a promising solution-processable alternative to PEDOT:PSS as the effective hole transport layers (HTLs) in OPV devices.
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The Relationship Between Physical Activity and Aging Symptoms Among Community-Dwelling Men Aged 40-70 Years in Shanghai, China.
J Phys Act Health
PUBLISHED: 04-16-2014
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Research on the relationship between physical activity and aging symptoms among men is limited in China.
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Relative quantitative expression of hypoxia-inducible factor 1? messenger ribonucleic acid in recurrent craniopharyngiomas.
Neurol India
PUBLISHED: 03-11-2014
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Craniopharyngioma is a benign tumor, but recurrences are common and prognosis is poor. The pathologic mechanism underlying the high recurrence is still unknown.
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Dickkopf-1 expression is downregulated along colorectal adenoma-carcinoma sequence and correlates with reduced microvessel density and VEGF expression.
Histopathology
PUBLISHED: 03-08-2014
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Dickkopf-1 (Dkk1), an antagonist of wnt/?-catenin signaling pathway, has been reported to play a role in cancer progression. However, little is known about the role of Dkk1 during colorectal adenoma-carcinoma sequence. This study aimed to elucidate the role of Dkk1 in tumorigenesis and angiogenesis in colorectal cancer.
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[Heparin for treatment of sepsis: a systemic review].
Zhonghua Wei Zhong Bing Ji Jiu Yi Xue
PUBLISHED: 03-07-2014
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To systemically review the efficacy and safety of heparin for treatment of sepsis.
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MiR-129-5p is down-regulated and involved in the growth, apoptosis and migration of medullary thyroid carcinoma cells through targeting RET.
FEBS Lett.
PUBLISHED: 02-28-2014
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Dysregulation of the REarranged during Transfection proto-oncogene (RET) pathway and microRNA (miRNAs) are crucial for the development of medullary thyroid carcinomas (MTC). Here we demonstrate that miR-129-5p is down-regulated in MTC tissues and cell lines and inhibits RET expression by directly binding its 3' untranslated regions. Ectopic expression of miR-129-5p significantly decreases cell growth, induces apoptosis and suppresses migration ability in MTC cells through decreasing the phosphorylated AKT, thus functioning as a tumor suppressor. These findings give new clues for understanding MTC carcinogenesis and may help in developing a therapeutic approach for the treatment of RET-activated MTC.
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Mandarin lexical tones identification among children with cochlear implants or hearing aids.
Int. J. Pediatr. Otorhinolaryngol.
PUBLISHED: 02-21-2014
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Mandarin Chinese is a lexical tone language that has four tones, with a change in tone denoting a change in lexical meaning. There are few studies regarding lexical tone identification abilities in deafened children using either cochlear implants (CIs) or hearing aids (HAs). Furthermore, no study has compared the lexical tone identification abilities of deafened children with their hearing devices turned on and off. The present study aimed to investigate the lexical tone identification abilities of deafened children with CIs or HAs.
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Azurocidin-induced inhibition of oxygen metabolism in mitochondria is antagonized by heparin.
Exp Ther Med
PUBLISHED: 02-17-2014
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Heparin is a potent blood anticoagulant that has been demonstrated to attenuate inflammatory responses in sepsis. Sepsis is considered to be a microcirculation-mitochondrial distress syndrome. Azurocidin (AZU), a protein with strong heparin-binding potential that induces inflammatory responses and apoptosis, has been shown to increase the permeability of endothelial cells and induce the prognosis of sepsis. However, the function of AZU in mitochondrial oxygen metabolism has yet to be reported. The aim of the present study was to investigate whether heparin exhibits an antagonistic effect on AZU-induced mitochondrial dysfunction in human umbilical vein endothelial cells (HUVECs) and to further investigate the possible underlying mechanisms. HUVECs were randomly assigned into blank control, AZU, heparin plus AZU and heparin groups. The blank control group were incubated with phosphate-buffered saline for 12 h, while the AZU group were incubated with 1 ?g/ml AZU for 12 h. The heparin plus AZU group were incubated with 100 ?g/ml heparin for 2 h, followed by the addition of 1 ?g/ml AZU and incubation for 12 h. The heparin group were incubated with 100 ?g/ml heparin for 12 h. Flow cytometry was used to determine the mitochondrial membrane potential, while electron microscopy was used to determine the mitochondrial morphology. Western blotting and quantitative polymerase chain reaction were used to determine the protein and mRNA expression levels of Cox II in the mitochondria, respectively. Western blotting was also used to evaluate the concentration of AZU in cytoplasm, along with immunofluorescence analysis. AZU was revealed to decrease the mitochondrial membrane potential, reduce cytochrome c oxidase subunit II expression and destroy the morphology of the mitochondria. Heparin exhibited an antagonistic function on these processes and inhibited the endocytosis of AZU by HUVECs. In conclusion, the results indicated that AZU inhibited the oxygen metabolic function in mitochondria, and this function was effectively antagonized by heparin via the inhibition of AZU endocytosis by HUVECs. Therefore, heparin may be a potential therapeutic agent for treating mitochondrial dysfunction in the future.
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A marker-free automatic alignment method based on scale-invariant features.
J. Struct. Biol.
PUBLISHED: 02-17-2014
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In electron tomography, alignment accuracy is critical for high-resolution reconstruction. However, the automatic alignment of a tilt series without fiducial markers remains a challenge. Here, we propose a new alignment method based on Scale-Invariant Feature Transform (SIFT) for marker-free alignment. The method covers the detection and localization of interest points (features), feature matching, feature tracking and optimization of projection parameters. The proposed method implements a highly reliable matching strategy and tracking model to detect a huge number of feature tracks. Furthermore, an incremental bundle adjustment method is devised to tolerate noise data and ensure the accurate estimation of projection parameters. Our method was evaluated with a number of experimental data, and the results exhibit an improved alignment accuracy comparable with current fiducial marker alignment and subsequent higher resolution of tomography.
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Spontaneous hair cell regeneration in the neonatal mouse cochlea in vivo.
Development
PUBLISHED: 02-06-2014
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Loss of cochlear hair cells in mammals is currently believed to be permanent, resulting in hearing impairment that affects more than 10% of the population. Here, we developed two genetic strategies to ablate neonatal mouse cochlear hair cells in vivo. Both Pou4f3(DTR/+) and Atoh1-CreER™; ROSA26(DTA/+) alleles allowed selective and inducible hair cell ablation. After hair cell loss was induced at birth, we observed spontaneous regeneration of hair cells. Fate-mapping experiments demonstrated that neighboring supporting cells acquired a hair cell fate, which increased in a basal to apical gradient, averaging over 120 regenerated hair cells per cochlea. The normally mitotically quiescent supporting cells proliferated after hair cell ablation. Concurrent fate mapping and labeling with mitotic tracers showed that regenerated hair cells were derived by both mitotic regeneration and direct transdifferentiation. Over time, regenerated hair cells followed a similar pattern of maturation to normal hair cell development, including the expression of prestin, a terminal differentiation marker of outer hair cells, although many new hair cells eventually died. Hair cell regeneration did not occur when ablation was induced at one week of age. Our findings demonstrate that the neonatal mouse cochlea is capable of spontaneous hair cell regeneration after damage in vivo. Thus, future studies on the neonatal cochlea might shed light on the competence of supporting cells to regenerate hair cells and on the factors that promote the survival of newly regenerated hair cells.
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Effect of surfactant protein A on lipopolysaccharide-induced tumor necrosis factor-? expression in human proximal tubular epithelial cells.
Chin. Med. J.
PUBLISHED: 01-21-2014
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Surfactant protein A (SP-A) contributes to the regulation of sepsis-induced acute lung injury. In a previous study, we demonstrated the expression and localization of SP-A in the kidneys. The present study evaluated the effect of SP-A on lipopolysaccharide (LPS)-induced tumor necrosis factor-a (TNF-?) expression and its underlying mechanisms in the human renal tubular epithelial (HK-2) cells.
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Elevated levels of plasma TNF-? are associated with microvascular endothelial dysfunction in patients with sepsis through activating the NF-?B and p38 mitogen-activated protein kinase in endothelial cells.
Shock
PUBLISHED: 01-17-2014
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Inflammatory responses can induce microvascular and endothelial dysfunction, which is associated with the development of sepsis. This study is aimed at examining the concentrations of plasma tissue factor (TF), von Willebrand factor (vWF), and tumor necrosis factor-? (TNF-?) in patients with sepsis and at determining how septic plasma (SP) regulates TF and vWF expression and p38 mitogen activated protein kinase (p38 MAPK)/nuclear factor-?B (NF-?B) pathways in human endothelial cells. The concentrations of plasma TF, vWF, and TNF-? in 22 septic patients and eight healthy controls (HCs) were examined by enzyme-linked immunosorbent assay, and their potential association with disease severity was analyzed. Human umbilical vein endothelial cells (HUVECs) were treated with SP from patients or normal plasma (NP) from the HCs, and the levels of TF and vWF were measured. The SP-induced ERK, p38 MAPK, and NF-?B activation was characterized by Western blot and immunofluorescent assays. The SP-induced HUVEC apoptosis was detected by flow cytometry. The concentrations of plasma TF, vWF, and TNF-? in the patients were significantly higher than that in the HCs and were positively correlated with the Acute Physiology and Chronic Health Evaluation II scores in the patients. Furthermore, treatment with SP, but not NP, induced TF and vWF production in HUVECs in a dose- and time-dependent manner, which was associated with sequential activation of the p38 MAPK and NF-?B pathways. Septic plasma induced HUVEC apoptosis, which was inhibited by activating the NF-?B pathway. The sepsis-related inflammatory factors promoted endothelial cell activation, dysfunction, and apoptosis through activation of the p38 MAPK pathway that was regulated by NF-?B signaling.
