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Find video protocols related to scientific articles indexed in Pubmed.
The complete mitochondrial genome of Vanellus vanellus (Charadriiformes: Charadriidae).
Mitochondrial DNA
PUBLISHED: 10-21-2014
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Abstract The northern lapwing, Vanellus vanellus (Charadriiformes: Charadriidae), is commonly found in temperate Eurasia. In this study, the complete mitogenome sequence of V. vanellus has been determined by polymerase chain reaction (PCR) method using 13 primer pairs. It was a circular molecule with 16,795?bp in length which contained 13 protein-coding genes, 2 ribosomal RNAs, 22 transfer RNAs and a control region. The composition and gene order are similar to most other vertebrates. There are 28 genes encoded on the H-strand, 9 genes (ND6 subunit and 8 tRNA genes) encoded on the L-strand. The overall base composition of the H-strand is A (31.44%), T (24.03%), G (13.76%), C (30.77%), with a slight A?+?T bias of 55.47%. This mitogenome sequence of V. vanellus could contribute to a better solution of its phylogenetic position and will contribute to further genetic researches on Charadriidae.
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Interaction of nNOS with PSD-95 negatively controls regenerative repair after stroke.
J. Neurosci.
PUBLISHED: 10-03-2014
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Stroke is a major public health concern. The lack of effective therapies heightens the need for new therapeutic targets. Mammalian brain has the ability to rewire itself to restore lost functionalities. Promoting regenerative repair, including neurogenesis and dendritic remodeling, may offer a new therapeutic strategy for the treatment of stroke. Here, we report that interaction of neuronal nitric oxide synthase (nNOS) with the protein postsynaptic density-95 (PSD-95) negatively controls regenerative repair after stroke in rats. Dissociating nNOS-PSD-95 coupling in neurons promotes neuronal differentiation of neural stem cells (NSCs), facilitates the migration of newborn cells into the injured area, and enhances neurite growth of newborn neurons and dendritic spine formation of mature neurons in the ischemic brain of rats. More importantly, blocking nNOS-PSD-95 binding during the recovery stage improves stroke outcome via the promotion of regenerative repair in rats. Histone deacetylase 2 in NSCs may mediate the role of nNOS-PSD-95 association. Thus, nNOS-PSD-95 can serve as a target for regenerative repair after stroke.
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Highly stereoselective synthesis of functionalized pyrrolo[3,2-c]quinolines via N-heterocyclic carbene catalyzed cascade sequence.
Org. Lett.
PUBLISHED: 09-23-2014
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An N-heterocyclic carbene-catalyzed stereoselective Michael-Mannich-lactamization cascade reaction of tosyl-protected o-amino aromatic aldimines and 2-bromoenals for the construction of functionalized pyrrolo[3,2-c]quinolines with three consecutive stereocenters was achieved in good yields with excellent diastereo- and enantioselectivities.
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Hippocampus and nitric oxide.
Vitam. Horm.
PUBLISHED: 09-06-2014
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Since it was first identified to play an important role in relaxation of blood vessels, nitric oxide has been demonstrated to regulate many biological processes, especially in the central nervous system. Of the three types of enzymes that produce nitric oxide in humans and rodents, neuronal type is found almost exclusively in the nervous system. This gaseous molecule is a nonclassical neurotransmitter, which maintains the activities of neural cells and regulates the normal functions of brain. It appears to play a role in promoting the transfer of nerve signals from one neuron to another, maintaining the synaptic strength. Meanwhile, nitric oxide is a unique regulator on neurogenesis and synaptogenesis, producing the positive or negative effects upon different signal pathways or cellular origins and locations. Based on its significant roles in neural plasticity, nitric oxide is involved in a number of central nervous diseases, such as ischemia, depression, anxiety, and Alzheimer's disease. Clarifying the profiles of nitric oxide in the brain tissues and its participation in pathophysiological processes opens a new avenue for development of new therapeutic strategies. Thus, this chapter specifies the effects of nitric oxide in the hippocampus, a key structure implicated in the modulation of mood and memories, exhibiting the trend of future research on nitric oxide.
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Systemic analysis on risk factors for breast cancer related lymphedema.
Asian Pac. J. Cancer Prev.
PUBLISHED: 08-30-2014
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To evaluate risk factors for upper extremity lymphedema due to breast cancer surgery.
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CAPON-nNOS coupling can serve as a target for developing new anxiolytics.
Nat. Med.
PUBLISHED: 08-17-2014
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Anxiety disorders are highly prevalent psychiatric diseases. There is need for a deeper understanding of anxiety control mechanisms in the mammalian brain and for development of new anxiolytic agents. Here we report that the coupling between neuronal nitric oxide synthase (nNOS) and its carboxy-terminal PDZ ligand (CAPON) can serve as a target for developing new anxiolytic agents. Augmenting nNOS-CAPON interaction in the hippocampus of mice by overexpressing full-length CAPON gave rise to anxiogenic-like behaviors, whereas dissociating CAPON from nNOS by overexpressing CAPON-125C or CAPON-20C (the C-terminal 125 or 20 amino acids of CAPON) or delivering Tat-CAPON-12C (a peptide comprising Tat and the 12 C-terminal amino acids of CAPON) in the hippocampus of mice produced anxiolytic-like effects. Mice subjected to chronic mild stress (CMS) displayed a substantial increase in nNOS-CAPON coupling in the hippocampus and a consequent anxiogenic-like phenotype. Disrupting nNOS-CAPON coupling reversed the CMS-induced anxiogenic-like behaviors. Moreover, small-molecule blockers of nNOS-CAPON binding rapidly produced anxiolytic-like effects. Dexamethasone-induced ras protein 1 (Dexras1)-extracellular signal-regulated kinase (ERK) signaling was involved in the behavioral effects of nNOS-CAPON association. Thus, nNOS-CAPON association contributes to the modulation of anxiety-related behaviors via regulating Dexras1-ERK signaling and can serve as a target for developing potential anxiolytics.
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The MBD4 Gene Plays an Important Role in Porcine Adipocyte Differentiation.
Cell. Physiol. Biochem.
PUBLISHED: 08-06-2014
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MBD4 (methyl-CpG binding domain protein 4) is an important G: T glycosylase that can identify T-G mismatches. It plays a role in active demethylation through base excision repair. Overexpression of MBD4 gene can cause the demethylation of numerous genes, and the remethylation of MBD4-associated genes can occur when the MBD4 gene is knocked out. To date, the functions and regulatory mechanisms of the MBD4 gene in the differentiation of porcine preadipocytes have not been clearly established.
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Structural and electronic properties of the Pt(n)-PAH complex (n = 1, 2) from density functional calculations.
Phys Chem Chem Phys
PUBLISHED: 07-31-2014
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A detailed density functional study of the Pt atom and the Pt dimer adsorption on a polyaromatic hydrocarbon (PAH) is presented. The preferred adsorption site for a Pt atom is confirmed to be the bridge site. Upon adsorption of a single Pt atom, however, it is found here that the electronic ground state changes from the triplet state (5d(9)6s(1) configuration) to the closed-shell singlet state (5d(10)6s(0) configuration), which consequently will affect the catalytic activity of Pt when single Pt atoms bind to a carbon surface. The preferred adsorption site for the Pt dimer in the upright configuration is the hollow site. In contrast to the adsorption of a single Pt atom, the formation of a Pt-C bond in the adsorption of a Pt dimer is not accompanied by a change in the spin state, so the most stable electronic state is still the triplet state. While the atomic charge on the Pt atoms and dimers (in parallel configuration) in the Ptn-PAH complex is positive, a negative charge is found on the upper Pt atom for the upright configuration, indicating that single layers of Pt atoms will have a different catalytic activity as compared to Pt clusters on a carbon surface. Comparing the Pt-C bond length and the charge transfer on different sites, the magnitude of the charge transfer decreases with bond elongation, indicating that the catalytic activity of the Pt atom and dimer can be changed by modifying its chemical surroundings. The adsorption energy for the Pt dimer on a PAH surface is larger than that for two individual Pt atoms on the surface indicating that aggregation of Pt atoms on the PAH surface is favorable.
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The biomechanical role of periodontal ligament in bonded and replanted vertically fractured teeth under cyclic biting forces.
Int J Oral Sci
PUBLISHED: 07-07-2014
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After teeth are replanted, there are two possible healing responses: periodontal ligament healing or ankylosis with subsequent replacement resorption. The purpose of this study was to compare the fatigue resistance of vertically fractured teeth after bonding the fragments under conditions simulating both healing modes. Thirty-two human premolars were vertically fractured and the fragments were bonded together with Super-Bond C&B. They were then randomly distributed into four groups (BP, CP, CA, BA). The BP and CP groups were used to investigate the periodontal ligament healing mode whilst the BA and CA groups simulated ankylosis. All teeth had root canal treatment performed. Metal crowns were constructed for the CP and CA groups. The BP and BA groups only had composite resin restorations in the access cavities. All specimens were subjected to a 260?N load at 4?Hz until failure of the bond or until 2×10(6) cycles had been reached if no fracture occurred. Cracks were detected by stereomicroscope imaging and also assessed via dye penetration tests. Finally, interfaces of the resin luting agent were examined by scanning electron microscope. The results confirmed that the fatigue resistance was higher in the groups with simulated periodontal ligament healing. Periodontal reattachment showed important biomechanical role in bonded and replanted vertically fractured teeth.International Journal of Oral Science advance online publication, 12 September 2014; doi:10.1038/ijos.2014.51.
