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Find video protocols related to scientific articles indexed in Pubmed.
Effect of deoxycholic acid on Ca(2+) movement, cell viability and apoptosis in human gastric cancer cells.
Toxicol. Mech. Methods
PUBLISHED: 11-20-2014
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Abstract Deoxycholic acid (DOA) is one of the secondary bile acids used as a mild detergent for the isolation of membrane associated proteins. This study examined whether the secondary bile acid, DOA, altered Ca(2+) movement, cell viability and apoptosis in SCM1 human gastric cancer cells. The Ca(2+)-sensitive fluorescent dye fura-2 was used to measure [Ca(2+)]i. DOA evoked [Ca(2+)]i rises concentration-dependently. The response was reduced by removing extracellular Ca(2+). DOA-evoked Ca(2+) entry was inhibited by store-operated Ca(2+) channel inhibitors (nifedipine, econazole and SKF96365), the protein kinase C (PKC) activator phorbol 12-myristate 13 acetate (PMA) and the PKC inhibitor GF109203X. In Ca(2+)-free medium, treatment with the endoplasmic reticulum Ca(2+) pump inhibitor thapsigargin (TG) abolished DOA-evoked [Ca(2+)]i rises. Conversely, treatment with DOA abolished TG-evoked [Ca(2+)]i rises. Inhibition of phospholipase C with U73122 abolished DOA-evoked [Ca(2+)]i rises. At 100-500 ?M, DOA decreased cell viability, which was not changed by chelating cytosolic Ca(2+) with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid-acetoxymethyl ester (BAPTA/AM). DOA between 100 ?M and 300 ?M also induced apoptosis. Collectively, in SCM1 cells, DOA induced [Ca(2+)]i rises by evoking phospholipase C-dependent Ca(2+) release from the endoplasmic reticulum and Ca(2+) entry via store-operated Ca(2+) channels. DOA also caused Ca(2+)-independent apoptosis.
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The discovery of the genusSpasskia Belokobylskij, 1989 (Hymenoptera: Braconidae) in China, with description of a new species.
J. Insect Sci.
PUBLISHED: 11-05-2014
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The genus Spasskia Belokobylskij, 1989 (Hymenoptera: Braconidae: Helconinae) is reported for the first time from China. Two species, namely Spasskia brevicarinata Yan et Chen sp. n.and Spasskia indica Singh, Belokobylskij et Chauhan, 2005 are described and illustrated. A key to the species of this genus is updated to include the new species.
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Ultrasonography and Computed Tomography Diagnostic Evaluation of Budd-Chiari Syndrome Based on Radical Resection Exploration Results.
Ultrasound Q
PUBLISHED: 11-04-2014
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The current study used exploration during radical resection, which reveals the vascular lesion directly, as a criterion standard to evaluate and compare sonography and computed tomography (CT) diagnosis.
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Double-Line Hammett Relationship Revealed through Precise Acidity Measurement of Benzenethiols in Neat Ionic Media: A Typical "Ionic Liquid Effect"?
Org. Lett.
PUBLISHED: 10-14-2014
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Equilibrium acidities (pKa) of 14 aromatic thiols in four pure room temperature ionic liquids (RTILs) were precisely determined and the corresponding acidity scales were established for the first time. Regression analyses show a distinct double-line Hammett relationship with similar slopes and excellent linearity (R(2) of 0.996-0.999) in all four ILs. This could be rationalized by a resonance enhanced effect of the IL cation to solvate the para substituent of feasible electronic structure (CSAR effect), revealing a typical and rarely seen "ionic liquid effect".
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Sc(OTf)3-Catalyzed Transfer Diazenylation of 1,3-Dicarbonyls with Triazenes via N-N Bond Cleavage.
Org. Lett.
PUBLISHED: 10-08-2014
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A new and efficient method for diazenylation reactions was developed with a Sc(OTf)3-catalyzed nitrogen-nitrogen bond cleavage process with triazenes. The transfer diazenylation reactions accommodate a diverse range of active methylene substrates including simple ketones to give aliphatic azo compounds that are of significant potential as azo prodrugs in high yields under mild conditions.
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Effect of Fluoxetine on [Ca²?]i and Cell Viability in OC2 Human Oral Cancer Cells.
Chin J Physiol
PUBLISHED: 09-23-2014
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Fluoxetine is a serotonin-specific reuptake inhibitor that has been used as an antidepressant. This study examined the effect of fluoxetine on cytosolic free Ca²? concentrations ([Ca2?]i) and viability in OC2 human oral cancer cells. The Ca²?-sensitive fluorescent dye fura-2 was used to measure [Ca²?]i, and the water soluble tetrazolium (WST-1) regent was used to measure viability. Fluoxetine induced [Ca²?]i rises concentration-dependently. The response was reduced by half by removing extracellular Ca²?. Fluoxetine-induced Ca²? entry was enhanced by activation of protein kinase C (PKC) with phorbol 12-myristate 13 acetate (PMA) but was inhibited by inhibition of the enzyme with GF109203X. In Ca²?-free medium, treatment with the endoplasmic reticulum Ca²? pump inhibitor 2,5-di-tert-butylhydroquinone (BHQ) or thapsigargin abolished fluoxetine-evoked [Ca²?]i rise. Conversely, treatment with fluoxetine inhibited BHQ/thapsigargin-evoked [Ca²?]i rise. Inhibition of phospholipase C (PLC) with U73122 abolished fluoxetine-induced [Ca²?]i rise. At 20-80 ?M, fluoxetine decreased cell viability concentration-dependently, which was not altered by chelating cytosolic Ca²? with 1,2-bis(2- aminophenoxy)ethane-N,N,N',N'-tetraacetic acid-acetoxymethyl ester (BAPTA/AM). At 20-60 ?M, fluoxetine induced apoptosis as detected by annexin V/propidium iodide (PI) staining. Together, in OC2 cells, fluoxetine induced [Ca²?]i rises by evoking PLC-dependent Ca²? release from the endoplasmic reticulum and Ca²? entry via PKC-regulated mechanisms. Fluoxetine also caused Ca²?-independent apoptosis.
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Direct intramolecular conjugate addition of simple alkenes to ?,?-unsaturated carbonyls catalyzed by Cu(OTf)2.
Org. Lett.
PUBLISHED: 09-22-2014
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An unprecedented intramolecular conjugate addition of simple alkenes to ?,?-unsaturated carbonyl compounds has been developed. A simple Lewis acid such as Cu(OTf)2 was found to effectively catalyze the reaction, and six- and five-membered cyclic products were obtained in moderate to high yields.
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Prognostic significance of plasma chemerin levels in patients with gastric cancer.
Peptides
PUBLISHED: 08-23-2014
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Chemerin is a novel adipokine, which is linked to adipogenesis and chemotaxis of the innate immune system. This study aimed to evaluate the relationship between preoperative plasma chemerin level and prognosis of gastric cancers. One hundred ninety-six patients and 196 age- and gender-matched healthy individuals were recruited. Fasting venous blood samples were collected 2 days prior to surgery for the gastric cancer patients and at the physical examination day for the healthy volunteers. Recorded clinicopathological information included invasion depth, lymph node metastasis, distant metastasis, peritoneal dissemination, tumor size and tumor-node metastasis stage. Plasma chemerin levels were determined using enzyme-linked immunosorbent assay. Plasma chemerin levels were statistically significantly in all patients than in healthy controls (53.1±19.0ng/mL vs. 31.3±11.3ng/mL; P<0.001). And it was identified as an independent predictor for 5-year mortality [odds ratio (OR), 2.718; 95% confidence interval (CI), 1.201-4.229; P=0.005) and adverse event (OR, 2.982; 95% CI, 1.223-4.879; P=0.003) of gastric cancer, and had high area under receiver operating characteristic curve (AUC) for prediction of 5-year mortality (AUC, 0.808; 95% CI, 0.745-0.860) and adverse event (AUC, 0.787; 95% CI, 0.723-0.842). It also emerged as an independent predictor for overall survival (Hazard ratios, 1.788; 95% CI, 1.200-2.663; P=0.002) and disease-free survival (Hazard ratios, 2.016; 95% CI, 1.312-3.125; P=0.004). Thus, our results suggest that high plasma chemerin levels are associated with long-term poor prognosis and survival of gastric cancer as well as may play a role as prognostic biomarker in gastric cancer survival.
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Asymmetric sulfa-Michael addition to ?-substituted vinyl ketones catalyzed by chiral primary amine.
Org. Lett.
PUBLISHED: 08-21-2014
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The first effective example of asymmetric conjugate addition-protonation reactions of thiols to ?-substituted vinyl ketones by chiral primary amine catalysis is reported. A simple chiral primary-tertiary diamine catalyst derived from l-phenylalanine was found to promote the sulfa-Michael addition-protonation reactions with good to excellent enantioselectivity.
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Copper-catalyzed aerobic oxidative cleavage of C-C bonds in epoxides leading to aryl ketones.
J. Org. Chem.
PUBLISHED: 08-15-2014
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A novel copper-catalyzed aerobic synthesis of ketones from epoxides via cleavage of C-C single bonds has been discovered. This reaction constitutes a new transformation from epoxides into ketones.
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Dying tumor cells stimulate proliferation of living tumor cells via caspase-dependent protein kinase C? activation in pancreatic ductal adenocarcinoma.