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Hypoxic preconditioning increases survival of cardiac progenitor cells via the pim-1 kinase-mediated anti-apoptotic effect.
Circ. J.
PUBLISHED: 01-08-2014
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?Stem cells transplanted to the ischemic myocardium usually encounter massive cell death within a few days after transplantation, and hypoxic preconditioning (HPC) is currently used as a strategy to prepare stem cells for increased survival and engraftment in the heart. The purpose of this study is to determine whether Pim-1 kinase mediates any beneficial effects of HPC for human cardiac progenitor cells (CPCs).
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Fine physical and genetic mapping of powdery mildew resistance gene MlIW172 originating from wild emmer (Triticum dicoccoides).
PLoS ONE
PUBLISHED: 01-01-2014
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Powdery mildew, caused by Blumeria graminis f. sp. tritici, is one of the most important wheat diseases in the world. In this study, a single dominant powdery mildew resistance gene MlIW172 was identified in the IW172 wild emmer accession and mapped to the distal region of chromosome arm 7AL (bin7AL-16-0.86-0.90) via molecular marker analysis. MlIW172 was closely linked with the RFLP probe Xpsr680-derived STS marker Xmag2185 and the EST markers BE405531 and BE637476. This suggested that MlIW172 might be allelic to the Pm1 locus or a new locus closely linked to Pm1. By screening genomic BAC library of durum wheat cv. Langdon and 7AL-specific BAC library of hexaploid wheat cv. Chinese Spring, and after analyzing genome scaffolds of Triticum urartu containing the marker sequences, additional markers were developed to construct a fine genetic linkage map on the MlIW172 locus region and to delineate the resistance gene within a 0.48 cM interval. Comparative genetics analyses using ESTs and RFLP probe sequences flanking the MlIW172 region against other grass species revealed a general co-linearity in this region with the orthologous genomic regions of rice chromosome 6, Brachypodium chromosome 1, and sorghum chromosome 10. However, orthologous resistance gene-like RGA sequences were only present in wheat and Brachypodium. The BAC contigs and sequence scaffolds that we have developed provide a framework for the physical mapping and map-based cloning of MlIW172.
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In vivo generation of immature inner hair cells in neonatal mouse cochleae by ectopic Atoh1 expression.
PLoS ONE
PUBLISHED: 01-01-2014
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Regeneration of auditory hair cells (HCs) is a promising approach to restore hearing. Recent studies have demonstrated that induced pluripotent stem cells/embryonic stem cells or supporting cells (SCs) adjacent to HCs can be converted to adopt the HC fate. However, little is known about whether new HCs are characteristic of outer or inner HCs. Here, we showed in vivo conversion of 2 subtypes of SCs, inner border cells (IBs) and inner phalangeal cells (IPhs), to the inner HC (IHC) fate. This was achieved by ectopically activating Atoh1, a transcription factor necessary for HC fate, in IBs/IPhs at birth. Atoh1+ IBs/IPhs first turned on Pou4f3, another HC transcription factor, before expressing 8 HC markers. The conversion rate gradually increased from ? 2.4% at 1 week of age to ? 17.8% in adult. Interestingly, new HCs exhibited IHC characteristics such as straight line-shaped stereociliary bundles, expression of Fgf8 and otoferlin, and presence of larger outward currents than those of outer HCs. However, new HCs lacked the terminal differentiation IHC marker vGlut3, exhibited reduced density of presynaptic Cbtp2 puncta that had little postsynaptic GluR2 specialization, and displayed immature IHC outward currents. Our results demonstrate that the conversion rate of IBs/IPhs in vivo by Atoh1 ectopic expression into the IHC fate was higher and faster and the conversion was more complete than that of the 2 other SC subtypes underneath the outer HCs; however, these new IHCs are arrested before terminal differentiation. Thus, IBs/IPhs are good candidates to regenerate IHCs in vivo.
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Mussel-Inspired Thermosensitive Polydopamine-graft-Poly(N-isopropylacrylamide) Coating for Controlled-Release Fertilizer.
J. Agric. Food Chem.
PUBLISHED: 12-09-2013
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A thermoresponsive release multi-element compound fertilizer was first reported on the basis of a polydopamine-graft-poly(N-isopropylacrylamide) bilayer coated on a salty core by a combination of dopamine chemistry and surface-initiated atom transfer radical polymerization techniques, and the control of nutrient release in response to the environmental temperature was investigated. The successful synthesized stimuli-responsive fertilizers were confirmed by transmission electron microscopy (TEM), Fourier transforms infrared spectroscopy (FTIR), thermogravimetric analysis (TGA) and gel permeation chromatography (GPC). The release of elements from fertilizer was determined by an inductively coupled plasma (ICP) emission spectrometer. The thermosensitive fertilizers exhibit outstanding stimuli-responsive permeability to encapsulated nutrients, and the release rate of coated elements can be tailored by the ambient temperature. They can release nutrients easily at T < lower critical solution temperature (LCST) but slow at T > LCST. This strategy of grafting thermoresponsive polymer brushes on polydopamine (Pdop)-coated substrates is useful to prepare a stimuli-responsive release system, which can adjust the release rate according to different conditions, and will be effective and promising in the research and development of a stimuli-sensitive controlled-release system.
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The influence of disease and age on human cardiac stem cells.
Ann. Clin. Biochem.
PUBLISHED: 11-06-2013
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Recent studies have found that cardiac stem cells (CSCs) are present in the adult heart. CSCs play an important role in maintaining the balance of the number of myocardial cells. The purpose of this study was to examine characteristics of human CSCs and their correlation with clinical characteristics of patients. We collected heart auricles of 105 patients (age range, 1-78 years; mean, 55.6?±?17.0 years) undergoing cardiac surgery to obtain CSCs. We assayed the percentage of c-kit positive (c-kit(+)) CSCs with flow cytometry. Plasma N(?)-(carboxymethyl)lysine (CML) concentrations were measured by enzyme-linked immunosorbent assay. The percentage of c-kit(+) CSCs was 4.96?±?3.12% (0.98-17.17%), and this was significantly negatively correlated with age, the presence of diabetes mellitus (DM) and coronary heart disease (CHD) (r values were -0.797 [P?
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The Study on Biocompatibility of Porous nHA/PLGA Composite Scaffolds for Tissue Engineering with Rabbit Chondrocytes In Vitro.
Biomed Res Int
PUBLISHED: 07-06-2013
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Objective. To examine the biocompatibility of a novel nanohydroxyapatite/poly[lactic-co-glycolic acid] (nHA/PLGA) composite and evaluate its feasibility as a scaffold for cartilage tissue engineering. Methods. Chondrocytes of fetal rabbit were cultured with nHA/PLGA scaffold in vitro and the cell viability was assessed by MTT assay first. Cells adhering to nHA/PLGA scaffold were then observed by inverted microscope and scanning electron microscope (SEM). The cell cycle profile was analyzed by flow cytometry. Results. The viability of the chondrocytes on the scaffold was not affected by nHA/PLGA comparing with the control group as it was shown by MTT assay. Cells on the surface and in the pores of the scaffold increased in a time-dependent manner. Results obtained from flow cytometry showed that there was no significant difference in cell cycle profiles between the coculture group and control (P > 0.05). Conclusion. The porous nHA/PLGA composite scaffold is a biocompatible and good kind of scaffold for cartilage tissue engineering.
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The in-vitro spheroid culture induces a more highly differentiated but tumorigenic population from melanoma cell lines.
Melanoma Res.
PUBLISHED: 06-12-2013
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Cancer stem cells (CSCs) have been identified in various malignancies, and different properties have been examined to characterize CSCs: tumorigenicity in immunocompromised mice, stem cell surface markers, label-retaining properties, and proliferation as nonadherent spheres. This study explored the consistency and efficiency among these methods. Among the melanoma cell lines examined (A375, A875, MUM-2b, and MUM-2c), only A375 and MUM-2c grew as nonadherent spheres and continuously propagated in a defined serum-free medium in vitro. Flow cytometry and immunofluorescence analysis indicated that sphere-derived cells contained a smaller proportion of cells expressing the candidate surface markers of melanoma stem cells such as ABCB5, CD133, CD20 and CD271, and a larger proportion of cells expressing melanocytic differentiation markers such as HMB45 and S100 protein, compared with adherent cells. Surprisingly, the more highly differentiated sphere-derived melanoma cells exhibited increased tumorigenic potential in vivo, as indicated by shorter tumor incubation (A375) and smaller number of cells required to initiate tumor formation (A375 and MUM-2c) compared with those of parental cells. Despite the similarity in histopathological characteristics, the expression profile indicated that xenografts derived from sphere-derived melanoma cells exhibited a more tumorigenic phenotype with respect to the stem or the differentiation markers detected by immunohistochemical analysis. Therefore, sphere formation in nonadherent cultures may not be a preferred surrogate in-vitro method for enriching melanoma stem cells according to candidate markers but may be a favorable condition for activating potential CSCs.
