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Find video protocols related to scientific articles indexed in Pubmed.
[Application of cervical triangulating stapled anastomosis in minimally invasive esophagectomy].
Zhonghua Wei Chang Wai Ke Za Zhi
PUBLISHED: 10-03-2014
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To explore the safety and feasibility of cervical triangulating stapled anastomosis(TSA) for esophagogastric anastomosis(EGA) after minimally invasive esophagectomy (MIE).
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Identification of reference miRNAs in human tumors by TCGA miRNA-seq data.
Biochem. Biophys. Res. Commun.
PUBLISHED: 09-18-2014
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Although the accuracy of detecting the expression of miRNAs by quantitative real-time polymerase chain reaction (qRT-PCR) is highly dependent on reliable reference miRNAs, many commonly used reference miRNAs are not stably expressed and as such are not suitable for quantification and normalization of qRT-PCR data. To solve this problem, we analyzed the global expression profiles of thousands of samples in 14 types of common human tumors released by The Cancer Genome Atlas (TCGA), and identified the most stably and highly expressed miRNAs as candidate reference miRNAs in each type of tumor. We found that miR-361-5p and let-7i-5p were the most recommended candidate reference miRNAs in nine and eight types of tumors, respectively, followed by let-7a-5p, mir-28-5p and miR-99b-5p. Our results are of important value to those researchers focused on miRNA; however, these candidate reference miRNAs still need to be validated prior to their use in qRT-PCR studies.
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Apollon modulates chemosensitivity in human esophageal squamous cell carcinoma.
Oncotarget
PUBLISHED: 09-13-2014
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Patients with esophageal squamous cell carcinoma (ESCC) are often diagnosed with advanced diseases that respond poorly to chemotherapy. Here we reported that Apollon, a membrane-associated inhibitor of apoptosis protein, was overexpressed in ESCC cell lines and clinical ESCC tissues, and Apollon overexpression clinically correlated with poor response to chemotherapy (P = 0.001), and short overall survival (P = 0.021). Apollon knockdown increased cisplatin/docetaxel-induced apoptosis, mitochondrial dysfunction and cytochrome c release in two ESCC cell lines. Apollon knockdown potentiated cisplatin/docetaxel-induced long-term cell growth inhibition, and enhanced chemosensitivity of ESCC cells to cisplatin/docetaxel in xenograft tumor models. Apollon knockdown also enhanced cisplatin/docetaxel-induced activation of caspase-8 (extrinsic pathway) and caspase-9 (intrinsic pathway) in ESCC cells and xenograft tumor models. Mechanism studies revealed that the effect of Apollon on chemosensitivity is mainly mediated by Smac. Apollon expression strongly and negatively correlated with Smac expression in clinical ESCC tissues (P = 0.001). Apollon targeted Smac for degradation in ESCC cells. The effect of Apollon on chemosensitivity was reversed by Smac knockdown in ESCC cells. Taken together, our data show association of Apollon expression with chemotherapeutic response in ESCC, and provide a strong rationale for combining Apollon antagonism with chemotherapy to treat ESCC.
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High-value utilization of lignin to synthesize Ag nanoparticles with detection capacity for Hg²?.
ACS Appl Mater Interfaces
PUBLISHED: 08-29-2014
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This study reports the rapid preparation of silver nanoparticles (AgNPs) from Tollens' reagent under microwave irradiation. In the synthesis, lignin with reducing groups and spatial three-dimensional structure was used as reducing and stabilizing agents without other chemical reagents, and the effects of the ratio of lignin to Ag(+), reaction temperature, and heating time on the synthesis of AgNPs were investigated. The obtained AgNPs were further characterized by UV-vis, Malvern particle size, TEM, XRD, and XPS analyses. The structural changes of lignin before and after reaction were also studied by FT-IR, (1)H NMR, (13)C NMR, and GC-MS. The results revealed that the obtained AgNPs were mostly spherical with diameters of around 24 nm. The optimum reaction conditions were a ratio 50 mg of lignin to 0.3 mM of Ag(+), a microwave irradiation temperature of 60 °C, and a heating time of 10 min. Moreover, AgNPs redispersed well in water and ethanol after centrifugation for the removal of lignin. During the formation of AgNPs, lignin was oxidized, and the side chains of lignin were partly disrupted into small molecules, such as hydrocarbon and alcohol. The resultant lignin-AgNPs showed highly selective sensing detection for Hg(2+), and the color of the lignin-AgNP solution containing Hg(2+) decreased gradually with increasing amounts of Hg(2+) within seconds, but the other 19 metal ions had little effect on the color and surface plasmon absorption band of the lignin-AgNPs. Also, there was a linear relationship between the absorbance and Hg(2+) concentration, with a limit of detection concentration of 23 nM. This study provides not only a new way to take advantage of agricultural and forestry residues, but also a green and rapid method for the synthesis of AgNPs to detect the toxic ion Hg(2+) selectively and sensitively.
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Advances in self-assembled chitosan nanomaterials for drug delivery.
Biotechnol. Adv.
PUBLISHED: 08-07-2014
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Nanomaterials based on chitosan have emerged as promising carriers of therapeutic agents for drug delivery due to good biocompatibility, biodegradability, and low toxicity. Chitosan originated nanocarriers have been prepared by mini-emulsion, chemical or ionic gelation, coacervation/precipitation, and spray-drying methods. As alternatives to these traditional fabrication methods, self-assembled chitosan nanomaterials show significant advantages and have received growing scientific attention in recent years. Self-assembly is a spontaneous process by which organized structures with particular functions and properties could be obtained without additional complicated processing or modification steps. In this review, we focus on recent progress in the design, fabrication and physicochemical aspects of chitosan-based self-assembled nanomaterials. Their applications in drug delivery of different therapeutic agents are also discussed in details.
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Clinical and molecular characteristics of community-acquired methicillin-resistant Staphylococcus aureus infections in Chinese neonates.
APMIS
PUBLISHED: 08-06-2014
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This study aims to characterize the clinical features of community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA) infections in Chinese neonates, as well as the molecular characteristics and expression of key virulence genes of isolates. Clinical information and molecular characteristics of 130 cases were analyzed. Up to 83.8% patients were affected with late-onset infection. Cesarean delivery was the main delivery route, accounting for 74.6% of the total deliveries. Pneumonia (69, 53.1%) was the most common infection. A total of 38 patients (29.2%) suffered from complications. Moreover, 35 cases (26.9%) were invasive infections, among which 88.6% involved multiple organs and 45.7% suffered from complications. Cesarean section and premature birth were the risk factors for invasive CA-MRSA infection. ST59-MRSA-SCCmecIVa-t437 (54, 41.5%) was the most predominant CA-MRSA clone. The hla expression in the ST59 isolates was higher than that in ST910 (p = 0.02) and the hla expression in ST59-SCCmecV-t437 was higher than that in ST59-SCCmecIVa-t437. Approximately, 46.4% (13/28) of the infections caused by ST59-SCCmecV were invasive. This value is higher than that of ST59-SCCmecVa caused infections (14/59, 23.7%) (p = 0.03). This study showed that neonatal CA-MRSA infections in China readily become invasive, involve multiple organs, and are often accompanied by complications. The SCCmec V clone may be more pathogenic than the SCCmecVIa clone.
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Synergistic cytotoxic effect of genistein and doxorubicin on drug-resistant human breast cancer MCF-7/Adr cells.
Oncol. Rep.
PUBLISHED: 07-29-2014
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The molecular mechanisms underlying genistein-mediated reversal of chemoresistance remains unknown. In the present study, we investigated the molecular mechanisms by which genistein overcomes chemoresistance and its effect on doxorubicin-induced cytotoxicity. Consistent with previous reports, genistein combined with doxorubicin had a synergistic effect on MCF-7/Adr cells, and genistein reduced the chemoresistance of these cells. Genistein treatment increased the intracellular accumulation of doxorubicin but did not influence P-gp function. The combination of genistein and doxorubicin significantly induced cell cycle arrest and apoptosis. Genistein treatment strongly inhibited HER2/neu but not MDR-1 expression at both the mRNA and protein levels. Therefore, our results demonstrated that genistein combined with doxorubicin had a synergistic effect on MCF-7/Adr cells, and the mechanisms likely involve an increase in the intracellular accumulation of doxorubicin and suppression of HER2/neu expression.
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Novel multipotent AChEI-CCB attenuates hyperhomocysteinemia-induced memory deficits and Neuropathologies in rats.
J. Alzheimers Dis.
PUBLISHED: 07-16-2014
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Alzheimer's disease (AD) has multiple etiopathogenic factors, yet the definitive cause remains unclear and the therapeutic strategies have been elusive. Combination therapy, as one of the promising treatments, has been studied for years and may exert synergistic beneficial effects on AD through polytherapeutic targets. In this study, we tested the effects of a synthesized juxtaposition (named SCR1693) composed of an acetylcholinesterase inhibitor (AChEI) and a calcium channel blocker (CCB) on the hyperhomocysteinemia (HHcy)-induced AD rat model, and found that SCR1693 remarkably improved the HHcy-induced memory deficits and preserved dendrite morphologies as well as spine density by upregulating synapse-associated proteins PSD95 and synapsin-1. In addition, SCR1693 attenuated HHcy-induced tau hyperphosphorylation at multiple AD-associated sites by regulating the activity of protein phosphatase-2A and glycogen synthase kinase-3?. Furthermore, SCR1693 was more effective than individual administration of both donepezil and nilvadipine which were used as AChEI and CCB, respectively, in the clinical practice. In conclusion, our data suggest that the polytherapeutic targeting juxtaposition SCR1693 (AChEI-CCB) is a promising therapeutic candidate for AD.
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Emerging chitin and chitosan nanofibrous materials for biomedical applications.