Mol Oncol
PUBLISHED: 08-07-2014
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Pancreatic cancer is one of the most lethal human cancers, and radiotherapy is often implemented for locally advanced pancreatic ductal adenocarcinoma. Tumor cell repopulation is a major challenge in treating cancers after radiotherapy. In order to address the problem of tumor repopulation, our previous studies have demonstrated that dying cells stimulate the proliferation of living tumor cells after radiotherapy. In particular, dying cells undergoing apoptosis also activate survival or proliferation signals and release growth factors to surrounding living cells. In the present study, we used an in vitro model to examine the possible mechanisms for dying cell stimulated tumor repopulation in pancreatic cancer. In this model, a small number of living, luciferase-labeled pancreatic cancer cells (reporter) were seeded onto a layer of a much larger number of irradiated, unlabeled pancreatic cancer cells and the growth of the living cells was measured over time as a gage of tumor repopulation. Our results indicate that irradiated, dying Panc1 feeder cells significantly stimulated the proliferation of living Panc1 reporter cells. Importantly, we identified that the percentage of apoptotic cells and the cleavage of caspases 3 and 7 and protein kinase C? (PKC?) were increased in irradiated Panc1 cells. We presumed that caspases 3 and 7 and PKC? as integral mediators in the process of dying pancreatic cancer cell stimulation of living tumor cell growth. In order to demonstrate the importance of caspases 3, 7 and PKC?, we introduced dominant-negative mutants of caspase 3 (DN_C3), caspase 7 (DN_C7), or PKC? (DN_PKC?) into Panc1 cells using lentiviral vectors. The stably transduced Panc1 cells were irradiated and used as feeders and we found a significant decrease in the growth of living Panc1 reporter cells when compared with irradiated wild-type Panc1 cells as feeders. Moreover, the role of PKC? in the growth stimulation of living tumor cells was further confirmed using a pan PKC inhibitor GF109203x and a specific PKC? inhibitor, rottlerin. Additionally, we found significantly increased phosphorylation of Akt, p38 mitogen-activated protein kinase (MAPK) and c-Jun N-terminal kinase/stress-activated protein kinase (JNK1/2) in the irradiated Panc1 cells. Mechanistically, PKC? cleavage was attenuated in both DN_C3 and DN_C7 transduced Panc1 cells, and both Akt and p38 MAPK phosphorylation were attenuated in DN_PKC? transduced Panc1 cells following radiation. Thus, this report suggests a novel finding that cellular signaling caspase 3/7-PKC?-Akt/p38 MAPK is crucial to the repopulation in Panc1 cells after radiotherapy.
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The mechanism of honokiol-induced intracellular Ca(2+) rises and apoptosis in human glioblastoma cells.
Chem. Biol. Interact.
PUBLISHED: 08-07-2014
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Honokiol, an active constituent of oriental medicinal herb Magnolia officinalis, caused Ca(2+) mobilization and apoptosis in different cancer cells. In vivo, honokiol crossed the blood-brain or -cerebrospinal fluid barrier, suggesting that it may be an effective drug for the treatment of brain tumors, including glioblastoma. This study examined the effect of honokiol on intracellular Ca(2+) concentration ([Ca(2+)]i) and apoptosis in DBTRG-05MG human glioblastoma cells. Honokiol concentration-dependently induced a [Ca(2+)]i rise. The signal was decreased partially by removal of extracellular Ca(2+). Honokiol-triggered [Ca(2+)]i rise was not suppressed by store-operated Ca(2+) channel blockers (nifedipine, econazole, SK&F96365) and the protein kinase C (PKC) activator phorbol 12-myristate 13 acetate (PMA), but was inhibited by the PKC inhibitor GF109203X. GF109203X-induced inhibition was not altered by removal of extracellular Ca(2+). In Ca(2+)-free medium, pretreatment with the endoplasmic reticulum Ca(2+) pump inhibitor thapsigargin (TG) or 2,5-di-tert-butylhydroquinone (BHQ) abolished honokiol-induced [Ca(2+)]i rise. Conversely, incubation with honokiol abolished TG or BHQ-induced [Ca(2+)]i rise. Inhibition of phospholipase C (PLC) with U73122 abolished honokiol-induced [Ca(2+)]i rise. Honokiol (20-80?M) reduced the cell viability, which was not reversed by prechelating cytosolic Ca(2+) with BAPTA-AM (1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid-acetoxymethyl ester). Honokiol (20-60?M) enhanced reactive oxygen species (ROS) production, decreased mitochondrial membrane potential, released cytochrome c, and activated caspase-9/caspase-3. Together, honokiol induced a [Ca(2+)]i rise by inducing PLC-dependent Ca(2+) release from the endoplasmic reticulum and Ca(2+) entry via PKC-dependent, non store-operated Ca(2+) channels. Moreover, honokiol activated the mitochondrial pathway of apoptosis in DBTRG-05MG human glioblastoma cells.
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Route to optimal generation of soft X-ray high harmonics with synthesized two-color laser pulses.
Sci Rep
PUBLISHED: 08-06-2014
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High harmonics extending to X-rays have been generated from gases by intense lasers. To establish these coherent broadband radiations as an all-purpose tabletop light source for general applications in science and technology, new methods are needed to overcome the present low conversion efficiencies. Here we show that the conversion efficiency may be drastically increased with an optimized two-color pulse. By employing an optimally synthesized 2-µm mid-infrared laser and a small amount of its third harmonic, we show that harmonic yields from sub- to few-keV energy can be increased typically by ten-fold over the optimized single-color one. By combining with favorable phase-matching and together with the emerging high-repetition MHz mid-infrared lasers, we anticipate efficiency of harmonic yields can be increased by four to five orders in the near future, thus paving the way for employing high harmonics as useful broadband tabletop light sources from the extreme ultraviolet to the X-rays, as well as providing new tools for interrogating ultrafast dynamics of matter at attosecond timescales.
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Comparison of proton pump inhibitor and histamine-2 receptor antagonist in the prevention of recurrent peptic ulcers/erosions in long-term low-dose aspirin users: a retrospective cohort study.
Biomed Res Int
PUBLISHED: 07-23-2014
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Proton pump inhibitor and histamine-2 receptor antagonist can prevent aspirin-related ulcers/erosions but few studies compare the efficacy of these two agents. Aims. We evaluated the efficacy of omeprazole and famotidine in preventing recurrent ulcers/erosions in low-dose aspirin users.
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Visible-light promoted catalyst-free imidation of arenes and heteroarenes.
Chemistry
PUBLISHED: 07-20-2014
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We described herein a catalyst-free visible-light photolytic protocol for the imidation of arenes and heteroarenes. N-Bromosaccharin was identified as a viable and chemoselective nitrogen radical precursor that undergoes controllable homolytic cleavage under ambient light irradiation. The reaction can be applied to a number of arenes and heteroarenes with good chemo- and regioselectivity. Mechanistic studies revealed that radical chain termination by electron transfer-proton transfer (ET-PT) is the leading productive pathway for the reaction.
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Universality of returning electron wave packet in high-order harmonic generation with midinfrared laser pulses.
Phys. Rev. Lett.
PUBLISHED: 07-15-2014
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We show that a returning electron wave packet in high-order harmonic generation (HHG) with midinfrared laser pulses converges to a universal limit for a laser wavelength above about 3 ?m. The results are consistent among the different methods: a numerical solution of the time-dependent Schrödinger equation, the strong-field approximation, and the quantum orbits theory. We further analyze how the contribution from different electron "trajectories" survives the macroscopic propagation in the medium. Our result thus provides a new framework for investigating the wavelength scaling law for the HHG yields.
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Chemical Acylation of Water-Soluble Antioxidant of Bamboo Leaves (AOB-w) and Functional Evaluation of Oil-Soluble AOB (cAOB-o).
J. Food Sci.
PUBLISHED: 07-14-2014
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Antioxidant of bamboo leaves (AOB) is a novel natural food antioxidant approved in China since 2004. Natural phenolics contained in the current AOB are usually polyhydroxy derivatives exhibiting hydrophilic character, which has been marked as water-soluble AOB (AOB-w). In order to broaden the application fields, oil-soluble AOB (cAOB-o) was obtained by chemical acylation of AOB-w with different chain-length fatty acids varying from C8 to C18. Results indicated that the yield and solubility of cAOB-o in 1-octanol solvent depended on the carbon chain length of acyl donor, and cAOB-o derived from C12 fatty acid exhibited the more powerful antioxidant activity evaluated by ?-carotene/linoleic acid bleaching assay. Total phenolic content decreased by Folin-Ciocalteu assay. Fourier transform infrared spectra showed the increase of a carbonyl (C = O) peak at 1701 cm(-1) and a decrease in the intensity of the absorption at 3400 cm(-1) (O-H stretching) in cAOB-o. Acylation was inferred to mainly occur on the hydroxyl groups of flavones C-glycosides according to the change of high-performance liquid chromatography spectra and the contents of total flavonoids and phenolic acids. cAOB-o with the addition of 0.02% significantly increased oxidative stability of palm oil 1.59 times, lard 3.74 times, and fried potato chips 2.08 times, which was better than the effect of oil-soluble tea polyphenol (P < 0.01). Moreover, cAOB-o was identified to be actually nontoxicity by an acute oral toxicity test. All the above results indicated that cAOB-o could be used as a novel and effective oil-soluble antioxidant in the food industry.
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Effect of melamine on [Ca(2+)]i and viability in PC3 human prostate cancer cells.
Environ. Toxicol. Pharmacol.
PUBLISHED: 06-20-2014
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Melamine is thought to be an endocrine disrupter that affects physiology in cells. This study examined the effect of melamine on cytosolic free Ca(2+) concentrations ([Ca(2+)]i) and viability in PC3 human prostate cancer cells. Melamine evoked [Ca(2+)]i rises concentration-dependently. Melamine-evoked Ca(2+) entry was inhibited by nifedipine, econazole, SKF96365, GF109203X and phorbol 12-myristate 13 acetate. In Ca(2+)-free medium, treatment with the endoplasmic reticulum Ca(2+) pump inhibitor thapsigargin inhibited melamine-evoked [Ca(2+)]i rise. Conversely, treatment with melamine abolished thapsigargin-evoked [Ca(2+)]i rise. Inhibition of phospholipase C with U73122 did not alter melamine-evoked [Ca(2+)]i rise. Melamine at 500-800?M decreased cell viability, which was not reversed by pretreatment with the Ca(2+) chelator 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid-acetoxymethyl ester (BAPTA/AM). Collectively, our data suggest that in PC3 cells, melamine induced [Ca(2+)]i rises by evoking phospholipase C-independent Ca(2+) release from the endoplasmic reticulum, and Ca(2+) entry via protein kinase C-regulated store-operated Ca(2+) entry. Melamine also caused Ca(2+)-independent cell death.