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The Accumulation of the Glycoxidation Product N(?)-carboxymethyllysine in cardiac tissues with age, diabetes mellitus and coronary heart disease.
Tohoku J. Exp. Med.
PUBLISHED: 05-17-2013
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Heart disease is one of the most important causes of death in developed countries. N(?)-carboxymethyllysine (CML) is a major advanced glycation end product formed by combined reactions of non-enzymatic glycation and oxidation (glycoxidation), and it represents a general marker of oxidative stress. CML has been suggested to be involved in the pathogenesis of heart disease. Plasma CML is elevated in aging, atherosclerosis and/or diabetes. In this study, we measured cardiac CML levels to elucidate its role in the pathogenesis of heart disease. Cardiac tissues were collected from 105 patients (55.6 ± 17.0 years old: age range, 1-78 years) undergoing cardiac surgery. The diseases comprised coronary heart disease (CHD), CHD associated with diabetes mellitus (DM), valvular heart disease and congenital heart disease. The concentration of CML in cardiac tissues of each group was 4.31 ± 0.66, 5.29 ± 0.59, 2.74 ± 1.05 and 1.75 ± 1.16 ?g/g, respectively. ELISA was used for measuring cardiac and plasma CML concentrations. Multiple linear regression analysis showed a significant positive correlation of CML concentrations with age (r = 0.803, p < 0.001), DM (r = 0.567, p < 0.001) and CHD (r = 0.523 p < 0.001). R(2) was 0.872 (p < 0.001); the three independent variables could explain 87.2% variation of CML concentrations. Cardiac CML concentrations exhibited a significant positive correlation with plasma CML (r = 0.983, p < 0.001). Our data indicate that cardiac CML concentrations increase with age, DM and/or CHD, and exhibit a positive correlation with plasma CML concentrations.
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Dual-responsive capsules with tunable low critical solution temperatures and their loading and release behavior.
Langmuir
PUBLISHED: 05-03-2013
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Dual-responsive capsules sensitive to pH and temperature changes were successfully prepared by grafting random copolymer brushes of 2-(2-methoxyethoxy)ethyl methacrylate (MEO2MA) and oligo(ethylene glycol) methacrylate (OEGMA) from polydopamine (Pdop)-coated SiO2 via a surface-initiated atom-transfer radical polymerization (SI-ATRP) method with subsequent removal of the SiO2 core. The uptake and release properties of the resulting capsules are highly affected by changes in the pH values and temperature of the solution. The capsules can take up cationic dye rhodamine 6G (Rh6G) at high pH and T < LCST but not at low pH and T > LCST. In contrast, the capsules can release Rh6G at pH < 7 and temperature below the LCST, but release is less efficient under the opposite conditions. This dual-responsive property was also observed for the anionic dye methyl orange.
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Hypoxia-induced senescence contributes to the regulation of microenvironment in melanomas.
Pathol. Res. Pract.
PUBLISHED: 04-18-2013
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Senescence, an irreversible state of cell cycle arrest, maintains metabolic activity. Although being a barrier against tumor development, senescence could also promote tumor progression by influencing the microenvironment. Necrosis is a common feature of various malignant tumors, which also has two opposing effects: pro-tumor by chronic inflammation and anti-tumor by effective cell clearance. However, the role of senescence in melanoma and whether it is associated with necrosis remain unclear. By detecting senescence-associated ?-galactosidase activity and pimonidazole (hypoxia probe), we found that senescent cells (SA-?-gal positive) are mainly located around the necrotic/hypoxic areas of melanoma from C57BL/6J mice. Moreover, treatment of hypoxia induced irreversibly cellular senescence in vitro. In addition, the senescent cells may facilitate microenvironment modulation and promote the invasion of melanoma cells by secreting matrix metalloproteinase-2(MMP-2). Moreover, Kaplan-Meier analysis showed that the presence of necrosis in melanomas had an inverse correlation with patient survival and may serve as an independent prognostic marker. Therefore, hypoxic stress imposed on melanomas may lead to cellular senescence surrounding necrotic areas, and the adverse effects of necrosis in tumor may be attributed to the adjacent senescent cells with senescence-associated secretion phenotype (SASP), including secretion of MMP-2.
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Guangxis rural health insurance scheme: evidence from an ethnic minority region in China.
Rural Remote Health
PUBLISHED: 04-11-2013
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Chinas New Rural Cooperative Medical Scheme (NCMS) is designed to improve health-service access for rural residents and prevent impoverishment due to medical expenses. Ethnic minority regions in China have specific socioeconomic conditions and policy environments worthy of exploration to assist in the development of rural health insurance schemes.
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Clinical significances and prognostic value of cancer stem-like cells markers and vasculogenic mimicry in renal cell carcinoma.
J Surg Oncol
PUBLISHED: 04-07-2013
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The aim of this study was to investigate the clinical significances and prognostic value of CD133 and CD44 (markers of cancer stem-like cells, CSCs), and vasculogenic mimicry (VM) in renal cell carcinoma (RCC).
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Protein resistance and pH-responsive controlled release from the modification of single-walled carbon nanotubes with a double polymer layer.
Macromol Biosci
PUBLISHED: 03-25-2013
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A reliable and easy method is developed to confer protein resistance and drug controlled release dual functions to single-walled carbon nanotubes (SWNTs) by modification with a polydopamine (Pdop) layer and poly(oligo(ethylene glycol) methacrylate) (POEGMA) brushes. The study of protein resistance demonstrates that POEGMA-modified SWNTs exhibit higher protein resistance than pristine SWNTs, and the protein resistance ability is dependant on the length of the POEGMA molecule. Furthermore, the Pdop layer can serve as a pH-responsive switch for controlled loading and release of rhodamine 6G. The integration of protein resistance and pH-responsive controlled release make the double-layer modified SWNTs attractive for drug delivery in cancer therapy.
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Long non-coding RNA metastasis associated in lung adenocarcinoma transcript 1 derived miniRNA as a novel plasma-based biomarker for diagnosing prostate cancer.
Eur. J. Cancer
PUBLISHED: 03-15-2013
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Examining plasma RNA is an emerging non-invasive diagnosis technique. However, whether tumour-derived long non-coding RNAs (lncRNAs) in plasma can be used as a novel approach to detect human prostate cancer (PCa) has not yet been established. The study was divided into three parts: (1) the characteristics of PCa-related lncRNA fragments were systematically studied in the plasma or serum of 25 patients; (2) the source of the circulating lncRNA fragments was explored in vitro and in vivo; and (3) the diagnostic performance of metastasis associated in lung adenocarcinoma transcript 1 (MALAT-1) derived (MD) miniRNA was validated in an independent cohort of 192 patients. The expression levels of lncRNAs were measured by quantitative real time polymerase chain reaction (qRT-PCR). The MD-miniRNA copies were calculated using a standard curve in an area under the ROC curve (AUC)-receiver operating characteristic (ROC) analysis. Genome-wide profiling revealed that MALAT-1 and prostate cancer gene 3 (PCA3) are overexpressed in PCa tissues. Plasma lncRNAs probably exist in the form of fragments in a stable form. MD-miniRNA enters cell culture medium at measurable levels, and MD-miniRNA derived from human PCa xenografts actually enters the circulation in vivo and can be measured to distinguish xenografted mice from controls. In addition, plasma MD-miniRNA levels are significantly elevated in PCa patients compared to non-PCa patients (p<0.001). At a cut-off of 867.8 MD-miniRNA copies per microlitre of plasma, the sensitivity is 58.6%, 58.6% and 43.5% and the specificity is 84.8%, 84.8% and 81.6% for discriminating PCa from non-PCa, positive biopsy from negative biopsy and positive biopsy from negative biopsy, respectively. We conclude that MD-miniRNA can be used as a novel plasma-based biomarker for PCa detection and can improve diagnostic accuracy by predicting prostate biopsy outcomes. Further large-scale studies are needed to confirm our findings.
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Chemistry and tumor cell growth inhibitory activity of 11,20-epoxy-3Z,5(6)E-diene briaranes from the South China Sea gorgonian Dichotella gemmacea.
Mar Drugs
PUBLISHED: 03-01-2013
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Eighteen new 11,20-epoxy-3Z,5E-dien briaranes, gemmacolides AA-AR (1-18), were isolated together with three known analogs, dichotellides F (19) and I (20), and juncenolide C (21), from the South China Sea gorgonian Dichotella gemmacea. The structures of the compounds were elucidated by detailed spectroscopic analysis and comparison with reported data. The absolute configuration was determined based on the ECD experiment. In the in vitro bioassay, compounds 1-3, 5, 6, 8-12, and 14-19 exhibited different levels of growth inhibition activity against A549 and MG63 cell lines. Preliminary structure-activity analysis suggests that 12-O-isovalerate may increase the activity whereas 13- or 14-O-isovalerate may decrease the activity. Contribution of substitutions at C-2 and C-16 remains uncertain.