Nanoscale
PUBLISHED: 07-08-2014
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Over the past several decades, we have witnessed significant progress in chitosan and chitin based nanostructured materials. The nanofibers from chitin and chitosan with appealing physical and biological features have attracted intense attention due to their excellent biological properties related to biodegradability, biocompatibility, antibacterial activity, low immunogenicity and wound healing capacity. Various methods, such as electrospinning, self-assembly, phase separation, mechanical treatment, printing, ultrasonication and chemical treatment were employed to prepare chitin and chitosan nanofibers. These nanofibrous materials have tremendous potential to be used as drug delivery systems, tissue engineering scaffolds, wound dressing materials, antimicrobial agents, and biosensors. This review article discusses the most recent progress in the preparation and application of chitin and chitosan based nanofibrous materials in biomedical fields.
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Diffusion-induced periodic transition between oscillatory modes in amplitude-modulated patterns.
Chaos
PUBLISHED: 07-03-2014
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We study amplitude-modulated waves, e.g., wave packets in one dimension, overtarget spirals and superspirals in two dimensions, under mixed-mode oscillatory conditions in a three-variable reaction-diffusion model. New transition zones, not seen in the homogeneous system, are found, in which periodic transitions occur between local 1(N-1) and 1(N) oscillations. Amplitude-modulated complex patterns result from periodic transition between (N?-?1)-armed and N-armed waves. Spatial recurrence rates provide a useful guide to the stability of these modulated patterns.
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Comparing the predictive abilities of different metabolic syndrome definitions for acute coronary syndrome: a case-control study in Chinese adults.
Metab Syndr Relat Disord
PUBLISHED: 06-24-2014
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Different institutions have proposed various definitions for metabolic syndrome, which is a combination of risk factors for cardiovascular diseases (CVD). This study aimed to compare the feasibilities and abilities of different metabolic syndrome definitions in predicting acute coronary syndrome (ACS) in Chinese adults.
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RNAi screening identifies HAT1 as a potential drug target in esophageal squamous cell carcinoma.
Int J Clin Exp Pathol
PUBLISHED: 06-15-2014
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Esophageal carcinoma (EC) is one of the most fatal carcinomas of the gastrointestinal tract. Aberrant activity of histone acetyltransferases (HATs)/deacetylases (HDACs) play a critical role in carcinogenesis through the regulation of the genes involved in cell differentiation, proliferation, and apoptosis. However, cellular functions of HATs/HDACs in esophageal cancer and its molecular mechanisms remain unclear. An RNAi screen was used in this study to identify the histone acetyltransferases (HATs) and deacetylases (HDACs) that could be critical for the survival of EC cells. We demonstrated that HAT1 (histone acetyltransferase 1) was an important determinant to regulate the proliferation of human EC Eca-109 cells. Furthermore, we showed that the knockdown of HAT1 induced a G2/M cell cycle arrest, which was associated with the disruption of cell cycle-related events, including the decrease of cyclinD1 as well as alteration in cyclinB1 expression. The expression of HAT1 was validated to be higher in the primary tumors and adjacent tissue as compared to that of the normal esophageal tissue. Furthermore, we found that HAT1 expression was directly correlated with the poor tumor differentiation of EC tissue, which suggested that HAT1 played an important role in esophageal carcinoma and that it could be a novel EC therapeutic target.
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Characterization and phylogenetic analysis of the mating-type loci in the asexual ascomycete genus Ulocladium.
Mycologia
PUBLISHED: 06-02-2014
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The genus Ulocladium is thought to be strictly asexual. Mating-type (MAT) loci regulate sexual reproduction in fungi and their study may help to explain the apparent lack of sexual reproduction in Ulocladium. We sequenced the full length of two MAT genes in 26 Ulocladium species and characterized the entire MAT idiomorphs plus flanking regions of Ulocladium botrytis. The MAT1-1 ORF encodes a protein with an alpha-box motif by the MAT1-1-1 gene and the MAT1-2 ORF encodes a protein with an HMG box motif by the MAT1-2-1 gene. Both MAT1-1-1 and MAT1-2-1 genes were detected in a single strain of every species. Moreover, the results of RT-PCR revealed that both MAT genes are expressed in all 26 Ulocladium species. This demonstrates that MAT genes of Ulocladium species might be functional and that they have the potential for sexual reproduction. Phylogenies based on MAT genes were compared with GAPDH and Alt a 1 phylograms in Ulocladium using maximum parsimony (MP) and Bayesian analysis. The MAT genealogies and the non-MAT trees displayed different topologies, indicating that MAT genes are unsuitable phylogenetic markers at the species level in Ulocladium. Furthermore, the conflicting topologies between MAT1-1-1 and MAT1-2-1 phylogeny indicate separate evolutionary events for the two MAT genes. However, the intergeneric phylogeny of four closely allied genera (Ulocladium, Alternaria, Cochliobolus, Stemphylium) based on MAT alignments demonstrated that MAT genes are suitable for phylogenetic analysis among allied genera.
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Differential diagnosis between hepatic metastases and benign focal lesions using DWI with parallel acquisition technique: a meta-analysis.
Tumour Biol.
PUBLISHED: 05-24-2014
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We aim to investigate the diagnostic capability of diffusion-weighted imaging using parallel acquisition technique for the differentiation between hepatic metastases and benign focal lesions with a meta-analysis. The meta-analysis included a total of 858 hepatic metastases and 440 benign liver lesions from nine studies. The pooled sensitivity and specificity of diffusion-weighted imaging (DWI) were 0.87 (95 % CI, 0.84-0.89) and 0.90 (95 % CI, 0.87-0.93), respectively. The positive likelihood ratio (PLR) and negative likelihood ratio (NLR) were 8.50 (95 % CI, 4.97-14.52) and 0.17 (95 % CI, 0.11-0.26), respectively. The P value for ? (2) heterogeneity for all pooled estimates was <0.05. From the fitted summary receiver operating characteristics (SROC), the area under the curve (AUC) and Q* index were 0.95 and 0.88, respectively. Publication bias is not present (t?=?-0.76, P?=?0.471). The meta-regression analysis indicated that evaluated covariates included patient number, patient population, mean age, maximum of b factor, number of cysts, number of hemangiomas, and field were not sources of heterogeneity (all P value >0.05). Diffusion-weighted imaging was useful for differentiation between hepatic metastases and benign focal lesions. The diffusion characteristics of the benign hepatocellular lesions, including cases of focal nodular hyperplasia (FNH) and adenoma, have rarely been reported and need further studies. The diagnostic capability of DWI with parallel acquisition technique for differentiation between metastases and benign hepatic focal lesions might be overestimated.
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Quantifying the expression of tumor marker genes in lung squamous cell cancer with RNA sequencing.
J Thorac Dis
PUBLISHED: 05-22-2014
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We measured the expression of some commonly used tumor markers with RNA sequencing (RNA-Seq) to identify any that might be useful for the evaluation of squamous cell lung cancer and identify possible correlations between these tumor markers and any clinical characteristics.
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The effect of narrowed gastric conduits on anastomotic leakage following minimally invasive oesophagectomy.
Interact Cardiovasc Thorac Surg
PUBLISHED: 05-20-2014
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Anastomotic leakage remains a major complication following minimally invasive oesophagectomy (MIO). In this study, our objective was to determine whether a narrower gastric conduit would lead to lower incidence of anastomotic leakage following MIO.
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Immuno-PCR for detection of Giardia lamblia cysts in water.
J AOAC Int
PUBLISHED: 05-17-2014
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Giardia lamblia cysts at low concentrations were detected in water samples using a highly sensitive immunological-PCR (IPCR) method. Magnetic gold particles were precoated with monoclonal anti-Giardia antibodies, and Giardia lamblia cysts ranging from 1 to 100 cysts were diluted in 500 microL of water. A biotinylated detection antibody bound to the G. lamblia cysts was then linked by a streptavidin bridge to biotinylated Giardia-reporter DNA. After extensive washing, reporter DNA was released by denaturation, transferred to PCR tubes, amplified, electrophoresed, and visualized. An optimized immuno-PCR method detected as little as five G. lamblia cysts. To further evaluate the specificity and the clinical application of this IPCR assay for G. lamblia cysts, Entamoeba histolytica and Cryptosporidium parvum were also collected and detected by IPCR. The data demonstrated that this monoclonal antibody-based IPCR method is particularly useful for analysis of environmental water samples in which the densities of G. lamblia cysts is very low, and provides a platform capable of rapid screening of samples from drinking water, wells, rivers, lakes, and recreational swimming pools for trace levels of G. lamblia cysts.
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Novel chitosan-based pH-sensitive and disintegrable polyelectrolyte nanogels.
Colloids Surf B Biointerfaces
PUBLISHED: 05-16-2014
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A novel approach to design pH-sensitive and disintegrable polyelectrolyte nanogels composed of citraconic-based N-(carboxyacyl) chitosan (polyanion) and quaternary chitosan (polycation) was reported. Firstly, the hydrolysis of citraconic-modified chitosan was monitored using fluorescamine assay and it could selectively dissociate in acidic media (e.g., pH ?5.0) due to the isomerization during the addition of citraconic anhydride to chitosan. Secondly, the self-assembly behaviors of different polyelectrolyte pairs between citraconic-based chitosan and quaternary chitosan were investigated via colloidal titration assay. It was indicated that the difference in molecular weight (MW) of opposite charged polyelectrolytes played an important role on the formation of polyelectrolyte nanogels. Results showed that polyelectrolyte nanogels (ca. 300nm in size) only formed when polyanion and polycation had a very large difference in MW. The pH-sensitive behavior of polyelectrolyte nanogels was comprehensively investigated by dynamic light scattering (DLS) and transmission electron microscope (TEM). The incorporation of charge-conversional citraconic-based chitosan into polyelectrolyte complexes has provided an effective approach to prepare polyelectrolyte nanogels which were very stable at neutral pH but disintegrated quickly in acidic media.
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[The correlation between the seasonal variation of house dust mite allergens exposure level in household and the level of asthma control in asthmatic children].