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Nicotine-induced upregulation of VCAM-1, MMP-2, and MMP-9 through the ?7-nAChR-JNK pathway in RAW264.7 and MOVAS cells.
Mol. Cell. Biochem.
PUBLISHED: 07-01-2014
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The ability of nicotine to induce aortic aneurysms has been shown in animal models; however, its underlying mechanisms remain elusive. In the present experiment, both the RAW264.7 and MOVAS cell lines were employed to examine the nicotine-induced modulation of VCAM-1, MMP-2, and MMP-9 expressions in macrophages and vascular smooth muscle cells. Our results showed that nicotine concentrations of both 0.5 and 5 ng/ml induced VCAM-1, MMP-2, and MMP-9 upregulation, while a concentration of 50 ng/ml had a slight inhibitory effect and a concentration of 500 ng/ml showed a significant inhibitory effect. When cells were pretreated with either SP600125 (JNK inhibitor) or PNU-282987 (?7-nAChR agonist) prior to nicotine exposure, the nicotine-induced upregulation of VCAM-1, MMP-2, MMP-9, and p-JNK was suppressed, with a joint treatment producing a more significant inhibitory effect. Moreover, PNU-282987 had a comparable inhibitory effect on VCAM-1, MMP-2, and MMP-9 expressions and JNK activation via phosphorylation as did SP600125. In conclusion, nicotine-induced VCAM-1, MMP-2, and MMP-9 expressions occur in a dose-dependent fashion in both of the cell lines tested. Furthermore, the nicotine exposure equivalent to plasma levels found in regular smokers can augment VCAM-1, MMP-2, and MMP-9 expressions through the ?7-nAChR-JNK pathway.
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A micromechanical model for estimating alveolar wall strain in mechanically ventilated edematous lungs.
J. Appl. Physiol.
PUBLISHED: 06-19-2014
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To elucidate the micromechanics of pulmonary edema has been a significant medical concern, which is beneficial to better guide ventilator settings in clinical practice. In this paper, we present an adjoining two-alveoli model to quantitatively estimate strain and stress of alveolar walls in mechanically ventilated edematous lungs. The model takes into account the geometry of the alveolus, the effect of surface tension, the length-tension properties of parenchyma tissue, and the change in thickness of the alveolar wall. On the one hand, our model supports experimental findings (Perlman CE, Lederer DJ, Bhattacharya J. Am J Respir Cell Mol Biol 44: 34-39, 2011) that the presence of a liquid-filled alveolus protrudes into the neighboring air-filled alveolus with the shared septal strain amounting to a maximum value of 1.374 (corresponding to the maximum stress of 5.12 kPa) even at functional residual capacity; on the other hand, it further shows that the pattern of alveolar expansion appears heterogeneous or homogeneous, strongly depending on differences in air-liquid interface tension on alveolar segments. The proposed model is a preliminary step toward picturing a global topographical distribution of stress and strain on the scale of the lung as a whole to prevent ventilator-induced lung injury.
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Evaluation of a Pyrenophora teres f. teres mapping population reveals multiple independent interactions with a region of barley chromosome 6H.
Fungal Genet. Biol.
PUBLISHED: 06-17-2014
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The necrotrophic fungal pathogen Pyrenophora teres f. teres causes the foliar disease net form net blotch (NFNB) on barley. To investigate the genetics of virulence in the barley- P. teres f. teres pathosystem, we evaluated 118 progeny derived from a cross between the California isolates 15A and 6A on the barley lines Rika and Kombar, chosen based on their differential reactions to isolates 15A and 6A for NFNB disease. Genetic maps generated with SNP, SSR, and AFLP markers were scanned for quantitative trait loci (QTL) associated with virulence in P. teres f. teres. Loci underlying two major QTL, VR1 and VR2, were associated with virulence on Rika barley, accounting for 35% and 20% of the disease reaction type variation, respectively. Two different loci, VK1 and VK2, were shown to underlie two major QTL associated with virulence on Kombar barley accounting for 26% and 19% of the disease reaction type variation, respectively. Progeny isolates harboring VK1, VK2, or VR2 alone were inoculated onto a Rika×Kombar recombinant inbred line mapping population and the susceptibility induced by each pathogen genotype corresponded to the same region on barley chromosome 6H as that identified for the parental isolates 15A and 6A. The data presented here indicate that the P. teres f. teres - barley interaction can at least partially be explained by pathogen-produced necrotrophic effectors (NEs) that interact with dominant barley susceptibility genes resulting in NE triggered susceptibility (NETS).
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Visible-light driven degradation of ibuprofen using abundant metal-loaded BiVO4 photocatalysts.
Chemosphere
PUBLISHED: 06-16-2014
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An efficient method for the degradation of ibuprofen as an aqueous contaminant was developed under visible-light irradiation with as-prepared bismuth vanadate (BiVO4) catalysts. The metal-loaded catalysts Cu-BiVO4 and Ag-BiVO4 were synthesized using a hydrothermal process and then a wet-impregnation method. All of the materials were fully characterized by X-ray diffraction, scanning electron microscopy, UV-vis diffuse reflectance spectroscopy, X-ray photoelectron spectroscopy and BET surface area. The results indicated that all of the prepared samples had monoclinic scheelite structures. In the metal-loaded catalysts, silver existed as a mixture of Ag and Ag2O on the surface of the catalysts. However, copper existed as Cu2O and CuO. Additionally, the band gap values of BiVO4, Ag-BiVO4, and Cu-BiVO4 were 2.38, 2.31, and 2.30eV, respectively. Compared to the BiVO4 catalyst, the metal-loaded BiVO4 catalysts showed superior photocatalytic properties for the degradation of ibuprofen.
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Improvement of ventilation-induced lung injury in a rodent model by inhibition of inhibitory ?B kinase.
J Trauma Acute Care Surg
PUBLISHED: 05-24-2014
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Inhibition of nuclear factor ?B (NF-?B) activation is a well-know strategy to ameliorate ventilation-induced lung injury (VILI). Inhibitory ?B kinase (IKK) plays a key role in the regulation of NF-?B activation. In this study, we determined whether inhibition of IKK by an IKK inhibitor exerts lung protection in a rat model of VILI.
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The synergetic effect of edaravone and borneol in the rat model of ischemic stroke.
Eur. J. Pharmacol.
PUBLISHED: 05-12-2014
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Free radical production contributes to the early ischemic response and the neuroinflammatory response to injury initiates the second wave of cell death following ischemic stroke. Edaravone is a free radical scavenger, and borneol has shown anti-inflammatory effect. We investigated the synergistic effect of these two drugs in the rat model of transient cerebral ischemia. Edaravone scavenged OH, NO and ONOO? concentration-dependently, and borneol inhibited ischemia/reperfusion-induced tumor necrosis factor-? (TNF-?), inducible nitric oxide synthase (iNOS), interleukin-1? (IL-1?) and cyclooxygenase-2 (COX-2) expressions. In the rat model of transient cerebral ischemia and reperfusion, the combination of edaravone and borneol significantly ameliorated ischemic damage with an optimal proportion of 4:1. Emax (% inhibition) of edaravone, borneol and two drugs in combination was 55.7%, 65.8% and 74.3% respectively. ED50 of edaravone and borneol was 7.17 and 0.36 mg/kg respectively. When two drugs in combination, ED50 was 0.484 mg/kg, in which edaravone was 0.387 mg/kg (ineffective dose) and borneol was 0.097 mg/kg (ineffective dose). Combination index (CI)<1 among effects observed in experiments, suggesting a significant synergistic effect. Reduced levels of pro-inflammatory mediators and free radicals were probably associated with the synergistic effect of edaravone and borneol. The combination exhibited a therapeutic time window of 6h in ischemia/reperfusion model, and significantly ameliorated damages in permanent ischemia model. Moreover, two drugs in combination promoted long-term effect, including improved elemental vital signs, sensorimotor functions and spatial cognition. Our results suggest that the combination of edaravone and borneol have a synergistic effect for treating ischemic stroke.
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Exosomes mediate drug resistance transfer in MCF-7 breast cancer cells and a probable mechanism is delivery of P-glycoprotein.
Tumour Biol.
PUBLISHED: 05-06-2014
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Acquired drug resistance is a major obstacle to chemotherapy of cancers. In this study, we aim to investigate the role of exosomes in drug-resistance transfer between breast cancer cells and detect the probable mechanism. A docetaxel-resistant variant of MCF-7 cell line (MCF-7/DOC) was established and then compared with the drug-sensitive variant (MCF-7/S). Exosomes were expelled from the cell supernatant using ultracentrifugation. Drug resistance was assessed by apoptosis assay and MTT examination. Expressions of P-glycoprotein (P-gp) were analyzed by flow cytometry. Stained exosomes were absorbed by receipt cells. MCF-7/S in the presence of exosomes extracted from the supernatant of MCF-7/DOC (DOC/exo) acquired drug resistance, while MCF-7/S exposed to their own exosomes (S/exo) did not. P-gp expression patterns of exosomes were similar as the originated cells. P-gp expression of MCF-7/S increased after incubation with DOC/exo and was affected by the amount of exosomes. Exosomes are effective in transferring drug resistance as well as P-gp from drug-resistant breast cancer cells to sensitive ones. The delivery of P-gp via exosomes may be a mechanism of exosome-mediated drug resistance transfer.