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All-trans retinoic acid inhibits craniopharyngioma cell growth: study on an explant cell model.
J. Neurooncol.
PUBLISHED: 02-09-2013
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The ratio between FABP5 and CRABPII determines cellular response to physiological level of retinoic acid; tumor cells undergo proliferation with high level of FABP5 and apoptosis with high level of CRABPII. We intended to study FABP5 and CRABPII expression in craniopharyngiomas, to establish craniopharyngioma cell model using explants method, and to study the effect of pharmacological dose of retinoic acid on craniopharyngioma cells. Expression of FABP5 and CRABPII in craniopharyngioma tissue from 20 patients was studied using immunohistochemistry. Primary craniopharyngioma cell cultures were established using tissue explants method. Craniopharyngioma cells were treated using various concentrations of all-trans retinoic acid, and cell growth curve, apoptosis, expression of FABP5, CRABPII and NF-?B were assayed in different groups. FABP5/CRABPII ratio was significantly higher in adamatinomatous group than that in papillary group. Cell cultures were established in 19 cases (95 %). Pharmacological level retinoic acid inhibited cell growth and induced cellular apoptosis in dose dependent manner, and apoptosis rate cells treated with 30 ?M retinoic acid for 24 h was 43 %. Also, retinoic acid increased CRABPII, and decreased FABP5 and NF-?B expression in craniopharyngioma cells. High FABP5/CRABPII ratio is observed in adamatinomatous craniopharyngioma. Retinoic acid at pharmacological level induced craniopharyngioma cell apoptosis via increasing FABP5/CRABPII ratio and inhibiting NF-?B signaling pathway. Our study demonstrated that all-trans retinoic acid might be a candidate for craniopharyngioma adjuvant chemotherapy in future.
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Risk factors for and prevalence of knee osteoarthritis in the rural areas of Shanxi Province, North China: a COPCORD study.
Rheumatol. Int.
PUBLISHED: 01-29-2013
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The purpose of this study is to investigate the prevalence of knee osteoarthritis (OA) in the rural areas of Shanxi Province, North China. A total of 7,126 permanent residents aged from 16 to 90 years were surveyed using Community Oriented Program for the Control of Rheumatic Diseases methodology. Diagnosis of knee OA was reached according to the examination results by 3 rheumatologists. Possible risk factors for knee OA were analyzed. Among the 7,126 participants, 983 cases were diagnosed with knee OA. Of the 983 cases, 446 were male (12.4%) and 537 were female (15.3%). The overall prevalence of knee OA was 13.8%. The prevalence rate of knee pain was significantly higher in women than in men. There was a tendency of increased knee OA prevalence with age, especially after 40 years old. Participants with higher body mass index (BMI) showed a higher prevalence rate of knee OA than those with lower BMI. Multivariable analysis indicates age, gender, dietary bias, underground work history, BMI, waist-to-hip ratio (WHR), and concomitant cardiovascular diseases (CSDs) are risk factors for knee OA in rural Shanxi. The prevalence of knee OA in the rural areas of Shanxi Province is high. Age, gender, dietary bias, underground work history, BMI, WHR, and CSDs are risk factors for knee OA. Primary and secondary prevention programs aimed at improving ventilation condition, reducing obesity, and treating concomitant cardiovascular diseases are required.
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Isothermal detection of multiple point mutations by a surface plasmon resonance biosensor with Au nanoparticles enhanced surface-anchored rolling circle amplification.
Biosens Bioelectron
PUBLISHED: 01-19-2013
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In this study, we developed a surface plasmon resonance (SPR) DNA biosensor method using surface-anchored rolling circle amplification (RCA) and Au nanoparticles modified probes (AuNPs) to isothermally detect multiple point mutations associated with drug-resistance in multidrug-resistant Mycobacterium Tuberculosis (MDRTB). A set of probes contains an allele-specific padlock probe (PLP), a capture probe and an AuNPs. The linear PLPs, circularized by ligation upon the recognition of the point mutation on DNA targets, hybridize to the capture probes via the specific tag/anti-tag recognition. Upon recognition each point mutation is identified by locating into the corresponding channel on the chip. Then the immobilized primer (capture probe)-template (circular PLP) complex are amplified isothermally as RCA and further amplified by AuNPs. The RCA products immobilized on the chip surface cause great SPR angle changes consequently. The 5 pM synthetic oligonucleotides and 8.2 pg uL(-1) of genomic DNA from clinical samples can be detected by the method. The positive mutation detection is achieved with a wild-type to mutant ratio of 5000:1. The method was demonstrated by targeting five clinically meaningful mutations in MDRTB. Thirty clinical samples were identified and they were in good agreement with the results from sequencing.
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Silencing of human phosphatidylethanolamine-binding protein 4 enhances rituximab-induced death and chemosensitization in B-cell lymphoma.
PLoS ONE
PUBLISHED: 01-15-2013
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Rituximab is the first line drug to treat non Hodgkins lymphoma (B-NHL) alone or in combination with chemotherapy. However, 30-40% of B-NHL patients are unresponsive to rituximab or resistant after therapy. Human phosphatidylethanolamine-binding protein 4 (hPEBP4) is a novel member of PEBP family and functions as an anti-apoptotic molecule. In this study, we found hPEBP4 to be expressed in up to 90% of B-cell lymphoma patients, but in only 16.7% of normal lymph nodes. Interestingly, hPEBP4 overexpression inhibited rituximab-mediated complement dependent cytotoxicity (R-CDC) and antibody-dependent cell-mediated cytotoxicity (ADCC) in B-NHL cells while downregulation of hPEBP4 augmented the therapeutic efficacy of rituximab both in vitro and in vivo. Furthermore, hPEBP4 silencing sensitized the primary B-acute lymphocytic leukemia (B-ALL) cells to R-CDC. During rituximab-mediated complement dependent cytotoxicity, hPEBP4 was recruited to the cell membrane in a PE-binding domain dependent manner and inhibited R-CDC induced calcium flux and reactive oxygen species (ROS) generation. These events contributed to the decrease of cell death induced by R-CDC in B-cell lymphomas. Meanwhile, hPEBP4 knockdown potentiated the chemosensitization of the rituximab in B-cell lymphoma cells by regulating the expression of Bcl-xl, Cycline E, p21(waf/cip1) and p53 and the activation of caspase-3 and caspase-9. Considering that hPEBP4 conferred cellular resistance to rituximab treatment and was preferentially expressed in lymphoma tissue, it could be a potential valuable target for adjuvant therapy for B-cell lymphoma.
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Platelet-derived growth factor-BB accelerates prostate cancer growth by promoting the proliferation of mesenchymal stem cells.
J. Cell. Biochem.
PUBLISHED: 01-09-2013
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Mesenchymal stem cells (MSCs) favor cancer growth by facilitating immunosuppression status in tumor microenvironment. However, the function and mechanism of MSCs in initiating and developing prostate cancer remains to be fully understood. In this study, we first found that MSCs promoted prostate cancer (PCa) tumor growth in vivo and cell proliferation in vitro by using PCs cell strain RM-1. Both exogenous and endogenous MSCs could be recruited into the tumor microenvironment by using bone-marrow transplantation model. We further demonstrated that PDGF-BB produced by RM-1 cell promoted MSCs proliferation in vivo and in vitro, which was abrogated by Si-RNA specific to PDGF-BB. And inflammatory cytokines, such as interferon gamma, tumor necrosis factor alpha, and anti-inflammatory cytokine transformation growth factor alpha, further increased the ability of RM-1 to produce PDGF-BB. Overall, PCa cells produced PDGF-BB favors the proliferation of MSCs, which may elicit immunosuppressive function and enable PCa cells to escape from the immunity surveillance in tumor inflammatory microenvironment.
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Comparative high-resolution mapping of the wax inhibitors iw1 and iw2 in hexaploid wheat.
PLoS ONE
PUBLISHED: 01-01-2013
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THE WAX (GLAUCOUSNESS) ON WHEAT LEAVES AND STEMS IS MAINLY CONTROLLED BY TWO SETS OF GENES: glaucousness loci (W1 and W2) and non-glaucousness loci (Iw1 and Iw2). The non-glaucousness (Iw) loci act as inhibitors of the glaucousness loci (W). High-resolution comparative genetic linkage maps of the wax inhibitors Iw1 originating from Triticum dicoccoides, and Iw2 from Aegilops tauschii were developed by comparative genomics analyses of Brachypodium, sorghum and rice genomic sequences corresponding to the syntenic regions of the Iw loci in wheat. Eleven Iw1 and eight Iw2 linked EST markers were developed and mapped to linkage maps on the distal regions of chromosomes 2BS and 2DS, respectively. The Iw1 locus mapped within a 0.96 cM interval flanked by the BE498358 and CA499581 EST markers that are collinear with 122 kb, 202 kb, and 466 kb genomic regions in the Brachypodium 5S chromosome, the sorghum 6S chromosome and the rice 4S chromosome, respectively. The Iw2 locus was located in a 4.1 to 5.4-cM interval in chromosome 2DS that is flanked by the CJ886319 and CJ519831 EST markers, and this region is collinear with a 2.3 cM region spanning the Iw1 locus on chromosome 2BS. Both Iw1 and Iw2 co-segregated with the BF474014 and CJ876545 EST markers, indicating they are most likely orthologs on 2BS and 2DS. These high-resolution maps can serve as a framework for chromosome landing, physical mapping and map-based cloning of the wax inhibitors in wheat.