Zhonghua Er Ke Za Zhi
PUBLISHED: 05-15-2014
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The prevalence of allergic asthma has been rising continually which is correlated with the increasingly closed living environment. House dust mites are the major sources of indoor aeroallergens which induce asthma in sensitized people. To monitor the seasonal variation of house dust mite (HDM)-allergens exposure level in the asthmatic children, which was evaluated to show its correlation with the level of asthma control, HDM allergic sensitization and fraction of exhaled nitric oxide, and to provide basic data for HDM environmental control.
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Clinical outcome of subcentimeter non-small cell lung cancer after surgical resection: single institution experience of 105 patients.
J Surg Oncol
PUBLISHED: 04-17-2014
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The detection of subcentimeter lung cancers has significantly improved with advances in computed tomography and the emergence of screening protocols. We reviewed the clinicopathological features and surgical outcomes of patients with subcentimeter non-small cell lung cancer (NSCLC) in our institution.
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Novel layer-by-layer structured nanofibrous mats coated by protein films for dermal regeneration.
J Biomed Nanotechnol
PUBLISHED: 04-17-2014
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Layer-by-layer coating technique is effective in modifying the surface of nanofibrous mats, but overmuch film-coating makes the mats less porous to hardly suit the condition for tissue engineering. We developed novel nanofibrous mats layer-by-layer coated by silk fibroin and lysozyme on the cellulose electrospun template via electrostatic interaction. The film-coating assembled on the mats was not excessive because the charge of the proteins varied in the coating process due to different pH value. In addition, pure nature materials made the mats nontoxic, biodegradable and low-cost. The morphology and composition variation during layer-by-layer coating process was investigated and the results showed that the structure and thickness of film-coatings could be well-controlled. The antibacterial assay and in vitro cell experiments indicated that the mats could actively inhibit bacteria and exhibit excellent biocompatibility. In vivo implant assay further verified the mats cultured with human epidermal cells could promote wound healing and avoid wound infection. Therefore, these mats showed promising prospects when performed for dermal reconstruction.
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Functional nanoparticles in targeting glioma diagnosis and therapies.
J Nanosci Nanotechnol
PUBLISHED: 04-16-2014
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Successful therapy and diagnosis of glioma is one of the biggest challenges in the biomedical fields. The incidence is growing gradually around the world. Annually, there are approximately 13,000 cases of glioblastoma multiforme diagnosed with historical 1-year and 5-year survival rates of 29.3% and 3.3%, respectively. The prognosis of patients with malignant glioma remains dismal. Due to its highly proliferative, infiltrative, and invasive property, development of effective preventative strategies to control the gliomas is in high demand. Meanwhile, the efficiency of drug delivery to glioma remains low owing to the non-specific, non-targeted nature of anti-tumor agents. Furthermore, the presence of the blood brain barrier and blood brain tumor barrier is another obstacle for gliomas treatments. Nanotechnology has brought a paradigm shift in the diagnosis and treatment of glioma. This review discusses the potential of various nanoparticles in the diagnosis of gliomas using some metal oxide, and in the therapy of gliomas including receptor-mediated, magnetic directing, and cell-mediated drug delivery systems. In this review, some physical techniques combined with nanoparticles (NPs) such as ultrasound therapy, thermochemotherapy, photodynamic therapy, and fluorescent magnetic NPs have also been summarized.
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Knockdown of phosphotyrosyl phosphatase activator induces apoptosis via mitochondrial pathway and the attenuation by simultaneous tau hyperphosphorylation.
J. Neurochem.
PUBLISHED: 04-02-2014
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Phosphotyrosyl phosphatase activator (PTPA) is decreased in the brains of Alzheimer's disease (AD) and the AD transgenic mouse models. Here, we investigated whether down-regulation of PTPA affects cell viability and the underlying mechanisms. We found that PTPA was located in the integral membrane of mitochondria, and knockdown of PTPA induced cell apoptosis in HEK293 and N2a cell lines. PTPA knockdown decreased mitochondrial membrane potential and induced Bax translocation into the mitochondria with a simultaneous release of Cyt C, activation of caspase-3, cleavage of poly (DNA ribose) polymerase (PARP), and decrease in Bcl-xl and Bcl-2 protein levels. Over-expression of Protein phosphatase 2A (PP2A) catalytic subunit (PP2AC ) did not rescue the apoptosis induced by PTPA knockdown, and PTPA knockdown did not affect the level of and their phosphorylation of mitogen-activated protein kinases (MAPKs), indicating that PP2A and MAPKs were not involved in the apoptosis induced by PTPA knockdown. In the cells with over-expression of tau, PTPA knockdown induced PP2A inhibition and tau hyperphosphorylation but did not cause significant cell death. These data suggest that PTPA deficit causes apoptotic cell death through mitochondrial pathway and simultaneous tau hyperphosphorylation attenuates the PTPA-induced cell death. Phosphotyrosyl phosphatase activator (PTPA) is decreased in the brains of Alzheimer's disease (AD) and AD transgenic mouse models. Here, we investigated whether down-regulation of PTPA affects cell viability. We found that PTPA located in the integral membrane of mitochondria, and knockdown of PTPA induced cell apoptosis in HEK293 and N2a cell lines by decreasing mitochondrial membrane potential, which leads to translocation of Bax and a simultaneous release of Cyt C. In the cells with tau over-expression, PTPA knockdown inactivated PP2A to phosphorylate tau to avoid cell apoptosis which induced by PTPA knockdown.
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Characteristic of PGDS potential regulation role on spermatogenesis in the Chinese mitten crab Eriocheir sinensis.
Gene
PUBLISHED: 04-01-2014
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Prostaglandin D synthase (PGDS) catalyzes the isomerization of PGH2 to produce PGD2 in the presence of sulfhydryl compounds. In this study, a full length PGDS gene comprising 1250 nucleotides from the Chinese mitten crab Eriocheir sinensis (Es-PGDS) was characterized, with a 615 bp open reading frame encoding 204 amino acid residues. Its deduced peptide has high homology with other species' PGDS protein. The Es-PGDS mRNA expression was tissue-related, with the highest expression observed in the hepatopancreas, accessory sex gland, testis and ovaries. We also detected the different stages of tissue expression and the enzyme activity for Es-PGDS in the testis and male crab hepatopancreas. The different expression patterns and its corresponding enzyme activity level indicated that PGDS is involving in the regulation of reproductive action during the period of rapid development in E. sinensis. Furthermore our research could arouse a heat debate on the PGDS reproductive function in invertebrate and further study will be needed to determine the molecular mechanism(s) linking PGDS functions to spermatogenesis and ontogenesis if this gene is to be exploited as a molecular biomarker in further studies of development.
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Humanin attenuates Alzheimer-like cognitive deficits and pathological changes induced by amyloid ?-peptide in rats.
Neurosci Bull
PUBLISHED: 03-19-2014
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Amyloid ?-peptide (A?) has been implicated as a key molecule in the neurodegenerative cascades of Alzheimer's disease (AD). Humanin (HN) is a secretory peptide that inhibits the neurotoxicity of A?. However, the mechanism(s) by which HN exerts its neuroprotection against A?-induced ADlike pathological changes and memory deficits are yet to be completely defined. In the present study, we provided evidence that treatment of rats with HN increases the number of dendritic branches and the density of dendritic spines, and upregulates pre- and post-synaptic protein levels; these effects lead to enhanced long-term potentiation and amelioration of the memory deficits induced by A?1-42. HN also attenuated A?1-42-induced tau hyperphosphorylation, apparently by inhibiting the phosphorylation of Tyr307 on the inhibitory protein phosphatase-2A (PP2A) catalytic subunit and thereby activating PP2A. HN also inhibited apoptosis and reduced the oxidative stress induced by A?1-42. These findings provide novel mechanisms of action for the ability of HN to protect against A?1-42-induced AD-like pathological changes and memory deficits.
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Is minimally invasive esophagectomy beneficial to elderly patients with esophageal cancer?
Surg Endosc
PUBLISHED: 02-26-2014
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Open esophagectomy (OE) in elderly patients with esophageal cancer is hazardous due to high surgical mortality and limited survival. The aim of this study was to explore whether minimally invasive esophagectomy (MIE) has perioperative or long-term benefits in elderly patients with esophageal cancer compared with OE.
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Corneal biomechanical assessment using corneal visualization scheimpflug technology in keratoconic and normal eyes.
J Ophthalmol
PUBLISHED: 02-25-2014
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Purpose. To compare the corneal biomechanical properties of keratoconic patients and age-matched controls using corneal visualization Scheimpflug technology (Corvis ST). Methods. Sixty keratoconic eyes from 47 keratoconus patients and 60 normal eyes from 60 controls were enrolled in this prospective study. Tomography and biomechanical parameters of all eyes were obtained with the Pentacam and Corvis ST, respectively. Intraocular pressure was measured using a Goldmann applanation tonometer. Results. The tomography and biomechanical parameters of the keratoconic corneas were significantly different from those of the normal corneas except for the anterior chamber angle, first applanation length, the highest concavity time, and peak distance. The deformation amplitude was the best predictive parameter (area under the curve: 0.882), with a sensitivity of 81.7%, although there was a significant overlap between keratoconic and normal corneas that ranged from 1.0 to 1.4?mm. In both the keratoconus and control groups, the deformation amplitude was negatively correlated with intraocular pressure, central corneal thickness, and corneal volume at 3 and 5?mm. Conclusions. Corvis ST offers an alternative method for measuring corneal biomechanical properties. The possibility of classifying keratoconus based on deformation amplitude deserves clinical attention.
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Impeded Nedd4-1-mediated Ras degradation underlies Ras-driven tumorigenesis.