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Nasopharyngeal adenoid cystic carcinoma: a case report and review of the literature.
Int J Clin Exp Pathol
PUBLISHED: 06-15-2014
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ACC derived from nasopharyngeal epithelial cells is rare, usually benign. In this article, we reported a nasopharyngeal adenoid cystic carcinoma (NACC) in a 31-year-old woman with a symptom of hoarseness, headache, epistaxis slightly, diplopia, facial numbness and dysphagia near 3 months. A tumor on the right side of the nasopharynx was confirmed by laryngoscope check and MRI of the skull base. Histopathological findings showed that tumor cells were arranged in cord-like or acinar-like by atypical hyperplastic epithelial cells forming a cribriform and tubular pattern, and immunohistochemical findings showed that tumor tissues were immunopositive for p63 (+), CK7 (+), CK19 (+), CK8 (+), CK18 (+), SMA (+), CK (+), p53 (++), S-100 (+) and Ki-67 (5%+), and negative for CD34 (-), CK5/6 (-), CEA (-) and CD117 (-). Patient was treated by surgical operation and radiotherapy, and was followed-up near 10 months, no local recurrence and distant metastasis.
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ATG4B promotes colorectal cancer growth independent of autophagic flux.
Autophagy
PUBLISHED: 06-12-2014
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Autophagy is reported to suppress tumor proliferation, whereas deficiency of autophagy is associated with tumorigenesis. ATG4B is a deubiquitin-like protease that plays dual roles in the core machinery of autophagy; however, little is known about the role of ATG4B on autophagy and proliferation in tumor cells. In this study, we found that ATG4B knockdown induced autophagic flux and reduced CCND1 expression to inhibit G 1/S phase transition of cell cycle in colorectal cancer cell lines, indicating functional dominance of ATG4B on autophagy inhibition and tumor proliferation in cancer cells. Interestingly, based on the genetic and pharmacological ablation of autophagy, the growth arrest induced by silencing ATG4B was independent of autophagic flux. Moreover, dephosphorylation of MTOR was involved in reduced CCND1 expression and G 1/S phase transition in both cells and xenograft tumors with depletion of ATG4B. Furthermore, ATG4B expression was significantly increased in tumor cells of colorectal cancer patients compared with adjacent normal cells. The elevated expression of ATG4B was highly correlated with CCND1 expression, consistently supporting the notion that ATG4B might contribute to MTOR-CCND1 signaling for G 1/S phase transition in colorectal cancer cells. Thus, we report that ATG4B independently plays a role as a positive regulator on tumor proliferation and a negative regulator on autophagy in colorectal cancer cells. These results suggest that ATG4B is a potential biomarker and drug target for cancer therapy.
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Phytoextraction potential of poplar (Populus alba L. var. pyramidalis Bunge) from calcareous agricultural soils contaminated by cadmium.
Int J Phytoremediation
PUBLISHED: 06-11-2014
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To investigate the phytoextraction potential of Populus alba L. var. pyramidalis Bunge for cadmium (Cd) contaminated calcareous soils, a concentration gradient experiment and a field sampling experiment (involving poplars of different ages) were conducted. The translocation factors for all experiments and treatments were greater than 1. The bioconcentration factor decreased from 2.37 to 0.25 with increasing soil Cd concentration in the concentration gradient experiment and generally decreased with stand age under field conditions. The Cd concentrations in P. pyramidalis organs decreased in the order of leaves > stems > roots. The shoot biomass production in the concentration gradient experiment was not significantly reduced with soil Cd concentrations up to or slightly over 50 mg kg(-1). The results show that the phytoextraction efficiency of P. pyramidalis depends on both the soil Cd concentration and the tree age. Populus pyramidalis is most suitable for remediation of slightly Cd contaminated calcareous soils through the combined harvest of stems and leaves under actual field conditions.
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Programmed cell death 2 protein induces gastric cancer cell growth arrest at the early S phase of the cell cycle and apoptosis in a p53-dependent manner.
Oncol. Rep.
PUBLISHED: 05-29-2014
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Programmed cell death 2 (PDCD2) is a highly conserved nuclear protein, and aberrant PDCD2 expression alters cell apoptosis. The present study aimed to investigate PDCD2 expression in gastric cancer. Tissue specimens from 34 gastric cancer patients were collected for analysis of PDCD2 expression using immunohistochemistry, western blotting and qRT-PCR. Gastric cancer cell lines (a p53-mutated MKN28 line and a wild-type p53 MKN45 line) were used to assess the effects of PDCD2 overexpression. p53-/- nude mice were used to investigate the effect of PDCD2 on ultraviolet B (UVB)-induced skin carcinogenesis. The data showed that PDCD2 expression was reduced in gastric cancer tissue specimens, and loss of PDCD2 expression was associated with the poor survival of patients. PDCD2 expression induced gastric cancer cell growth arrest at the early S phase of the cell cycle and apoptosis. The antitumor effects of PDCD2 expression were dependent on p53 expression in gastric cancer cells. Moreover, PDCD2 expression inhibited activity of the ATM/Chk1/2/p53 signaling pathway. In addition, PDCD2 expression suppressed UVB-induced skin carcinogenesis in p53+/+ nude mice, but not in p53-/- mice. The data from the present study demonstrated that loss of PDCD2 expression could contribute to gastric cancer development and progression and that PDCD2-induced gastric cancer cell growth arrest at the early S phase of the cell cycle and apoptosis are p53-dependent.
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Gastrointestinal hemorrhage in warfarin anticoagulated patients: incidence, risk factor, management, and outcome.
Biomed Res Int
PUBLISHED: 04-14-2014
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Warfarin reduces the incidence of thromboembolism but increases the risk of gastrointestinal bleeding (GIB). GIB during warfarin anticoagulation is rarely evaluated in Asian patients.
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Eight Weeks of Esomeprazole Therapy Reduces Symptom Relapse, Compared with 4 Weeks, in Patients with Los Angeles Grade A or B Erosive Esophagitis.
Clin. Gastroenterol. Hepatol.
PUBLISHED: 04-11-2014
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There is disagreement over the ideal duration of initial proton pump inhibitor (PPI) therapy for gastroesophageal reflux disease, and whether prolonged therapy increases healing of the esophagitis and prevents symptom relapse. We performed a multicenter, prospective, randomized, controlled study to compare the efficacies of 4 weeks vs 8 weeks PPI therapy in reducing reflux symptoms and preventing symptom relapse in patients with Los Angeles grade A or B erosive esophagitis.
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Transition-metal-free, visible-light induced cyclization of arylsulfonyl chlorides with 2-isocyanobiphenyls to produce phenanthridines.
Chem. Commun. (Camb.)
PUBLISHED: 03-28-2014
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6-Aryl substituted phenanthridines were synthesized via a visible-light-catalyzed cyclization of 2-isocyanobiphenyls with arylsulfonyl chlorides under oxidant-free and transition-metal-free conditions. This transformation represents an efficient and attractive synthetic utilization of arylsulfonyl chlorides.
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Ardisia crenata extract stimulates melanogenesis in B16F10 melanoma cells through inhibiting ERK1/2 and Akt activation.
Mol Med Rep
PUBLISHED: 03-17-2014
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Melanin protects the skin against ultraviolet radiation by scattering incoming light and absorbing diverse free radicals. Agents that increase melanin synthesis in melanocytes may reduce the risk of photodamage and skin cancer. The present study investigated the effect of a methanol extract of Ardisia crenata (AC) on melanogenesis in B16F10 cells. Treatment of cultured B16F10 cells with AC extract (10, 20 and 40 µg/ml) stimulated an increase in melanin levels in a concentration?dependent manner, without cytotoxicity. Tyrosinase is key in the regulation of melanin production, thus the effect of AC extract on tyrosinase activity and protein expression was analyzed. AC extract was observed to significantly increase tyrosinase activity and protein expression in B16F10 cells. Furthermore, AC extract was found to markedly increase the protein expression of microphthalmia?associated transcription factor, which is an important transcription factor involved in tyrosinase gene expression. In addition, AC extract (40 µg/ml) was observed to suppress the activation of extracellular signal?regulated kinase (ERK) and Akt, which negatively regulate melanin synthesis in B16F10 cells. In conclusion, to the best of our knowledge, the present study is the first to show that a methanol extract of AC stimulates melanogenesis by increasing tyrosinase expression via the inhibition of ERK and Akt. Thus, methanol extract of AC may be a potential treatment for hypopigmentation diseases and may be a candidate for skin?tanning cosmetic products.
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Investigation of variations in the acrylamide and N(?) -(carboxymethyl) lysine contents in cookies during baking.
J. Food Sci.
PUBLISHED: 02-18-2014
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Baking processing is indispensable to determine special sensory prosperities of cookies and induces the formation of some beneficial components such as antioxidants. However, the formation of some Maillard reaction-derived chemical hazards, such as acrylamide (AA) and N(?) -(carboxymethyl) lysine (CML) in cookies is also a significant consequence of baking processing from a food safety standpoint. This study investigated the effects of baking conditions on the formation of AA and CML, as well as the antioxidant activity (AOA) of cookies. Cookies were baked at various baking temperatures (155 to 230 °C) and times (1.5 to 31 min). AA and CML contents were determined by ultra-performance liquid chromatography-tandem mass spectrometry, respectively. The highest level of AA was obtained in the cookies baked at 155 °C/21 min and 205 °C/11 min (328.93 ± 3.10 ?g/kg and 329.29 ± 5.29 ?g/kg), while the highest level of CML was obtained in the cookies baked at 230 °C/1.5 min (118.05 ± 0.21 mg/kg). AA was prone to form at relatively low temperature range (155 to 205 °C), however, CML at relatively high temperature range (205 to 230 °C). The CML content was much higher than the AA content in the same set of cookies, by about 2 to 3 orders of magnitude. The AOA of cookies increased at more severe baking conditions. According to the AA and CML content, AOA and sensory properties of cookies, the temperature-time regime of 180 °C/16 min might be a compromised selection. However, only optimizing the baking condition was not enough for manufacture of high-quality cookies.