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The Maize glossy13 Gene, Cloned via BSR-Seq and Seq-Walking Encodes a Putative ABC Transporter Required for the Normal Accumulation of Epicuticular Waxes.
PLoS ONE
PUBLISHED: 01-01-2013
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Aerial plant surfaces are covered by epicuticular waxes that among other purposes serve to control water loss. Maize glossy mutants originally identified by their "glossy" phenotypes exhibit alterations in the accumulation of epicuticular waxes. By combining data from a BSR-Seq experiment and the newly developed Seq-Walking technology, GRMZM2G118243 was identified as a strong candidate for being the glossy13 gene. The finding that multiple EMS-induced alleles contain premature stop codons in GRMZM2G118243, and the one knockout allele of gl13, validates the hypothesis that gene GRMZM2G118243 is gl13. Consistent with this, GRMZM2G118243 is an ortholog of AtABCG32 (Arabidopsis thaliana), HvABCG31 (barley) and OsABCG31 (rice), which encode ABCG subfamily transporters involved in the trans-membrane transport of various secondary metabolites. We therefore hypothesize that gl13 is involved in the transport of epicuticular waxes onto the surfaces of seedling leaves.
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In vivo visualization of Notch1 proteolysis reveals the heterogeneity of Notch1 signaling activity in the mouse cochlea.
PLoS ONE
PUBLISHED: 01-01-2013
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Mechanosensory hair cells (HCs) and surrounding supporting cells (SCs) in the mouse cochlea are important for hearing and are derived from the same prosensory progenitors. Notch1 signaling plays dual but contrasting and age-dependent roles in mouse cochlear development: early lateral induction and subsequent lateral inhibition. However, it has been difficult to directly visualize mouse cochlear cells experiencing various levels of Notch1 activity at single cell resolution. Here, we characterized two knock-in mouse lines, Notch1(Cre (Low)/+) and Notch1(Cre (High)/+) , with different Cre recombinase activities, that can detect Notch1 receptor proteolysis or Notch1 activity at high and low thresholds, respectively. Using both lines together with a highly sensitive Cre reporter line, we showed that Notch1 activity is nearly undetectable during lateral induction but increases to medium and high levels during lateral inhibition. Furthermore, we found that within the neonatal organ of Corti, the vast majority of cells that experience Notch1 activity were SCs not HCs, suggesting that HCs kept undetectable Notch1 activity during the entire lineage development. Furthermore, among SC subtypes, ?85-99% of Deiters and outer pillar cells but only ?19-38% of inner pillar cells experience medium and high levels of Notch1 activity. Our results demonstrate that Notch1 activity is highly heterogeneous: 1) between lateral induction and inhibition; 2) between HC and SC lineages; 3) among different SC subtypes; 4) among different cells within each SC subtype. Such heterogeneity should elucidate how the development of the cochclear sensory epithelium is precisely controlled and how HC regeneration can be best achieved in postnatal cochleae.
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NONCODE v3.0: integrative annotation of long noncoding RNAs.
Nucleic Acids Res.
PUBLISHED: 12-01-2011
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Facilitated by the rapid progress of high-throughput sequencing technology, a large number of long noncoding RNAs (lncRNAs) have been identified in mammalian transcriptomes over the past few years. LncRNAs have been shown to play key roles in various biological processes such as imprinting control, circuitry controlling pluripotency and differentiation, immune responses and chromosome dynamics. Notably, a growing number of lncRNAs have been implicated in disease etiology. With the increasing number of published lncRNA studies, the experimental data on lncRNAs (e.g. expression profiles, molecular features and biological functions) have accumulated rapidly. In order to enable a systematic compilation and integration of this information, we have updated the NONCODE database (http://www.noncode.org) to version 3.0 to include the first integrated collection of expression and functional lncRNA data obtained from re-annotated microarray studies in a single database. NONCODE has a user-friendly interface with a variety of search or browse options, a local Genome Browser for visualization and a BLAST server for sequence-alignment search. In addition, NONCODE provides a platform for the ongoing collation of ncRNAs reported in the literature. All data in NONCODE are open to users, and can be downloaded through the website or obtained through the SOAP API and DAS services.
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Transurethral seminal vesiculoscopy using a 6F vesiculoscope for ejaculatory duct obstruction: initial experience.
J. Androl.
PUBLISHED: 11-17-2011
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Ejaculatory duct obstruction (EDO) is a surgically correctable condition that occurs in some infertile men. The standard therapy is transurethral resection of ejaculatory ducts (TURED). However, TURED has been associated with a high risk of complications, including the impairment of semen parameters and retrograde ejaculation. In our clinical practice, vesiculoscopy has demonstrated potential as a minimally invasive alternative technique for the diagnosis and treatment of EDO. Very few studies have examined transurethral seminal vesiculoscopy (TRU-SVS) in recent years, and no study has examined 6F vesiculoscopes. Therefore, we performed a retrospective study of TRU-SVS using a 6F vesiculoscope and its effect on the diagnosis and treatment of EDO. A total of 21 patients who underwent this procedure were included in the study. The mean patient age was 28.8 years (range, 23-36 years). The procedure was completed successfully in all patients within a mean time of 31.5 minutes and a mean hospital stay of 1.17 days. All patients had EDO. Calculi were found in the ejaculatory ducts or in the seminal vesicles of 5 patients. Sperm was detected in 11 patients 1-3 months postsurgery and in another 8 patients 3-12 months postsurgery. No sperm was detected in the remaining 2 patients by 12 months postsurgery. Epididymitis, retrograde ejaculation, urinary incontinence, and rectal injury were not observed. These data indicate that TRU-SVS using a 6F vesiculoscope affords direct access to the seminal vesicle and offers the advantages of fewer complications and more optimal sperm recovery as well as direct, dynamic video imaging.
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Improving photovoltaic properties by incorporating solution-processable functionalized graphene into poly(3-octylthiophene)/phenyl c61 butyric acid methyl ester photovoltaic devices.
J Nanosci Nanotechnol
PUBLISHED: 11-15-2011
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We report the optoelectronic properties occurring in solution-processable functionalized graphene (SPFGraphene)-PCBM/poly(3-octylthiophene) (P3OT) composites. The structural configuration of the devices is ITO/PEDOT:PSS/P3OT:PCBM-SPFGraphene/LiF/Al. The best results were obtained with a P3OT/PCBM (1:1) mixture with 8 wt% of SPFGraphene in an open-circuit voltage (V(OC)) of 0.65 V, a short-circuit current density (J(SC)) of 4.2 mA/cm2, and a fill factor (FF) of 0.35, which led to a power conversion efficiency of 0.95% at illumination at 100 mW/cm2 AM 1.5. In the P3OT:PCBM-SPFGraphene composite, the SPFGraphene acted as exciton dissociation sites and provided the transport pathway of the lowest unoccupied molecular orbital (LUMO)-graphene-Al. Doping SPF-Graphene into P3OT resulted in appropriate energetic distance between the highest occupied molecular orbital (HOMO) and LUMO of the donor/acceptor for a high open circuit voltage and provided higher exciton dissociation volume mobility of carrier transport for a large short-circuit current density.
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Graphene layers from reduction of exfoliated graphite oxide.
J Nanosci Nanotechnol
PUBLISHED: 11-15-2011
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Graphene oxide (GO) of approximately 1 nm was generated from exfoliated graphitic oxide using a modified Hummers method through ultrasonic treatment in water, and the GO film was reduced under protection of Ar/H2 flow at 800 degrees C. Moreover, the obtained graphene film has a high conductivity of 383 S/cm at 10-20 nm thickness.
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Differential expression and function of AR isoforms in prostate cancer.
Oncol. Rep.
PUBLISHED: 10-08-2011
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The androgen receptor (AR) plays a key role in prostate cancer (PCa). Two isoforms of AR, are AR-A and -B, which differ by a lack of the first 187 amino acids in the NH2-terminal transactivation domain of AR-A. Since little is known about the expression and basic function of the AR-A/B isoforms in prostate cancer, the aim of this study was to analyze this possible association. The AR-A, -B and AR-A/B ratio was determined in the tissues of healthy controls, benign prostate hyperplasia (BPH) and PCa by means of Western blot analysis and immunofluorescence. The elevation of AR-A, and -B, as well as the AR-A/B ratio with regard to Gleason scores, were assessed in prostate cancer compared to BPH and normal prostate. In order to further investigate the role of AR A/B isoform function, we transfected PC3 cells with an AR or AR-A expression vector. The overexpression of AR-A and -B significantly decreased the invasion and proliferation of PC3 cells. However, the overexpression of AR-A further decreased proliferation but accelerated the invasion of PC3 cells compared to AR-B. In conclusion, the elevation of AR-A and -B, and the AR-A/B ratio, is associated with prostate cancer occurrence and progression. Furthermore, AR-A could provide a new potential therapy with regard to the decrease in the invasion and proliferation of prostate cancer cells. Our study provides insight into further understanding the biological role of AR-A in its interaction with AR-B and its impact on PCa clinical treatment.