Cell Rep
PUBLISHED: 02-22-2014
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RAS genes are among the most frequently mutated proto-oncogenes in cancer. However, how Ras stability is regulated remains largely unknown. Here, we report a regulatory loop involving the E3 ligase Nedd4-1, Ras, and PTEN. We found that Ras signaling stimulates the expression of Nedd4-1, which in turn acts as an E3 ubiquitin ligase that regulates Ras levels. Importantly, Ras activation, either by oncogenic mutations or by epidermal growth factor (EGF) signaling, prevents Nedd4-1-mediated Ras ubiquitination. This leads to Ras-induced Nedd4-1 overexpression, and subsequent degradation of the tumor suppressor PTEN in both human cancer samples and cancer cells. Our study thus unravels the molecular mechanisms underlying the interplay of Ras, Nedd4-1, and PTEN and suggests a basis for the high prevalence of Ras-activating mutations and EGF hypersignaling in cancer.
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p27(kip1) deficiency accelerates dentin and alveolar bone formation.
Clin. Exp. Pharmacol. Physiol.
PUBLISHED: 02-19-2014
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To assess the role of p27(kip1) in regulating dental formation and alveolar bone development, we compared the teeth and mandible phenotypes of homozygous p27(kip1) -deficient (p27(-/-) ) mice with their wild-type littermates at 2 weeks of age. At 2 weeks of age, dental mineral density, dental volume and dentin sialoprotein-immunopositive areas were increased significantly, whereas the predentin area : total dentin area and biglycan-immunopositive area : dentin area ratios were decreased significantly in p27(-/-) mice compared with their wild-type (WT) littermates. Mandible mineral density, cortical thickness, alveolar bone volume, type I collagen and osterix-immunopositive areas, osteoblast number and activity and mRNA expression of Runt-related transcription factor 2 (Runx2), alkaline phosphatase (ALP), osteocalcin and bone morphogenetic protein (bmp2) were all significantly increased in the mandibles, as was the number and surface of tartrate-resistant acid phosphatase-positive osteoclasts in the alveolar bone of p27(-/-) mice compared with their WT littermates. Furthermore, the percentage of proliferating cell nuclear antigen-positive cells in Hertwig's epithelial root sheath and protein expression of cyclin E and cyclin-dependent kinase 2 were increased significantly in p27(-/-) mice relative to their WT littermates. The results from this study indicate that p27 plays a negative regulatory role in dentin formation and alveolar bone development.
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Establishment and characterization of esophageal squamous cell carcinoma patient-derived xenograft mouse models for preclinical drug discovery.
Lab. Invest.
PUBLISHED: 02-15-2014
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The purpose of this study was to establish and characterize patient-derived esophageal squamous cell carcinoma xenograft (PDECX) mice for utilization in antitumor drug discovery. A total of 96 esophageal squamous cell carcinoma (ESCC) tissues from Chinese patients were transplanted subcutaneously into immunodeficient mice. Histology, EGFR, K-ras, B-raf, and PIK3CA mutations, and HER2 gene amplifications were analyzed in both patient tumors and mouse xenograft tissues using immunohistochemistry, mutant-enriched liquid chip sequencing and fluorescence in situ hybridization assays, respectively. Furthermore, in vivo efficacy studies using five PDECX mice harboring a variety of genetic aberrations were performed using the chemotherapy agents 5-fluorouracil (5-FU) and cisplatin. Thirty-seven PDECX mouse models were successfully established in immunodeficient mice. Pathological analysis revealed similar histological architecture and degrees of differentiation between patient ESCC and xenografted tumors. No mutations were identified in EGFR, K-ras, and B-raf genes in either xenograft models or patient ESCC tissues. In contrast, PIK3CA gene mutations were detected in 12.5% (12/96) ESCC patients and 18.9% (7/37) PDECX models. Interestingly, patient ESCC tissues exhibiting HER2 overexpression or gene amplification were unable to survive in immunodeficient mice. Further analysis showed that PDECX models carrying HER2 2+ expression had no response to 5-FU/cisplatin, compared with HER2-negative models. In conclusion, a panel of PDECX mouse models, which include PIK3CA mutant and HER2-positive models, was established and characterized thus mimicking the current clinical genetic setting of esophageal carcinoma. The sensitivity of HER2-negative ESCC models to chemotherapy supports stratification approaches in the treatment of esophageal carcinoma patients and warrants further investigation of the impact of PI3KCA on treatment response.
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Waste Biogas residue from cassava dregs as carbon source to produce Galactomyces sp. Cczu11-1 cellulase and its enzymatic saccharification.
Appl. Biochem. Biotechnol.
PUBLISHED: 01-27-2014
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In the conversion of cassava starch dregs to biogas by anaerobic fermentation, the biogas residue (BR) containing lignocellulosic materials still remained in the environment. In order to effectively utilize BR, the complexed 1-methyl-3-methylimidazolium dimethyl phosphate ([Mmim]DMP) media were used for pretreating cellulosic materials. After the optimization of pretreatment, the IL [Mmim]DMP-HCl-water (78.5:1.5:20, w/w/w) pretreament media were used for pretreating BR at 130 °C for 30 min. Furthermore, BR pretreated could be effectively saccharified by cellulase of Galactomyces sp. CCZU11-1. Moreover, BR could be used as a cheap carbon source for the production of Galactomyces sp. CCZU11-1 cellulase. After the culture optimization, the optimal culture conditions were obtained as follows: BR 5 g/L, (NH4)2SO4 5 g/L, K2HPO4 2 g/L, MgSO4 0.2 g/L, NaCl 1 g/L, PEG6000 4 g/L, pH 5.5, and culture temperature 30 °C. After the fermentation for 6 days, the FPA and CMCase were 26.2 and 52.8 U/mL, respectively. In conclusion, waste BR could be chosen as a promising feedstock for biofuels.
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Biosynthesis of benzoylformic acid from benzoyl cyanide with a new bacterial isolate of Brevibacterium sp. CCZU12-1.
Appl. Biochem. Biotechnol.
PUBLISHED: 01-27-2014
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Brevibacterium sp. CCZU12-1 with high nitrilase activity could effectively hydrolyze benzoyl cyanide into benzoylformic acid. After the culture optimization, the preferred carbon sources, nitrogen sources, and inducer were glucose (10 g/L), a composite of peptone (10 g/L) plus yeast extract (2.5 g/L), and ?-caprolactam (2.0 mM), respectively. After the reaction optimization, the optimum reaction temperature, reaction pH, organic cosolvent, and metal ion were 30 °C, 7.0, ethanol (2%, v/v), and Ca(2+) (0.1 mM), respectively. At biotransformation of 120-mM benzoyl cyanide for 24 h, the yield of benzoylformic acid reached 91.8%. Moreover, the microbial nitrilase from Brevibacterium sp. CCZU12-1 could hydrolyze various nitriles, and it significantly exhibited high nitrilase activity against benzoyl cyanide, 3-cyanopyridine, and ?-cyclohexyl-mandelonitrile.
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ATR/Chk1/Smurf1 pathway determines cell fate after DNA damage by controlling RhoB abundance.
Nat Commun
PUBLISHED: 01-24-2014
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ATM- and RAD3-related (ATR)/Chk1 and ataxia-telangiectasia mutated (ATM)/Chk2 signalling pathways play critical roles in the DNA damage response. Here we report that the E3 ubiquitin ligase Smurf1 determines cell apoptosis rates downstream of DNA damage-induced ATR/Chk1 signalling by promoting degradation of RhoB, a small GTPase recognized as tumour suppressor by promoting death of transformed cells. We show that Smurf1 targets RhoB for degradation to control its abundance in the basal state. DNA damage caused by ultraviolet light or the alkylating agent methyl methanesulphonate strongly activates Chk1, leading to phosphorylation of Smurf1 that enhances its self-degradation, hence resulting in a RhoB accumulation to promote apoptosis. Suppressing RhoB levels by overexpressing Smurf1 or blocking Chk1-dependent Smurf1 self-degradation significantly inhibits apoptosis. Hence, our study unravels a novel ATR/Chk1/Smurf1/RhoB pathway that determines cell fate after DNA damage, and raises the possibility that aberrant upregulation of Smurf1 promotes tumorigenesis by excessively targeting RhoB for degradation.
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Cervical triangulating stapled anastomosis: technique and initial experience.
J Thorac Dis
PUBLISHED: 01-23-2014
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To explore the safety and efficacy of modified cervical triangulating stapled anastomosis (TSA) for gastroesophageal anastomosis (GEA) in minimally invasive esophagectomy (MIE).
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Thoracoscopic esophagectomy in prone versus decubitus position: ergonomic evaluation from a randomized and controlled study.
Ann. Thorac. Surg.
PUBLISHED: 01-21-2014
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The prone position (PP) and decubitus position (DP) have both been used for thoracoscopic esophagectomy. However, which of these positions is ergonomically better for the operating surgeon is unknown. In this randomized controlled trial (NCT01144325), we aimed to assess the surgeon's physical and mental stress in operating on patients in the PP compared with that in the DP.
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Selection of Reference Genes for Quantitative Real-Time PCR Normalization in Panax ginseng at Different Stages of Growth and in Different Organs.
PLoS ONE
PUBLISHED: 01-01-2014
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Quantitative real-time reverse transcription PCR (qRT-PCR) has become a widely used method for gene expression analysis; however, its data interpretation largely depends on the stability of reference genes. The transcriptomics of Panax ginseng, one of the most popular and traditional ingredients used in Chinese medicines, is increasingly being studied. Furthermore, it is vital to establish a series of reliable reference genes when qRT-PCR is used to assess the gene expression profile of ginseng. In this study, we screened out candidate reference genes for ginseng using gene expression data generated by a high-throughput sequencing platform. Based on the statistical tests, 20 reference genes (10 traditional housekeeping genes and 10 novel genes) were selected. These genes were tested for the normalization of expression levels in five growth stages and three distinct plant organs of ginseng by qPCR. These genes were subsequently ranked and compared according to the stability of their expressions using geNorm, NormFinder, and BestKeeper computational programs. Although the best reference genes were found to vary across different samples, CYP and EF-1? were the most stable genes amongst all samples. GAPDH/30S RPS20, CYP/60S RPL13 and CYP/QCR were the optimum pair of reference genes in the roots, stems, and leaves. CYP/60S RPL13, CYP/eIF-5A, aTUB/V-ATP, eIF-5A/SAR1, and aTUB/pol IIa were the most stably expressed combinations in each of the five developmental stages. Our study serves as a foundation for developing an accurate method of qRT-PCR and will benefit future studies on gene expression profiles of Panax Ginseng.