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[Research on spectral characteristic of miniature X-ray tube and determination of beryllium window thickness].
Guang Pu Xue Yu Guang Pu Fen Xi
PUBLISHED: 05-03-2014
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Applying Monte Carlo method, the present paper simulates the emitted X-ray spectrum of miniature X-ray tube with thirteen thickness of beryllium window in the range from 50 to 500 microm. By analyzing the characteristic of the spectrums, the reasonable choice of thickness of beryllium window relies on the application and for the beryllium window it is not the thinner the better. Taking in-situ EDXRF as an example, though the emission X-ray intensity is higher as the thickness of the beryllium window becomes thinner, the proportion of useless low-energy X-ray (<5 keV) intensity to all energy X-ray intensity also is higher (>20%). The accuracy of in-situ EDXRF will be reduced when the high-throughput low-energy X-ray enters the detector. Therefore, this paper puts forward several parameters as judgment index for beryllium window thickness, which is described as follows: 1)The intensity ratios of the K-series X-ray to middle-energy (5-25 keV) bremsstrahlung and middle-high-energy (5-50 keV) bremsstrahlung (F1 and F3); 2)The intensity ratios of useless low-energy X-ray (<5 keV) to middle-energy (5-25 keV) X-ray and middle-high-energy (5-50 keV) X-ray (F2 and F4), it can reflect the relative intensity of useless low-energy X-ray. The simulation results demonstrate that with the increase in the beryllium window thickness, the value of F1 (F3) improves slowly, and the value of F2 (F4) decreases rapidly. In addition to the judgment index discussed above, and considering the X-ray shielded by beryllium window, the beryllium window of miniature X-ray tube can be determined. Based on simulation analysis, the thickness of around 250 microm is appropriate to miniature X-ray tube applied in the in-situ EDXRF. Comparing the emitted spectrum with 50 microm-thick beryllium window, 71.66% of low-energy X-rays are shielded, only 21.31% of X-rays with energy from 5 to 50 keV is shielded, the intensity ratio of low-energy X-ray to total energy X-ray is less than 10%, and the intensity proportion of K-series X-ray to middle-high energy X-ray maintains a high level. In other words, when the mobile X-ray source with 250 microm beryllium window is used in the in-situ EDXRF, proportion of effective signal is higher, and effect of energy resolution of the detection is least; Moreover, the relative intensity of the excitation spectral scattering background, which is obtained by detection for specimen excitation analysis, will remain at low level, thus to ensure the precision of the result of element analysis. For the beryllium window in the application of radiation therapy, the thicker the better. At this time, low-energy X-ray flux maintains a high level, and it can ensure that radiation dose is concentrated on treatment tissue.
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Galangin dampens mice lipopolysaccharide-induced acute lung injury.
Inflammation
PUBLISHED: 04-19-2014
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Galangin, an active ingredient of Alpinia galangal, has been shown to possess anti-inflammatory and antioxidant activities. Inflammation and oxidative stress are known to play vital effect in the pathogenesis of acute lung injury (ALI). In this study, we determined whether galangin exerts lung protection in lipopolysaccharide (LPS)-induced ALI. Male BALB/c mice were randomized to receive galangin or vehicle intraperitoneal injection 3 h after LPS challenge. Samples were harvested 24 h post LPS administration. Galangin administration decreased biochemical parameters of oxidative stress and inflammation, and improved oxygenation and lung edema in a dose-dependent manner. These protective effects of galangin were associated with inhibition of nuclear factor (NF)-?B and upregulation of heme oxygenase (HO)-1. Galangin reduces LPS-induced ALI by inhibition of inflammation and oxidative stress.
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Characterization of integrons among Escherichia coli in a region at high incidence of ESBL-EC.
Pak J Med Sci
PUBLISHED: 03-19-2014
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Objective : The aim of study was to investigate the distribution of the integrons in Escherichia coli (E. coli) isolates, and analyze the possible relationship between the antimicrobial resistance profiles and the integrons. Methods : The antimicrobial profiles of 376 E. coli strains were analysed by disk diffusion test. The integron genes and variable regions were detected by PCR. Some amplicons were sequenced to determine the gene cassettes style. Results : Of 376 isolates, 223 isolates (59.3%) were confirmed as ESBL-EC. Comparison to ESBL-negative E. coli, the high rates of resistance to the third and fourth generation of cephalosporins, penicillins and amikacin were found in ESBL-EC. Only class 1 was integron detected in the isolates, and the prevalence of it was 66.5%. It was commonly found in ESBL-EC (77.6%, 173/223), which was higher than that of ESBL-negative E. coli (50.3%, 77/153) (p<0.001). Six different genes cassettes were detected in this study and were classified into three groups: dfr17-aadA5, dfrA12-aadA2 and aacA4-CmlA1. Additionally, more than one gene array harboured in 13.9% isolates of ESBL-EC, while in 9.1% isolates of ESBL-negative E.coli. Conclusion : The high incidence of ESBL-EC with resistance to multiple antibiotics were detected in the isolates from Blood stream infection (BSI). More resistant gene cassettes in ESBL-EC may partially underlie the high resistance to amikacin, while no relation exists between the high incidence of ESBL-EC and classes 1~ 3 integrons in this region.
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Development of reverse-transcription loop-mediated isothermal amplification assay for rapid detection of novel avian influenza A (H7N9) virus.
BMC Microbiol.
PUBLISHED: 03-16-2014
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BackgroundThe emerged human infection with avian influenza A (H7N9) virus in China since 2013 has aroused global concerns. There is great demand for simple and rapid diagnostic method for early detection of H7N9 to provide timely treatment and disease control. The aim of the current study was to develop a rapid, accurate and feasible reverse-transcription loop-mediated isothermal amplification (RT-LAMP) assay for detection of H7N9 virus.ResultsThe detection limits of the H7- and N9-specific RT-LAMP assay were both approximately 0.2 PFU per reaction. No cross-reactivity was observed with other subtype of influenza viruses or common respiratory viral pathogens. The assay worked well with clinical specimens from patients and chickens, and exhibited high specificity and sensitivity.ConclusionsThe H7/N9 specific RT-LAMP assay was sensitive and accurate, which could be a useful alternative in clinical diagnostics of influenza A (H7N9) virus, especially in the hospitals and laboratories without sophisticated diagnostic systems.
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[Expression of death-associated protein kinase gene and methylation status of promoter region in acute leukemia].
Zhongguo Shi Yan Xue Ye Xue Za Zhi
PUBLISHED: 03-07-2014
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This study was purpose to investigate the expression of death-associated protein kinase (DAPK) gene in acute leukemia (AL) patients and the methylation status of its promoter region through experiments of DAPK methylation and expression, and to analyze the relation between them. The expression of DAPK gene in leukemia cells and normal bone marrow cells was detected by RT-PCR; the methylation status of DAPK gene promoter region in cells from AL patients and leukemia cell lines HL-60 and U937 was detected by nested methylation specific PCR (n-MSP); 2 randomly primers selected from randomly amplified products of second round nMS-PCR were cloned and sequenced in professional company. The results showed that the DAPK gene expressed in bone marrow specimens of all 10 normal controls, with average value of expression 0.92 ± 0.18, while the average value of DAPK expression in bone marrow specimens of AL patients was 0.61 ± 0.40 which was lower than that in normal controls (P < 0.05). The low or deletion of DAPK mRNA expression were found in bone marrow specimens of 9/17 (52.94%) cases of ALL and 42/102 (41.18%) cases of AML. The cell line U937 showed normal expression of DAPK gene, while cell line HL-60 showed the expression detection of DAPK gene. The methylation of DAPK promoter region existed in 33 out of bone marrow specimens of 102 AML patients and in 8 out of bone marrow specimens of 17 ALL patients, the methylation rates were 32.4% (33/102) and 47% respectively. The DAPK promoter region in bone marrow of 7 normal controls was unmethylated, while DAPK promoter region in U937 cells and HL-60 cells were unmethylated and methylated respectively. The DAPK mRNA expression in ALL and AML patients significantly negatively correlated with the methylation of its promoter region (r = -0.855, P < 0.05, in AML patients and r = -0.343, P < 0.05, in AML patients) suggesting the close relationship between them. It is concluded that the methylation of DAPK gene promoter region relates with abnormal expression or detection of DAPK mRNA in AL patients.
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NGF accelerates cutaneous wound healing by promoting the migration of dermal fibroblasts via the PI3K/Akt-Rac1-JNK and ERK pathways.