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Comparison of high-intensity ultraviolet and NB-UVB on the maturation of melanocytes derived from hair follicle neural crest stem cells.
Lasers Med Sci
PUBLISHED: 02-13-2014
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Both high-intensity ultraviolet and narrowband ultraviolet B (NB-UVB) are important therapeutic options for vitiligo management, but high-intensity ultraviolet is more effective than NB-UVB. However, the underlying mechanisms have not been well investigated. Herein, we compare the effects of high-intensity ultraviolet and NB-UVB on the pigmentation of melanocytes derived from hair follicle-derived neural crest stem cells (HF-NCSCs) in vitro and study the underlying mechanisms. The HF-NCSCs were isolated from mouse whisker follicles. After radiation with high-intensity ultraviolet and NB-UVB, respectively, the cell viability by the CCK-8 assay showed gradual inhibitory effects in a dose-dependent manner, which has no apparent difference between the two modalities. The mRNA for melanogenesis factors such as tyrosinase and tyrp1 of the differentiated melanocytes increased significantly with high-intensity ultraviolet compared to the same dose of NB-UVB exposure. Furthermore, the expression of Mc1r was significantly increased by high-intensity ultraviolet in contrast to NB-UVB at the dosage of 0.5 J. By and large, these data suggest that high-intensity ultraviolet exhibited greater efficiency on the maturation of the melanocyte lineage differentiated from HF-NCSCs compared to NB-UVB with the same dose, which was probably due to the stronger stimulatory action of Mc1r. This may provide new insights into the different efficacies of high-intensity ultraviolet and NB-UVB in the treatment of vitiligo repigmentation.
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Role of endoscopic papillary balloon dilation in patients with recurrent bile duct stones after endoscopic sphincterotomy.
J Chin Med Assoc
PUBLISHED: 02-12-2014
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Endoscopic sphincterotomy (ES) is an established treatment for patients with choledocholithiasis or common bile duct stones (CBDS), but further management of patients after ES with recurrent CBDS remains controversial. Endoscopic papillary large balloon dilation (EPLBD) has been used safely and effectively for stone removal in patients after ES with recurrent CBDS. The aim of this study was to evaluate the clinical efficacy of EPLBD in patients after complete ES with recurrent CBDS.
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Waveforms for optimal sub-keV high-order harmonics with synthesized two- or three-colour laser fields.
Nat Commun
PUBLISHED: 02-07-2014
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High-order harmonics extending to the X-ray region generated in a gas medium by intense lasers offer the potential for providing tabletop broadband light sources but so far are limited by their low conversion efficiency. Here we show that harmonics can be enhanced by one to two orders of magnitude without an increase in the total laser power if the laser's waveform is optimized by synthesizing two- or three-colour fields. The harmonics thus generated are also favourably phase-matched so that radiation is efficiently built up in the gas medium. Our results, combined with the emerging intense high-repetition MHz lasers, promise to increase harmonic yields by several orders to make harmonics feasible in the near future as general bright tabletop light sources, including intense attosecond pulses.
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USP21 negatively regulates antiviral response by acting as a RIG-I deubiquitinase.
J. Exp. Med.
PUBLISHED: 02-03-2014
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Lys63-linked polyubiquitination of RIG-I is essential in antiviral immune defense, yet the molecular mechanism that negatively regulates this critical step is poorly understood. Here, we report that USP21 acts as a novel negative regulator in antiviral responses through its ability to bind to and deubiquitinate RIG-I. Overexpression of USP21 inhibited RNA virus-induced RIG-I polyubiquitination and RIG-I-mediated interferon (IFN) signaling, whereas deletion of USP21 resulted in elevated RIG-I polyubiquitination, IRF3 phosphorylation, IFN-?/? production, and antiviral responses in MEFs in response to RNA virus infection. USP21 also restricted antiviral responses in peritoneal macrophages (PMs) and bone marrow-derived dendritic cells (BMDCs). USP21-deficient mice spontaneously developed splenomegaly and were more resistant to VSV infection with elevated production of IFNs. Chimeric mice with USP21-deficient hematopoietic cells developed virus-induced splenomegaly and were more resistant to VSV infection. Functional comparison of three deubiquitinases (USP21, A20, and CYLD) demonstrated that USP21 acts as a bona fide RIG-I deubiquitinase to down-regulate antiviral response independent of the A20 ubiquitin-editing complex. Our studies identify a previously unrecognized role for USP21 in the negative regulation of antiviral response through deubiquitinating RIG-I.
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Axitinib sensitization of high Single Dose Radiotherapy.
Radiother Oncol
PUBLISHED: 01-14-2014
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Single Dose Radiation Therapy (SDRT) provides remarkably high rates of control even for tumors resistant to fractionated radiotherapy. SDRT tumor control depends on acute acid sphingomyelinase-mediated endothelial cell injury and monoclonal antibodies targeting Vascular Endothelial Cell Growth Factor (VEGF) signaling radiosensitized tumor endothelium when delivered immediately prior to irradiation. Here we evaluate the ability of the oral VEGF receptor inhibitor, axitinib, to sensitize tumor endothelium and increase tumor control with SDRT.
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Promotion of quality standard of herbal medicine by constituent removing and adding.
Sci Rep
PUBLISHED: 01-14-2014
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To identify major active constituents and measure their levels in a typical medicinal herb-Rhizoma coptidis, we applied the concept of removing and adding, taking inspiration from functional genetic methods. As this herb has bacteriostatic properties and is used to treat bacterial diarrhea, we examined the effects of individual constituents (berberine, palmatine, coptisine, epiberberine, jateorrhizine and columbamine) on the growth of Shigella dysenteriae with microcalorimetry. The removing and adding procedures revealed that berberine and coptisine were the main antibacterial constituents of R. coptidis, with bacteriostatic activities of 54.10% and 39.75%, respectively. The relative levels of berberine and coptisine in R. coptidis were 8.08%-31.92% and 4.05%-14.45%, respectively. On the basis of whole effect, the method of constituents removing and adding, coupled with a bioassay, is a useful strategy to identify the active constituents and measure their levels in herbal medicines, which may provide reference to other natural products.
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Early entecavir treatment for chronic hepatitis B with severe acute exacerbation.
Antimicrob. Agents Chemother.
PUBLISHED: 01-13-2014
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A previous study found that lamivudine, if started early enough, may improve the chance of survival in chronic hepatitis B virus (HBV) with severe acute exacerbation (SAE). The aim of this study was to investigate the effect of early entecavir treatment before the bilirubin level exceeds 20 mg/dl for chronic HBV with SAE. Consecutive patients with chronic HBV with SAE and a serum bilirubin level of <20 mg/dl who received lamivudine or entecavir were enrolled. Short-term (4 months) survival was evaluated. One hundred fourteen patients received lamivudine, and 53 patients received entecavir. The baseline characteristics were similar for the two groups except that the entecavir group was older and had a lower alanine aminotransferase (ALT) level. Three patients (8.0%) in the entecavir group and 9 patients (7.9%) in the lamivudine group died (P=1.000). If only patients who started antiviral treatment before serum bilirubin level rose to more than 15 mg/dl were included, 3 patients (8.3%) in the entecavir group and 3 patients (3.0%) in the lamivudine group died (P=0.189). If only patients with an HBV DNA level higher than 10(5) copies/ml and a bilirubin level lower than 15 mg/dl were included, 5 out of 40 patients (12.5%) in the entecavir group died and 1 out of 59 patients (1.7%) in the lamivudine group died. Multivariate analysis found that entecavir treatment was associated with more mortality than lamivudine (P=0.035). Early entecavir treatment in patients with high viral load is associated with more short-term mortality than lamivudine for chronic HBV with severe acute exacerbation.
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Photoluminescence properties of the high-brightness Eu(3+)-doped KNaCa2(PO4)2 phosphors.
Spectrochim Acta A Mol Biomol Spectrosc
PUBLISHED: 01-07-2014
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A series of red-emitting phosphors Eu(3+)-doped KNaCa2(PO4)2 were synthesized by solid-state reaction, and the photoluminescence (PL) properties were also investigated. The excitation spectrum is composed of charge-transfer (CT) of Eu-O and excitation lines of Eu(3+) ions. The strongest excitation lines appeared at 393 nm. The emission spectra of KNaCa2(PO4)2:Eu(3+) phosphors exhibit five peaks assigned to the (5)D0-(7)FJ (J=0, 1, 2, 3, 4) transitions of Eu(3+) and have dominating emission peak at 621 nm under 393 nm excitation. The luminescence intensity enhanced with increasing Eu(3+) content and the emission reached the maximum intensity at x=0.02 in KNaCa2-x(PO4)2:xEu(3+). The effect of the charge compensators on the emission intensity of the phosphors was investigated. The integral intensity of the emission spectrum of KNaCa1.96(PO4)2:0.02Eu(3+), 0.02Na(+) excited at 393 nm is about 2.4 times as strong as that of Y2O3:0.05Eu(3+) commercial red phosphor. The color coordinates for KNaCa1.96(PO4)2:0.02Eu(3+),0.02Na(+) were measured. The results indicate that KNaCa2(PO4)2:Eu(3+) might be a promising phosphor for w-LEDs.
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BTLA exhibits immune memory for ?? T cells in patients with active pulmonary tuberculosis.