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A fast mapping method in the ISAF reconstruction algorithm.
Conf Proc IEEE Eng Med Biol Soc
PUBLISHED: 08-29-2011
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ISAF (icosahedral symmetry-adapted functions) algorithm is the new high-resolution algorithm of icosahedral molecules. But its running speed is very slow because of the time-consuming operations of mapping sampling points into 3D space. In this paper, a fast mapping method is proposed to increase the running speed of this stage. First of all, the angle corresponding to one pixel arc in the maximum Fourier ring was taken as the sampling angle and the same angle sampling was applied in every rings. After that, the sampling points in ring R=1 were mapped into 3D space. Finally, the 3D spatial positions of radial sampling points in other rings were deduced according to the rotate angle invariability of radial sampling points. The simulated data of PSV-F (Penicillium stoloniferum virus F) and experimental Cryo-EM data of CPV (cytoplasmic polyhedrosis virus) were used for validating this method. The results show that the whole speedup reaches to an order of magnitude at the premise of assuring accuracy. In addition, the speedup is increasing with the increase of the maximum Fourier radius and the number of projections.
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Characterization of highly antimicrobial-resistant clinical pneumococcal isolates recovered in a Chinese hospital during 2009-2010.
J. Med. Microbiol.
PUBLISHED: 08-26-2011
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Ninety-one consecutive pneumococcal isolates (primarily from sputum), recovered in Chongqing Southwest Hospital during a 12 month period in 2009-2010 from individuals of all ages with suspected cases of pneumococcal disease, were subjected to PCR-serotyping, Quellung reaction serotyping, antimicrobial-susceptibility testing and multilocus sequence typing (MLST). Although 20 different serotypes were observed, most isolates (69, 75.8 %) were of serotypes included in the pneumococcal 13-valent conjugate vaccine (PCV13), including 33 of the 46 (71.7 %) isolates recovered from individuals less than 5 years of age. The prevalent serotypes were 19F (34 %), 19A (9.9 %), 6B (9.9 %), 23F (7.7 %), 14 (6.6 %) and 6A (4.4 %). PCR-determined serotypes were in agreement with Quellung testing, with the exception of two serotype 33C isolates. Most or all isolates within each PCV13 serotype were represented by one genotype, with the globally disseminated MLST sequence types (STs) ST271, ST320, ST90 and ST81 each accounting for the highly resistant isolates within serotypes 19F, 19A, 6B and 23F, respectively. Sixty-six (72.5 %) isolates were resistant to combinations of ?-lactam antibiotics (BLAs). A total of 63 of these 66 (95.5 %) BLA-resistant isolates were of serotypes included in PCV13; however, 3 serogroup 15 isolates were also BLA-resistant. Most isolates (88/91 = 96.7 %) were resistant to erythromycin and clindamycin. The majority of isolates were also resistant to tetracycline (76, 84 %) and to cotrimoxazole (67, 74 %). This work revealed that the majority of antimicrobial-resistant isolates (50/91 = 54.9 %) recovered in this Chinese hospital were represented by four global clones. Serotypes for these as well as more obscure strains were readily determined by using PCR.
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[Comparative study on repairing rabbit radius segmental defects with two different proportions of chitosan combined with allogeneic morselized bone].
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi
PUBLISHED: 08-09-2011
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To give a preliminary experimental evidence and to prove chitosan and allogeneic morselized bone as potential bone substitutions in repairing rabbit radius segmental defect.
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Extracelluar matrix metalloproteinase as a novel target for pancreatic cancer therapy.
Anticancer Drugs
PUBLISHED: 07-07-2011
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The objective of this study was to evaluate extracellular matrix metalloproteinase (EMMPRIN) as a novel target in orthotopic pancreatic cancer murine models. MIA PaCa-2 human pancreatic tumor cells were implanted in groups 1 and 3-7, whereas MIA PaCa-2 EMMPRIN knockdown cells were implanted in group 2. Dosing with anti-EMMPRIN antibody started immediately after implantation for groups 1-3 (residual tumor model) and at 21 days after cell implantation for groups 4-7 (established tumor model). Groups 3, 5, and 7 were treated with anti-EMMRPIN antibody (0.2-1.0 mg) twice weekly for 2-3 weeks, whereas the other groups served as the control. In the residual tumor model, tumor growth of anti-EMMPRIN-treated group was successfully arrested for 21 days (15 ± 4 mm(3)), which was significantly lower than that of the EMMPRIN knockdown group (80 ± 15 mm(3); P=0.001) or the control group (240 ± 41 mm(3); P<0.001). In the established tumor model, anti-EMMPRIN therapy lowered tumor volume increase by approximately 40% compared with the control, regardless of the dose amount. Ki67-expressed cell density of group 5 was 939 ± 150 mm(-2), which was significantly lower than that of group 4 (1709 ± 145 mm(-2); P=0.006). Microvessel density of group 5 (30 ± 6 mm(-2)) was also significantly lower than that of group 4 (53 ± 5 mm(-2); P=0.014), whereas the microvessel size of group 5 (191 ± 22 ?m(2)) was significantly larger than that of group 4 (113 ± 26 ?m(2); P=0.049). These data show the high potential of anti-EMMPRIN therapy for pancreatic cancer and support its clinical translation.
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Targeted therapy in breast cancer: whats new?
Swiss Med Wkly
PUBLISHED: 06-28-2011
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Breast cancer is the most commonly diagnosed malignancy and one of the major causes of death among women. Breast cancer is also one of the most investigated diseases but whose biological features are still not well understood, several effective treating strategies having been explored in dealing with different types of advanced breast cancer, such as endocrine therapy and molecular targeted therapy. Trastuzumab is the first approved targeted anti-cancer agent to show an attractive response rate and outcomes in treating HER-2 positive metastatic breast cancer patients. However, primary or acquired trastuzumab resistance usually occurs some time into the use of trastuzumab and leads to treatment resistance or tumour progression. The promising results with trastuzumab targeted therapy encouraged further investigations in this area exploring several novel targeted agents aiming to overcome the resistance drawback of trastuzumab. In this review we discuss the major newly developed targeted agents in breast cancer treatment, including the novel anti-HER-2 monoclonal antibody pertuzumab or ertumaxomab, small molecular tyrosine inhibitor lapatinib, selective PARP1 inhibitor olaparib, mTOR inhibitor rapamycin analogues, and sheddase inhibitors. Many of these novel targeted drugs or molecules showed additional or complementary effects to trastuzumab therapy that need further and wider investigation.
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Identification and comparative mapping of a powdery mildew resistance gene derived from wild emmer (Triticum turgidum var. dicoccoides) on chromosome 2BS.
Theor. Appl. Genet.
PUBLISHED: 06-21-2011
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Powdery mildew, caused by Blumeria graminis f. sp. tritici, is an important foliar disease of wheat worldwide. Wild emmer (Triticum turgidum var. dicoccoides) is a valuable genetic resource for improving disease resistance in common wheat. A powdery mildew resistance gene conferring resistance to B. graminis f. sp. tritici isolate E09 at the seedling and adult stages was identified in wild emmer accession IW170 introduced from Israel. An incomplete dominant gene, temporarily designated MlIW170, was responsible for the resistance. Through molecular marker and bulked segregant analyses of an F(2) population and F(3) families derived from a cross between susceptible durum wheat line 81086A and IW170, MlIW170 was located in the distal chromosome bin 2BS3-0.84-1.00 and flanked by SSR markers Xcfd238 and Xwmc243. MlIW170 co-segregated with Xcau516, an STS marker developed from RFLP marker Xwg516 that co-segregated with powdery mildew resistance gene Pm26 on 2BS. Four EST-STS markers, BE498358, BF201235, BQ160080, and BF146221, were integrated into the genetic linkage map of MlIW170. Three AFLP markers, XPaacMcac, XPagcMcta, XPaacMcag, and seven AFLP-derived SCAR markers, XcauG2, XcauG3, XcauG6, XcauG8, XcauG10, XcauG20, and XcauG25, were linked to MlIW170. XcauG3, a resistance gene analog (RGA)-like sequence, co-segregated with MlIW170. The non-glaucousness locus Iw1 was 18.77 cM distal to MlIW170. By comparative genomics of wheat-Brachypodium-rice genomic co-linearity, four EST-STS markers, CJ658408, CJ945509, BQ169830, CJ945085, and one STS marker XP2430, were developed and MlIW170 was mapped in an 2.69 cM interval that is co-linear with a 131 kb genomic region in Brachypodium and a 105 kb genomic region in rice. Four RGA-like sequences annotated in the orthologous Brachypodium genomic region could serve as chromosome landing target regions for map-based cloning of MlIW170.