Biomed Res Int
PUBLISHED: 03-06-2014
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As a well-known neurotrophic factor, nerve growth factor (NGF) has also been extensively recognized for its acceleration of healing in cutaneous wounds in both animal models and randomized clinical trials. However, the underlying mechanisms accounting for the therapeutic effect of NGF on skin wounds are not fully understood. NGF treatment significantly accelerated the rate of wound healing by promoting wound reepithelialization, the formation of granulation tissue, and collagen production. To explore the possible mechanisms of this process, the expression levels of CD68, VEGF, PCNA, and TGF-?1 in wounds were detected by immunohistochemical staining. The levels of these proteins were all significantly raised in NGF-treated wounds compared to untreated controls. NGF also significantly promoted the migration, but not the proliferation, of dermal fibroblasts. NGF induced a remarkable increase in the activity of PI3K/Akt, JNK, ERK, and Rac1, and blockade with their specific inhibitors significantly impaired the NGF-induced migration. In conclusion, NGF significantly accelerated the healing of skin excisional wounds in rats and the fibroblast migration induced by NGF may contribute to this healing process. The activation of PI3K/Akt, Rac1, JNK, and ERK were all involved in the regulation of NGF-induced fibroblast migration.
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Bradyrhizobium ganzhouense sp. nov., an effective symbiotic bacterium isolated from Acacia melanoxylon R. Br. nodules.
Int. J. Syst. Evol. Microbiol.
PUBLISHED: 02-28-2014
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Three slow-growing rhizobial strains, designated RITF806(T), RITF807 and RITF211, isolated from root nodules of Acacia melanoxylon grown in Ganzhou city, Jiangxi Province, China, had been previously defined, based on amplified 16S rRNA gene restriction analysis, as a novel group within the genus Bradyrhizobium. To clarify their taxonomic position, these strains were further analysed and compared with reference strains of related bacteria using a polyphasic approach. According to 16S rRNA gene sequence analysis, the isolates formed a group that was closely related to 'Bradyrhizobium rifense' CTAW71, with a similarity value of 99.9%. In phylogenetic analyses of the housekeeping and symbiotic gene sequences, the three strains formed a distinct lineage within the genus Bradyrhizobium, which was consistent with the results of DNA-DNA hybridization. In analyses of cellular fatty acids and phenotypic features, some differences were found between the novel group and related species of the genus Bradyrhizobium, indicating that these three strains constituted a novel group distinct from any recognized species of the genus Bradyrhizobium. Based on the data obtained in this study, we conclude that our strains represent a novel species of the genus Bradyrhizobium, for which the name Bradyrhizobium ganzhouense sp. nov. is proposed, with RITF806(T) (?=?CCBAU 101088(T)?=?JCM 19881(T)) as the type strain. The DNA G+C content of strain RITF806(T) is 64.6 mol% (T(m)).
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Characterization of live-attenuated Japanese encephalitis vaccine virus SA14-14-2.
Vaccine
PUBLISHED: 02-14-2014
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The live attenuated Japanese encephalitis (JE) vaccine SA14-14-2 was licensed decades ago and now approved for clinical use in most JE endemic countries. Large-scale clinical trials demonstrate ideal safety and efficacy profile of this Chinese vaccine. The SA14-14-2 vaccine was derived from a virulent strain SA14 after hundreds of serial passaging in cells and animals, concern about virulence reversion remains exist. In the present study, to study the in vitro and in vivo genetic and attenuation stability of the vaccine virus, SA14-14-2 was serially passaged in Vero cells and mouse brain followed by sequence comparison and attenuation phenotype analysis. The results showed that no significant mutation was acquired after serial passaging in Vero cells except a single Ser66Leu mutation within capsid protein, which had no effect on viral virulence in mice. Importantly, serial passaging of SA14-14-2 in suckling mouse brain resulted in emergence of adaptive mutations and increased virulence in mice. Population and plaque-purified clone consensus sequence analysis showed four adaptive mutations in envelope (E) protein, F107L, K138E, T226R and I270T, sequentially occurred and become predominant during serial passaging in suckling mouse brain. Especially, these adaptive mutations were close related with the enhanced neurovirulence and neuroinvasiveness in mice. Our results provide experimental evidence of highly genetic and attenuation stability of SA14-14-2 following passaging in Vero cells, and reveal the potential virulence reversion during passaging in mouse brain in association with critical adaptive mutations in E protein. These findings are important for quality control and evaluation of live JE vaccines and will help understand the attenuation mechanism of flavivirus vaccine.
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Development and characterization of 16 polymorphic microsatellite markers from Taiwan cow-tail fir, Keteleeria davidiana var. formosana (Pinaceae) and cross-species amplification in other Keteleeria taxa.
BMC Res Notes
PUBLISHED: 02-13-2014
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Keteleeria davidiana var. formosana (Pinaceae), Taiwan cow-tail fir, is an endangered species listed on the IUCN Red List of Threatened Species and only two populations remain, both on the Taiwan Island. Sixteen polymorphic microsatellite loci were developed in an endangered and endemic gymnosperm species, Keteleeria davidiana var. formosana, and were tested in an additional 6 taxa, K. davidiana var. calcarea, K. davidiana var. chienpeii, K. evelyniana, K. fortunei, K. fortunei var. cyclolepis, and K. pubescens, to evaluate the genetic variation available for conservation management and to reconstruct the phylogeographic patterns of this ancient lineage.
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A novel reporter system for neutralizing and enhancing antibody assay against dengue virus.
BMC Microbiol.
PUBLISHED: 02-12-2014
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Dengue virus (DENV) still poses a global public health threat, and no vaccine or antiviral therapy is currently available. Antibody plays distinct roles in controlling DENV infections. Neutralizing antibody is protective against DENV infection, whereas sub-neutralizing concentration of antibody can increase DENV infection, termed antibody-dependent enhancement (ADE). Plaque-based assay represents the most widely accepted method measuring neutralizing or enhancing antibodies.
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Role of cytoskeleton in axonal regeneration after neurodegenerative diseases and CNS injury.
Rev Neurosci
PUBLISHED: 02-09-2014
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Abstract With the occurrence of neurodegenerative diseases, such as Alzheimer's and Parkinson's disease, a number of well-functioning neurons need to be developed to make up for the loss of neurons and to restore the brain functions. Unfortunately, because the axons cannot regenerate well, brain function cannot be well compensated for even with the increasing number of newborn neurons, let alone the reformation of neural network. Cytoskeletal proteins play a crucial role in regeneration of axon. In this review, we summarize some cytoskeletal proteins, for instance, actin and actin-binding proteins, as well as tubulin and microtubule-associated proteins, and more importantly, their roles in the regulation of axonal regeneration in the brain. It will provide new opportunities for axonal regeneration after brain damage and will even bring new treatments to patients with neurodegenerative diseases.
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Cucurbitane-type triterpenoids from the leaves of Momordica charantia.
J Asian Nat Prod Res
PUBLISHED: 02-05-2014
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Phytochemical investigation of the ethanol extract from the leaves of Momordica charantia L. led to the isolation of two new (1, 2) and four known (3-6) cucurbitane-type triterpenoids. Their structures were elucidated on the basis of extensive analyses of spectroscopic data including IR, UV, MS, 1D, and 2D NMR. Also the absolute configurations of momordicines I (3) and II (4) were determined for the first time by application of the modified Mosher's method, acid hydrolysis, and GC analysis.
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Modeling of progesterone-induced intracellular calcium signaling in human spermatozoa.
J. Theor. Biol.
PUBLISHED: 01-23-2014
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Calcium ion is a secondary messenger of mammalian spermatozoa. The dynamic change of its concentration plays a vital role in the process of sperm motility, capacitation, acrosome and fertilization. Progesterone released by the cumulus cells, as a potent stimulator of fertilization, can activate the calcium channels on the plasma membrane, which in turn triggers the dynamic change of intracellular calcium concentration. In this paper, a mathematical model of calcium dynamic response in mammalian spermatozoa induced by progesterone is proposed and numerical simulation of the dynamic model is conducted. The results show that the dynamic response of calcium concentration predicted by the model is in accordance with experimental evidence. The proposed dynamic model can be used to explain the phenomena observed in the experiments and predict new phenomena to be revealed by experimental investigations, which will provide the basis to quantitatively investigate the fluid mechanics and biochemistry for the sperm motility induced by progesterone.
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Effects of deferoxamine on the repair ability of dental pulp cells in vitro.
J Endod
PUBLISHED: 01-17-2014
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In previous studies, we found that hypoxia promoted the mineralization of dental pulp cells (DPCs). However, the clinical application of hypoxia as a therapy is questionable or unfeasible. Deferoxamine (DFO), a medication for iron overload, has also been shown to induce hypoxia. The purpose of this study was to investigate the effects of DFO on the repair ability of DPCs.
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Doxorubicin induces drug resistance and expression of the novel CD44st via NF-?B in human breast cancer MCF-7 cells.
Oncol. Rep.
PUBLISHED: 01-17-2014
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CD44, a major receptor for hyaluronan (HA), is a member of a class of adhesion molecules of unknown classification involved in cell proliferation, differentiation, migration, angiogenesis, and the presentation of specific cytokines to the corresponding receptors as well as in cell signaling transduction. It has recently been discovered that CD44, a marker of tumor stem cells, is involved in the drug resistance and invasion of multiple types of tumors. The 20 exons in the CD44 gene that are alternatively spliced, give rise to many CD44 isoforms, possibly including tumor-specific sequences. Dozens of CD44 isoforms have been found, to date, and the standard CD44 (CD44s) isoform is the most common. We recently showed that a novel short-tail isoform of CD44 (CD44st) was expressed in multidrug-resistant human breast cancer MCF-7/Adr cells. Moreover, the novel CD44st was able to interact with HA and regulate the expression of matrix metalloproteinase (MMP)-2 and MMP-9, which increased the invasive capability of MCF-7 cells through the Ras/MAPK signaling pathway. In the present study, we verified that MCF-7 cells subjected to drug pressure develop multidrug resistance to doxorubicin, and the expression levels of multidrug resistance protein 1 (MDR1), CD44st and nuclear factor-?B (NF-?B) mRNA and protein were gradually upregulated in a dose?dependent manner in MCF-7 cells treated with doxorubicin. HA increases the secretion of MMP-2 and MMP-9 in multidrug-resistant MCF-7 cells and affected the invasive ability of MCF-7 cells through the upregulation of CD44st expression, and such an effect was blocked by the NF-?B-specific inhibitor BMS-345541.