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Spatial variation in carbon and nitrogen in cultivated soils in henan province, china: potential effect on crop yield.
PLoS ONE
PUBLISHED: 01-01-2014
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Improved management of soil carbon (C) and nitrogen (N) storage in agro-ecosystems represents an important strategy for ensuring food security and sustainable agricultural development in China. Accurate estimates of the distribution of soil C and N stores and their relationship to crop yield are crucial to developing appropriate cropland management policies. The current study examined the spatial variation of soil organic C (SOC), total soil N (TSN), and associated variables in the surface layer (0-40 cm) of soils from intensive agricultural systems in 19 counties within Henan Province, China, and compared these patterns with crop yield. Mean soil C and N concentrations were 14.9 g kg-1 and 1.37 g kg-1, respectively, whereas soil C and N stores were 4.1 kg m-2 and 0.4 kg m-2, respectively. Total crop production of each county was significantly, positively related to SOC, TSN, soil C and N store, and soil C and N stock. Soil C and N were positively correlated with soil bulk density but negatively correlated with soil porosity. These results indicate that variations in soil C could regulate crop yield in intensive agricultural systems, and that spatial patterns of C and N levels in soils may be regulated by both climatic factors and agro-ecosystem management. When developing suitable management programs, the importance of soil C and N stores and their effects on crop yield should be considered.
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Engineering Aspergillus oryzae A-4 through the chromosomal insertion of foreign cellulase expression cassette to improve conversion of cellulosic biomass into lipids.
PLoS ONE
PUBLISHED: 01-01-2014
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A genetic modification scheme was designed for Aspergillus oryzae A-4, a natural cellulosic lipids producer, to enhance its lipid production from biomass by putting the spotlight on improving cellulase secretion. Four cellulase genes were separately expressed in A-4 under the control of hlyA promoter, with the help of the successful development of a chromosomal genetic manipulation system. Comparison of cellulase activities of PCR-positive transformants showed that these transformants integrated with celA gene and with celC gene had significantly (p<0.05) higher average FPAase activities than those strains integrated with celB gene and with celD gene. Through the assessment of cellulosic lipids accumulating abilities, celA transformant A2-2 and celC transformant D1-B1 were isolated as promising candidates, which could yield 101%-133% and 35.22%-59.57% higher amount of lipids than the reference strain A-4 (WT) under submerged (SmF) conditions and solid-state (SSF) conditions, respectively. Variability in metabolism associated to the introduction of cellulase gene in A2-2 and D1-B1 was subsequently investigated. It was noted that cellulase expression repressed biomass formation but enhanced lipid accumulation; whereas the inhibitory effect on cell growth would be shielded during cellulosic lipids production owing to the essential role of cellulase in substrate utilization. Different metabolic profiles also existed between A2-2 and D1-B1, which could be attributed to not only different transgene but also biological impacts of different integration. Overall, both simultaneous saccharification and lipid accumulation were enhanced in A2-2 and D1-B1, resulting in efficient conversion of cellulose into lipids. A regulation of cellulase secretion in natural cellulosic lipids producers could be a possible strategy to enhance its lipid production from lignocellulosic biomass.
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SAMD9L inactivation promotes cell proliferation via facilitating G1-S transition in hepatitis B virus-associated hepatocellular carcinoma.
Int. J. Biol. Sci.
PUBLISHED: 01-01-2014
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Hepatocellular carcinoma (HCC) is a highly malignant cancer with poor prognosis, and driver genes harboring genetic lesions and/or expression dysregulation contribute to hepatocarcinogenesis. Sterile Alpha Motif Domain-containing 9-like (SAMD9L) was a novel identified mutated gene in our previous study on exome sequencing of hepatitis B virus (HBV)-associated HCC, but its expression and role in HCC remain unknown. Here, we demonstrated that SAMD9L was frequently inactivated by somatic mutations, and that its expression was deregulated in HCC patients with hepatitis B virus (HBV) infection. SAMD9L knockdown significantly promoted cell proliferation, colony formation of SK-hep-1, QGY-7701, BEL-7721 and MHCC-97H HCC cells. Furthermore, SK-hep-1 and MHCC-97H cells with stable SAMD9L knockdown exhibited enhanced tumorigenicity in athymic mice. Interestingly, SAMD9L silence facilitated G1-S transition of cell cycle progression and led to the elevated activity of Wnt/?-catenin pathway. Collectively, these findings highlight a novel tumor-suppressive role of SAMD9L inactivation by somatic mutation and decreased expression in human HBV-related HCC.
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Silencing [Formula: see text] Rescues Tau Pathologies and Memory Deficits through Rescuing PP2A and Inhibiting GSK-3? Signaling in Human Tau Transgenic Mice.
Front Aging Neurosci
PUBLISHED: 01-01-2014
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Increase of inhibitor-2 of protein phosphatase-2A [Formula: see text] is associated with protein phosphatase-2A (PP2A) inhibition and tau hyperphosphorylation in Alzheimer's disease (AD). Down-regulating [Formula: see text] attenuated amyloidogenesis and improved the cognitive functions in transgenic mice expressing amyloid precursor protein (tg2576). Here, we found that silencing [Formula: see text] by hippocampal infusion of [Formula: see text] down-regulated [Formula: see text] (~45%) with reduction of tau phosphorylation/accumulation, improvement of memory deficits, and dendritic plasticity in 12-month-old human tau transgenic mice. Silencing [Formula: see text] not only restored PP2A activity but also inhibited glycogen synthase kinase-3? (GSK-3?) with a significant activation of protein kinase A (PKA) and Akt. In HEK293/tau and N2a/tau cells, silencing [Formula: see text] by [Formula: see text] also significantly reduced tau hyperphosphorylation with restoration of PP2A activity and inhibition of GSK-3?, demonstrated by the decreased GSK-3? total protein and mRNA levels, and the increased inhibitory phosphorylation of GSK-3? at serine-9. Furthermore, activation of PKA but not Akt mediated the inhibition of GSK-3? by [Formula: see text] silencing. We conclude that targeting [Formula: see text] can improve tau pathologies and memory deficits in human tau transgenic mice, and activation of PKA contributes to GSK-3? inhibition induced by silencing [Formula: see text]in vitro, suggesting that [Formula: see text] is a promising multiple target of AD.
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Helicobacter pylori filtrate impairs spatial learning and memory in rats and increases ?-amyloid by enhancing expression of presenilin-2.
Front Aging Neurosci
PUBLISHED: 01-01-2014
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Helicobacter pylori (H. pylori) infection is related with a high risk of Alzheimer's disease (AD), but the intrinsic link between H. pylori infection and AD development is still missing. In the present study, we explored the effect of H. pylori infection on cognitive function and ?-amyloid production in rats. We found that intraperitoneal injection of H. pylori filtrate induced spatial learning and memory deficit in rats with a simultaneous retarded dendritic spine maturation in hippocampus. Injection of H. pylori filtrate significantly increased A?42 both in the hippocampus and cortex, together with an increased level of presenilin-2 (PS-2), one key component of ?-secretase involved in A? production. Incubation of H. pylori filtrate with N2a cells which over-express amyloid precursor protein (APP) also resulted in increased PS-2 expression and A?42 overproduction. Injection of Escherichia coli (E.coli) filtrate, another common intestinal bacterium, had no effect on cognitive function in rats and A? production in rats and cells. These data suggest a specific effect of H. pylori on cognition and A? production. We conclude that soluble surface fractions of H. pylori may promote A?42 formation by enhancing the activity of ?-secretase, thus induce cognitive impairment through interrupting the synaptic function.
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Targeting SHCBP1 Inhibits Cell Proliferation in Human Hepatocellular Carcinoma Cells.
Asian Pac. J. Cancer Prev.
PUBLISHED: 12-03-2013
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Src homology 2 domain containing (SHC) is a proto-oncogene which mediates cell proliferation and carcinogenesis in human carcinomas. Here, the SHC SH2-domain binding protein 1 (SHCBP1) was first established to be up-regulated in human hepatocellular carcinoma (HCC) tissues by array-base comparative genome hybridization (aCGH). Meanwhile, we examine and verify it by quantitative real-time PCR and western blot. Our current data show that SHCBP1 was up-regulated in HCC tissues. Overexpression of SHCBP1 could significantly promote HCC cell proliferation, survival and colony formation in HCC cell lines. Furthermore, knockdown of SHCBP1 induced cell cycle delay and suppressed cell proliferation. Furthermore, SHCBP1 could regulate the expression of activate extracellular signal-regulated kinase 1/2 (ERK1/2) and cyclin D1. Together, our findings indicate that SHCBP1 may contribute to human hepatocellular carcinoma by promoting cell proliferation and may serve as a molecular target of cancer therapy.
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Enantioselective Synthesis of Indoloquinolizidines via Asymmetric Catalytic Hydrogenation/Lactamization of Imino Diesters.
J. Org. Chem.
PUBLISHED: 11-12-2013
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We have developed a highly efficient cascade sequence for asymmetric synthesis of indoloquinolizidines with absolute control of cis-H2/H12b relative geometry in good to excellent yields and excellent enantioselectivities. This cascade was triggered by the Ru(II)-TsDPEN-catalyzed asymmetric transfer hydrogenation of imino diesters, with subsequent spontaneous lactamization with discrimination between the two diastereotopic 2-alkoxy-2-oxoethyl groups. The synthetic utility of this strategy was demonstrated by the asymmetric preparation of dihydrocorynantheol, geissoschizol, and isogeissoschizol.
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Generation of nano-sized core-shell particles using a coaxial tri-capillary electrospray-template removal method.