Am J Transl Res
PUBLISHED: 01-01-2014
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Despite past extensive studies, the role of B and T lymphocyte attenuator (BTLA) in ?? T cells in patients with active pulmonary tuberculosis (ATB) remains poorly understood. Here we demonstrate that BTLA expression on ?? T cells is decreased in patients with M. tuberculosis (Mtb) infection. Particularly, BTLA expression levels are likely critical for ?? T cells to manifest and maintain an active central memory phenotype with high capacity for secretion of IFN-? and perforin, which are important for immune memory against TB infection. BTLA(high) ?? T cells also exhibited higher capacity in response to Mtb peptide stimulation. In contrast to the role of BTLA played for negative regulation of immune responses, our data in the current studies suggest that BTLA expression on ?? T cells is likely associated with protective immune memory against Mtb infection in the setting of patients with active pulmonary tuberculosis. This previous unappreciated role for BTLA may have implications for prevention and treatment of patients with Mtb infection.
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Clinicopathological features of an ascending colon mixed adenoneuroendocrine carcinoma with clinical serosal invasion.
Int J Clin Exp Pathol
PUBLISHED: 01-01-2014
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Mixed adenoneuroendocrine carcinoma (MANEC) is exceedingly rare with a poor outcome. In this article, we reported a MANEC in a 68-year-old woman with a symptom of abdominal pain and distension. MANEC derived from the ascending colon with highly aggressive behavior. The diagnosis and distinguish of MANEC must base on histological findings and immunohistochemical findings. In this case, microscopic observation showed tumor cells were arranged in conglobate and nested by fibrous tissue with a visible cell atypia and mitotic. NEC-like and exocrine glandular cells were also been seen in a single neoplasm. MANEC tissues were immunopositive for CK, CK20, P53, CK7, CDX-2, Ki-67 (70%+), E-cad, CD56, CEA, Syn, villin and CgA, and immunonegative for CA125, NSE, ER and PR. Here, the patient was treated by surgical operation and was followed-up near 3 months, no local recurrence and distant metastasis.
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Modulation of ErbB2 blockade in ErbB2-positive cancers: the role of ErbB2 Mutations and PHLDA1.
PLoS ONE
PUBLISHED: 01-01-2014
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We set out to study the key effectors of resistance and sensitivity to ErbB2 tyrosine kinase inhibitors, such as lapatinib in ErbB2-positive breast and lung cancers. A cell-based in vitro site-directed mutagenesis lapatinib resistance model identified several mutations, including the gatekeeper ErbB2 mutation ErbB2-T798I, as mediating resistance. ErbB2-T798I engineered cell models indeed show resistance to lapatinib but remain sensitive to the irreversible EGFR/ErbB2 inhibitor, PD168393, suggestive of potential alternative treatment strategies to overcome resistance. Gene expression profiling studies identified a select group of downstream targets regulated by ErbB2 signaling and define PHLDA1 as an immediately downregulated gene upon oncogenic ErbB2 signaling inhibition. We find significant down-regulation of PHLDA1 in primary breast cancer and PHLDA1 is statistically significantly less expressed in ErbB2 negative compared with ErbB2 positive tumors consistent with its regulation by ErbB2. Lastly, PHLDA1 overexpression blocks AKT signaling, inhibits cell growth and enhances lapatinib sensitivity further supporting an important negative growth regulator function. Our findings suggest that PHLDA1 might have key inhibitory functions in ErbB2 driven lung and breast cancer cells and a better understanding of its functions might point at novel therapeutic options. In summary, our studies define novel ways of modulating sensitivity and resistance to ErbB2 inhibition in ErbB2-dependent cancers.
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A case of mucoepidermoid carcinoma located in the left forearm of a middle-aged pregnant woman.
Int J Clin Exp Med
PUBLISHED: 01-01-2014
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In this article, we described a mucoepidermoid carcinoma (MEC) located in the left forearm of a 39-year-old pregnant woman. Here, the patient had a superficial tip size apophysis nearly 3 years, which begin to sustained growth after pregnant in 2012, and stopped growth after childbirth. MEC is a rare malignant tumor. Previously reports showed it mainly arise from the salivary, bronchial, thyroid, breast, lacrimal gland and conjunctiva. Here, we reported a case of MEC arising from the forearm gland for the first time. Histological finding showed a cystic and solid tumor in fibrous tissue below the squamous epithelium, and some columnar or cuboidal mucous cells covering on the epidermal cells or mixed with epidermal cells included in the tumor tissues. Also, Focal hyperplasia epidermal cells with round or oval nucleus in center were distributed in small pieces but no keratosis. The tumor tissues were immunopositive for CEA, P63, ki-67 (10%), CK7 and CK5/6, and immunonegative for CK20 and GCDFP-15. This case is a low-grade MEC and the patient's postoperative recovery is smooth.
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Embryonal rhabdomyosarcoma of the paranasal sinuses: a case report and review of literature.
Int J Clin Exp Med
PUBLISHED: 01-01-2014
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Embryonal rhabdomyosarcoma (ERMS) is a rare malignancy with a poor outcome. In this article, we describe a case of ERMS in the paranasal sinuses from a 60-year-old male patient. ERMS derived from the paranasal sinuses is extremely rare. The diagnosis of ERMS must be based on histological findings and immunohistochemical findings. In this case, microscopic observation showed tumor cells were arranged in flocked sheets, cord-like and acinar-like by hyperplastic fibrous tissue. And ERMS tissues were immunopositive for myogenin, desmin, MSA, CD56, vimentin, CD99, Syn and Ki-67 (40%+), and immunonegative for CK, EMA, LCA, GFAP, NSE, S-100, HMB-45 and Melan-A. Here, the patient was treated with multimodal therapy including endoscopic surgery, chemotherapy and radiation, but the patient's postoperative recovery is not too smooth.
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Mutations of C-reactive protein (CRP) -286 SNP, APC and p53 in colorectal cancer: implication for a CRP-Wnt crosstalk.
PLoS ONE
PUBLISHED: 01-01-2014
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C-reactive protein (CRP) is an established marker of inflammation with pattern-recognition receptor-like activities. Despite the close association of the serum level of CRP with the risk and prognosis of several types of cancer, it remains elusive whether CRP contributes directly to tumorigenesis or just represents a bystander marker. We have recently identified recurrent mutations at the SNP position -286 (rs3091244) in the promoter of CRP gene in several tumor types, instead suggesting that locally produced CRP is a potential driver of tumorigenesis. However, it is unknown whether the -286 site is the sole SNP position of CRP gene targeted for mutation and whether there is any association between CRP SNP mutations and other frequently mutated genes in tumors. Herein, we have examined the genotypes of three common CRP non-coding SNPs (rs7553007, rs1205, rs3093077) in tumor/normal sample pairs of 5 cancer types (n?=?141). No recurrent somatic mutations are found at these SNP positions, indicating that the -286 SNP mutations are preferentially selected during the development of cancer. Further analysis reveals that the -286 SNP mutations of CRP tend to co-occur with mutated APC particularly in rectal cancer (p?=?0.04; n?=?67). By contrast, mutations of CRP and p53 or K-ras appear to be unrelated. There results thus underscore the functional importance of the -286 mutation of CRP in tumorigenesis and imply an interaction between CRP and Wnt signaling pathway.
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Malignant myoepithelioma of the breast: a case report and review of literature.
Int J Clin Exp Pathol
PUBLISHED: 01-01-2014
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In this article, we described a malignant myoepithelioma of the breast (MMB) in a 69-year-old woman. Breast cancer derived from myoepithelial cells is very rare, usually benign. The diagnosis of MMB based on histological and immunohistochemical finding. In this case, the author diagnosed the tumor as MMB, because tumor tissues were immunopositive for 34?E12, P63, SMA, S-100, CD10, E-Cad and Ki-67, and immunnegative for CK5/6, desmin, ER, PR and C-erbB-2, because tumor tissue showed invasive growth and local hemorrhage or necrosis, suggesting malignant, and also because there was a transition between the tumor cells and hyperplastic myoepithelium of non-tumorous ducts. The patient's postoperative recovery is smooth and regular following of patient is essential.
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Hepatic arterial infusion chemotherapy for patients with huge unresectable hepatocellular carcinoma.
PLoS ONE
PUBLISHED: 01-01-2014
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The optimal treatment for huge unresectable hepatocellular carcinoma (HCC) remains controversial. The outcome of transcatheter arterial chemoembolization (TACE) for patients huge unresectable HCC is generally poor and the survival benefit of TACE in these patients is unclear. The aim of the study is to compare the effect of hepatic arterial infusion chemotherapy (HAIC) versus symptomatic treatment in patients with huge unresectable HCC.
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Multi-spectroscopic analysis and molecular modeling on the interaction of curcumin and its derivatives with human serum albumin: a comparative study.
Spectrochim Acta A Mol Biomol Spectrosc
PUBLISHED: 01-01-2014
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The comparative study about the interaction between curcumin and its derivatives (demothxycurcumin and bisdeoxycurcumin) with human serum albumin (HSA) has been carried out using multi-spectroscopic analysis and molecular modeling method. The characteristic of fluorescence quenching and the thermodynamic parameters have been studied by state emission fluorescence experiments under different temperatures with an interval of 6 K. Curcumin shows largest quenching constant and bisdeoxycurcumin shows the smallest at the temperature of 298 K. However, the quenching constant of curcumin drops quickly with the increase of temperature. Demothxycurcumin gives the largest quenching efficiency at the temperature of 310 K. An average distance of 6.7 nm for energy transfer has been determined based on förster resonance energy theory (FRET). The site competitive replacement experiments illustrate three compounds mainly binding on site I (Subdomain IIA) of the protein, and show tendency of binding on site II (Subdomain IIIA) with the removing of methoxyl groups. Circular dichroism spectra and Fourier transform infrared spectroscopy (FTIR) have been used to investigate the influence on protein secondary structure. Content of the ?-helix increases at low concentrations of the compounds, while unfolding occurs at high concentrations. Docking simulation reveals possible mechanism for different quenching behavior and binding sites preferred by three compounds. The binding modes have effectively supported the conclusion of the experiments.
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Ovarian sclerosing stromal tumor in a young woman with ectopic pregnancy: clinical, pathological, and immunohistochemical studies.