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Fibroblast growth factor receptor mediates fibroblast-dependent growth in EMMPRIN-depleted head and neck cancer tumor cells.
Mol. Cancer Res.
PUBLISHED: 06-10-2011
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Head and neck squamous cell carcinoma tumors (HNSCC) contain a dense fibrous stroma which is known to promote tumor growth, although the mechanism of stroma-mediated growth remains unclear. As dysplastic mucosal epithelium progresses to cancer, there is incremental overexpression of extracellular matrix metalloprotease inducer (EMMPRIN) which is associated with tumor growth and metastasis. Here, we present evidence that gain of EMMPRIN expression allows tumor growth to be less dependent on fibroblasts by modulating fibroblast growth factor receptor-2 (FGFR2) signaling. We show that silencing EMMPRIN in FaDu and SCC-5 HNSCC cell lines inhibits cell growth, but when EMMPRIN-silenced tumor cells were cocultured with fibroblasts or inoculated with fibroblasts into severe combined immunodeficient mice, the growth inhibition by silencing EMMPRIN was blunted by the presence of fibroblasts. Coculture experiments showed fibroblast-dependent tumor cell growth occurred via a paracrine signaling. Analysis of tumor gene expression revealed expression of FGFR2 was inversely related to EMMPRIN expression. To determine the role of FGFR2 signaling in EMMPRIN-silenced tumor cells, ligands and inhibitors of FGFR2 were assessed. Both FGF1 and FGF2 enhanced tumor growth in EMMPRIN-silenced cells compared with control vector-transfected cells, whereas inhibition of FGFR2 with blocking antibody or with a synthetic inhibitor (PD173074) inhibited tumor cell growth in fibroblast coculture, suggesting the importance of FGFR2 signaling in fibroblast-mediated tumor growth. Analysis of xenografted tumors revealed that EMMPRIN-silenced tumors had a larger stromal compartment compared with control. Taken together, these results suggest that EMMPRIN acquired during tumor progression promotes fibroblast-independent tumor growth.
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Three-dimensional reconstruction using an adaptive simultaneous algebraic reconstruction technique in electron tomography.
J. Struct. Biol.
PUBLISHED: 06-03-2011
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Three-dimensional (3D) reconstruction of electron tomography (ET) has emerged as an important technique in analyzing structures of complex biological samples. However most of existing reconstruction methods are not suitable for extremely noisy and incomplete data conditions. We present an adaptive simultaneous algebraic reconstruction technique (ASART) in which a modified multilevel access scheme and an adaptive relaxation parameter adjustment method are developed to improve the quality of the reconstructed 3D structure. The reconstruction process is facilitated by using a column-sum substitution approach. This modified multilevel access scheme is adopted to arrange the order of projections so as to minimize the correlations between consecutive views within a limited angle range. In the adaptive relaxation parameter adjustment method, not only the weight matrix (as in the existing methods) but the gray levels of the pixels are employed to adjust the relaxation parameters so that the quality of the reconstruction is improved while the convergence process of the reconstruction is accelerated. In the column-sum substitution approach, the computation to obtain the reciprocal of the sum for the columns in each view is avoided so that the needed computations for each iteration can be reduced. Experimental results show that the proposed technique ASART is better based on objective quality measures than other methods, especially when data is noisy and limited in tilt angles. At the same time, the reconstruction by ASART outperforms that of simultaneous algebraic reconstruction technique (SART) in speed.
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A high throughput barley stripe mosaic virus vector for virus induced gene silencing in monocots and dicots.
PLoS ONE
PUBLISHED: 04-28-2011
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Barley stripe mosaic virus (BSMV) is a single-stranded RNA virus with three genome components designated alpha, beta, and gamma. BSMV vectors have previously been shown to be efficient virus induced gene silencing (VIGS) vehicles in barley and wheat and have provided important information about host genes functioning during pathogenesis as well as various aspects of genes functioning in development. To permit more effective use of BSMV VIGS for functional genomics experiments, we have developed an Agrobacterium delivery system for BSMV and have coupled this with a ligation independent cloning (LIC) strategy to mediate efficient cloning of host genes. Infiltrated Nicotiana benthamiana leaves provided excellent sources of virus for secondary BSMV infections and VIGS in cereals. The Agro/LIC BSMV VIGS vectors were able to function in high efficiency down regulation of phytoene desaturase (PDS), magnesium chelatase subunit H (ChlH), and plastid transketolase (TK) gene silencing in N. benthamiana and in the monocots, wheat, barley, and the model grass, Brachypodium distachyon. Suppression of an Arabidopsis orthologue cloned from wheat (TaPMR5) also interfered with wheat powdery mildew (Blumeria graminis f. sp. tritici) infections in a manner similar to that of the A. thaliana PMR5 loss-of-function allele. These results imply that the PMR5 gene has maintained similar functions across monocot and dicot families. Our BSMV VIGS system provides substantial advantages in expense, cloning efficiency, ease of manipulation and ability to apply VIGS for high throughput genomics studies.
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Knowledge, attitudes, and practices on malaria prevention among Chinese international travelers.
J Travel Med
PUBLISHED: 04-06-2011
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To address the lack of understanding in malaria prevention among Chinese international travelers, we have conducted knowledge, attitudes, and practices (KAP) study in five different Chinese geographic areas. This survey represents one part of the background information needed to analyze imported malaria.
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Pu and 137Cs in the Yangtze River estuary sediments: distribution and source identification.
Environ. Sci. Technol.
PUBLISHED: 02-09-2011
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Pu isotopes and (137)Cs were analyzed using sector field ICP-MS and ? spectrometry, respectively, in surface sediment and core sediment samples from the Yangtze River estuary. (239+240)Pu activity and (240)Pu/(239)Pu atom ratios (>0.18) shows a generally increasing trend from land to sea and from north to south in the estuary. This spatial distribution pattern indicates that the Pacific Proving Grounds (PPG) source Pu transported by ocean currents was intensively scavenged into the suspended sediment under favorable conditions, and mixed with riverine sediment as the water circulated in the estuary. This process is the main control for the distribution of Pu in the estuary. Moreover, Pu is also an important indicator for monitoring the changes of environmental radioactivity in the estuary as the river basin is currently the site of extensive human activities and the sea level is rising because of global climate changes. For core sediment samples the maximum peak of (239+240)Pu activity was observed at a depth of 172 cm. The sedimentation rate was estimated on the basis of the Pu maximum deposition peak in 1963-1964 to be 4.1 cm/a. The contributions of the PPG close-in fallout Pu (44%) and the riverine Pu (45%) in Yangtze River estuary sediments are equally important for the total Pu deposition in the estuary, which challenges the current hypothesis that the riverine Pu input was the major source of Pu budget in this area.
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RH knowledge and service utilization among unmarried rural-to-urban migrants in three major cities, China.
BMC Public Health
PUBLISHED: 02-02-2011
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Large numbers of unmarried migrants are on the continuous move from rural-to-urban areas within China mainland, meanwhile their Reproductive Health (RH) is underserved when it is compared with the present urban RH policies. The purpose of this study is to investigate the RH knowledge and the utilization of RH services among unmarried migrants.
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Polymorphism analysis of csd gene in six Apis mellifera subspecies.
Mol. Biol. Rep.
PUBLISHED: 01-30-2011
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The complementary sex determination (csd) gene is the primary gene determining the gender of honey bees (Apis spp). In this study we analyzed the polymorphism of csd gene in six Apis mellifera subspecies. The genomic region 3 of csd gene in these six A. mellifera was cloned, and identified. A total of 79 haplotypes were obtained from these six subspecies. Analysis showed that region 3 of csd gene has a high level of polymorphism in all the six A. mellifera subspecies. The A. m. anatolica subspecies has a slightly higher nucleotide diversity (?) than other subspecies, while the ? values showed no significant difference among the other five subspecies. The phylogenetic tree showed that all the csd haplotypes from different A. mellifera subspecies are scattered throughout the tree, without forming six different clades. Population differentiation analysis showed that there are significant genetic differentiations among some of the subspecies. The NJ phylogenetic tree showed that the A. m. caucasica and A. m. carnica have the closest relationship, followed by A. m. ssp, A. m. ligustica, A. m. carpatica and A. m. anatolica that were gathered in the tree in turn.
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Dynamic expression pattern of Sonic hedgehog in developing cochlear spiral ganglion neurons.
Dev. Dyn.
PUBLISHED: 05-27-2010
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Sonic hedgehog (Shh) signaling plays important roles in the formation of the auditory epithelium. However, little is known about the detailed expression pattern of Shh and the cell sources from which Shh is secreted. By analyzing Shh(CreEGFP/+) mice, we found that Shh was first expressed in all cochlear spiral ganglion neurons by embryonic day 13.5, after which its expression gradually decreased from base to apex. By postnatal day 0, it was not detected in any spiral ganglion neurons. Genetic cell fate mapping results also confirmed that Shh was exclusively expressed in all spiral ganglion neurons and not in surrounding glia cells. The basal-to-apical wave of Shh decline strongly resembles that of hair cell differentiation, supporting the idea that Shh signaling inhibits hair cell differentiation. Furthermore, this Shh(CreEGFP/+) mouse is a useful Cre line in which to delete floxed genes specifically in spiral ganglion neurons of the developing cochlea.