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Recombinant tandem multi-linear neutralizing epitopes of human enterovirus 71 elicited protective immunity in mice.
Virol. J.
PUBLISHED: 01-16-2014
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Human Enterovirus 71 (EV71) has emerged as the leading cause of viral encephalitis in children, especially in the Asia-Pacific regions. EV71 vaccine development is of high priority at present, and neutralization antibodies have been documented to play critical roles during in vitro and in vivo protection against EV71 infection.
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Growth performance and stress responses of larval Mississippi Paddlefish Polyodon spathula to hypoxia under different diet treatments.
Biomed Res Int
PUBLISHED: 01-12-2014
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A growth trial was conducted to detect the effects of different diets on the growth performance and hypoxia adaptation capacity of Mississippi Paddlefish (Polyodon spathula) larvae. The larvae were fed with live food, formulated diets, and 1/2 live food with 1/2 formulated diets. After a 15-d growth trial, final body weight and total body length were measured, and five larvae from each dietary group were subjected to 1 h of hypoxia treatment. Serum total antioxidant capacity (T-AOC), serum superoxide dismutase (SOD), and liver malondialdehyde (MDA) were measured. Final body weight and weight gain of the fish fed live food were significantly higher than the values for the other two groups. Total body length of the fish fed live food and 1/2 live food with 1/2 formulated diets exhibited no significant difference. After hypoxia treatment, serum T-AOC and SOD activities of the fish fed formulated diets were significantly lower than those of the other two groups. Liver MDA content of the fish fed with live food was significantly higher than that of the other two groups. In conclusion, larval paddlefish fed with an appropriate proportion of live food and formulated diets exhibit improved adaptive capacity to hypoxia.
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Comparison between tenocutaneous suture and Kessler suture techniques in treating acute closed Achilles tendon rupture.
Foot Ankle Surg
PUBLISHED: 01-09-2014
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To compare the effectiveness of tenocutaneous suture and conventional Kessler suture techniques in treating acute closed Achilles tendon rupture.
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In vitro characterization of human adenovirus type 55 in comparison with its parental adenoviruses, types 11 and 14.
PLoS ONE
PUBLISHED: 01-01-2014
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Human adenovirus type 55 (HAdV-B55) represents a re-emerging human pathogen, and this adenovirus has been reported to cause outbreaks of acute respiratory diseases among military trainees and in school populations around the world. HAdV-B55 has been revealed to have evolved from homologous recombination between human adenovirus type 14 (HAdV-B14) and type 11 (HAdV-B11), but it presents different clinical manifestations from parental virus HAdV-B11. In the present paper, we report the distinct biological features of HAdV-B55 in comparison with the parental viruses HAdV-B11 and HAdV-B14 in cell cultures. The results showed that HAdV-B55 replicated well in various cells, similar to HAdV-B11 and HAdV-B14, but that its processing had a slower and milder cytopathic effect in the early stages of infection. Viral fitness analysis showed that HAdV-B55 exhibited higher levels of replication in respiratory cells than did either of its parents. Cytotoxicity and apoptosis analyses in A549 cells indicated that HAdV-B55 was less cytotoxic than HAdV-B11 and HAdV-B14 were and induced milder apoptosis. Finally, thermal sensitivity analysis revealed that HAdV-B55 exhibited lower thermostability than did either HAdV-B11 or HAdV-B14, which may limit the transmission of HAdV-B55 in humans. Together, the findings described here expand current knowledge about this re-emerging recombinant HAdV, shedding light on the pathogenesis of HAdV-B55.
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The different roles of glucocorticoids in the hippocampus and hypothalamus in chronic stress-induced HPA axis hyperactivity.
PLoS ONE
PUBLISHED: 01-01-2014
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Hypothalamus-pituitary-adrenal (HPA) hyperactivity is observed in many patients suffering from depression and the mechanism underling the dysfunction of HPA axis is not well understood. Chronic stress has a causal relationship with the hyperactivity of HPA axis. Stress induces the over-synthesis of glucocorticoids, which will arrive at all the body containing the brain. It is still complicated whether glucocorticoids account for chronic stress-induced HPA axis hyperactivity and in which part of the brain the glucocorticoids account for chronic stress-induced HPA axis hyperactivity. Here, we demonstrated that glucocorticoids were indispensable and sufficient for chronic stress-induced hyperactivity of HPA axis. Although acute glucocorticoids elevation in the hippocampus and hypothalamus exerted a negative regulation of HPA axis, we found that chronic glucocorticoids elevation in the hippocampus but not in the hypothalamus accounted for chronic stress-induced hyperactivity of HPA axis. Chronic glucocorticoids exposure in the hypothalamus still exerted a negative regulation of HPA axis activity. More importantly, we found mineralocorticoid receptor (MR) - neuronal nitric oxide synthesis enzyme (nNOS) - nitric oxide (NO) pathway mediated the different roles of glucocorticoids in the hippocampus and hypothalamus in regulating HPA axis activity. This study suggests that the glucocorticoids in the hippocampus play an important role in the development of HPA axis hyperactivity and the glucocorticoids in the hypothalamus can't induce hyperactivity of HPA axis, revealing new insights into understanding the mechanism of depression.
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Parallel mRNA and microRNA profiling of HEV71-infected human neuroblastoma cells reveal the up-regulation of miR-1246 in association with DLG3 repression.
PLoS ONE
PUBLISHED: 01-01-2014
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Human enterovirus 71 (HEV71) has emerged as the leading cause of viral encephalitis in children in most Asian countries. The roles of host miRNAs in the neurological pathogenesis of HEV71 infection remain unknown. In the present study, comprehensive miRNA expression profiling in HEV71-infected human neuroblastoma SH-SY5Y cells was performed using the Affymetrix Gene Chip microarray assay and was validated using real-time RT-PCR. Among the 69 differentially expressed miRNAs, miR-1246 was specifically induced by HEV71 infection in human neuroblastoma cells, but inhibition of miR-1246 failed to affect HEV71 replication. Parallel mRNA and microRNA profiling based on the 35 K Human Genome Array identified 182 differentially regulated genes. Target prediction of miR-1246 and network modeling revealed 14 potential target genes involved in cell death and cell signaling. Finally, a combined analysis of the results from mRNA profiling and miR-1246 target predication led to the identification of disc-large homolog 3 (DLG3), which is associated with neurological disorders, for further validation. Sequence alignment and luciferase reporter assay showed that miR-1246 directly bound with the 3'-UTR of DLG3 gene. Down-regulation of miR-1246 induced significant changes in DLG3 expression levels in HEV71-infected SHSY5Y cells. Together, these results suggested that miR-1246 might play a role in neurological pathogenesis of HEV71 by regulating DLG3 gene in infected cells. These findings provide new information on the miRNA and mRNA profiles of HEV71-infected neuroblastoma cells. The biological significance of miR-1246 and DLG3 during the course of HEV71 infection deserves further investigation.
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GE11-modified liposomes for non-small cell lung cancer targeting: preparation, ex vitro and in vivo evaluation.
Int J Nanomedicine
PUBLISHED: 01-01-2014
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Non-small cell lung cancer (NSCLC) is a serious threat to human health, and 40%-80% of NSCLCs express high levels of epidermal growth factor receptor (EGFR). GE11 is a novel peptide and exhibits high affinity for EGFR binding. The aim of this study was to construct and evaluate GE11-modified liposomes for targeted drug delivery to EGFR-positive NSCLC. Doxorubicin, a broad-spectrum antitumor agent, was chosen as the payload. GE11 was conjugated to the distal end of DSPE-PEG2000-Mal by an addition reaction with a conjugation efficiency above 90%. Doxorubicin-loaded liposomes containing GE11 (GE11-LP/DOX) at densities ranging from 0% to 15% were prepared by combination of a thin film hydration method and a post insertion method. Irrespective of GE11 density, the physicochemical properties of these targeted liposomes, including particle size, zeta potential, and drug entrapment efficiency, were nearly identical. Interestingly, the cytotoxic effect of the liposomes on A549 tumor cells was closely related to GE11 density, and liposomes with 10% GE11 had the highest tumor cell killing activity and a 2.6-fold lower half maximal inhibitory concentration than that of the nontargeted counterpart (PEG-LP/DOX). Fluorescence microscopy and flow cytometry analysis revealed that GE11 significantly increased cellular uptake of the liposomes, which could be ascribed to specific EGFR-mediated endocytosis. It was found that multiple endocytic pathways were involved in entry of GE11-LP/DOX into cells, but GE11 assisted in cellular internalization mainly via the clathrin-mediated endocytosis pathway. Importantly, the GE11-modified liposomes showed enhanced accumulation and prolonged retention in tumor tissue, as evidenced by a 2.2-fold stronger mean fluorescence intensity in tumor tissue than the unmodified liposomes at 24 hours. In summary, GE11-modified liposomes may be a promising platform for targeted delivery of chemotherapeutic drugs in NSCLC.