Colloids Surf B Biointerfaces
PUBLISHED: 10-29-2013
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This study proposed a new strategy based on a coaxial tri-capillary electrospray-template removal process for producing nanosized polylactide-b-polyethylene glycol (PLA-PEG) particles with a core-shell structure. Microparticles with core-shell-corona structures were first fabricated by coaxial tri-capillary electrospray, and core-shell nanoparticles less than 200nm in size were subsequently obtained by removing the PEG template from the core-shell-corona microparticles. The nanoparticle size could be modulated by adjusting the flow rate of corona fluid, and nanoparticles with an average diameter of 106±5nm were obtained. The nanoparticles displayed excellent dispersion stability in aqueous media and very low cytotoxicity. Paclitaxel was used as a model drug to be incorporated into the core section of the nanoparticles. A drug loading content in the nanoparticles as high as 50.7±1.5wt% with an encapsulation efficiency of greater than 70% could be achieved by simply increasing the feed rate of the drug solution. Paclitaxel exhibited sustained release from the nanoparticles for more than 40 days. The location of the paclitaxel in the nanoparticles, i.e., in the core or shell layer, did not have a significant effect on its release.
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The juxtamembrane and carboxy-terminal domains of Arabidopsis PRK2 are critical for ROP-induced growth in pollen tubes.
J. Exp. Bot.
PUBLISHED: 10-17-2013
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Polarized growth of pollen tubes is a critical step for successful reproduction in angiosperms and is controlled by ROP GTPases. Spatiotemporal activation of ROP (Rho GTPases of plants) necessitates a complex and sophisticated regulatory system, in which guanine nucleotide exchange factors (RopGEFs) are key components. It was previously shown that a leucine-rich repeat receptor-like kinase, Arabidopsis pollen receptor kinase 2 (AtPRK2), interacted with RopGEF12 for its membrane recruitment. However, the mechanisms underlying AtPRK2-mediated ROP activation in vivo are yet to be defined. It is reported here that over-expression of AtPRK2 induced tube bulging that was accompanied by the ectopic localization of ROP-GTP and the ectopic distribution of actin microfilaments. Tube depolarization was also induced by a potentially kinase-dead mutant, AtPRK2K366R, suggesting that the over-expression effect of AtPRK2 did not require its kinase activity. By contrast, deletions of non-catalytic domains in AtPRK2, i.e. the juxtamembrane (JM) and carboxy-terminal (CT) domains, abolished its ability to affect tube polarization. Notably, AtPRK2K366R retained the ability to interact with RopGEF12, whereas AtPRK2 truncations of these non-catalytic domains did not. Lastly, it has been shown that the JM and CT domains of AtPRK2 were not only critical for its interaction with RopGEF12 but also critical for its distribution at the plasma membrane. These results thus provide further insight into pollen receptor kinase-mediated ROP activation during pollen tube growth.
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Imaging and estimation of human abdominal fat by electrical impedance tomography using multiple voltage measurement patterns.
Conf Proc IEEE Eng Med Biol Soc
PUBLISHED: 10-11-2013
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A measuring device for human abdominal fat from the conductivity image derived by electrical impedance tomography (EIT) is rarely found. This study was aimed to reconstruct precise conductivity images from multiple voltage measurements in different patterns of the combination of current and voltage electrodes. We examined two voltage measuring patterns using electrodes located at upper and lower levels around the abdomen of a subject. In the experiment, after 1024 voltage data were taken from one specified voltage measurement pattern, another 1024 data were also taken continuously using another pattern. The reconstruction of conductivity image was made using entire data. As a result, the tomography image was improved compared with the image obtained from single voltage measurement pattern. We then obtained the histogram of the conductivities and estimated the area of abdominal fat. The present method using multiple voltage measurement patterns would be effective, if the measuring time can be much reduced through future modification of the tomography device.
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[Clinical analysis of 83 cases undergoing esophageal leiomyoma enucleation by video-assisted thoracoscopy].
Zhonghua Wei Chang Wai Ke Za Zhi
PUBLISHED: 09-25-2013
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To investigate the feasibility of video-assisted thoracoscopic surgery in the treatment of esophageal leiomyoma.
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Involvement of the single Cul4 gene of Chinese mitten crab Eriocheir sinensis in spermatogenesis.
Gene
PUBLISHED: 09-22-2013
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The Cullin-RING finger ligases (CRLs) are involved in the ubiquitin-mediated degradation of cell cycle regulators and play an important role in gametogenesis. Cullin 4 (CUL4) is a conserved core component of a new class of ubiquitin E3 ligase, and participates in the proteolysis of several regulatory proteins through the ubiquitin-proteasome pathway. The mammals encode two paralogs of CUL4, CUL4A and CUL4B, and the two Cul4 genes are functionally redundant. However, Drosophila or other metazoans only contain one Cul4 gene. Here we cloned the Cul4 gene and confirmed that there is only one protein of CUL4 in Eriocheir sinensis, a full length Cul4 comprised of 2777 nucleotides, an open-reading frame of 2373bp encoding 790 amino acid residues. The expression level of Cul4 mRNAs, as demonstrated by quantitative real-time PCR, varied significantly during testis development, with the greatest transcript levels found at an early stage. Localization analysis using antibodies against CUL4A/4B in the reproductive system showed that EsCUL4 mainly distribute in spermatogonia and primary spermatocytes, and gradually reduced during the development and maturation of sperm. The results indicated that a single CUL4 protein may play a role in spermatogenesis in E. sinensis.
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Anterior dislocation after total hip replacement - effects of pelvic rotation and femoral head size.
Hip Int
PUBLISHED: 09-10-2013
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We investigated the effects of pelvic rotation and femoral head diameter on the anterior stability of the hip joint after total hip replacement. Computer navigation and cadaveric bone were used to simulate the range of motion after total hip replacement. The hip was put at 0 degrees of flexion and it was gradually externally rotated until the hip dislocated. The degree of external rotation when the hip was dislocated was recorded. The test was repeated with the hip at +10, -10, -20, -30, -40 and -50 degrees of flexion. The acetabular component was positioned with abduction of 45 degrees and anteversion at 20 degrees. There was a significant difference amongst each group of pelvic rotation from 0 to -50 degrees in the degree of external rotation when the hip was dislocated. The degree became insignificant when pelvic rotation was increased from 0 to 10 degrees. From -10 to -50 degree of pelvic rotation, 36mm head had significant better stability compared with 32mm and 28mm femoral heads. The presence of significant pelvic sagittal malrotation can increase the risk of anterior dislocation. A larger femoral head is more stable than smaller heads. When pelvic sagittal malrotation is not present, there is no difference in stability between large and small femoral heads.
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fMRI measurement of the integrative effects of visual and chemical senses stimuli in humans.
J. Integr. Neurosci.
PUBLISHED: 09-05-2013
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The purpose of this study was to evaluate the integrative effects of visual stimuli with chemical senses (olfactory and gustatory) stimuli in humans. Noninvasive measurement tools such as magnetoencephalography (MEG) and functional magnetic resonance imaging (fMRI) are used to describe the mechanism of olfactory information processing in the human brain, and the neurophysiological properties of olfactory-related neurons are described. The first study analyzed the interaction between visual and olfactory stimuli. Two odors (lemon and iso-valeric acid) were selected as pleasant and unpleasant odors, respectively and pleasant and unpleasant images were also selected. These cross-modal stimulus combinations were presented to the subject at random, and responses were measured by fMRI using an event related task. These results revealed that active brain areas with pleasant/unpleasant stimuli and matched/mismatched stimuli were different for memory and cognition. The second study analyzed the interaction between visual and gustatory stimuli. Total four conditions (hunger-not hunger, and intake-not intake of monosodium glutamate (MSG)) were tested. Visual stimuli were food-related and nonfood-related photos. A visual analog scale (VAS) was also used to evaluate before and at regular time intervals after intake of MSG, and responses were measured using fMRI. Brain activity related to feeding desire after intake of MSG occurred near the insula cortex, and orbito-frontal cortex, among other areas. These results on the integrative effects of visual stimuli with olfactory and gustatory stimuli, cross-modal and complex effects on olfaction and gustation were suggested to be obtained as an emotional response such as "pleasantness/unpleasantness" and as cognitive and memory responses such as "matching/mismatching" or the responses such as "feeding desire" afterwards intake of foods.
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Programmed cell death-4 deficiency prevents diet-induced obesity, adipose tissue inflammation, and insulin resistance.
Diabetes
PUBLISHED: 08-29-2013
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Programmed cell death-4 (PDCD4), a selective protein translation inhibitor, has shown proinflammatory effect in some inflammatory diseases, but its roles in obesity remain unestablished. This study aims to investigate the effects of PDCD4 on obesity, inflammation, and insulin resistance. Surprisingly, high-fat diet (HFD)-fed PDCD4-deficient (PDCD4(-/-)) mice exhibited an absolutely lean phenotype together with improved insulin sensitivity. Compared with wild-type obese mice, HFD-fed PDCD4(-/-) mice showed higher energy expenditure, lower epididymal fat weight, and reduced macrophage infiltration inflammatory cytokine secretion in white adipose tissue (WAT). Alleviated hepatic steatosis along with decreased plasma levels of triglyceride and cholesterol was also observed in these mice. Importantly, PDCD4 appeared to disturb lipid metabolism via inhibiting the expression of liver X receptor (LXR)-?, a master modulator of lipid homeostasis, which was elevated in HFD-fed PDCD4(-/-) mice accompanied by upregulation of its target genes and relieved endoplasmic reticulum stress in WAT. These data demonstrate that PDCD4 deficiency protects mice against diet-induced obesity, WAT inflammation, and insulin resistance through restoring the expression of LXR-?, thereby proposing PDCD4 as a potential target for treating obesity-associated diseases.
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Human tumor necrosis factor (TNF)-alpha-induced protein 8-like 2 suppresses hepatocellular carcinoma metastasis through inhibiting Rac1.