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A novel function for vimentin: the potential biomarker for predicting melanoma hematogenous metastasis.
J. Exp. Clin. Cancer Res.
PUBLISHED: 05-03-2010
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The incidence of malignant melanoma (MM) was occurring at a faster rate than for most neoplasm worldwide, and melanoma metastasis is still the most formidable problem. So it is necessarily to find some biomarkers associated with melanoma metastasis.
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In vivo proliferation of postmitotic cochlear supporting cells by acute ablation of the retinoblastoma protein in neonatal mice.
J. Neurosci.
PUBLISHED: 04-30-2010
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Cochlear hair cells (HCs) are mechanosensory receptors that transduce sound into electrical signals. HC damage in nonmammalian vertebrates induces surrounding supporting cells (SCs) to divide, transdifferentiate and replace lost HCs; however, such spontaneous HC regeneration does not occur in the mammalian cochlea. Here, we acutely ablate the retinoblastoma protein (Rb), a crucial cell cycle regulator, in two subtypes of postmitotic SCs (pillar and Deiters cells) using an inducible Cre line, Prox1-CreER(T2). Inactivation of Rb in these SCs results in cell cycle reentry of both pillar and Deiters cells, and completion of cell division with an increase in cell number of pillar cells. Interestingly, nuclei of Rb(-/-) mitotic pillar and Deiters cells migrate toward the HC layer and divide near the epithelial surface in a manner similar to the SCs in the regenerating avian auditory epithelium. In contrast to postmitotic Rb(-/-) HCs which abort cell division, postmitotic Rb(-/-) pillar cells can proliferate, maintain their SC fate and survive for more than a week. However, no newly formed HCs are detected and SC death followed by HC loss occurs. Our studies accomplish a crucial step toward functional HC regeneration in the mammalian cochlea in vivo, demonstrating the critical role of Rb in maintaining quiescence of postmitotic pillar and Deiters cells and highlighting the heterogeneity between these two cell types. Therefore, the combination of transient Rb inactivation and further manipulation of transcription factors (i.e., Atoh1 activation) in SCs may represent an effective therapeutic avenue for HC regeneration in the mammalian cochlea.
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Genetic and comparative genomics mapping reveals that a powdery mildew resistance gene Ml3D232 originating from wild emmer co-segregates with an NBS-LRR analog in common wheat (Triticum aestivum L.).
Theor. Appl. Genet.
PUBLISHED: 03-27-2010
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Powdery mildew caused by Blumeria graminis f. sp. tritici is one of the most important wheat diseases worldwide and breeding for resistance using diversified disease resistance genes is the most promising approach to prevent outbreaks of powdery mildew. A powdery mildew resistance gene, originating from wild emmer wheat (Triticum turgidum var. dicoccoides) accessions collected from Israel, has been transferred into the hexaploid wheat line 3D232 through crossing and backcrossing. Inoculation results with 21 B. graminis f. sp. tritici races indicated that 3D232 is resistant to all of the powdery mildew isolates tested. Genetic analyses of 3D232 using an F(2) segregating population and F(3) families indicated that a single dominant gene, Ml3D232, confers resistance in the host seedling stage. By applying molecular markers and bulked segregant analysis (BSA), we have identified polymorphic simple sequence repeats (SSR), expressed sequence tags (EST) and derived sequence tagged site (STS) markers to determine that the Ml3D232 is located on chromosome 5BL bin 0.59-0.76. Comparative genetic analyses using mapped EST markers and genome sequences of rice and Brachypodium established co-linearity of the Ml3D232 genomic region with a 1.4 Mb genomic region on Brachypodium distachyon chromosome 4, and a 1.2 Mb contig located on the Oryza sativa chromosome 9. Our comparative approach enabled us to develop new EST-STS markers and to delimit the genomic region carrying Ml3D232 to a 0.8 cM segment that is collinear with a 558 kb region on B. distachyon. Eight EST markers, including an NBS-LRR analog, co-segregated with Ml3D232 to provide a target site for fine genetic mapping, chromosome landing and map-based cloning of the powdery mildew resistance gene. This newly developed common wheat germplasm provides broad-spectrum resistance to powdery mildew and a valuable resource for wheat breeding programs.
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Protective effects of xyloketal B against MPP+-induced neurotoxicity in Caenorhabditis elegans and PC12 cells.
Brain Res.
PUBLISHED: 03-16-2010
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Parkinsons disease (PD) is the second most common neurodegenerative disease, affecting 2% of the population over age 65years. Mitochondrial defect and oxidative stress actively participate in the dopaminergic (DA) neuron degeneration in PD. Xyloketal B is a novel marine compound with unique chemical structure isolated from mangrove fungus Xylaria sp. (no. 2508). Recently, we have demonstrated that Xyloketal B can directly scavenge DPPH free radicals and protects mitochondria against oxidative insult. In the present study, we investigate the neuroprotective action of xyloketal B against MPP+-induced neurotoxicity in Caenorhabditis elegans and PC12 cells. The viability and DA neurodegeneration was assessed in C. elegans selectively expressing green fluorescent protein (GFP) in DA neurons. PC12 cell damage was measured using MTT and nuclear morphology. Intracellular reactive oxygen species (ROS), mitochondrial membrane potential and total GSH were assessed. Xyloketal B dose-dependently protected against MPP+-induced loss of viability and DA neurodegeneration in C. elegans. Similar neuroprotection was replicated in MPP+ PC12 cell model. In addition, xyloketal B attenuated MPP+-induced intracellular ROS accumulation, loss of mitochondrial membrane potential and restored total GSH level in PC12 cells. All together, the present study demonstrates that xyloketal B protects against MPP+-induced neurotoxicity in C. elegans and PC12 cells mainly through its antioxidant property and restoration of total GSH level.
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Cloning of the papaya chromoplast-specific lycopene beta-cyclase, CpCYC-b, controlling fruit flesh color reveals conserved microsynteny and a recombination hot spot.
Plant Physiol.
PUBLISHED: 02-24-2010
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Carotenoid pigments in fruits are indicative of the ripening process and potential nutritional value. Papaya (Carica papaya) fruit flesh color is caused by the accumulation of lycopene or beta-carotenoids in chromoplasts. It is a distinct feature affecting nutritional composition, fruit quality, shelf life, and consumer preference. To uncover the molecular basis of papaya flesh color, we took map-based cloning and candidate gene approaches using integrated genetic and physical maps. A DNA marker tightly linked to flesh color colocalized on a contig of the physical map with a cDNA probe of the tomato (Solanum lycopersicum) chromoplast-specific lycopene beta-cyclase, CYC-b. Candidate gene sequences were obtained from amplified fragments and verified by sequencing two bacterial artificial chromosomes containing the two alleles. Sequence comparison revealed a 2-bp insertion in the coding region of the recessive red flesh allele resulting in a frame-shift mutation and a premature stop codon. A color complementation test in bacteria confirmed that the papaya CpCYC-b is the gene controlling fruit flesh color. Sequence analysis of wild and cultivated papaya accessions showed the presence of this frame-shift mutation in all red flesh accessions examined. Evaluation of DNA markers near CpCYC-b revealed a recombination hot spot, showing that CpCYC-b is located in a gene-rich region with a recombination rate at 3.7 kb per centimorgan, more than 100-fold higher than the genome average at 400 kb per centimorgan. Conserved microsynteny of the CpCYC-b region is indicated by colinearity of two to four genes between papaya, Arabidopsis (Arabidopsis thaliana), grape (Vitis vinifera), and tomato. Our results enhanced our understanding of papaya flesh color inheritance and generated new tools for papaya improvement.
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Influence of soil washing with a chelator on subsequent chemical immobilization of heavy metals in a contaminated soil.
J. Hazard. Mater.
PUBLISHED: 01-14-2010
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To assess the influence of soil washing with a chelator on the chemical immobilization of heavy metals, batch experiments were performed on the fine fraction of a contaminated soil under various operating conditions. Results show that pre-washing with EDTA facilitated the chemical immobilization of Cu and Cr, while an opposite effect for Pb and Zn was observed, in particular when Ca(OH)(2) was added as the immobilizing agent. Metal fraction analyses of the soils indicate that soil washing can reduce the metal mobility by removing the labile fractions, while it may also destabilize some strongly bound fractions, reversely increasing the mobility and thus compromising the subsequent immobilization performance to some extents. To secure an effective combination of soil washing and chemical immobilization for the remediation of heavy metal-contaminated sites, a comprehensive study on metal fraction distribution in the soil is needed.
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In developing our video relationships, we compare around 5 million PubMed articles to our library of over 4,500 methods videos. In some cases the language used in the PubMed abstracts makes matching that content to a JoVE video difficult. In other cases, there happens not to be any content in our video library that is relevant to the topic of a given abstract. In these cases, our algorithms are trying their best to display videos with relevant content, which can sometimes result in matched videos with only a slight relation.