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Induction of neutralizing antibodies against four serotypes of dengue viruses by MixBiEDIII, a tetravalent dengue vaccine.
PLoS ONE
PUBLISHED: 01-01-2014
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The worldwide expansion of four serotypes of dengue virus (DENV) poses great risk to global public health. Several vaccine candidates are under development. However, none is yet available for humans. In the present study, a novel strategy to produce tetravalent DENV vaccine based on envelope protein domain III (EDIII) was proposed. Tandem EDIIIs of two serotypes (type 1-2 and type 3-4) of DENV connected by a Gly-Ser linker ((Gly4Ser)3) were expressed in E. coli, respectively. Then, the two bivalent recombinant EDIIIs were equally mixed to form the tetravalent vaccine candidate MixBiEDIII, and used to immunize BALB/c mice. The results showed that specific IgG and neutralizing antibodies against all four serotypes of DENV were successfully induced in the MixBiEDIII employing Freund adjuvant immunized mice. Furthermore, in the suckling mouse model, sera from mice immunized with MixBiEDIII provided significant protection against four serotypes of DENV challenge. Our data demonstrated that MixBiEDIII, as a novel form of subunit vaccine candidates, might have the potential to be further developed as a tetravalent dengue vaccine in the near future.
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[Chemical constituents of Aconitum bulleyanum].
Zhong Yao Cai
PUBLISHED: 12-31-2013
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To study the chemical constituents of chloroform fraction from Aconitum bulleyanum.
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[Association between matrix metalloproteinase-10 gene polymorphisms and instability of carotid plaque].
Zhonghua Yi Xue Yi Chuan Xue Za Zhi
PUBLISHED: 12-12-2013
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To assess the association between 2 single nucleotide polymorphisms (SNPs) located in exonic regions of matrix metalloproteinase-10 (MMP-10) gene and instability of carotid plaques in a Han Chinese population.
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Copy number variation at 6q13 is associated with lung cancer risk in a Han Chinese population.
Exp. Lung Res.
PUBLISHED: 11-18-2013
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ABSTRACT Copy number variations (CNVs), a major source of human genetic polymorphism, have been suggested to have an important role in genetic susceptibility to common diseases such as cancer, immune diseases, and neurological disorders. Lung cancer is a multifactorial tumor closely associated with genetic background. Previous genome-wide association studies have identified single nucleotide polymorphisms (SNPs) that are associated with lung cancer susceptibility. This study examined the CNVR2966.1 at 6q13 and its association with lung cancer susceptibility. The CNVR2966.1 was found to be a 10,379 bp nucleotides deletion/insertion within the uniform boundaries chromosome 6: 74,648,791-74,659,169. The risk of lung cancer observed in 503 cases and 623 controls was significantly associated with copy number of CNVR2966.1, with the odds ratio (OR) being 1.38 [95% confidence interval (CI) = 1.05-1.79; P = .007] for one copy genotype compared with two copies genotype. These results suggest that CNVR2966.1 is associated with lung cancer risk.
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[Evidence-based quality assessment of Chinese clinical trials on the effects of stabilization of permanent anterior dental trauma].
Shanghai Kou Qiang Yi Xue
PUBLISHED: 11-16-2013
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To appraise the quality of Chinese clinical trials during recent 10 years about the treatment of permanent anterior dental trauma with evidence-based quality control methods.
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A Chimeric Dengue Virus Vaccine using Japanese Encephalitis Virus Vaccine Strain SA14-14-2 as Backbone Is Immunogenic and Protective against Either Parental Virus in Mice and Nonhuman Primates.
J. Virol.
PUBLISHED: 10-09-2013
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The development of a safe and efficient dengue vaccine represents a global challenge in public health. Chimeric dengue viruses (DENV) based on an attenuated flavivirus have been well developed as vaccine candidates by using reverse genetics. In this study, based on the full-length infectious cDNA clone of the well-known Japanese encephalitis virus live vaccine strain SA14-14-2 as a backbone, a novel chimeric dengue virus (named ChinDENV) was rationally designed and constructed by replacement with the premembrane and envelope genes of dengue 2 virus. The recovered chimeric virus showed growth and plaque properties similar to those of the parental DENV in mammalian and mosquito cells. ChinDENV was highly attenuated in mice, and no viremia was induced in rhesus monkeys upon subcutaneous inoculation. ChinDENV retained its genetic stability and attenuation phenotype after serial 15 passages in cultured cells. A single immunization with various doses of ChinDENV elicited strong neutralizing antibodies in a dose-dependent manner. When vaccinated monkeys were challenged with wild-type DENV, all animals except one that received the lower dose were protected against the development of viremia. Furthermore, immunization with ChinDENV conferred efficient cross protection against lethal JEV challenge in mice in association with robust cellular immunity induced by the replicating nonstructural proteins. Taken together, the results of this preclinical study well demonstrate the great potential of ChinDENV for further development as a dengue vaccine candidate, and this kind of chimeric flavivirus based on JE vaccine virus represents a powerful tool to deliver foreign antigens.
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Structure of the CCR5 chemokine receptor-HIV entry inhibitor maraviroc complex.
Science
PUBLISHED: 09-12-2013
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The CCR5 chemokine receptor acts as a co-receptor for HIV-1 viral entry. Here we report the 2.7 angstrom-resolution crystal structure of human CCR5 bound to the marketed HIV drug maraviroc. The structure reveals a ligand-binding site that is distinct from the proposed major recognition sites for chemokines and the viral glycoprotein gp120, providing insights into the mechanism of allosteric inhibition of chemokine signaling and viral entry. A comparison between CCR5 and CXCR4 crystal structures, along with models of co-receptor-gp120-V3 complexes, suggests that different charge distributions and steric hindrances caused by residue substitutions may be major determinants of HIV-1 co-receptor selectivity. These high-resolution insights into CCR5 can enable structure-based drug discovery for the treatment of HIV-1 infection.
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[A prevalence study on mild cognitive impairment among elderly populations in Zhejiang province].
Zhonghua Liu Xing Bing Xue Za Zhi
PUBLISHED: 09-11-2013
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To understand the prevalence of older people with mild cognitive impairment (MCI) in Zhejiang province and to provide the basis for elderly early detection and diagnosis of Alzheimers disease (AD).
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[Correlation between androgen receptor expression and hepatitis B virus X protein and its clinical significance in hepatocellular carcinoma].
Zhonghua Zhong Liu Za Zhi
PUBLISHED: 08-30-2013
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To investigate the expression of androgen receptor (AR) and hepatitis B virus X protein (HBx) in hepatocellular carcinoma (HCC), and analyze the relationship between AR and HBx expressions.
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L_RNA_scaffolder: scaffolding genomes with transcripts.
BMC Genomics
PUBLISHED: 08-28-2013
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Generation of large mate-pair libraries is necessary for de novo genome assembly but the procedure is complex and time-consuming. Furthermore, in some complex genomes, it is hard to increase the N50 length even with large mate-pair libraries, which leads to low transcript coverage. Thus, it is necessary to develop other simple scaffolding approaches, to at least solve the elongation of transcribed fragments.
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Virus-like particles produced in Saccharomyces cerevisiae elicit protective immunity against Coxsackievirus A16 in mice.
Appl. Microbiol. Biotechnol.
PUBLISHED: 07-16-2013
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Hand, foot, and mouth disease (HFMD) has caused significant morbidity and mortality in the Asia-Pacific regions, particularly in infants and young children. Coxsackievirus A16 (CA16) represents one of the major causative agents for HFMD, and the development of a safe and effective vaccine preventing CA16 infections has become a public health priority. In this study, we have developed a yeast system for the production of virus-like particles (VLPs) for CA16 by co-expressing P1 and 3CD of CA16 in Saccharomyces cerevisiae. These VLPs exhibit similarity in both protein composition and morphology as empty particles from CA16-infected cells. Immunization with CA16 VLPs in mice potently induced CA16-specific IgG and neutralization antibodies in a dose-dependent manner. IgG subclass isotyping revealed that IgG1 and lgG2b were dominantly induced by VLPs. Meanwhile, cytokine profiling demonstrated that immunization with VLPs significantly induced the secretion of IFN-?, indicating potent cellular immune response. Furthermore, in vivo challenge experiments showed that passive immunization with anti-VLPs sera conferred full protection against lethal CA16 challenge in neonate mice. Taken together, our data demonstrated that VLPs produced in yeast might have the potential to be further developed as a vaccine candidate against HFMD.
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In situ determination mechanisms for the depuration of polycyclic aromatic hydrocarbons adsorbed onto the leaf surfaces of living mangrove seedlings.