Int J Clin Exp Pathol
PUBLISHED: 01-01-2014
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In this article, we described an ovarian sclerosing stromal tumor (SST) in a young woman with ectopic pregnancy. It is important to distinguish SST from fibroma, thecoma, and lipoid cell tumors clinically and histologically. Several unique histologic features including pseudolobulation, sclerosis and prominent vascularity are clearly reflected at histopathological findings. The SST cells were immunopositive for CD34, Desmin and SMA, and negative for factor VIII-related antigen, CD31, S-100, ER and PR. The patient's postoperative recovery was smooth and she was discharged after 21 days.
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A novel strategy for the construction of substituted benzoxazoles via a tandem oxidative process.
Chem. Commun. (Camb.)
PUBLISHED: 10-18-2013
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A practical and simple synthesis of benzoxazoles from easily available substrates was developed. The protocol is triggered by an iron-catalyzed tandem oxidative process from simple toluene derivatives and 2-aminophenols. This method represents a straightforward approach to access substituted benzoxazoles.
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P53-participated cellular and molecular responses to irradiation are cell differentiation-determined in murine intestinal epithelium.
Arch. Biochem. Biophys.
PUBLISHED: 10-15-2013
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Cells respond differently to DNA damaging agents, which may related to cell context and differentiation status. The aim of present study was to observe the cellular and molecular responses of cells in different differentiation status to ionizing irradiation (IR).
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Pathways of [Ca(2+)]i rise evoked by angiotensin II in MDCK renal tubular cells.
J. Recept. Signal Transduct. Res.
PUBLISHED: 09-25-2013
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Abstract The effect of angiotensin II (Ang II) on cytosolic Ca(2+) concentrations ([Ca(2+)]i) in MDCK renal tubular cells was explored. The Ca(2+)-sensitive fluorescent dye fura-2 was applied to measure [Ca(2+)]i. Ang II at concentrations of 5-40?µM induced a [Ca(2+)]i rise in a concentration-dependent manner. The response was reduced partly by removing Ca(2+). Ang II evoked store-operated Ca(2+) entry that was inhibited by La(3+) and Gd(3+). In the absence of extracellular Ca(2+), incubation with the endoplasmic reticulum Ca(2+) pump inhibitor 2,5-di-tert-butylhydroquinone (BHQ) or thapsigargin abolished Ang II-induced Ca(2+) release. Inhibition of phospholipase C with U73122 abolished Ang II-induced [Ca(2+)]i rise. Three Ang II analogues [(ASN1,VAL5)-Ang II acetate, (SAR1,THR8)-Ang II acetate, (VAL5)-Ang II acetate] failed to induce a [Ca(2+)]i rise. Together, in MDCK cells, Ang II induced a [Ca(2+)]i rise via Ca(2+) entry through store-operated Ca(2+) channels and phospholipase C-dependent Ca(2+) release from the endoplasmic reticulum. Moreover, Ang IIs amino acid sequence is important in its stimulatory effect on [Ca(2+)]i.
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Long-term alcohol consumption is an independent risk factor of hypertension development in northern China: evidence from Kailuan study.
J. Hypertens.
PUBLISHED: 09-14-2013
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The purpose of the present study was to determine the impact of alcohol consumption on the incidence of hypertension.
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Erythropoietin promotes the repair effect of acute kidney injury by bone-marrow mesenchymal stem cells transplantation.
Exp. Biol. Med. (Maywood)
PUBLISHED: 08-07-2013
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Bone-marrow mesenchymal stem cells (BMSCs) transplantation is effective for acute kidney injury (AKI) repair but with limited efficiency. In the present study, BMSCs marked by bromodeoxyuridine (BrdU) were transplanted to the AKI mouse model with erythropoietin (EPO) being subcutaneously injected. The blood urea nitrogen (BUN) and serum creatinine (Scr) levels, pathological changes, distribution of BMSCs, expressions of the cytokeratin 18 (CK18) and the stromal cell-derived factor 1 (SDF-1) in the nephridial tissues were measured. The directional migration of BMSCs to the AKI microenvironment in vitro was also tested. The results showed that BMSCs transplantation or EPO injection alone decreased the BUN and Scr levels and the acute tubular necrosis (ATN) scoring in varied degrees. The combination of these decreased the above indicators levels significantly. BrdU(+) cells (BMSCs) were observed in the AKI nephridial tissues, and CK18 expressed in the cytoplasm of these cells. EPO injection increased the proportion of BrdU(+) cells with the enhanced expression of SDF-1 in the AKI nephridial tissues. EPO increased the migrating number of BMSCs to the AKI microenvironment in vitro, and additional anti-SDF-1 treatment with SDF-1 antibody neutralized this effect. Our results showed that EPO increased the number of the transplanted BMSCs in the injured nephridial tissues and enhanced the AKI repair effect of BMSCs transplantation. The enhanced kidney-homing efficiency for BMSCs mediated by the SDF-1/CXCR4 pathway is one of the possible mechanisms for EPO performance.
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[Expression and clinical significance of Muc1, p63 protein in diffuse sclerosing variant of papillary thyroid carcinoma and conventional papillary thyroid carcinoma].
Lin Chung Er Bi Yan Hou Tou Jing Wai Ke Za Zhi
PUBLISHED: 08-01-2013
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To investigate the expression and clinical significance of Muc1, p63 protein in diffuse sclerosing variant of papillary thyroid carcinoma and conventional papillary thyroid carcinoma.
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Genetic and structural validation of Aspergillus fumigatus N-acetylphosphoglucosamine mutase as an antifungal target.
Biosci. Rep.
PUBLISHED: 07-13-2013
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Aspergillus fumigatus is the causative agent of invasive aspergillosis in immunocompromised patients. It possesses a cell wall composed of chitin, glucan and galactomannan, polymeric carbohydrates synthesized by processive glycosyltransferases from intracellular sugar nucleotide donors. Here we demonstrate that A. fumigatus possesses an active N-acetylphosphoglucosamine mutase (AfAGM1), a key enzyme in the biosynthesis of UDP-GlcNAc, the nucleotide sugar donor for chitin synthesis. A conditional agm1 mutant revealed the gene to be essential. Reduced expression of agm1 resulted in retarded cell growth and altered cell wall ultrastructure and composition. The crystal structure of AfAGM1 revealed an amino acid change in the active site compared to the human enzyme, which could be exploitable in the design of selective inhibitors. AfAGM1 inhibitors were discovered by high-throughput screening, inhibiting the enzyme with IC50s in the low µM range. Together, these data provide a platform for the future development of AfAGM1 inhibitors with antifungal activity.
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Genetic and structural validation of Aspergillus fumigatus?UDP-N-acetylglucosamine pyrophosphorylase as an antifungal target.
Mol. Microbiol.
PUBLISHED: 06-05-2013
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The sugar nucleotide UDP-N-acetylglucosamine (UDP-GlcNAc) is an essential metabolite in both prokaryotes and eukaryotes. In fungi, it is the precursor for the synthesis of chitin, an essential component of the fungal cell wall. UDP-N-acetylglucosamine pyrophosphorylase (UAP) is the final enzyme in eukaryotic UDP-GlcNAc biosynthesis, converting UTP and N-acetylglucosamine-1-phosphate (GlcNAc-1P) to UDP-GlcNAc. As such, this enzyme may provide an attractive target against pathogenic fungi. Here, we demonstrate that the fungal pathogen Aspergillus fumigatus possesses an active UAP (AfUAP1) that shows selectivity for GlcNAc-1P as the phosphosugar substrate. A conditional mutant, constructed by replacing the native promoter of the A.?fumigatus?uap1 gene with the Aspergillus nidulans?alcA promoter, revealed that uap1 is essential for cell survival and important for cell wall synthesis and morphogenesis. The crystal structure of AfUAP1 was determined and revealed exploitable differences in the active site compared with the human enzyme. Thus AfUAP1 could represent a novel antifungal target and this work will assist the future discovery of small molecule inhibitors against this enzyme.
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TAK1 inhibitor 5Z-7-oxozeaenol sensitizes neuroblastoma to chemotherapy.
Apoptosis
PUBLISHED: 05-24-2013
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Treatment failure in high risk neuroblastoma is largely due to development of chemoresistance. NF-?B activation is one of the resistance mechanisms for cancer cells to escape from chemotherapy-induced cell-death. TAK1 is an essential component in genotoxic stresses-induced NF-?B activation; however, the role of TAK1 in the development of chemoresistance in neuroblastoma remains unknown. Using a panel of neuroblastoma cell lines, we found that TAK1 inhibitor 5Z-7-oxozeaenol significantly augmented the cytotoxic effects of doxorubicin (Dox) and etoposide (VP-16) on neuroblastoma cell lines. TAK1 inhibition also enhanced the inhibitory effect of Dox and VP-16 on anchorage-independent growth. Treatment of neuroblastoma cells with 5Z-7-oxozeaenol blocked Dox- and VP16-induced NF-?B activation and enhanced Dox- and VP16-induced apoptosis. Moreover, 5Z-7-oxozeaenol was able to overcome the established chemoresistance in LA-N-6 neuroblastoma cells. Using an orthotopic neuroblastoma mouse model, we found that 5Z-7-oxozeaenol significantly enhanced chemotherapeutic efficacy in vivo. Together, our results provide a proof-of-concept that TAK1 inhibition significantly increases the sensitivity of neuroblastoma cells to chemotherapy-induced cell-death and can serve as an effective adjunct to current chemotherapeutic regimens for high risk diseases.
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Relationship between total antioxidant capacities of cereals measured before and after in vitro digestion.