Mol. Cancer
PUBLISHED: 08-27-2013
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Tumor invasion and metastasis are the major reasons for leading death of patients with hepatocellular carcinoma (HCC). Therefore, to identify molecules that can suppress invasion and metastasis of tumor will provide novel targets for HCC therapies. Tumor necrosis factor (TNF)-alpha-induced protein 8-like 2, TIPE2, is a novel immune negative molecule and an inhibitor of the oncogenic Ras in mice but its function in human is unclear. Our previous research has shown that TIPE2 is downregulated in human primary HCC compared with the paired adjacent non-tumor tissues.
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A rare collision tumor of squamous carcinoma and small cell carcinoma in esophagus involved with separate lymph nodes: a case report.
J Thorac Dis
PUBLISHED: 08-19-2013
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We report a case of an esophageal collision tumor composed of squamous cell carcinoma and small cell carcinoma (SmCC). A 66-year-old man complained of chest pain after oral intake for nearly one month. The patient received two cycles of neoadjuvant platinum-based combination chemotherapy and enhanced computed tomography showed a partial response of the tumor. He then underwent a thoracolaparoscopic esophagectomy with extensive mediastinal lymphadenectomy. Two cycles of chemotherapy and prophylactic irradiation of the lymphatic drainage region were sequentially achieved after surgery. The patient has survived for more than 18 months with no evidence of recurrent disease since surgical resection.
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Video-assisted thoracic surgery versus open thoracotomy for non-small-cell lung cancer: a propensity score analysis based on a multi-institutional registry.
Eur J Cardiothorac Surg
PUBLISHED: 08-15-2013
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Comparative long-term survival and oncological outcomes for patients with non-small-cell lung cancer (NSCLC) who undergo video-assisted thoracic surgery (VATS) or conventional open lobectomy remain uncertain. We conducted a multi-institutional propensity-matched study to stratify potential differences in these outcomes.
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Parallelization of MODFLOW Using a GPU Library.
Ground Water
PUBLISHED: 08-12-2013
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A new method based on a graphics processing unit (GPU) library is proposed in the paper to parallelize MODFLOW. Two programs, GetAb_CG and CG_GPU, have been developed to reorganize the equations in MODFLOW and solve them with the GPU library. Experimental tests using the NVIDIA Tesla C1060 show that a 1.6- to 10.6-fold speedup can be achieved for models with more than 10(5) cells. The efficiency can be further improved by using up-to-date GPU devices.
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Hybrid Hydroxyapatite Nanoparticle Colloidal Gels are Injectable Fillers for Bone Tissue Engineering.
Tissue Eng Part A
PUBLISHED: 08-12-2013
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Injectable bone fillers have emerged as an alternative to the invasive surgery often required to treat bone defects. Current bone fillers may benefit from improvements in dynamic properties such as shear thinning during injection and recovery of material stiffness after placement. Negatively charged inorganic hydroxyapatite (HAp) nanoparticles (NPs) were assembled with positively charged organic poly(d,l-lactic-co-glycolic acid) (PLGA) NPs to create a cohesive colloidal gel. This material is held together by electrostatic forces that may be disrupted by shear to facilitate extrusion, molding, or injection. Scanning electron micrographs of the dried colloidal gels showed a well-organized, three-dimensional porous structure. Rheology tests revealed that certain colloidal gels could recover after being sheared. Human umbilical cord mesenchymal stem cells were also highly viable when seeded on the colloidal gels. HAp/PLGA NP colloidal gels offer an attractive scheme for injectable filling and regeneration of bone tissue.
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Multifunctional nanoparticles for targeted delivery of immune activating and cancer therapeutic agents.
J Control Release
PUBLISHED: 08-07-2013
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Nanoparticles (NPs) have been extensively investigated for applications in both experimental and clinical settings to improve delivery efficiency of therapeutic and diagnostic agents. Most recently, novel multifunctional nanoparticles have attracted much attention because of their ability to carry diverse functionalities to achieve effective synergistic therapeutic treatments. Multifunctional NPs have been designed to co-deliver multiple components, target the delivery of drugs by surface functionalization, and realize therapy and diagnosis simultaneously. In this review, various materials of diverse chemistries for fabricating multifunctional NPs with distinctive architectures are discussed and compared. Recent progress involving multifunctional NPs for immune activation, anticancer drug delivery, and synergistic theranostics is the focus of this review. Overall, this comprehensive review demonstrates that multifunctional NPs have distinctive properties that make them highly suitable for targeted therapeutic delivery in these areas.
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Telocytes in human oesophagus.
J. Cell. Mol. Med.
PUBLISHED: 08-06-2013
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Telocytes (TCs), a new type of interstitial cells, were identified in many different organs and tissues of mammalians and humans. In this study, we show the presence, in human oesophagus, of cells having the typical features of TCs in lamina propria of the mucosa, as well as in muscular layers. We used transmission electron microscopy (TEM), immunohistochemistry (IHC) and primary cell culture. Human oesophageal TCs present a small cell body with 2-3 very long Telopodes (Tps). Tps consist of an alternation of thin segments (podomers) and thick segments (podoms) and have a labyrinthine spatial arrangement. Tps establish close contacts (stromal synapses) with other neighbouring cells (e.g. lymphocytes, macrophages). The ELISA testing of the supernatant of primary culture of TCs indicated that the concentrations of VEGF and EGF increased progressively. In conclusion, our study shows the existence of typical TCs at the level of oesophagus (mucosa, submucosa and muscular layer) and suggests their possible role in tissue repair.
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Epitope Mapping of Monoclonal Antibodies using Synthetic Oligosaccharides Uncovers Novel Aspects of Immune Recognition of the Psl Exopolysaccharide of Pseudomonas aeruginosa.
Chemistry
PUBLISHED: 07-24-2013
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Pseudomonas aeruginosa is an opportunistic Gram-negative bacterium that can cause life-threatening infections in critically ill and cystic fibrosis patients. The Psl exopolysaccharide of P.?aeruginosa offers an attractive serotype-independent antigen for the development of immunotherapies. Here, the first chemical synthesis of a panel of oligosaccharides derived from the exopolysaccharide of P.?aeruginosa by a synthetic strategy that efficiently deals with the stereoselective installation of several ?-mannosides and the formation of a mannoside that is extended by saccharide moieties at C-1, C-2, and C-3 in a crowded 1,2,3-cis configuration is described. The approach was employed to prepare tetra-, penta-, and hexa- and decasaccharide part structures. The compounds were employed to define the epitope requirements of several functionally active monoclonal antibodies (mAbs) that can bind three distinct epitopes of Psl (class?I, II, and III). The class?II mAb reacted potently with each oligosaccharide indicating its epitope resides within the tetrasaccharide and does not require the branched mannoside of Psl. The class?III antibody did not bind the tetra- or pentasaccharide; however, it did react potently with the hexasaccharide and weakly with the decasaccharide, suggesting a terminal glucoside is required for optimal binding. Unexpectedly, the class?I mAb did not bind any of the oligosaccharides indicating that Psl contains a yet to be elucidated sub-stoichiometric isoform. This study demonstrates that functional activity of a mAb does not only depend on the avidity of binding but also on the location of an epitope within a bacterial polysaccharide. The results also provide a strong impetus to analyze further the structure of Psl to identify the class?I epitope, that is expected to provide an attractive target for the development of a synthetic vaccine for P.?aeruginosa.
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A simple method minimizes chylothorax after minimally invasive esophagectomy.
J. Am. Coll. Surg.
PUBLISHED: 07-19-2013
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Postoperative chylothorax is a rare, but potentially fatal complication after esophagectomy. Preventive measures aimed at decreasing the incidence of chyle leakage after minimally invasive esophagectomy (MIE) could potentially reduce the high postoperative mortality associated with this complication. However, previous techniques are traumatic and time consuming. We present a simple method in the prophylaxis of chylothorax after MIE.
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Molecular cloning and expression analysis of a dorsal homologue from Eriocheir sinensis.
Dev. Comp. Immunol.
PUBLISHED: 07-10-2013
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Dorsal as a crucial component of Toll signaling pathway, played important roles in induction and regulation of innate immune responses. In this study, we cloned a NF-?B-like transcription factor Dorsal from Eriocheir sinensis and designated it as EsDorsal. The full-length cDNA of EsDorsal was 2493 bp with a 2022-bp open reading frame (ORF) encoding a 673-amino acid protein. This protein contained a 171-residue conserved Rel homology domain (RHD) and a 102-residue Ig-like, plexins and transcription factors domain (IPT). By phylogenetic analysis, EsDorsal was clustered into one group together with other invertebrate Dorsals or NF-?Bs, and then clustered with vertebrate NF-?Bs. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis results showed that (a) EsDorsal had higher expression level in immune organs; (b) EsDorsal differentially induced after injection of lipopolysaccharides (LPS), peptidoglycan (PG) or zymosan (GLU). Importantly, EsDorsal was more responsive to LPS than GLU and PG. Collectively, EsDorsal was differentially inducibility in response to various PAMPs, suggesting its involvement in a specific innate immune regulation in E. sinensis.
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Cathepsin A protein from the accessory sex gland of the Chinese mitten crab (Eriocheir sinensis) plays a key role in spermatophore digestion.
J. Insect Physiol.
PUBLISHED: 07-08-2013
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Accessory sex gland (ASG) secretory proteins of the Chinese mitten crab (Eriocheir sinensis) can effectively digest the spermatophore wall. In order to identify which proteins participate in spermatophore wall digestion, a 50-kDa protein secreted from the ASG was purified to homogeneity by a series of isolation steps, including ammonium sulfate fractionation, Sephadex G-25 S gel-filtration, ion exchange chromatography on a DEAE-Sephacel column and Sephacryl S-200 gel-filtration. The purified protein was effective in spermatophore wall rupture, and the subsequent HPLC-ESI-MS/MS shotgun analysis showed the digestive protein to be cathepsin A (cathA). This finding was also confirmed by Western blot analysis and a cathA inhibitor digestion experiment. ELISA analysis showed that cathA enzymatic activity from ASG secretions increased during its purification process. Furthermore, enzymatic activity was significantly higher in the mating period of E. sinensis parallel to the latest developmental stage of the gland. Moreover, analysis from a cathA inhibitor that inhibits spermatophore wall digestion showed that cathA is the main enzyme involved. Hence, we first report the characterization of cathA from the ASG, which might play a key role in digesting the spermatophore wall of E. sinensis.