J. Hazard. Mater.
PUBLISHED: 07-09-2013
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To further increase understanding of the mechanisms responsible for air-surface exchange processes, the depuration of adsorbed individual fluorene (Flu), anthracene (Ant), phenanthrene (Phe), fluoranthene (Fla), and pyrene (Pyr) from the leaf surfaces of living Aegiceras corniculatum (Ac) and Kandelia obovata (Ko) seedlings were in situ investigated in real time using laser-induced nanosecond time-resolved fluorescence (LITRF) system. Depuration of the PAHs from the leaf surfaces of the two mangrove seedlings included a rapid and a slow phase, and both of them followed first-order kinetics. Furthermore, significant inter-species and inter-chemical variability existed in terms of the elimination rates and the remaining PAHs residues during the two phases. The rapid phase mainly represented a fast volatilization, of which the volatilization rates moderately correlate with PAH molecular weight, while combined effect of volatilization and photolysis was the dominant mechanism for the slow phase. The retainment of PAHs on the leaf surfaces was associated with the plant species and physicochemical properties of PAHs, especially logKOA.
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Trefoil factor 3 peptide regulates migration via a Twist-dependent pathway in gastric cell.
Biochem. Biophys. Res. Commun.
PUBLISHED: 06-20-2013
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Trefoil factor 3 (TFF3) is a member of the TFF-domain peptide family and essential in regulating cell migration and maintaining mucosal integrity in gastrointestinal tract. However, the role of TFF3 and its downstream regulating mechanisms in cancer cell migration remain unclear. We previously reported that TFF3 prolonged the up-regulation of Twist protein to modulate IL-8 secretion in intestinal epithelial cells. In this study, we investigated the role of Twist protein in TFF3-induced migration of SGC7901 cells. While Twist was activated by TFF3, siRNA-mediated knockdown of Twist abolished TFF3-induced cell migration. Furthermore, the migration related marker CK-8 as well as ZO-1 and MMP-9 was also regulated by TFF3 via a Twist-dependent mechanism. Our study suggests that Twist, as an important potential downstream effector, plays a key role in TFF3-modulated metastasis in gastric cancer and can be a promising therapeutic target against intestinal-type gastric cancer.
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Glomus tumor of uncertain malignant potential arising in the bronchus.
J Cardiothorac Surg
PUBLISHED: 06-06-2013
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Glomus tumor is usually a small, benign tumor and typically occurs in the dermis or subcutis or soft tissue of the extremities and rarely in the visceral locations. Its occurrence in the main bronchus is extremely rare. The current case reported a 30-year-old woman with dyspnea on exertion and hemoptysis, she had a glomus tumor which has large size, deep location and exhibits an infiltrative margin as well as increased atypical mitotic figures. These characteristics suggest malignant behavior. However, there is little data regarding glomus tumors arising in the bronchus, the need for caution in diagnosing this case as a malignant glomus tumor must be highlighted. Therefore, the diagnosis of bronchial glomus tumor of uncertain malignant potential was favored. To the best of our knowledge, both the type and the location of this glomus tumor are extremely rare. Accumulation of more cases are needed to clarify their diagnosis and significance since there is little data regarding glomus tumors arising in the bronchus.
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Synthesis, characterization and cytocompatibility of a degradable polymer using ferric catalyst for esophageal tissue engineering.
J Mater Sci Mater Med
PUBLISHED: 06-05-2013
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This study focused on the synthesis, characterization and cytocompatibility of a biodegradable polymer by the cross-linking from poly(ethylene glycol-co-lactide) dimethacrylate (PLEGDMA), polyethylene glycol diacrylate (PEGDA) and N-isopropylacrylamide, where PLEGDMA was synthesized by ring-opening oligomerization of poly(ethylene glycol) with different molecular weights (Mn = 400, 600, 1000, 2000 Da) and L-lactide using low toxic iron(III) acetylacetonate (Fe(acac)3) as the catalyst and subsequently being terminated with dimethacrylate. The product, PLEGDMA, was analyzed to confirm its chemistry using FTIR spectroscopy, (1)H NMR spectra and gel permeation chromatography etc. The thermodynamic properties, mechanical behaviors, surface hydrophilicity, degradability and cytotoxicity of the cross-linked product were evaluated by differential scanning calorimetry, tensile tests, contact angle measurements and cell cultures. The effects of reaction variables such as PEGDA content and reactants ratio were optimized to achieve a material with low glass transition temperature (Tg), high wettability and preferable mechanical characteristics. Using a tubular mould which has been patented in our group, a tubular scaffold with predetermined dimension and pattern was fabricated, which aims at guiding the growth and phenotype regulation of esophageal primary cells like fibroblast and smooth muscle cell towards fabricating tissue engineered esophagus in future.
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[Effects of increased precipitation on the water use of Nitraira tangutorum at southeast edge of Baddain Jaran Desert in China].
Ying Yong Sheng Tai Xue Bao
PUBLISHED: 05-31-2013
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This paper studied the threshold value of the water use of Nitraria tanturorum shrubs at the southeast edge of Baddain Jiran Desert. From the early May to late September in 2009, an irrigation simulating increased precipitation was conducted once every month. Three ratios of increased precipitation (0, 50% and 100%) were designed, based on the local mean annual precipitation (115 mm). On the 1 day before irrigation and the 1, 3 and 7 days after irrigation in May, July and September, the deltaD in the xylem water of N. tangutorum, the soil water at the depths 10 and 30 cm, and the well water and natural rainfall, and the variations of the soil water content were measured. Under natural condition, the N. tangutorum mainly utilize ground water in May and September, and utilize the soil water at the depths 10 and 30 cm in July. After irrigation, the ground water use rate of the N. tangutorum decreased, while the soil water use rate increased. In the treatment of 100% increased precipitation, the deltaD ratio of the water in N. tangutorum xylem was affected significantly, and the water use of the N. tangutorum in May, July and September increased. In the treatment of 50% increased precipitation, the soil water condition in May and July was improved, but the water use rate had little improvement. Only when the increased precipitation reached 100% of the local mean annual precipitation, could the water use rate of the N. tangutorum have an obvious increase.
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Hydrogen peroxide induces apoptosis in human dental pulp cells via caspase-9 dependent pathway.
J Endod
PUBLISHED: 05-22-2013
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Reactive oxygen species are a group of metabolic intermediates produced during oxidative metabolism in eukaryotic cells. They include superoxide anion (O2(-)), hydrogen peroxide (H2O2), hydroxyl radical (·OH), and (1)O2. Of these intermediates, H2O2 is the most stable. Dental pulp cells can be invaded by tooth bleaching, laser radiation, and dental materials. This can influence the intracellular level of reactive oxygen species. Apoptosis, which is the best-known form of programmed cell death, is pivotal to tissue development and regeneration. Little information is available regarding the relationship between H2O2 and apoptosis of human dental pulp cells (hDPCs). The purpose of this study was to investigate whether H2O2 can induce apoptosis in hDPCs and its signaling way.
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Primary pulmonary adenocarcinoma mimicking papillary thyroid carcinoma.
J Cardiothorac Surg
PUBLISHED: 05-14-2013
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We herein reported a primary pulmonary papillary carcinoma with colloid-like luminal content in the glandular cavity and classic nuclear features such as pseudo-inclusions, intranuclear grooves in the tumor cell nuclei and ground glass nuclei which closely mimics papillary thyroid carcinoma. Meanwhile, lymph node in the left pulmonary hilum was involved and showed similar features to the primary pulmonary papillary carcinoma. This specific histopathological presentation caused a diagnostic dilemma.The patient didnt show previous concomitant or subsequent evidence of a thyroid tumor. Immunohistochemistry further confirmed pulmonary origin and excluded a metastasis from the thyroid, as it was thyroglobulin negative, thyroid transcription factor 1 and surfactant apoprotein A positive, which was consistent with the imageology and history.Based on the above features, the diagnosis of primary pulmonary papillary carcinoma was confirmed. Understanding the existence of papillary thyroid carcinoma-like pulmonary papillary carcinoma will avoid misdiagnosis or unnecessary clinical and radiologic investigations in future.
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Phenotypic and genomic characterization of human coxsackievirus A16 strains with distinct virulence in mice.
Virus Res.
PUBLISHED: 05-08-2013
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Human coxsackievirus A16 (CA16) infection results in hand, foot, and mouth disease (HFMD) along with other severe neurological diseases in children and poses an important public health threat in Asian countries. During an HFMD epidemic in 2009 in Guangdong, China, two CA16 strains (GD09/119 and GD09/24) were isolated and characterized. Although both strains were similar in plaque morphology and growth properties in vitro, the two isolates exhibited distinct pathogenicity in neonatal mice upon intraperitoneal or intracranial injection. Complete genome sequences of both CA16 strains were determined, and the possible virulence determinants were analyzed and predicted. Phylogenetic analysis revealed that these CA16 isolates from Guangdong belonged to the B1b genotype and were closely related to other recent CA16 strains isolated in mainland China. Similarity and bootscanning analyses of these CA16 strains detected homologous recombination with the EV71 prototype strain BrCr in the non-structural gene regions and the 3-untranslated regions. Together, the phenotypic and genomic characterizations of the two clinical CA16 isolates circulating in China were compared in detail, and the potential amino acid residues responsible for CA16 virulence in mice were predicted. These findings will help explain the evolutionary relationship of the CA16 strains circulating in China, warranting future studies investigating enterovirus virulence.
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[Preliminary study of establishing clinical effect evaluation methods of chinese medicine based on combination of disease and syndrome, systematic staging, and multi-dimension index].