Int J Food Sci Nutr
PUBLISHED: 05-07-2013
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Total antioxidant capacity (TAC), which is the cumulative capacity of different antioxidants to reduce oxidative stress, proved to be very useful to correlate the antioxidant capacity of the diet with the incidence of some disease or with modification of disease biomarkers. Therefore, it is important to estimate the real antioxidant potential of cereals. Most methods are based mainly on extractive methods which may largely underestimate the TAC of cereals. Several recent articles have reported a direct approach, the Quencher procedure, which produces a higher TAC compared to extractive methods. However, both the extractive methods and the Quencher procedure are performed prior to digestion. In this article, an in vitro approach using enzymatic digestion, designed to mimic digestion in the gastrointestinal tract (physiological conditions), had been used to release antioxidants from cereals. Seven whole cereals were employed for the determination of DPPH as well as ABTS radical scavenging activity before and after simulated digestion. The objective was to compare the in vitro procedure of antioxidant extraction with the Quencher procedure and water extraction. The values of the TACs from the cereal grains obtained from the in vitro procedure were 1.8-10.3 times higher than the Quencher procedure and 3.5-10.5 times higher than water extraction. Correlation between the results of the TAC obtained using the three different procedures was also investigated. The in vitro gastrointestinal digestion procedure is more useful in the screening of grains, assessing their beneficial health effects compared to the Quencher procedure or water extraction.
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N-Glycosylation of Gel1 or Gel2 is vital for cell wall ?-glucan synthesis in Aspergillus fumigatus.
Glycobiology
PUBLISHED: 05-06-2013
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Fungal cell wall is a dynamic structure that communicates with and protects the cell from outside stress. In Aspergillus fumigatus, the cell wall ?-glucans are mainly elongated by ?-1,3-glucanosyltransferases Gels, which consist of seven family members (Gel1-7) utilizing ?-1,3-glucan chains as substrates. Previously, we have shown that the mutant deficient of N-glycan processing displays a reduction in the cell wall ?-glucans, suggesting that N-glycosylation is required for the proper function of ?-1,3-glucanosyltransferase. To verify this hypothesis, in this study, the gene encoding ?-1,3-glucanosyltransferase Gel1 or Gel2 was deleted in the ?cwh41 mutant to construct a double-mutant ?gel1?cwh41 or ?gel2?cwh41. The growth phenotypes of both double mutants were similar to the single-mutant ?cwh41, suggesting that Gel1 and Gel2 are proteins that are mainly affected by deficient N-glycan processing in ?cwh41. Furthermore, the mutant ?gel1(Gel1-NM) or ?gel2(Gel2-NM), in which all potential N-glycosylation sites on Gel1 or Gel2 were removed by site-directed mutagenesis, showed phenotypes similar to the single-mutant ?gel1 or ?gel2. Biochemical analysis revealed that N-glycosylation was essential for the function of Gel1 or Gel2 and thus required for ?-glucan synthesis in A. fumigatus.
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Cytochrome P450 2A6 deletion polymorphism and risk of lung cancer: a meta-analysis.
Mol. Biol. Rep.
PUBLISHED: 04-30-2013
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Previous studies concerning the association between cytochrome P450 2A6 (CYP2A6) deletion polymorphism and lung cancer risk provided controversial results. To clarify the precise association, a meta-analysis was performed. The electronic databases PubMed, Chinese Biomedical Database and Chinese National Knowledge Infrastructure Database were searched for case-control studies last updated on June 3, 2012 that investigated CYP2A6 deletion polymorphism and lung cancer risk. The odds ratio (OR) and its respective 95 % confidence interval (95 % CI) were used to measure the strength of association by means of a genetic model free approach. A total of 8 studies including 2,607 cases and 2,595 controls met the inclusion criteria and were subjected to the final analysis. The most appropriate co-dominant model was adopted. Overall, we found that CYP2A6 *1/*1 genotype was associated with an increased risk of lung cancer relative to *4/*4 genotype (OR = 2.65, 95 % CI: 1.84-3.81, P < 0.001). Significant association was also detected among Asians. Publication bias was absent in this meta-analysis. Therefore, our data suggested that the presence of the CYP2A6 *1/*1 might be associated with an increased lung cancer risk, especially for Asians. Further studies well-designed among different ethnicity populations are required.
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[Construction of a cell-surface expression system in Trichoderma reesei].
Wei Sheng Wu Xue Bao
PUBLISHED: 04-26-2013
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AfMp1p is a glycosylphosphatidylinositol (GPI) anchored cell wall protein (GPI-CWP) identified in filamentous fungus Aspergillus fumigatus, which contains a specific C terminal signal for cell wall localization. Trichoderma reesei is known as a safe fungal species (GRAS) and widely used in the industry. Thus, developing of the cell-surface expression systems in T. reesei is of industrial interest.
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One single basic amino acid at the ?-1 or ?-2 site is a signal that retains glycosylphosphatidylinositol-anchored protein in the plasma membrane of Aspergillus fumigatus.
Eukaryotic Cell
PUBLISHED: 04-12-2013
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Although the plasma membrane is the terminal destination for glycosylphosphatidylinositol (GPI) proteins in higher eukaryotes, cell wall-attached GPI proteins (GPI-CWPs) are found in many fungal species. In yeast, some of the cis-requirements directing localization of GPI proteins to the plasma membrane or cell wall are now understood. However, it remains to be determined how Aspergillus fumigatus, an opportunistic fungal pathogen, signals, and sorts GPI proteins to either the plasma membrane or the cell wall. In this study, chimeric green fluorescent proteins (GFPs) were constructed as fusions with putative C-terminal GPI signal sequences from A. fumigatus Mp1p, Gel1p, and Ecm33p, as well as site-directed mutations thereof. By analyzing cellular localization of chimeric GFPs using Western blotting, electron microscopy, and fluorescence microscopy, we showed that, in contrast to yeast, a single Lys residue at the ?-1 or ?-2 site alone could retain GPI-anchored GFP in the plasma membrane. Although the signal for cell wall distribution has not been identified yet, it appeared that the threonine/serine-rich region at the C-terminal half of AfMp1 was not required for cell wall distribution. Based on our results, the cis-requirements directing localization of GPI proteins in A. fumigatus are different from those in yeast.
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Human adipose-derived stem cells modified by HIF-1? accelerate the recovery of cisplatin-induced acute renal injury in vitro.
Biotechnol. Lett.
PUBLISHED: 04-06-2013
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Human adipose-derived stem cells (hASCs) improve renal function in acute kidney injury. Hypoxia-inducible factor-1? (HIF-1?) was transfected into hASCs. hASCs modified by lentivirus-mediated empty-vector and HIF-1? maintained their stem cell characteristics. The expression of the renal-protective gene, heme oxygenase-1 and vascular endothelial growth factor were significantly increased in hASCs modified by HIF-1?, compared to hASCs modified by empty-vector. Cellular ultra-structure and TUNEL staining revealed that hASCs modified by HIF-1? promoted the recovery of apoptotic morphology in cisplatin-treated human kidney-2 cells (HK-2 cells) when compared to hASCs modified by empty-vector. Additionally, hASCs modified by empty-vector inhibited caspase-3 expression and up-regulated Bcl-2 expression in cisplatin-treated HK-2 cells, an effect even more pronounced with hASCs modified by HIF-1?. Thus, HIF-1? gene-modified ASCs could be an effective way to enhance the renal-protective effect.
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Migration of CXCR4 gene-modified bone marrow-derived mesenchymal stem cells to the acute injured kidney.
J. Cell. Biochem.
PUBLISHED: 03-27-2013
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Bone marrow-derived mesenchymal stem cells (BMSCs) can migrate to the injured kidney after acute kidney injury (AKI) with limited efficiency. This study investigated the effect of CXCR4 overexpression on BMSC migration to the AKI kidney and the possible mechanisms. CXCR4 gene-modified BMSCs (CXCR4-BMSCs) and null-BMSCs were prepared and transplanted into the AKI mice. Blood indicators, histology, expression of stromal cell-derived factor 1 (SDF-1), and BMSC migration were investigated. Hypoxia/re-oxygenation-pretreated renal tubular epithelial cells (HR-RTECs) were prepared to generate AKI in vitro. The chemotaxis experiment was performed using the transwell chamber. The phosphorylation of AKT and MAPK in the BMSCs was also investigated. The CXCR4-BMSCs showed a remarkable expression of CXCR4. The SDF-1 expression in the AKI renal tissue was increased. CXCR4-BMSCs transplantation sharply increased the accumulation of BMSCs in the renal tissue, which was consistent with a greater improvement of renal function. The in vitro experiments showed that the migration of BMSCs to the HR-RTEC culturing chamber was CXCR4-dependent, and could be fully inhibited by AMD3100, a CXCR4-specific antagonist. The migration could also be partly blocked by either LY294002 (PI3K inhibitor) or PD98059 (MAPK inhibitor). Phosphorylated Akt and MAPK were increased in the BMSCs co-cultured with HR-RTECs and their expression was the highest in the CXCR4-BMSCs, which could be recovered by AMD3100. Overexpression of CXCR4 gene could enhance BMSC migration to the kidney area after AKI. The SDF-1/CXCR4 axis via its activation of PI3K/AKT and MAPK in BMSCs could be the possible mechanisms underlying this function.
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Characterization of genes for chitin catabolism in Haloferax mediterranei.
Appl. Microbiol. Biotechnol.
PUBLISHED: 03-21-2013
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Chitin is the second most abundant natural polysaccharide after cellulose. But degradation of chitin has never been reported in haloarchaea. In this study, we revealed that Haloferax mediterranei, a metabolically versatile haloarchaeon, could utilize colloidal or powdered chitin for growth and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) accumulation, and the gene cluster (HFX_5025-5039) for the chitin catabolism pathway was experimentally identified. First, reverse transcription polymerase chain reaction results showed that the expression of the genes encoding the four putative chitinases (ChiAHme, ChiBHme, ChiCHme, and ChiDHme, HFX_5036-5039), the LmbE-like deacetylase (DacHme, HFX_5027), and the glycosidase (GlyAHme, HFX_5029) was induced by colloidal or powdered chitin, and chiA Hme, chiB Hme, and chiC Hme were cotranscribed. Knockout of chiABC Hme or chiD Hme had a significant effect on cell growth and PHBV production when chitin was used as the sole carbon source, and the chiABCD Hme knockout mutant lost the capability to utilize chitin. Knockout of dac Hme or glyA Hme also decreased PHBV accumulation on chitin. These results suggested that ChiABCDHme, DacHme, and GlyAHme were indeed involved in chitin degradation in H. mediterranei. Additionally, the chitinase assay showed that each chitinase possessed hydrolytic activity toward colloidal or powdered chitin, and the major product of colloidal chitin hydrolysis by ChiABCDHme was diacetylchitobiose, which was likely further degraded to monosaccharides by DacHme, GlyAHme, and other related enzymes for both cell growth and PHBV biosynthesis. Taken together, this study revealed the genes and enzymes involved in chitin catabolism in haloarchaea for the first time and indicated the potential of H. mediterranei as a whole-cell biocatalyst in chitin bioconversion.