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Targeting therapy of hepatocellular carcinoma with doxorubicin prodrug PDOX increases anti-metastatic effect and reduces toxicity: a preclinical study.
J Transl Med
PUBLISHED: 06-26-2013
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This study was to investigate the effects and safety of cathepsin B-cleavable doxorubicin (DOX)-prodrug (PDOX) for targeting therapy of metastatic human hepatocellular carcinoma (HCC) using DOX as a positive control drug.
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A novel C-type lectin from Eriocheir sinensis functions as a pattern recognition receptor with antibacterial activity.
Fish Shellfish Immunol.
PUBLISHED: 06-25-2013
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As pattern recognition receptors (PRRs), C-type lectins (CTLs) play significant roles in recognizing and eliminating pathogens in innate immunity. In this study, a novel CTL (EsLecD) was identified from the crustacean Eriocheir sinensis. The cloning of full-length EsLecD cDNA was based on the initial expressed sequence tags (ESTs) isolated from a hepatopancreatic cDNA library. The full-length EsLecD cDNA of 686 bp with an open reading frame of 468 bp encodes a putative protein of 155 aa residues, including an N-terminal signal peptide and a single carbohydrate-recognition domain (CRD). By quantitative RT-PCR analysis, the EsLecD transcript was mainly detected in the hepatopancreas but rarely in other tissues, and it was significantly upregulated in the hepatopancreas after immune challenge with lipopolysaccharides. The recombinant EsLecD protein (rEsLecD) exhibited the ability to bind to all tested microorganisms, including bacteria and yeast. Meanwhile, calcium significantly increased the binding affinity of rEsLecD toward microorganisms, but it was not essential. The binding of rEsLecD induced the aggregation of microbial pathogens. Moreover, rEsLecD was capable of inhibiting the growth of microorganisms and even directly killing bacteria. Interestingly, rEsLecD could stimulate cellular encapsulation in vitro. In conclusion, results of this study suggest that EsLecD acts as an antibacterial PRR participating in the innate immunity of invertebrates.
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Comparative phylogeography of two sympatric beeches in subtropical China: Species-specific geographic mosaic of lineages.
Ecol Evol
PUBLISHED: 06-19-2013
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In subtropical China, large-scale phylogeographic comparisons among multiple sympatric plants with similar ecological preferences are scarce, making generalizations about common response to historical events necessarily tentative. A phylogeographic comparison of two sympatric Chinese beeches (Fagus lucida and F. longipetiolata, 21 and 28 populations, respectively) was conducted to test whether they have responded to historical events in a concerted fashion and to determine whether their phylogeographic structure is exclusively due to Quaternary events or it is also associated with pre-Quaternary events. Twenty-three haplotypes were recovered for F. lucida and F. longipetiolata (14 each one and five shared). Both species exhibited a species-specific mosaic distribution of haplotypes, with many of them being range-restricted and even private to populations. The two beeches had comparable total haplotype diversity but F. lucida had much higher within-population diversity than F. longipetiolata. Molecular dating showed that the time to most recent common ancestor of all haplotypes was 6.36 Ma, with most haplotypes differentiating during the Quaternary. [Correction added on 14 October 2013, after first online publication: the timeunit has been corrected to 6.36.] Our results support a late Miocene origin and southwards colonization of Chinese beeches when the aridity in Central Asia intensified and the monsoon climate began to dominate the East Asia. During the Quaternary, long-term isolation in subtropical mountains of China coupled with limited gene flow would have lead to the current species-specific mosaic distribution of lineages.
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Two novel Toll genes (EsToll1 and EsToll2) from Eriocheir sinensis are differentially induced by lipopolysaccharide, peptidoglycan and zymosan.
Fish Shellfish Immunol.
PUBLISHED: 06-11-2013
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Tolls/Toll-like receptors (TLRs) play an essential role in initiating innate immune responses against pathogens and are found throughout the insect kingdom but have not yet been reported in the crustacean, Eriocheir sinensis. For this purpose, we cloned two novel Toll genes from E. sinensis, EsToll1 and EsToll2. The full-length cDNA of EsToll1 was 3963 bp with a 3042-bp open reading frame (ORF) encoding a 1013-amino acid protein. The extracellular domain of this protein contains 17 leucine-rich repeats (LRRs) and a 139-residue cytoplasmic Toll/interleukin-1 receptor (TIR) domain. The cDNA full-length of EsToll2 was 4419 bp with a 2667-bp ORF encoding an 888-amino acid protein with an extracellular domain containing 10 LRRs and a 139-residue cytoplasmic TIR domain. By phylogenetic analysis, EsToll1 and EsToll2 clustered into one group together with Tolls from other crustaceans. Quantitative RT-PCR analysis demonstrated that a) both EsToll1 and EsToll2 were constitutively expressed in all tested crab tissues; b) EsToll1 and EsToll2 were differentially induced after injection of lipopolysaccharides (LPS), peptidoglycan (PG) or zymosan (GLU). Importantly, EsToll2 expression was significantly upregulated at almost all time intervals post-challenge with LPS, PG and GLU. Our study indicated that EsToll1 and EsToll2 are differentially inducibility in response to various PAMPs, suggesting their involvement in a specific innate immune recognition mechanism in E. sinensis.
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Two antibacterial C-type lectins from crustacean, Eriocheir sinensis, stimulated cellular encapsulation in vitro.
Dev. Comp. Immunol.
PUBLISHED: 05-30-2013
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The first step of host fighting against pathogens is that pattern recognition receptors recognized pathogen-associated molecular patterns. However, the specificity of recognition within the innate immune molecular of invertebrates remains largely unknown. In the present study, we investigated how invertebrate pattern recognition receptor (PRR) C-type lectins might be involved in the antimicrobial response in crustacean. Based on our previously obtained completed coding regions of EsLecA and EsLecG in Eriocheir sinensis, the recombinant EsLectin proteins were produced via prokaryotic expression system and affinity chromatography. Subsequently, both rEsLecA and rEsLecG were discovered to have wide spectrum binding activities towards microorganisms, and their microbial-binding was calcium-independent. Moreover, the binding activities of both rEsLecA and rEsLecG induced the aggregation against microbial pathogens. Both microorganism growth inhibitory activities assays and antibacterial activities assays revealed their capabilities of suppressing microorganisms growth and directly killing microorganisms respectively. Furthermore, the encapsulation assays signified that both rEsLecA and rEsLecG could stimulate the cellular encapsulation in vitro. Collectively, data presented here demonstrated the successful expression and purification of two C-type lectins proteins in the Chinese mitten crab, and their critical role in the innate immune system of an invertebrate.
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Arabidopsis RopGEF4 and RopGEF10 are important for FERONIA-mediated developmental but not environmental regulation of root hair growth.
New Phytol.
PUBLISHED: 05-19-2013
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We investigated a genetic pathway in root hair development in Arabidopsis thaliana, involving the receptor-like kinase FERONIA (FER), two guanine nucleotide exchange factors for ROPs (RopGEF4 and RopGEF10), and the small GTPase Rho of plants (ROPs). Loss- and gain-of-function analyses demonstrated distinct roles of RopGEF4 and RopGEF10 such that RopGEF4 is only important for root hair elongation, while RopGEF10 mainly contributes to root hair initiation. Domain dissection by truncation and domain-swapping experiments indicated that their functional distinctions were mainly contributed by the noncatalytic domains. Using fluorescent ratio imaging, we showed that functional loss of RopGEF4 and RopGEF10 additively reduced reactive oxygen species (ROS) production. Bimolecular fluorescence complementation experiments demonstrated that RopGEF4 and RopGEF10 had the same interaction specificity as ROPs, suggesting common downstream components. We further showed that the promoting effects of environmental cues such as exogenous auxin and phosphate limitation on root hair development depended on FER. However, although functional loss of RopGEF4 and RopGEF10 largely abolished FER-induced ROS production, it did not compromise the responses to FER-mediated environmental cues on root hair development. Our results demonstrated that RopGEF4 and RopGEF10 are genetic components in FER-mediated, developmentally (but not environmentally) regulated, root hair growth.
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Live/dead state is not the factor influencing adhesion ability of Bifidobacterium animalis KLDS2.0603.
J. Microbiol.
PUBLISHED: 04-22-2013
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Two essential requirements for probiotic bifidobacteria are that they be "live" and have "colonization" ability, following FAO/WHO guideline recommendations. The amount of research on the adhesion ability of bifidobacteria compares poorly with that of other probiotic bacteria, such as lactobacilli. The aim of the present study was to determine how gastrointestinal conditions affect the adhesion ability of bifidobacteria, and to investigate the relationship between the adhesion ability and the live/dead state of bifidobacteria. The adhesion ability of Bifidobacterium animalis KLDS2.0603 that had been subjected to the digestive enzymes, pepsin, trypsin, and proteinase K, was decreased significantly, but these treatments did not significantly change the strains survival rates, which were 98.78%, 97.60%, and 97.63% respectively. B. animalis KLDS2.0603 subjected to LiCl retained its adhesion ability but had a lower survival rate (59.28%) than the control group (P<0.01). B. animalis KLDS 2.0603 subjected to sodium metaperiodate exhibited higher adhesion ability than the control group (P<0.01), but the bacterial cells were killed totally. The results of transmission electron microscopy and laser scanning confocal microscopy showed that live/dead state of bifidobacteria was not one of the main factors that affected the adhesion ability of bifidobacteira, and that the substances affecting the adhesion ability of bifidobacteria were on the outer surface layer of the bifidobacterial cells. Our results also indicated that the substances related to the adhesion ability of bifidobacteria are proteinaceous. The above results will help us to understand the adhesion and colonization processes of bifidobacteria in the human gastrointestinal tract.
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