Zhongguo Zhong Xi Yi Jie He Za Zhi
PUBLISHED: 05-08-2013
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The clinical effect evaluation of Chinese medicine (CM) has been the bottleneck restricting its development. Based on the current situation, in this study, we integrated and combined previous results of clinical effect evaluation of CM, and proposed the clinical effect evaluation method of CM based on combination of disease and syndrome, systematic staging, and multi-dimension index. We also made a specific exposition on the connotation, establishment methods and practice of the clinical effect evaluation methods of CM based on combination of disease and syndrome, systematic staging, and multi-dimension index.
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[Analysis of risk factors for anastomotic infectious complications following bowel resection for Crohn disease].
Zhonghua Wei Chang Wai Ke Za Zhi
PUBLISHED: 04-24-2013
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To investigate the risk factors for anastomotic infectious complications after bowel resection in patients with Crohn disease.
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Effect of WFDC 2 silencing on the proliferation, motility and invasion of human serous ovarian cancer cells in vitro.
Asian Pac J Trop Med
PUBLISHED: 04-24-2013
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To investigate effect and possible mechanisms of silencing human WFDC2 (HE4) gene on biological behavior changes as cell proliferation, apoptosis, movement and invasion of human serous ovarian cancer cell line SKOV3.
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Rational design of thermostable vaccines by engineered peptide-induced virus self-biomineralization under physiological conditions.
Proc. Natl. Acad. Sci. U.S.A.
PUBLISHED: 04-15-2013
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The development of vaccines against infectious diseases represents one of the most important contributions to medical science. However, vaccine-preventable diseases still cause millions of deaths each year due to the thermal instability and poor efficacy of vaccines. Using the human enterovirus type 71 vaccine strain as a model, we suggest a combined, rational design approach to improve the thermostability and immunogenicity of live vaccines by self-biomineralization. The biomimetic nucleating peptides are rationally integrated onto the capsid of enterovirus type 71 by reverse genetics so that calcium phosphate mineralization can be biologically induced onto vaccine surfaces under physiological conditions, generating a mineral exterior. This engineered self-biomineralized virus was characterized in detail for its unique structural, virological, and chemical properties. Analogous to many exteriors, the mineral coating confers some new properties on enclosed vaccines. The self-biomineralized vaccine can be stored at 26 °C for more than 9 d and at 37 °C for approximately 1 wk. Both in vitro and in vivo experiments demonstrate that this engineered vaccine can be used efficiently after heat treatment or ambient temperature storage, which reduces the dependence on a cold chain. Such a combination of genetic technology and biomineralization provides an economic solution for current vaccination programs, especially in developing countries that lack expensive refrigeration infrastructures.
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Novel cis-acting element within the capsid-coding region enhances flavivirus viral-RNA replication by regulating genome cyclization.
J. Virol.
PUBLISHED: 04-10-2013
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cis-Acting elements in the viral genome RNA (vRNA) are essential for the translation, replication, and/or encapsidation of RNA viruses. In this study, a novel conserved cis-acting element was identified in the capsid-coding region of mosquito-borne flavivirus. The downstream of 5 cyclization sequence (5CS) pseudoknot (DCS-PK) element has a three-stem pseudoknot structure, as demonstrated by structure prediction and biochemical analysis. Using dengue virus as a model, we show that DCS-PK enhances vRNA replication and that its function depends on its secondary structure and specific primary sequence. Mutagenesis revealed that the highly conserved stem 1 and loop 2, which are involved in potential loop-helix interactions, are crucial for DCS-PK function. A predicted loop 1-stem 3 base triple interaction is important for the structural stability and function of DCS-PK. Moreover, the function of DCS-PK depends on its position relative to the 5CS, and the presence of DCS-PK facilitates the formation of 5-3 RNA complexes. Taken together, our results reveal that the cis-acting element DCS-PK enhances vRNA replication by regulating genome cyclization, and DCS-PK might interplay with other cis-acting elements to form a functional vRNA cyclization domain, thus playing critical roles during the flavivirus life cycle and evolution.
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Identification and characterization of a linearized B-cell epitope on the pr protein of dengue virus.
J. Gen. Virol.
PUBLISHED: 04-04-2013
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The four serotypes of dengue virus (DENV) represent one of the major mosquito-borne pathogens globally; so far no vaccine or specific antiviral is available. During virion maturation, the pr protein is cleaved from its precursor form the prM protein on the surface of immature DENV by host protease. Recent findings have demonstrated that the pr protein not only played critical roles in virion assembly and maturation, but was also involved in antibody-dependent enhancement of DENV infection. However, the B-cell epitopes on the pr protein of DENV have not been well characterized. In this study, a set of 11 partially overlapping peptides spanning the entire pr protein of DENV-2 were fused with glutathione S-transferase and expressed in Escherichia coli. ELISA screening with murine hyperimmune antiserum against immature DENV identified the P8 peptide (??KQNEPEDIDCWCNST?¹) in the pr protein as the major immunodominant epitope. Fine mapping by truncated protein assays confirmed the 8-e peptide ??KQNEPEDI?? was the smallest unit capable of antibody binding. Importantly, the 8-e epitope reacted with sera from dengue fever patients. Site-directed mutagenesis revealed the asparagine residue at position 59 was important for epitope recognition. The 8-e epitope coincided well with the B-cell epitopes predicted by Immune Epitope Database analysis, and 3D structural modelling mapped the 8-e peptide on the surface of prM-E heterodimers. Overall, our findings characterized a linearized B-cell epitope on the pr protein of DENV, which will help to understand the life cycle of DENV and pathogenesis of dengue infections in human.
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An efficacy analysis of surgical timing and procedures for high-energy complex tibial plateau fractures.
Orthop Surg
PUBLISHED: 03-19-2013
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To identify the most effective treatment for application to high-energy complex plateau fractures.
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Progress risk assessment of oral premalignant lesions with saliva miRNA analysis.
BMC Cancer
PUBLISHED: 03-13-2013
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Oral cancer develops through multi-stages: from normal to mild (low grade) dysplasia (LGD), moderate dysplasia, and severe (high grade) dysplasia (HGD), to carcinoma in situ (CIS) and finally invasive oral squamous cell carcinomas (OSCC). Clinical and histological assessments are not reliable in predicting which precursor lesions will progress. The aim of this study was to assess the potential of a noninvasive approach to assess progress risk of oral precancerous lesions.
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Rational design of a flavivirus vaccine by abolishing viral RNA 2-O methylation.
J. Virol.
PUBLISHED: 03-13-2013
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Viruses that replicate in the cytoplasm cannot access the host nuclear capping machinery. These viruses have evolved viral methyltransferase(s) to methylate N-7 and 2-O cap of their RNA; alternatively, they "snatch" host mRNA cap to form the 5 end of viral RNA. The function of 2-O methylation of viral RNA cap is to mimic cellular mRNA and to evade host innate immune restriction. A cytoplasmic virus defective in 2-O methylation is replicative, but its viral RNA lacks 2-O methylation and is recognized and eliminated by the host immune response. Such a mutant virus could be rationally designed as a live attenuated vaccine. Here, we use Japanese encephalitis virus (JEV), an important mosquito-borne flavivirus, to prove this novel vaccine concept. We show that JEV methyltransferase is responsible for both N-7 and 2-O cap methylations as well as evasion of host innate immune response. Recombinant virus completely defective in 2-O methylation was stable in cell culture after being passaged for >30 days. The mutant virus was attenuated in mice, elicited robust humoral and cellular immune responses, and retained the engineered mutation in vivo. A single dose of immunization induced full protection against lethal challenge with JEV strains in mice. Mechanistically, the attenuation phenotype was attributed to the enhanced sensitivity of the mutant virus to the antiviral effects of interferon and IFIT proteins. Collectively, the results demonstrate the feasibility of using 2-O methylation-defective virus as a vaccine approach; this vaccine approach should be applicable to other flaviviruses and nonflaviviruses that encode their own viral 2-O methyltransferases.
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Optimized pregelatinized starch technique for cell block preparation in cell cultures.
Exp. Mol. Pathol.
PUBLISHED: 03-11-2013
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The aim of the present study was to optimize the pregelatinized starch technique for cell block preparation and apply this approach in cultured cells of all types of growing forms, suspension and adherent. In order to evenly mix the starch powder and the cell suspension, we crafted a special plastic dropper. To prove the effectiveness of this optimized technique we used different cell lines, NCI-H69, NCI-H345, HCT-116, SKBR3 and MDA-MB-231. The morphology features, immunocytochemistry (ICC) and fluorescent/chromogenic in-situ hybridization (FISH/CISH) on the cell block sections were evaluated. The morphology features, the ICC and ISH results of cell block sections prepared by the new method were satisfactory comparing with the results obtained in biopsies, the gold standard test for this kind of analysis. The most attractive advantage of our optimized pregelatinized starch technique is that this new method is based on cell suspensions instead of cell sediment, so with our technique every section will contain cells due to the even distribution of the starch powder and the cells forming a homogeneous cell block. To the authors knowledge, this is the first description on cell block preparation based on cell suspension.
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Development and characterization of the replicon system of Japanese encephalitis live vaccine virus SA14-14-2.
Virol. J.
PUBLISHED: 02-22-2013
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Viral self-replicating sub-genomic replicons represent a powerful tool for studying viral genome replication, antiviral screening and chimeric vaccine development. Many kinds of flavivirus replicons have been developed with broad applications.
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