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Synthesis and microwave absorption properties of yolk-shell microspheres with magnetic iron oxide cores and hierarchical copper silicate shells.
ACS Appl Mater Interfaces
PUBLISHED: 03-21-2013
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Yolk-shell microspheres with magnetic Fe3O4 cores and hierarchical copper silicate shells have been successfully synthesized by combining the versatile sol-gel process and hydrothermal reaction. Various yolk-shell microspheres with different core size and shell thickness can be readily synthesized by varying the experimental conditions. Compared to pure Fe3O4, the as-synthesized yolk-shell microspheres exhibit significantly enhanced microwave absorption properties in terms of both the maximum reflection loss value and the absorption bandwidth. The maximum reflection loss value of these yolk-shell microspheres can reach -23.5 dB at 7 GHz with a thickness of 2 mm, and the absorption bandwidths with reflection loss lower than -10 dB are up to 10.4 GHz. Owing to the large specific surface area, high porosity, and synergistic effect of both the magnetic Fe3O4 cores and hierarchical copper silicate shells, these unique yolk-shell microspheres may have the potential as high-efficient absorbers for microwave absorption applications.
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Serotonin facilitates peripheral pain sensitivity in a manner that depends on the nonproton ligand sensing domain of ASIC3 channel.
J. Neurosci.
PUBLISHED: 03-08-2013
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Tissue acidosis and inflammatory mediators play critical roles in inflammatory pain. Extracellular acidosis activates acid-sensing ion channels (ASICs), which have emerged as key sensors for extracellular protons in the central and peripheral nervous systems and play key roles in pain sensation and transmission. Additionally, inflammatory mediators, such as serotonin (5-HT), are known to enhance pain sensation. However, functional interactions among protons, inflammatory mediators, and ASICs in pain sensation are poorly understood. In the present study, we show that 5-HT, a classical pro-inflammatory mediator, specifically enhances the proton-evoked sustained, but not transient, currents mediated by homomeric ASIC3 channels and heteromeric ASIC3/1a and ASIC3/1b channels. Unexpectedly, the effect of 5-HT on ASIC3 channels does not involve activation of 5-HT receptors, but is mediated via a functional interaction between 5-HT and ASIC3 channels. We further show that the effect of 5-HT on ASIC3 channels depends on the newly identified nonproton ligand sensing domain. Finally, coapplication of 5-HT and acid significantly increased pain-related behaviors as assayed by the paw-licking test in mice, which was largely attenuated in ASIC3 knock-out mice, and inhibited by the nonselective ASIC inhibitor amiloride. Together, these data identify ASIC3 channels as an unexpected molecular target for acute actions of 5-HT in inflammatory pain sensation and reveal an important role of ASIC3 channels in regulating inflammatory pain via coincident detection of extracellular protons and inflammatory mediators.
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Fast and direct quantification of underivatized muscone by ultra performance liquid chromatography coupled with evaporative light scattering detection.
J Sep Sci
PUBLISHED: 03-04-2013
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A new reversed phase ultra performance liquid chromatography coupled with evaporative light scattering detection is developed for the fast and direct quantification of underivatized muscone in precious herbal medicine musk. Separation of muscone was achieved on a Waters Acquity BEH C18 (50 × 2.1 mm id, 1.7 ?m) column. The runtime was as short as 5 min. The mode of evaporative light scattering detection was set at Impact On. The influence of evaporative light scattering detection condition on sensitivity was investigated. The optimized condition was: drift tube temperature at 30°C, gas flow rate 4.2 L/min. The method was validated with respect to the precision, sensitivity, accuracy, linearity, stability, and robustness were measured in this paper. The calibration curves showed good linear regression (r = 0.9914) within the test range. The recovery rate was 98.6%. The limit of detection for muscone was 2.0 ng. The validated method was rapid, simple, reproducible, and convenient for the quantification of muscone in musk and the related products.
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Lys63-linked polyubiquitination of BRAF at lysine 578 is required for BRAF-mediated signaling.
Sci Rep
PUBLISHED: 02-25-2013
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The RAF kinase family is essential in mediating signal transduction from RAS to ERK. BRAF constitutively active mutations correlate with human cancer development. However, the precise molecular regulation of BRAF activation is not fully understood. Here we report that BRAF is modified by Lys63-linked polyubiquitination at lysine 578 within its kinase domain once it is activated by gain of constitutively active mutation or epidermal growth factor (EGF) stimulation. Substitution of BRAF lysine 578 with arginine (K578R) inhibited BRAF-mediated ERK activation. Furthermore, ectopic expression of BRAF K578R mutant inhibited anchorage-independent colony formation of MCF7 breast cancer cell line. Our studies have identified a previously unrecognized regulatory role of Lys63-linked polyubiquitination in BRAF-mediated normal and oncogenic signalings.
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PyrG is required for maintaining stable cellular uracil level and normal sporulation pattern under excess uracil stress in Aspergillus nidulans.
Sci China Life Sci
PUBLISHED: 02-25-2013
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Tight control of the intracellular uracil level is believed to be important to reduce the occurrence of uracil incorporation into DNA. The pyrG gene of Aspergillus nidulans encodes orotidine 5-phosphate decarboxylase, which catalyzes the conversion of orotidine monophosphate (OMP) to uridine monophosphate (UMP). In this study, we found that pyrG is critical for maintaining uracil at a low concentration in A. nidulans cells in the presence of exogenous uracil. Excess uracil and its derivatives had a stronger inhibitory effect on the growth of the pyrG89 mutant with defective OMP decarboxylase activity than on the growth of wild type, and induced sexual development in the pyrG89 mutant but not in wild type. Analysis of transcriptomic responses to excess uracil by digital gene expression profiling (DGE) revealed that genes related to sexual development were transcriptionally activated in the pyrG89 mutant but not in wild type. Quantitative analysis by HPLC showed that the cellular uracil level was 6.5 times higher in the pyrG89 mutant than in wild type in the presence of exogenous uracil. This study not only provides new information on uracil recycling and adaptation to excess uracil but also reveals the potential effects of OMP decarboxylase on fungal growth and development.
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Transcriptome and biochemical analysis reveals that suppression of GPI-anchor synthesis leads to autophagy and possible necroptosis in Aspergillus fumigatus.
PLoS ONE
PUBLISHED: 02-08-2013
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Previously, it has been shown that GPI proteins are required for cell wall synthesis and organization in Aspergillus fumigatus, a human opportunistic pathogen causing life-threatening invasive aspergillosis (IA) in immunocompromised patients. Blocking GPI anchor synthesis leads to severe phenotypes such as cell wall defects, increased cell death, and attenuated virulence. However, the mechanism by which these phenotypes are induced is unclear. To gain insight into global effects of GPI anchoring in A. fumigatus, in this study a conditional expression mutant was constructed and a genome wide transcriptome analysis was carried out. Our results suggested that suppression of GPI anchor synthesis mainly led to activation of phosphatidylinositol (PtdIns) signaling and ER stress. Biochemical and morphological evidence showed that autophagy was induced in response to suppression of the GPI anchor synthesis, and also an increased necroptosis was observed. Based on our results, we propose that activation of PtdIns3K and increased cytosolic Ca(2+), which was induced by both ER stress and PtdIns signaling, acted as the main effectors to induce autophagy and possible necroptosis.
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Effect of erythropoietin on the migration of bone marrow-derived mesenchymal stem cells to the acute kidney injury microenvironment.
Exp. Cell Res.
PUBLISHED: 02-04-2013
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Bone marrow-derived mesenchymal stem cells (BMSCs) preferentially migrate to the injured tissue but with limited efficiency. Here we investigated the effect of erythropoietin (EPO) treatment on the BMSC migration to the acute kidney injury (AKI) microenvironment. The possible mechanisms were also discussed. A hypoxia/re-oxygenation (HR) model of renal tubular epithelial cells (RTECs) was established to generate AKI in vitro, and a chemotaxis experiment was conducted using the transwell chamber. EPO treatment enhanced the BMSC migration to the HR-RTEC culturing chamber in a SDF-1 level-dependent manner, which was fully inhibited by the treatment of anti-SDF-1 antibody. The BMSC migration could also be partly blocked by LY294002 (phosphoinositide 3-kinase (PI3K) inhibitor) and PD98059 (MAPK inhibitor). Western blot analysis showed that phosphorylated Akt and phosphorylated MAPK in BMSCs were enhanced by EPO treatment. In the in vivo experiment, BMSCs were transplanted into the AKI mice and EPO was subcutaneously injected. The results showed that EPO injection increased the SDF-1 protein expression and BMSC accumulation in the renal tissue, which was consistent with a decent improvement of renal function. In addition, the BMSC accumulation in the renal tissue was blocked by anti-SDF-1 antibody, LY294002 or PD98059. Our data suggest that AKI microenvironment had a directional chemotactic effect on BMSCs, which could be further enhanced by the EPO treatment. The increased SDF-1 level in the AKI microenvironment and the activations of PI3K/AKT and MAPK in BMSCs were the possible mechanisms for the effect of EPO. Therefore, BMSC transplantation combined with EPO injection can be a novel and effective approach for AKI repair.
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